Note: Descriptions are shown in the official language in which they were submitted.
W096/09836 22 ~ ~ 3 4 6 PCT/US95/12319
TITLE OF THE INVENTION
INHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
BACKGROUND OF THE INVENTION
The Ras gene is found activated in many human cancers,
including colorectal carcinoma, exocrine pancreatic carcinoma, and
myeloid leukemias. Biological and biochemical studies of Ras action
indicate that Ras functions like a G-regulatory protein, since Ras must
be localized in the plasma membrane and must bind with GTP in
order to transfonn cells (Gibbs, J. et al., Microbiol. Rev. 53:171-286
(1989). Forms of Ras in cancer cells have mutations that distinguish
the protein from Ras in normal cells.
At least 3 post-translational modifications are involved
with Ras membrane loc~li7~tion, and all 3 modifications occur at the
C-terminus of Ras. The Ras C-terminus contains a sequence motif
termed a "CAAX" or "Cys-Aaa1-Aaa2-Xaa" box (Aaa is an aliphatic
amino acid, the Xaa is any amino acid) (Willumsen et al., Nature
310:~83-~86 (1984)). Other proteins having this motif include the
Ras-related GTP-binding proteins such as Rho, fungal mating factors,
the nuclear lamins, and the gamma subunit of transducin.
Farnesylation of Ras by the isoprenoid farnesyl
pyrophosphate (FPP) occurs in vivo on Cys to form a thioether linkage
(Hancock et al., Cell 57:1 167 (1989), Casey et al., Proc. Natl. Acad.
Sci. USA 86:8323 (1989)). In addition, Ha-Ras and N-Ras are
palmitoylated via formation of a thioester on a Cys residue near a C-
terminal Cys farnesyl acceptor (Gutierrez et al., EMBO J. 8:1093-
1098 (1989), Hancock ef al., Cell 57:1167-1177 (1989)). Ki-Ras
lacks the palmitate acceptor Cys. The last 3 amino acids at the Ras C-
terminAl end are removed proteolytically, and methyl esterification
occurs at the new C-telminus (Hancock et al., ibid). Fungal mating
J factor and m~mm~ n nuclear lamins undergo identical modification
steps (Anderegg et al., J. Biol. Chem. 263:18236 (1988), Farnsworth
et al., J. Biol. Chenl. 264:20422 (1989)).
W096/09836 22 ~ ~ 3 ~ 6 P~usgs/l23l9
Inhibition of Ras farnesylation in 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 limiting enzyme for the production of polyisoprenoids and the
farnesyl pyrophosphate precursor. It has been shown that a farnesyl-
protein transferase using farnesyl pyrophosphate as a precursor is
responsible for Ras farnesylation. (Reiss et al., Cell, 62:81-88 (1990);
Schaber et al., J. Biol. Chem., 265:14701-14704 (1990); Schafer et al.,
0 Science, 249:1133-1139 (1990); Manne et al., Proc. Natl. Acad. Sci
USA, 87:7541 -7545 (1990)).
Inhibition of farnesyl-protein transferase and, thereby, of
farnesylation of the Ras protein, blocks the ability of Ras to transform
normal cells to cancer cells. The compounds of the invention inhibit
Ras farnesylation and, thereby, generate soluble Ras which, as
indicated infra, can act as a dominant negative inhibitor of Ras
function. While soluble Ras in cancer cells can become a domin~nt
negative inhibitor, soluble Ras in normal cells would not be an
inhibitor.
A cytosol-localized (no Cys-Aaal-Aaa2-Xaa box
membrane domain present) and activated (impaired GTPase activity,
staying bound to GTP) form of Ras acts as a domin~nt negative Ras
inhibitor of membrane-bound Ras function (Gibbs et al., Proc. Natl.
Acad. Sci. USA 86:6630-6634(1989)). Cytosollocalized forms of Ras
with normal GTPase activity do not act as inhibitors. Gibbs et al.,
ibid, showed this effect in Xenopus oocytes and in m~mm~ n cells.
A~lmini~tration of compounds of the invention to block
Ras farnesylation not only decreases the amount of Ras in the
membrane but also generates a cytosolic pool of Ras. In tumor cells
having activated Ras, the cytosolic pool acts as another antagonist of
membrane-bound Ras function. In normal cells having normal Ras,
the cytosolic pool of Ras does not act as an antagonist. In the absence
of complete inhibition of farnesylation, other farnesylated proteins are
able to continue with their ~unctions.
W096/09836 2~ ~ ~ 3 ~ 6 PCT/US95/12319
Farnesyl-protein transferase activity may be reduced or
completely inhibited by adjusting the compound dose. Reduction of
farnesyl-protein transferase enzyme activity by adjusting the
compound dose would be useful for avoiding possible undesirable
5 side effects resulting from interference with other metabolic processes
which utilize the enzyme.
These compounds and their analogs are inhibitors of
farnesyl-protein transferase. Farnesyl-protein transferase utilizes
farnesyl pyrophosphate to covalently modify the Cys thiol group of
0 the Ras CAAX box with a farnesyl group. Inhibition of farnesyl
pyrophosphate biosynthesis by inhibiting HMG-CoA reductase blocks
Ras membrane localization in vivo and inhibits Ras function.
Inhibition of farnesyl-protein transferase is more specific and is
attended by fewer side effects than is the case for a general inhibitor
15 of isoprene biosynthesis.
Previously, it has been demonstrated that tetrapeptides
cont~ining cysteine as an amino terminal residue with the CAAX
sequence inhibit Ras farnesylation (Schaber et al., ibid, Reiss et. al.,
ibid; Reiss et al., PNAS, 88:732-736 (1991)). Such inhibitors may
20 inhibit while serving as alternate substrates for the Ras farnesyl-
transferase enzyme, or may be purely competitive inhibitors (U.S.
Patent 5,141,85 l, University of Texas).
It has also been demonstrated that certain inhibitors of
farnesyl-protein transferase selectively bloc~ the processing of Ras
25 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 transferase blocks the growth of ras-dependent tumors in nude
mice (N.E. Kohl et al., Proc. Natl. Acad. Sci U.S.A., 91 :9141 -9145
30 (1994)-
Inhibitors of Ras 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 substrate for the enzyme, Ras. Almost all of the
W096/09836 22 0 ~ 3 4 6 PCTJUS95~12319
peptide derived inhibitors that have been described are cysteine
cont~ining molecules that are related to the CAAX motif that is the
signal for protein prenylation. The exception to this generalization is
a class of natural products known as the pepticinn;~min~ (Omura, et
al., J. Antibiotics 46:222 (1993). In general, deletion of the thiol from
a CAAX derivative dramatically reduces the inhibitory potency of
these compounds. However, the thiol group potentially places
limitations on the therapeutic application of FPTase inhibitors with
respect to ph~ cokinetics, ph~rm~codynamics and toxicity.
Therefore, a functional replacement for the thiol is desirable. With the
exception of the pepticinn~mins, non-thiol FPTase inhibitors that are
competitive with the Ras substrate have not been described and are the
subject of this invention.
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~li7~tion of the oncogene Ras protein. It is a further object of
this invention to develop chemotherapeutic compositions cont~ining
the compounds af this invention and methods for producing the
compounds of this invention.
SUMMARY OF THE INVENTION
The present invention comprises analogs of the CAAX
motif of the protein Ras that is modified by farnesylation in vivo.
2s These CAAX analogs inhibit the farnesylation of Ras. Furthermore,
the.se CAAX analogues differ from those previously described as
inhibitors of Ras farnesyl transfera,se in that they do not have a thiol
moiety. The lack of the thiol offers unique advantages in terms of
improved pharmacokinetic behavior in ~nim~ls, prevention of thiol-
3 o 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
cont~ining these farnesyl transferase inhibitors and methods for their
production.
W096/09836 ~2 ~ PCT/US95/12319
The compounds of ~is invention are illustrated by the
formulae:
V (CR1a ) -W (CR1b ) (CR1bR9)J~N~x~ ~N~
R3 R4
(R8)t ZR2aR21~ Z R5a p5b
V (CR1a2) - W- (CR1b2)p(CR1bR9)r R12 ~ HN
Il R3 R4
HOCH2(/CH2)q
V - (CR1a2)n - W- (cR1b2)p(cR1bR9)rJ~N1~2~x~ ~N
R3 R4
111
and
25 (R8)t z R2a~R2b z f~q
v - (CR1a2)n - w - (cR1b2)p(cR1bR9)r/l~N~x~ ~ N~O
IV
W096/09836 ~ 0 1 ~ 4 6 PCT/US95/12319
DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention inhibit the farnesylation
of Ras. In a first embodiment of this invention, the Ras farnesyl
transferase inhibitors are illustrated by the formula I:
(R8)t Z ~z R ~R
R1a2) - W - (CR1b2)p(CR1bR9), NR12 ~ H~
R3 R4
wherein:
Rla is selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
(R10)2N-C(NR10)-, R10C(O)-, or R10OC(O)-, and
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R10O-,
R 1 l S(O)m, R 10c(o)NR l 0-~ CN, (R1 0)2N-C(NR l 0)-,
R10C(O)-, R10OC(O)-, N3, -N(Rl0)2, or
Rl 1Oc(o)NRlo;
Rlb is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, (R10)2N-C(NR10)-, R10C(O)-, or R10OC(O)-,
and
c) Cl-C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, R10O-, Rl lS(O)m-, R10C(O)NR10-, CN,
(R10)2N C(NR10), R10C(O)-~ R10OC(O)-~ N3,
-N(R10)2, or
R 1 1 oc(o)NRlo;
provided that Rlb is not RlOC(O)NR10- when Rla is alkenyl,
V is hydrogen and X-Y is -C(o)NR7a-;
W096t09836 ~2 ~ 13 ~ 6 PcTluS95/12319
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,
o wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, R1 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
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o 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-c2o
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2 R1OO, Rl 1S(o)m, R10c(o)NRlo-~ CN,
(R10)2N-C(NR 10), R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, R1 1OC(O)NR10- and Cl-c2o alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl;
W096/09836 2a ~13 4 6 PCI'tUS95/12319
R3 and R4 are combined to form - (CH2)s -,
RSa and RSb independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized forrn of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
o c) substituted or unsubstituted Cl-c2o alkyl, C2-c2o
aLkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, RlOO-, Rl 1S(O)m-, R1OC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(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
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, -NC(O)-, and -N(COR 1 O)-;
W0 96/09836 ~ 6 PCT/US95/12319
X-Y is
~7a
a) ~ssS~N~5s
0
R7b
b) \sss~N~ss
c) ~ O~
(, )m
d) ~5ss~S~ss
H
e) ~SSs~ , or
H
f) -CH2-CH2-;
R7a is selected from
a) hydrogen,
2s 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
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
W096/09836 22 0 1 ~ 4 S PCT/US95/12319
- 10-
c) unsubstituted or substituted heterocyclic,
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 C1-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
o heterocyclic and cycloaIkyl, 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 lS(O)m-,
R1OC(O)NR10-, CN, NO2, R102N-C(NR10)-,
R1OC(O)-, R1OOC(O)-, N3, -N(R10)2, or
R1 loc(o)NRlo-~ and
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl,
F, Cl, Br, R1OO-, R1 1S(O)m-, R1OC(O)NH-, CN, H2N-
C(NH)-, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
R 1 1 OC(O)NH-;
0 R9 is selected from:
hydrogen, C 1 -C6 alkyl, R 1OO-, R l l S(O)m-,
R1OC(O)NR10-, CN, NO2, N3, -N(R10)2, and
Rl lOC(O)NR10;
~2 V ~ 3 ~ 6
PCIIUS95/12319
-- WO 96/09836
provided that R9 is not RlOC(O)NR10- when Rla is aL~enyl, V
is hydrogen and X-Y is -C(o)NR7a-;
R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl
5 and
aryl;
R1 l is independently selected from C1-C6 alkyl and aryl;
0 Rl2 is hydrogen or Cl-C6 alkyl;
V is selected from:
a) aryl;
b) heterocycle; or
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-, -S02NH-,
-N(R7a)- or -N[C(o)R7a]-;
20 Z is independently H2 or O;
misO, l or2;
n is O, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
25 p is 0, l, 2, 3 or 4, provided that p ~ 0 when R9 is not hydrogen or
Cl-C6 lower alkyl;
r is O or l ;
s is 4 or 5; and
t is 0, l or 2, provided that t = O when V is hydrogen;
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:
W096/09836 22 0 ~ 3 4 6 PCT/US95/12319
(R8)t Z R2a R2b z R5a R~b
(CR1a2) - W - (CR1b2)p(CR1bR ), R12 ~ ~ ` 6
R3 R4
wherem:
R1a is selected from:
o a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
(R10)2N-C(NR10)-, RlOC(O)-, or R1OOC(O)-, and
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-,
R1 1S(O)m, R1OC(O)NR10-~ CN, (R10)2N-C(NR10)-
R1OC(O)-, R1OOC(O)-, N3, -N(R10)2, or
Rl lOC(O)NR10;
Rlb is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, (R1 0)2N-C(NR10)-, R 1 C(O)-, or R 1 OC(O)-,
and
c) Cl -C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, RlOO-, Rl lS(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, RlOOC(O)-, N3,
-N(R10)2, or Rl lOC(O)NR10-;
provided that R1b is not R1OC(O)NR10- when Rla is alkenyl,
V is hydrogen and X-Y is -C(o)NR7a-;
R2a and R2b are independently selected from:
a) a side chain of a naturally occu~ing amino acid,
W096109836 ~2 ~ ~ ~ 4 6 PCT/US95/12319
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-C10 cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, C1, Br,
NO2, R1OO-, Rl 1S(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
o -N(R10)2, R11OC(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 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
ii) methionine sulfone,
c) substituted or unsubstituted C1 -C20 alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, NO2, RlOO, 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 -;
W096/09836 22 n 13 4 6 PCTIUS95/12319
- 14-
RSa and RSb 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 -c2o alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2 R10O-, R1 lS(O)m, R10C(O)NR10-~ CN,
(Rl0)2N-c(NRlo)-~ R10C(O)-~ R10OC(O)-~ N3,
-N(R10)2, RllOC(O)NR10- and C1-C20 alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1Q cycloalkyl; or
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)-;
20 R6 is
a) substituted or unsubstituted C1 -C~ alkyl, wherein the
substituent on the alkyl is selected from:
1) aryl,
2) heterocycle,
3) -N(R1 1)2,
4) -OR 10, or
b)
R12 0
~loJI~ R13
3 4 ~ `
WO 96/09836 - PCT/US95/12319
X-Y is
R7a
a) ~SSs~Nsss
O
R7b
b) \5sS~ N~ss
c) ~ O~
(I )m
d) ~5ss~S~ss
e) ~5SS~ , 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) Cl-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,
W 0 96/09836 22 0 ~ ~ 4 PC~rAUS95/12319
- 16 -
c) unsubstituted or substituted heterocyclic,
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,
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,
cycloalkyl and Cl-C6 aL~yl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R~S is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(O)m-,
R 1 0C(O)NR 10 , CN, NO2, R 1 02N-C(NR 10
R10C(O)-, R10OC(O)-, N3, -N(R10)2, or
R 1 1 OC(O)NR 10-, and
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyL alkynyl, perfluoroalkyl,
F, Cl, Br, R10O-, Rl lS(O)m-, R10C(O)NH-, CN, H2N-
C(NH)-, RlOC(O)-, RlOOC(O~-, N3, -N(R10)2, or
R 1 1 OC(O)NH-;
0 R9 is selected from:
hydrogen, C1-C6 alkyl, R10O-, Rl lS(O)m-,
R10C(O)NR10-, CN, NO2, N3, -N(R10)2, and
R 1 1 OC(O)NR 10 ;
W096/09836 ~2 fl ~ 3 4 6 PCI/US95/12319
provided that R9 is not RlOC(O)NR10- when Rla is aLkenyl, V
is hydrogen and X-Y is -C(o)NR7a-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
5 and
aryl;
Rl 1 is independently selected from Cl-C6 alkyl and aryl;
o R12 is independently selected from hydrogen and Cl-C6 alkyl;
R13 is Cl -C6 alkyl;
V is selected from:
a) aryl;
b) heterocycle; or
c) hydrogen,
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-, -S02NH-,
-N(R7a)- or-N[C(O)R7a];
Z is independently H2 or 0;
m is O, 1 or 2;
25 n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
C 1 -C6 lower alkyl;
r is O or l ;
30 sis40r5;and
`J t is 0, 1 or 2, provided that t = O when V is hydrogen;
or the pharmaceutically acceptable salts thereof.
W096/09836 2~ ~ ~ 3 ~ 6 PCT/US95/12319 ~
- 18-
In a third embodiment of this invention, the inhibitors of
farnesyl transferase are illustrated by the formula m
HOCH2(CH2)q
a ) - W - (CRlb2)p(CR1bR )r NR12 ~ H
R3 R4
111
wherein:
Rla is selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
(R10)2N-C(NR10)-, RlOC(O)-, or RlOOC(O)-, and
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-,
R1 lS(O)m-~ RlOC(O)NR10-~ CN, (Rl0)2N-C(NR10)-,
R1OC(O)-, RlOOC(O)-, N3, -N(R10~2, or
Rl lOC(O)NR10;
Rlb is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, cycloalkyl, alkenyl,
2 s alkynyl, (R l 0)2N-C(NR 1 0)-, R l OC(O)-, or R l OC(O)-,
and
c) Cl-C6 alkyl unsubstituted or .substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, RlOO-, Rl lS(O)m-, RlOC(O)NRl0-, CN,
(R10)2N-C(NRlO)-~ R1OC(O)-, RlOOC(O)-, N3,
-N(R l )2, or
R l l OC(O)NR l 0 ;
provided that Rlb is not Rl OC(O)NR10- when Rla is alkenyl,
V is hydrogen and X-Y is -C(o)NR7a-;
W0 96/09836 ~ ti PCT/US95112319
- 19- .
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-C10 cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
0 NO2 R100-, Rl lS(O)m, R10C(O)NR10-~ CN,
(R10)2N C(NR10), R10C(O)-~ R100C(o)-, N3,
-N(R10)2, Rl 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; 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-c2o alkyl, C2-c20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R100-, Rl lS(O)m-, R10C(O)NR10-, CN,
(R10)2N C(NR10), Rloc(o)-~ R10oc(o)-~ N3,
3 o -N(R 1)2, R 1 1 OC(O)NR 10 and C 1 -C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
W0 96/09836 2 ~ 6 PCI/US95/12319
- 20 -
R3 and R4 are combined to form - (CH2)s -;
X-Y is
R7a
a) ~sss~N~ss
o
R7b
b)\5sS~ N~ss
c) ~ O~
1 5 ()m
d) ~SsS~s~Ss
e) ~555~5sS , or
H
f) -C H2-C H2-
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
3 o e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
~ 2 Q ~
WO 96/09836 PCT/US95/12319
- 21 -
R7b is selected from
a) hydrogen,
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
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 aIkyl 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-
Rl0C(o)NRl0-, CN, NO2, Rl02N-C(NR10)-,
R10C(O)-, Rl0OC(O)-, N3, -N(Rl0)2, or
R l l OC(O)NR l 0-, and
c) Cl -C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl,
F, Cl, Br, Rl0O-, Rlls(o)m-~ Rl0C(O)NH-, CN, H2N-
C(NH)-, Rl0C(O)-, Rl0OC(O)-, N3, -N(Rl0)2, or
3 R l l OC(O)NH-;
W 0 96/09836 ~ ~ a ~ ~ 4 6 PC~rnuS95/12319
- 22 -
R9 is selected from:
hydrogen, C1-C6 alkyl, R100-, Rl 1S(O)m-,
R1OC(O)NR10-, CN, N02, N3, -N(R10)2, and
R1 1 OC(O)NR1 -;
provided that R9 is not RlOC(O)NR10- when Rla is alkenyl, V
is hydrogen and X-Y is -C(o)NR7a-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
and
o aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
R12 is hydrogen or Cl-C6 alkyl;
V is selected from:
a) aryl;
b) heterocycle; or
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-,
-S02NH-, -N(R7a)- or-N[C(O)R7a]-;
Z is independently H2 or 0;
misO, 1 or2;
n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
3 o C1-C6 lower alkyl;
qisO, 1 or2;
risOor l;
sis40r5;and
t is 0, 1 or 2, provided that t = O when V is hydrogen;
W0 96t09836 ~ 6 PCT/US95/12319
- 23 -
or the ph~ ceutically acceptable salts thereof.
In a fourth embodiment of this invention the prodrugs of
5 compounds of formula III are illustrated by the formula IV:
(I 8)t J~ ~" Z
V - (CR 2)n - W - (CR1b2)p(CR1bR9) N X'Y~ H O
R3 R4
IV
wherein:
R l a is selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
(R10)2N-C(NR10)-, R1OC(O)-, or R1OOC(O)-, and
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R1OO-,
Rl lS(o)m, RlOC(o)NRlo-~ CN, (R10)2N-C(NR10)-,
R 1 C(O)-, R 1 OC(O)-, N3, -N(R 1 )2, or
R 1 1 OC(O)NR 10 ;
R1b is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, (R10)2N-C(NR10)-, R1OC(O)-, or R1OOC(O)-,
and
c) C1-C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
alkynyl, RlOO-, R1 1S(O)m-, R1OC(O)NR10-, CN,
(RlO)2N C(NR10), R1OC(O)-, RlOOC(O)-~ N3,
-N(R 1)2, or R 1 1 OC(O)NR10-;
W096/09836 ~a~ll34~i PCT/US95/12319
- 24 -
provided that Rlb is not RlOC(O)NR10- when Rla is aLkenyl,
V is hydrogen and X-Y is -C(o)NR7a-;
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-c2o alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R10O-, Rl 1S(O)m-, R10C(O)NRl0-~ CN,
(R10)2N C(NR10), Rloc(o)-~ R10oc(o)-~ N3,
-N(R 1 0)2, R 1 1 OC(O)NR 1 0- 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,
3 wherein the substituent is selected from F, Cl, Br,
NO2, R10O-, Rl lS(O)m-, R10C(O)NR10-~ CN,
(R 1 0)2N-C(NR 10) , R 1 C(O)-, R 1 OC(O)-, N3,
-N(R1)2, R11OC(O)NR10 and Cl-C20 alkyl, and
~ W096/09836 ~2 ~ ~ ~ 4 ~ PCT/US95/12319
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 -;
X-Y is
R7a
a) ~S5s NSsS
R7b
b) \sSs~N~ss
c) ~,
(I )m
d) ~Sss~S~ss
e) ~sss~ , or
H
f) -C H2-C H2-
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloaLkyl, and
W096/09836 ~ 4 6 PCT/US95/12319
- 26 -
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,
o 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 C1-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, RlOO, RllS(o)m
R 10C(O)NR 10, CN, NO2, R 102N-C(NR10)-,
R1OC(O)-, R1OOC(O)-, N3, -N(R10)2, or
R1 lOC(O)NR10-, and
c) C1-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-
~ W091~ 1i`36 ~111346 PC~US95/1~319
C(NH)-, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
Rl lOC(O)NH-;
~,
R9 is selected from:
hydrogen, C1 -C6 alkyl, R100-, R1 1S(O)m-,
R1OC(O)NR10-, CN, N02, N3, -N(R10)2, and
Rl lOC(O)NR10;
provided that R9 is not R1OC(O)NR10- when R1a is alkenyl, V
is hydrogen and X-Y is -C(o)NR7a-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
and
aryl;
5 Rl 1 is independently selected from C1-C6 alkyl and aryl;
R12 is hydrogen or C1-C6 alkyl;
V is selected from:
a) aryl;
b) heterocycle; or
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHso2
-S02NH-, -N(R7a)- or-N~C(O)R7a]-;
Z is independently H2 or 0;
m is 0, 1 or2;
3 n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
Cl-C6 lower alkyl;
qisO, 1 or2;
~ n ~ 3 ~ 6
WO 96/09836 - PCT/US95/12319
- 28 -
r is O or 1 ;
s is 4 or 5, and
t is 0, 1 or 2, provided that t = O when V is hydrogen;
5 or the pharmaceutically acceptable salts thereof.
In a more preferred embodiment of the invention, the Ras
farnesyl transferase inhibitors are illustrated by the formula I:
10 (R8)t z R ~R z R ~R
(CRla2) -W-(CR1b2)p(CR1 R )r NRl2 ~N~ OH
R3 R4
I
wherein:
R 1 a is selected from:
a) hydrogen, and
b) Cl-C6 alkyl;
Rlb iS independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl or cycloalkyl, and
c) Cl -C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
Rl- 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,
3 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,
N02, R100-, Rl 1S(O)m-, R1OC(O)NR10-~ CN,
~2 ~ 6
WO 96/09836 PCI/US95/12319
- 29 -
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, RllOC(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-
Clo 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 C1 -Clo alkyl, C2-C10
aL~enyl, C3-C1o cycloalkyl, aryl or heterocycle group,
wherein the substituent is selected from F, Cl, Br,
NO2, RlOO-, R1 1S(O)m-, R1OC(O)NR10-, CN,
(R10)2N C(NR10), RlOc(o)-~ RlOOC(O)-, N3,
-N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and
d) Cl-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o 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,
W096109836 ~!~ fl 11 3 4 6 PCI/US95/12319
- 30 -
c) substituted or unsubstituted Cl-clo alkyl, C2-C10
alkenyl, C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02, R100-, Rl lS(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, RllOC(O)NR10- and Cl-C20 alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl;
RSb is selected from:
a) hydrogen, and
b) Cl-C3 alkyl, or
5 X-Y is
R7a
a) ~ N5ss
o
R7b
b) \sSs~N~sss
c)~ ~O~ss
d) ~5sS ~55S~ , or
H
e) -CH2-CH2-;
W096/09836 ~2 ~ ~ ~ 4 ~ PCT/US95/12319
- 31 -
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
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
wherein heterocycle is selected from pyrrolidinyl,
o imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
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) 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
3 0 an unsubstituted or substituted group selected from aryl,
- heterocyclic and cycloalkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
W096/09836 2~ ~ 11 3 4 6 PCI/US95/12319
- 32 -
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R8 is independently selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-C6 alkenyl, C2-c6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, R1OO-, R1OC(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2,
or R l lOC(O)NR1 0-, and
o c) Cl-C6 alkyl substituted by Cl-C6 perfluoroaLkyl,
R100-, RlOC(O)NR10-~ (R10)2N-c(NRlo)-~ RlOC(O)-,
R1OOC(O)-, -N(R10)2, or Rl lOC(O)NR10-;
R9 is selected from:
hydrogen, Cl-C6 lower alkyl, R100-, and -N(R10)2;
R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl
and
aryl;
K11 is independently selected from C1-C6 alkyl and aryl;
R12 iS hydrogen or Cl-C6 aLkyl;
25 V is selected from:
a) aryl;
b) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl,
quinolinyl, isoquinolinyl, and thienyl; and
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-,
-S02NH-, -N(R7a)- or-N~C(O)R7a]-;
WO 9G~r~3~ ~ 2 ~ ~ 3 4 ~ P~ 5S/12319
- 33 -
Z is independently H2 or 0;
misO, 1 or2;
n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
C1-C6 lower aL~yl;
r is O or l ;
sis40r5;and
10 t is O or 1, provided that t = O when V is hydrogen;
or the pharmaceutically acceptable salts thereof.
In a second more preferred embodiment of the invention,
15 the prodrugs of the Ras farnesyl transferase inhibitors of the formula I
are illustrated by the formula I:I:
(R8)t z R2a R2b z R5a R5b
(CR1a2) -W-(CR1b2)p(CR1bR ), R12 ~H
Il R3 R
wherein:
Rla is selected from:
a) hydrogen, and
b) Cl-C6 alkyl;
Rlb is independently selected from:
a) hydrogen,
3 b) unsubstituted or substituted aryl or cycloalkyl, and
c) Cl -C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, cycloalkyl, alkenyl,
R100- or -N(R10)2;
wog~Y~ 3 4 6 PCT/US95/12319 ~
- 34 -
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, R1OO-, Rl 1S(O)m-, R1OC(O)NR10-, CN,
(R10)2N C(NR10), R1OC(O)-, R1OOC(O)-~ N3,
o -N(R10)2, Rl lOC(O)N~10- and Cl-C20 alkyl, and
c) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; and
15 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 Cl-clo alkyl, C2-C10
alkenyl, C3-C1o cycloalkyl, aryl or heterocycle group,
wherein the sub.stituent is selected from F, Cl, Br,
NO2 RlOO, Rl lS(o)m, R10c(o)NRlo-~ CN,
(R10)2N C(NR10), R10c(o)-~ R1OOC(O)-, N3,
-N(R10)2, R1 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;
WO 96/09836 ~ 2 ~ ~ 3 ~ ~ PCTIUS95/12319
R5a is selected from:
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from
methionine and gl~ le~
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-C10
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2 R10O-, R1 1S(O)m, R10C(O)NR10-~ CN,
(R10)2N C(NR10), R10C(O)-, R10OC(O)-~ N3,
-N(R10)2, R1 1OC(O)NR10- and C1-C20 alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl;
R5b is selected from:
a) hydrogen, and
b) C1 -C3 alkyl; or
R6 is
a) substituted of unsubstituted Cl-c8 alkyl, wherein the
substituent on the alkyl is selected from:
l)aryl,
2) heterocycle,
3) -N(R1 1)2,
4) -OR 10, or
b)
R12 O
WO 96/09836 PCI/US95/12319
~a ~ 7 3 4 6
- 36 -
X-Y is
R7a
a) ~sss~N~g
0
R7b
b) \SsS~ N~SsS
c) ~ O~s~
d) ~~
H
e) -CH2-CH2-;
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsub,stituted or substituted cycloalkyl, and
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
a) hydrogen,
-
W096/09836 ~ ~ 9 ~ ~ 6 PCT/US95/12319
- 37 -
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 C 1 -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 C1-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 independently selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN, N02,
(RlO)2N C(NR10), RlOC(O)-~ RlOOC(O)-, -N(R10)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)-,
RlOOC(O)-, -N(R10)2, or R1 1OC(O)NR10-;
R9 is selected from:
hydrogen, C1-C6 lower alkyl, R100-, and -N(R10)2;
W 0 96/09836 22 Q ~ 3 ~ 6 PC~r~US95/12319
- 38 -
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
and
aryl;
5 R11 is independently selected from Cl-C6 alkyl and aryl;
R12 is independently selected from hydrogen and Cl-C6 alkyl;
R 1 3 is C l -C6 alkyl;
V is selected from:
a) aryl;
b) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl,
quinolinyl, isoquinolinyl, and thienyl; and
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHSO2-,
-S0~2NH-, -N(R7a)- or-N[C(O)R7a]-;
Z is independently H2 or O;
misO, 1 or2;
n is 0, 1, 2, 3 or 4, provided that n ~ 0 when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ 0 when R9 is not hydrogen or
C 1 -C6 lower alkyl;
r is 0 or 1 ;
s is 4 or 5; and
3 t is 0 or 1, provided that t = 0 when V is hydrogen;
or the pharmaceutically acceptable salts thereof.
~ Q q 3~6
WO 96/09836 L_ PCTlUS9~i/12319
- 39 -
In a third more preferred embodiment of the invention,
the Ras farnesyl transferase inhibitors are illustrated by the formula
III:
HOCH2(CH2)q
V - (CR1 a2)n - W - (CR1 b2)p(CR1 bR9),/l~N~X' ~ NH
R3 R4
wherein:
Rla is selected from:
a) hydrogen, and
b) C1-C6 alkyl;
R1b are independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl or cycloalkyl, and
c) C 1 -C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, 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-, R1 1S(O)m-, Rloc(o)NRlo- CN
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(RlO)æ Rl 1OC(O)NR10- and Cl-C20 alkyl, and
WO 96109836 2 2 a ~ ~ 4 6 PCT/US95/12319
- 40 -
c) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; and
5 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:
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 -Clo alkyl, C2-clo
alkenyl, C3-C1o cycloalkyl, aryl or heterocycle group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, Rl lS(O)m-, R1OC(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) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl;
WO 96/09836 2 ~ PCT/US95/12319
- 41 -
X-Y is
- R7a
a) ~ Nsss
0
R7b
b) \sSS~ N~ss
c) ~ O~
d
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
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,
22 0 ~ 3 4 6 ~
WO 96/09836 PCT/US95/12319
- 42 -
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) Cl -C6 aLkyl substituted with hydrogen or an
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,
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-u-nsubstituted 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;
R~ is independently ~selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-C6
2 5 perfluoroalkyl, F, Cl, R l Oo , R l OC(O)NR 1 0-, CN, NO2,
(R 1 0)2N-C(NR 10) , R 1 C(O)-, R 1 OC(O)-, -N(R 1 )2,
or Rl lOC(O)NR10-, and
c) Cl-C6 alkyl ,substituted by Cl-C6 perfluoroalkyl,
R l Oo , R 1 OC(O)NR 10, (R 1 0)2N-C(NR 1 0), R 1 C(O)-,
3 o R 10oC(O~-, -N(R 1)2, or R 1 lOC(O)NR10-;
R9 is selected from:
hydrogen, Cl -C6 lower alkyl, RlOO-, and -N(R10)2;
4 6 PCT/US95/12319
WO 96/09836
- 43 -
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
and
aryl;
5 R l 1 is independently selected from C1 -C6 alkyl and aryl;
R l 2 is hydrogen or Cl -C6 alkyl;
V is selected from:
o a) aryl;
b) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl,
quinolinyl, isoquinolinyl, and thienyl; and
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-,
-S02NH-, -N(R7a)- or-N[C(O)R7a]-;
Z is independently H2 or 0;
misO, 1 or2;
n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
Cl -C6 lower aL~yl;
qisO, 1 or2;
r is O or 1 ;
s is 4 or S; and
t is O or 1, provided that t = O when V is hydrogen;
- or the pharmaceutically acceptable salts thereof.
W096t09836 ~!~ Q ~ 3 ~ 6 PCT/US95/12319
- 44 -
In a fourth more preferred embodiment of the invention,
the prodrugs of the Ras farnesyl transferase inhibitors of the formula
m are illustrated by the formula IV:
(R8)t z R~R2~ z I q
V - (CR 2)n - W- (CRlb2)p(CR1bR9) N X ~N O
R~ R4
IV
wherein:
Rla is selected from:
a) hydrogen, and
b) C1-C6 alkyl;
Rlb are independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl or cycloalkyl, and
c) C l-C6 alkyl unsubstituted or substituted by
unsubstituted or substituted aryl, 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-clo alkyl, C2-C10
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,
(R10)2N C(NR10), RlOc(o)-~ RlOOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- ~nd Cl-C20 alkyl, and
WO 96/09836 ~ 3 ~ PCT/US95112319
- 45 -
c) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycioaLkyi; and
5 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:
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 C1-Clo alkyl, C2-C10
alkenyl, C3-C1o cycloalkyl, aryl or heterocycle group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, Rl 1S(O)m-, RlOC(O)NR10-, CN,
(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-
C1o cycloalkyl;
W096/09836 ~aQ~'3~6 PCI~/US95112319 1-
- 46 -
X-Y is
R7a
a) ~Sssb,N~s5s
0
R7b
b) \sSS~ N~SsS
0
c) ~ O~
lS d) 55S~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
2s e) C1-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,
W0 96,0g836 ~ 6 PCI/US95/12319
- 47 -
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloaL~yl,
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 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;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R8 is independently selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, C1-C6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN, N02,
(R10)2N C(NR10), RlOC(O)-~ RlOOC(O)-, -N(RlO)2
or R 1 1 OC(O)NR 10, and
c) Cl-C6 alkyl substituted by Cl-C6 per~uoroalkyl,
R10O-, RlOC(O)NR10-~ (R10)2N-C(NR10)-, RlOC(O)-,
R10OC(O)-, -N(R10)2, or R1 1OC(O)NR10-;
R9 is selected from:
hydrogen, Cl -C6 lower alkyl, R100-, and -N(R10)2;
22~346 ` ~
WO 96/09836 PCT/US95/12319
- 48 -
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl
and
aryl;
5 Rl 1 is independently selected from Cl-C6 alkyl and aryl;
R12 is hydrogen or Cl-C6 aL~yl;
V is selected from:
o a) aryl;
b) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl,
quinolinyl, isoquinolinyl, and thienyl; and
c) hydrogen;
W is -S(O)m-, -O-, -NHC(O)-, -C(O)NH-, -NHS02-,
-S02NH-, -N(R7a)- or-N~C(O)R7a]-;
Z is independently H2 or 0;
m is 0, 1 or2;
n is 0, 1, 2, 3 or 4, provided that n ~ O when V is hydrogen and W is
-S(O)m-;
p is 0, 1, 2, 3 or 4, provided that p ~ O when R9 is not hydrogen or
C1-C6 lower alkyl;
qisO, 1 or2;
risOor 1;
s is 4 or 5; and
t is O or 1, provided that t= O when V is hydrogen;
or the pharmaceutically acceptable salts thereof.
The preferred compounds of this invention are as
follows:
WO ~ 36 PCTIUS95/12319
- 49 -
N- { 2(S)-[(4-Nitrobenzylthio)acetamido] -3 (S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
s N- { 2(S)-[(4-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl-methionine
N- { 2(S)-[(Benzylthio)acetamido] -3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl-methionine
N-{2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl}-N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(Phenylthio)acetamido]-3(S)-methylpentyl } -N-(1-
s naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(Phenylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
20 N- { 2(S)-[(3-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl-methionine
N- { 2(S)-[(3-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthyimethyl)-glycyI-methionlne methyl ester
2S
N- { 2(S)-[(2-Nitrobenzylthio)acetamido]-3(S)-methylpentyl ) -N-(1-
naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(2-Nitrobenzylthio)acetamido] -3 (S)-methylpentyl } -N-(1-
30 naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-Cyanobenzylthio)acetamido]-3 (S)-methylpentyl } -N-( 1-
naphthylmethyl) -glycyl-methionine
~a~346 ~,
WO 96/09836 PCT/US95/12319
- 50 -
N- ~ 2(S)-[(4-Cyanobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-Trifluoromethylbenzylthio)acetamido]-3(S)-methyl-
5 pentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Trifluoromethylbenzylthio)acetamido]-3(S)-methyl-
pentyl}-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
o N- { 2(S)-[(4-Methoxybenzylthio)acetamido]-3(S)-methylpentyl } -N-
( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Methoxybenzylthio)acetamido]-3(S)-methylpentyl } -N-
(1-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-Methylsulfonylbenzylthio)acetamido]-3(S)-methyl-
pentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Methylsulfonylbenzylthio)acetamido]-3(S)-methyl-
20 pentyl)-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-Phenylbenzylthio)acetamido] -3(S)-methylpentyl } -N-( 1-
naphthylmethyl) -glycyl-methionine
25 N- ~ 2(S)-[(4-Phenylbenzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-(1 -H-Tetrazol-5-yl)benzylthio)acetamido]-3(S)-methyl-
pentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-( 1 -H-Tetrazol-5-yl)benzylthio)acetamido]-3(S)-methyl-
pentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine methyl ester
WO 96/09836 PCTIUS95/12319
N-{2(S)-[(4-Methylbenzylthio)acetamido]-3(S)-methylpentyl}-N-(1-
naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Methylbenzylthio)acetamido] -3(S)-methylpentyl } -N-( 1-
5 naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(8-Chloronaphth-1 -ylthio)acetamido]-3(S)-methylpentyl } -
N-(1-naphthylmethyl)-glycyl-methionine
o N-{2(S)-[(8-Chloronaphth-l-ylthio)acetamido]-3(S)-methylpentyl}-
N-(l-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[((2-Methylindol-3-yl)thio)acetamido]-4-methylpentyl } -N-
( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[((2-Methylindol-3-yl)thio)acetamido]-4-methylpentyl } -N-
(1-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(4-Pyridylthio)acetamido]-3(S)-methylpentyl } -N-(1-
2 0 naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Pyridylthio)acetamido]-3(S)-methylpentyl } -N-(l -
naphthylmethyl)-glycyl-methlonine methyl ester
2 S N- { 2(S)-[(4-Picolinylthio)acetamido] -3 (S)-methylpentyl } -N-( 1-
naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(4-Picolinylthio)acetamido] -3 (S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[3 -(Benzylthio)propionamido] -3 (S)-methylpentyl } -N-( 1 -
naphthylmethyl) -glycyl-methionine
WO 96/09836 2 2 Q lT 3 4 6 PCT/US95/12319
- 52 -
N- { 2(S)-[(Benzyloxy)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl-methionine
N- { 2(S)-[(Benzyloxy)acetamido] -3(S)-methylpentyl } -N-(1-
5 naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(N'-Benzylglycyl)amino]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine
10 N- { 2(S)-[(N'-Benzylglycyl)amino]-3(S)-methylpentyl } -N-( 1-
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(N'-Acetyl-N~-benzylglycyl)amino]-3(S)-methylpentyl } -N-
( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(N'-Acetyl-N'-benzylglycyl)amino]-3(S)-methylpentyl } -N-
(l-naphthylmethyl)-glycyl-methionine methyl ester
N- ~ 2(S)-r(N'-Benzyl-N'-t-butoxycarbonylglycyl)amino]-3(S)-
2 o methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(N'-Benzyl-N'-t-butoxycarbonylglycyl)arnino]-3(S)-methyl-
pentyl}-N-(I-naphthylmethyl)-glycyl-methionine methyl ester
25 N- { 2(S)-[(Benzylthio-S-oxide)acetamido]-3(S)-methylpentyl } -N-(I -
naphthylrnethyl) -glycyl-methionine
N- { 2(S)-[(Benzylthio-S-oxide)acetamido] -3(S)-methylpentyl } -N-(I -
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(Benzylthio-S,S-dioxide)acetamido]-3(S)-methylpentyl } -N-
( 1 -naphthylmethyl)-glycyl-methionine
WO 96/09836 - PCTIUS9S/12319
- 53 -
N- { 2(S)-[(Benzylthio-S,S-dioxide)acetamido]-3(S)-methylpentyl } -N-
(1-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
5 naphthylmethyl)-glycyl-methionine sulfone
N- { 2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine sulfone methyl ester
0 N- { 2(S)-[(N'-4-Nitrobenzoylglycyl)amino]-3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl-methionine
N-{2(S)-[(N'-4-Nitrobenzoylglycyl)amino]-3(S)-methylpentyl}-N-(1 -
naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(N'-Methyl-N'-4-nitrobenzoylglycyl)amino]-4-methyl-
pentyl } -N-(1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(N'-Methyl-N'-4-nitrobenzoylglycyl)amino]-4-methyl-
2 o pentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine methyl ester
N- ~ 2(S)-[(S-Benzyl-L-cysteinyl]-3(S)-methylpentyl } -N-(1-
naphthylmethyl) -glycyl -methionine
2s N- { 2(S)-[S-Benzyl-L-cysteinyl]-3(S)-methylpentyl }-N-(1 -naphthyl-
methyl)-glycyl-methionine methyl ester
N- { 2(S)-[S-Benzyl-D-cysteinyl]-3(S)-methylpentyl } -N-(1 -naphthyl-
methyl)-glycyl-methionine
- N- { 2(S)-[S-Benzyl-D-cysteinyl]-3 (S)-methylpentyl } -N-( 1 -naphthyl-
methyl)-glycyl-methionine methyl ester
3 -Benzylthiopropionyl-valyl-isoleucyl-methionine
W 0 96/09836 22 0 1 3 4 ~ PC~rnUS95/12319
- 5 4 -
N-[2(S)-(2(S),3-Diaminopropionyl)amino-3(S)-methylpentyl] -N-(1 -
naphthylmethyl)glycyl-methionine
N-[2(S)-(2(S),3-Diaminopropionyl)amino-3(S)-methylpentyl]-N-(l -
5 naphthylmethyl)glycyl-methionine methyl ester
N-[2(S)-(3-Aminopropionyl)amino-3(S)-methylpentyl]-N-(1 -
naphthylmethyl)glycyl-methionine
o N-[2(S)-(3-Aminopropionyl)amino-3 (S)-methylpentyl] -N-( 1 -
naphthylmethyl)glycyl-methionine methyl ester
N-(2(S)-L-Glllt~minylamino-3(S)-methylpentyl)-N-(l -naphthyl-
methyl)glycyl-methionine
N-(2(S)-L-Glutaminylamino-3(S)-methylpentyl)-N-( 1 -naphthyl-
methyl)glycyl-methionine methyl ester
N-[2(S)-( 1,1 -Dimethylethoxycarbonylamino)-3S)-methylpentyl] -N-
2 o ( 1 -naphthylmethyl)glycyl-methionine
N-~2(S)-( 1,1 -Dimethylethoxycarbonylamino)-3S)-methylpentyl]-N-
(l-naphthylmethyl)glycyl-methionine methylester
25 N-[2(S)-(l,l-Dimethylethoxycarbonylamino)-3(S)-methylpentyl]-N-
benzylglycyl-methionine
N-[2(S)-( 1,1 -Dimethylethoxycarbonylamino)-3(S)-methylpentyl] -N-
benzylglycyl-methionine methyl ester
N- ~ [ 1 -(4-Nitrobenzylthio)acetylamino]cyclopent-l -ylmethyl } -N-(l -
naphthylmethyl)-glycyl-methionine
N- { [ 1 -(4-Nitrobenzylthio)acetylamino]cyclopent- 1 -ylmethyl } -N-( 1-
naphthylmethyl)-glycyl-methionine methyl ester
W096/09836 ~ n ~ PCT/US95/12319`
N- { 2(S)-[2-(4-Nitrophenyl)ethylcarbamoylamino] -4-methylpentyl } -
N-( 1 -naphthylmethyl)-glycyl-methionine
5 N- { 2(S)-[2-(4-Nitrophenyl)ethylcarbamoylamino]-4-methylpentyl } -
N-(l-naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(3(S)-tetrahydrofuryloxy)carbonylamino]-3(S)-
methylpentyl } -N-(l -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(3(S)-tetrahydrofuryloxy)carbonylamino]-3(S)-
methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine methyl ester
N- { 2(S)-[(cis-4-benzyloxytetrahydrofur-3-yloxy)carbonylamino] -
3(S)-methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(cis-4-benzyloxytetrahydrofur-3-yloxy)carbonylamino]-
3 (S)-methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine methyl
ester
N- { 2(S)-[(N'-Methyl-N'-4-nitrophenylacetylglycyl)amino]-4-
methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
N- { 2(S)-[(N'-Methyl-N'-4-nitrophenylacetylglycyl)amino]-4-
2 5 methylpentyl } -N-~ 1 -naphthylmethyl)-glycyl-methionine methyl ester
or the pharmaceutically acceptable salts thereof.
The most preferred compounds of the invention are:
N- { 2(S)-[(4-Methoxybenzylthio)acetamido]-3(~)-methylpentyl } -N-
( 1 -naphthylmethyl)-glycyl-methionine
W096/09836 2~ 4 Ç PCI/US95/12319
- 56 -
o
~ ~ N N~ NJ~oH
H3C~ o~ O /\ ~ SCH3
N- ~ 2(S)-[(4-Methoxybenzylthio)acetamido]-3(S)-methylpentyl } -N-
o (1-naphthylmethyl)-glycyl-methionine methyl ester
H H
~N--N~NJ~oCH3
H3C oJ\~ /`1 ~3 SCH3
N-{2(S)-[(4-Cyanobenzylthio)acetamido]-3(S)-methylpentyl~-N-(1-
naphthylmethyl) -glycyl-methionine
~S~N N~ NJI~OH
- ~3 SCH3
N- { 2(S)-[(4-Cyanobenzylthio)acetamido] -3 (S)-methylpentyl ~ -N-( 1 -
naphthylmethyl)-glycyl-methionine methyl ester
H H
~S~N N~NJ~OCH3
~ /`1 ~3 SCH3
WO 96/09836 PCT/US95112319
~2 013 4 6
N-(2(S)-L-Glllt~minylamino-3(S)-methylpentyl)-N-(1 -naphthyl-
methyl)glycyl-methionine
H2N~,
H H
H2N~N N~N~oH
'`I ~3 SCH3
N-(2(S)-L-Glutaminylamino-3(S)-methylpentyl)-N-(1 -naphthyl-
methyl)glycyl-methionine methyl ester
H2N O
H H
H2N ~ N N~ NJ~ OCH3
O /~ ~3 SCH3
N- { 2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
25 naphthylmethyl)-glycyl-methionine sulfone
H H
~S~ N~, ~
N- { 2(S)-~(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine sulfone methyl ester
2ao~i346
WO 96/09836 PCT/US95112319
3--s~ --N~
or the pharmaceutically acceptable salts thereof.
WO 96/09836 PCT/US95/12319
~2 0 ~ 3 4 6
59
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
Asparticacid Asp D
Asparagine or
Aspartic acid Asx B
Cysteine Cys C
Glllt~mine Gln Q
Glutamic acid Glu E
Glutamine or
Glutamic acid Glx Z
Glycine Gly G
Histidine His H
Isoleucine Ile
Leucine Leu L
Lysine Lys K
Methionine Met M
Phenyl~l~nine Phe F
Proline Pro P
Serine Ser S
Threonine Thr T
Tryptophan Trp W
Tyrosine Tyr Y
Valine Val V
s
The compounds of the present invention may have
asymmetric centers and occur as racemates, racemic mixtures, and as
individual diastereomers, with all possible isomers, including optical
isomers, being included in the present invention.
W0 96/09836 2 ~ 6 PCT/US95112319
- - 60 -
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, "cycloaL~yl" 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
o bonds. Examples of alkenyl groups include vinyl, allyl, isopropenyl,
pentenyl, hexenyl, heptenyl, cyclopropenyl, cyclobutenyl,
cyclopentenyl, cyclohexenyl, l-propenyl, 2-butenyl, 2-methyl-2-
butenyl, 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, phenanthrenyl and the like.
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 R5a and R5b are combined to form - (CH2)s -,
cyclic moieties as described hereinabove for R2a and R'~b and R3 and t
30 R4 are formed. ln addition, such cyclic moieties may optionally
include a heteroatom(s). Examples of such heteroatom-cont~ining
cyclic moieties include, but are not limited to:
- = ~
W096/09836 ~ ~) 'I 3 4 6 PCTtUS95/12319
- 61 -
oJ ~5J
o O O H O N
The term heterocycle or heterocyclic, as used herein,
represents a stable 5- to 7-membered monocyclic or stable 8- to 11-
5 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
group consisting of N, O, and S, and including any bicyclic group in
which any of the above-defined heterocyclic rings is fused to a
20 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,
25 benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydro-benzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothio-pyranyl sulfone, furyl, imidazolidinyl, imidazolinyl,
imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl,
isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl,
3~ 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,
W096/09836 Zan134~ PCT/US95tl2319
- 62 -
tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydro-quinolinyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl,
thienofuryl, thienothienyl, and thienyl.
As used herein, the terms "substituted aryl", "substituted
5 heterocycle" and "substituted cycloalkyl" are intended to include the
cyclic group which is substituted with 1 or 2 substitutents selected
from the group which includes but is not limited to F, Cl, Br, NH2,
N(C1-C6 alkyl)2, NO2, CF3, (cl-c6 alkyl)O-, -OH, (C1-C6
alkyl)S(O)m-, (Cl-C6 alkyl)C(O)NH-, CN, H2N-C(NH)-, (Cl-C6
alkyl)C(O)-, (Cl-C6 alkyl)OC(O)-, N3~(Cl-C6 alkyl)OC(O)NH- and
Cl-C20 aL~cyl.
The pharmaceutically 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 inorganic
15 or organic acids. For example, such conventional non-toxic salts
include those derived from inorganic acids such as hydrochloric,
hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and
the salts prepared from organic acids such as acetic, propionic,
succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,
20 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
25 variable (e.g., Rl, Z, n, etc.) at a particular location in a molecule be
independent of its definitions elsewhere in that molecule. Thus,
-N(R 1 )2 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 ordinary skill in the art to
30 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 pharmaceutically acceptable salts of the compounds
of this invention can be synthesized from the compounds of this
W0961~9836 ~2 0 1 3 ~S 6 PCI/US95/12319
- 63 -
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 salt-
forming inorganic or organic acid in a suitable solvent or various
5 combinations of solvents.
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
10 following works: Schroeder et al., "The Peptides", Vol. I, Academic
Press 1965, or Bod~n.s7.ky et al., "Peptide Synthesis", Interscience
Publishers, 1966, or McOmie (ed.) "P7 0tective Groups in Organic
Che,~,lis.t7y", Plenurrl Press, 1973, or B~ally et al., "The Pepiides:
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;
DME 1,2-Dimethoxyethane;
DMF Dimethylformamide;
EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide-
hydrochloride;
HOBT l-Hydroxybenzotriazole hydrate;
3 o Et3N Triethylamine;
EtOAc Ethyl acetate;
FAB Fa.st atom bombardment;
HOOBT 3-Hydroxy- 1 ,2,2-benzotriazin-4(3H)-one;
HPLC High-performance liquid chromatography;
W096/09836 ~ ; 4 ~ ~Cr/US95/12319
- 64 -
MCPBA m-Chloroperoxybenzoic acid;
MsCl Methanesulfonyl chloride;
NaHMDS Sodium bis(trimethylsilyl)amide;
Py Pyridine;
TFA Trifluoroaceticacid;
THF Tetrahydrofuran.
Compounds of this invention are prepared by employing
the reactions shown in the following Reaction Schemes A-J, in
o 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
reductive alkylation of an amine by an aldehyde using sodium
cyanoborohydride or other reducing agents.
Reaction C. Alkylation 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.
W096/09836 ~2 ~ 1 3 4 6 PCT/US95/12319
- 65 -
REACTION SCHEME A
Reac~ion A. Coupiing of residues to form an amide bond
A RB
OH + H2N '~f
o
or HOOBT >loJ~ N J~OR
Et3N, DMFr RB
TFA H N$NJ~oR
O RB
REACTION SCHEME B
Reaction B. Preparation of reduced peptide subunits by reductive
alkylation
>~OJ~ N ~H + ~OR
R
22 0 ~ ~ 4 ~ ~
WO 96/09836 PCT/US95/12319
- 66 -
REACTION SCHEME C
Reaction C. Alkylation/reductive alkylation of reduced peptide
subunit~
~O~N~NJ~oR R XL, base
RB 1l
RCCH, NaCNBH3
RA R7b
H OR
RB
W096t09836 2~ 0 ~ 3 4 6 PCT/US95/12319
- 67 -
REACTION SCHEME D
Reaction D. Coupling of residues to form an amide bond
RA EDC, HOBT
O I ~ or HOOBT
>~oJ~N~oH + H2N~ Et3N, DMF
>~ J~ 7 NJ~HCl or TFA
~A
H2N ~ J~o
O -~,J
REACTION SCHEME E
Reaction E. Preparation of reduced dipeptides from peptides
2s >`O H~ J~OR
>loJ~N~NJl~oR
H RB
W 0 96/09836 2~ ~ 7 3 4 6 PC~r~US95/12319
- 68 -
where RA and RB are R2a, R2b, R3, R4, R5a or R5b as previously
defined; XL is a leaving group, e.g., Br-, I- or MsO-; and RC is
defined such that R7b is generated by the reductive alkylation process.
Certain compounds of this invention wherein X-Y is an
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 lltili7ing standard
manipulations such as Weinreb amide formation, Grignard reaction,
acetylation, ozonolysis, Wittig reaction, ester hydrolysis, peptide
o 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
corresponding syn amino-alcohol (Scheme F, Step B, Part 1), and
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
20 defined. In Step H of Scheme F, Rx, which represents the amino
terminus side chain substituent of the instant invention, is
incorporated using coupling reaction A and RXCOOH; the alkylation
reaction C using RXCHO and a reducing agent; or alkylation reaction
C using RxcH2xL.
2s The alkane analogs are prepared in a similar manner by
including an additional catalytic hydrogenation step a,s outlined in
Reaction Scheme G.
2~ 4 6 pCTrus95/12319
WO 96/09836
- 69 -
REACTION SCHEME F
- 1. ClCO2i-Bu o
BocNHJJ`OH MeONHMe BocNH
R2a2. I R2a
BrMg
Step A
1. NaBH4 OAc
~ BocN H ~J~
2. Ac20, Py R2a
Step B
1. O3, Me2S OAc
2. Ph3P=CHCO2Me BocNH ~CO2Me
Step C R2a
W096/098362~7 ~ 7 3 4 6 PCT/US95/12319 ~
- 70 -
REACTION SCHEME F (CONT'D)
Step D
1. LiOH OH H MsCI, py
2 EPDC, HOBT BocNH~ JlW~ Step F
amino acid (ester)
W"
W'=OMe,W"=SMe
W' -W"=O
OMs O
BocNH~ R3MgCuCNClBF3
R2a O ~ Step G
W"
1. HCI
R3 o 2. NaCNBH3
H ll RXCHO
BocNH ~ N ~~W'
R2a O ~ Step H
W"
R CH2N ~ ~ J~W'
R2a O W"
wherein Rx is:
R8 V - (CR12)n ~ W ~ (CR12)p(CRlR9)r-
W096/09836 2~n~4~ PCT/US95112319
REACTION SCHEME F (CONT'D)
NaOH
H R3 H O
RXC H2N ~~ ~OH
2 ~W"
or
1. HCI Alternate
1l Step H
2. RxcOH
EDC, HOBT
~f ~NJ~W~
O R2a o
W"
NaOH
2 5 RX N ~ OH
O R2a 0 ~'\W"
W096/09836 22 ~ ~ 3 4 6 PCT/US95112319
REACTION SCHEME G
0 1. ClCO2i-Bu o
BocNH J~OH . BocNH
R2aBrMg ~ R2a
1. NaBH4 OAc 1. 03, Me2S
BocNH~
2. Ac20, py R2a 2. Ph~P=CHCO2Me
OAc O
BocNH~cO Me + H2N~ 2 EDC, HOBT
OH H
2 0
R2a o - ~ MsCI, py
WO 96/09836 ~ PCT/US95112319
- 73 -
REACTION SCHEME G (CONT'D)
OMs O
H ll
BocNH~N
R2a O
1. R3 MgCuCNCl-BF3
2. H2, 5% Pd/C
Step K
BocNH~ ,o
R2a o -~
1. HCI
2. NaCNBH3,
RXCHO
R3 H
RXCH2NH~ o
R2a o -~
NaOH
RXCH2NH ~ OH
R2a O OH
2~ ~3~B
WO 9~ ?~ PCT/US9~/12319
- 74 -
REACTION SCHEME G (CONT'D)
or O
Il
1. HCI 2. RXCOH
EDC, HOBT
O R''CNH~~
R2a o
NaOH
RXCNH~ ~ J~ OH
R2a O OH
The oxa isostere compounds of this invention are
prepared according to the route outlined in Scheme H. An
25 aminoalcohol 1 is acylated with alpha-chloroacetyl chloride in the
presence of trialkylamines to yield amide 2. Subsequent reaction of 2
with a deprotonation reagent (e.g., sodium hydride or potassium t-
butoxide) in an ethereal solvent such as THF provides morpholinone
3. The N-Boc derivative 4 is then obtained by the treatment of ~ with
30 BOC anhydride and DMAP (4-dimethylaminopyridine) in methylene
chloride. Alkylation of 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 5, which is retreated with NaHMDS
followed by either protonation or the addition of an alkyl halide R4X
W096/09836 ~n~46 PCT/US9S/12319
- 75 -
to give 6a or 6b, respectively. Alternatively, 6a can be prepared from
4 via an aldol condensation approach. Namely, deprotonation of 4
with NaHMDS followed by the addition of a carbonyl compound
RYRZCO gives the adduct 7. Dehydration of 7 can be effected by
5 mesylation and subsequent elimin~ion catalyzed by DBU (1,8-
diazabicyclo[5.4.0]undec-7-ene) or the direct treatment of 7 with
phosphorus oxychloride in pyridine to give olefin 8. Then, catalytic
hydrogenation of ~ yields 6a. Direct hydrolysis of 6 with lithium
hydrogen peroxide in aqueous THF will produce acid 9b. Sometimes,
10 it is more efficient to carry out this conversion via a 2-step sequence,
namely, hydrolysis of 6 in hydrochloric acid to afford 9a, which is
then derivatized with BOC-ON or BOC anhydride to give 9b. The
peptide coupli~g of aGid 9b with çither an alpha-arl~inolacto~e (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 10. Treatment of 10 with gaseous
hydrogen chloride gives 11, which undergoes reductive alkylation in
the presence of an aldehyde RXCHO (12) and a reducing agent (e.g.,
sodium cyanoboro-hydride); or acylation in the presence of RXCOOH
20 (13) and a peptide coupling reagent affording the products 14a and _.
Hydrolysis of compounds 14 to the corresponding hydroxy acids and
acids, respectively, is accomplished by standard methods such as
treatment with NaOH in alcoholic or aqueous milieux followed by
careful acidifcation with dilute HCl.
W096109836 ~2 0 1 3 4 ~ PCTIUS95/12319
- 76 -
~CHEME H
HO) Cl~ Cl~ HO) base
H2N R2a o Cl O~ N ~R2a
~N~"'R2a ~~
BOC
3 4
4 Base o~N~ R2a Base ~kN~ R2a
R3X I R4X or
BOC H+ BOC
Base 5 6
RYRZCO
HO~o~ -H20 ~o H2, Pd/C
O IN ~ R2a O N "' R2a
BOC BOC
7 8
2~ ~ ~ 3 4 ~
WO 96/09836 PCTIUS95/12319
SCHEME H (CONT'D)
LiOOH; ,~R4
oraq. HCI, RW-NH o CO2H
then BOC20 R2a
a Rw = H
b, Rw = BOC
EDC IR~R4
9 + H-A ~ BOCNH ~A
R2a o
HCI R3~R4
HCI NH2 o~A
R2a o
1 1
W096/09836 2~ 4 6 PCI/US95112319
- 78 -
SCHEME H (CONT'D)
RXCHO, NaCNBH3 ~ R
12 'RXCH2NH O A
11 R2a 0
14a
RXCOOH, EDC, HOBT
13 ll R3~R4
RXCNH O A
R2a o
14b
A
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
Scheme I. Aminoalcohol 1 is derivatized with BOC2O to give 15.
Mesylation of 15 followed by reaction with methyl alpha-
mercaptoacetate in the presence of cesium carbonate gives sulfide 16.
Removal of the BOC group in 16 with TFA followed by neutralization
with di-isopropylethylamine leads to lactam 17. N-BOC derivative lf~
30 iS obtained via the reaction of 17 with BOC anhydride in THF
catalyzed by DMAP. Sequential alkylation of 1 g with the alkyl
halides R3X and R4X in THF/DME using NaHDMS as the
deprotonation reagent produces 19. Hydrolysis of 19 in hydro-
chloride to yield 20a, which is derivatized with Boc anhydride to yield
20b. The coupling of 20b with an alpha-aminolactone (e.g.,
W096/09836 ~2 a ~ ~ 4 ~ PCTIUS95/12319
- 79 -
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 21. Sulfide 21 is readily oxidized to
sulfone 22 by the use of MCPBA (m-chloroperoxybenzoic acid). The
5 N-BOC group of either 21 or 22 is readily removed by treatment with
gaseous hydrogen chloride. The resultant amine hydrochloride 23
undergoes reductive alkylation in the presence of an aldehyde RXCHO
(12) and a reducing agent (e.g., sodium cyanoborohydride); or
acylation in the presence of RXCOOH (13) and a peptide coupling
10 reagent to afford the products 24 and 25.
WO 96/09836 ~ PCT/US95tl2319
2U~3~ .
- 80 -
SCHEME I
HO HO
~ ~ 1) MsCI
H2N 'R2a BOC20 HN "' 2 2) Cs2CO3
HSCH2CO2CH3
- BOC 15
S
CH302CHN~"'R2a 1)TFA ~ ~ BOC20
1 2)(i-Pr)2- N "'R2a
BOC NEt H
16 17
R4
)"' R2a Base r ~ 2a
BOC Base BOC
18 19
R3 R4 R3 R4
~ ~ S CO2H H-A,EDC BOCNH S ~ A
25 Rw R2a HOBT R2a O
21
a,RW=H ~ BOC20 NH ~ or NH ~ 6
30 b,RW=BOC ~ R5a
q
~ W096t09836 22 ~ ~ ~ 4 6 PCI~/US95112319
- 81 -
SCHEME I (CONrr'D)
R3 R4
BOCNH s(o)?~A HCI
R2a O
m=0, 20 ) MCPBA
m=2, 22
R3 R4 R ~R
1 2
-- ,
23 NaCNBH3 24
m = 0 or 2
RXCOOH
EDC, HOBT
1l R3 R4
R CNH S(O)?~A
R2a o
2S
W0 96/09836 2 ~ 6 PCT/US95/12319
- 82 -
The compounds of this invention inhibit Ras famesyl
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 hllm~n~. 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 may be ~lmini~tered to
m~mm~l~, preferably hllm~n~, either alone or, preferably, in
combination with pharmaceutically acceptable carriers or diluents,
optionally with known adjuvants, such as alum, in a ph~ eutical
composition, according to standard pharmaceutical practice. The
compounds can be ~lministered orally or parenterally, including the
intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and
topical routes of ~lmini~tration.
For oral use of a chemotherapeutic compound according
to this invention, the selected compound may be a~lministered, for
20 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
commonly used include lactose and corn starch, and lubricating
agents, such as magnesium stearate, are commonly added. For oral
~clmini~tration in capsule form, useful diluents include lactose and
25 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
3 o 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.
W096/09836 2~ Q ~ 3 ~ 6 PCT/US95/12319
- 83 -
The present invention also encompasses a ph~rm~ceuticalcomposition useful in the treatment of cancer, comprising the
~lmini~tration of a therapeutically effective amount of the compounds
of this invention, with or without pharmaceutically acceptable carriers
5 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
a-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. A~lmini~tration occurs in an amount between about 0.1 mg/kg
of body weight to about 20 mg/kg of body weight per day, preferably
of between 0.5 mg/kg of body weight to about lO mg/kg of body
20 weight per day.
The compounds of the instant invention are also useful as
a component in an assay to rapidly determine the presence and
quantity of farnesyl-protein transferase (FPTase) in a composition.
Thus the composition to be tested may be divided and the two
25 portions contacted with mixtures which comprise a known substrate
of FPTase (for example a tetrapeptide having a cysteine at the amine
terminus) 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
3 o allow the FPTase to farnesylate the substrate, the chemical content of
the assay mixtures may be determined by well known immunological,
radiochemical or chromatographic techniques. Because the
compounds of the instant invention are selective inhibitors of FPTase,
absence or quantitative reduction of the amount of substrate in the
WO ~ 836 PCT/US95/12319
- - 84 -
assay mixture without the compound of the instant invention relative
to the presence of the unchanged substrate in the assay con~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 quantit~tin~ the
enzyme. Thus, potent inhibitor compounds of the instant invention may
be used in an active site titration assay to determine the quantity of
10 enzyme in the sample. A series of samples composed of aliquots of a
tissue extract containing 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 appropriate 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 substantially 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
20 that particular sample.
EXAMPLES
Examples provided are intended to assist in a further
25 understanding of the invention. Particular materials employed,
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
30 acid, l 0% sodium bicarbonate and brine as appropriate. Solutions
were dried over sodium sulfate and evaporated in vacuo on a rotary
evaporator.
W0 96/09836 ~ 6 PCT/US95tl2319
- - 85 -
EXAMPLE 1
Preparation of N- { 2(S)-[(4-Nitrobenzylthio)acetamido] -3-methylpentyl } -
N-(l-Naphthvlmethyl)-~lycyl-methionine methyl ester
Step A: Preparation of N-(2(S)-t-butoxycarbonylamino)-3-
methylpentvl)~lvcine methyl ester
Glycine methyl ester hydrochloride (23.1 g, 0.1 8mol)
was dissolved in methanol (700 mL) and treated with N-t-
butoxycarbonyl-isoleucinal (40 g, 0.18 mol) with stirring at 0C.
Sodium cyanoborohydride (17.3 g, 0.28 mol) was added, and the pH
of the mixture was adjusted to 7 with HOAc . After stirring for 3 h,
aqueous saturated NaHCO3 (50 mL) was added to the mixture which
was then concentrated to 250 mL. The solution was 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 (MgSO4).
Filtration and concentration provided the title compound after
purification by chromatography (sio27 EtOAc: hexane, 1:3). 1H
NMR (CDC13); ~ 4.69 (m, lH), 3.72 (s, 3H), 3.48-3.62 (m, lH), 3.42
(ABq, 2H), 2.65 (d, 2H, J=6Hz), 1.4-1.6 (m, 2H), 1.48 (s, 9H), 1.04-
1.2 (m, lH), 0.85-0.95 (m, 6H).
Step B: Preparation of N-~2(S)-t-Butoxycarbonylamino)-3-
methylpentyll-N-(l-naphthylmethyl) Iycine methyl ester
N-~2(S)-t-Butoxycarbonylamino)-3-methylpentyl]glycine
methyl ester (2.00 g, 6.97 mmol) was dissolved in 1,2-dichloroethane
(56 ml) and 3A molecular sieves were added followed by 1-
naphthaldehyde (1.~9 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.
NaHCO3 (100 ml/25 ml). The aqueous layer was washed with EtOAc
(3x50 ml). The organic layers were combined, dried with Na2SO4,
filtered, and concentrated to give crude product which was purified by
W0 96/09836 ~ PCT/US95/12319
- 86 -
chromatography (silica gel 1 :6 to 1 :3 ethyl acetate in hexane) to
provide the title compound. lH NMR (CD30D); ~ 8.44-8.38 (d, lH,
J=6Hz), 7.88-7.77 (m, 2H), 7.55-7.35 (m, 4H), 6.34-6.27 (m, lH),
4.25 (ABq, 2H), 3.66 (s, 3H), 3.40-3.23 (m, lH), 2.95-2.85 (dd, lH,
J=6, 15Hz), 2.68-2.57 (dd, lH, J=6, 15Hz), 1.57-1.46 (m, lH), 1.43 (s,
9H), 1.34-1.18 (m, 2H), 1.06-0.85 (m, lH), 0.85-0.71 (m, 6H).
Step C: Preparation of N-[2(S)-t-Butoxycarbonylamino)-3-
methylpentyll-N-( l -naphthylmethyl) lycine
N-[2(S)-t-Butoxycarbonylamino)-3-methylpentyl]-N-(1-
naphthylmethyl)glycine methyl ester (2.61 g, 6.10 mmol) was
dissolved in MeOH (50 ml) and lN NaOH (24.4 ml, 24.4 mmol) was
added. The mixture was stirred at ambient temperature for 4 h and
concentrated. The resulting residue was dissolved in H2O (25 ml) and
neutralized with lN HCl (24.4 ml). The aqueous layer was washed
with EtOAc (3x50 ml). The organic layers were combined, dried with
Na2SO4, filtered, and concentrated to provide the title compound. 1H
NMR (CD3OD)2 ~ g.4~-8.39 (d, lH, J=6Hz), 8.03-7.91 (t, 2H,
J=6Hz), 7.75-7.48 (m, 4H), 5.00-4.93 (d, lH, J=12Hz), 4.78-4.66 (d,
lH, J=12Hz), 3.80-3.5~ (m, 3H), 3.49-3.40 (dd, lH, J=3, 12Hz), 3.09-
2.98 (dd, lH, J=3, 12Hz), 1.42 (,s, 9H), 1.37-1.28 (m, 2H), 1.80-1.00
(m, lH), 0.94-0.78 (m, 6H).
Step D: Preparation of N-[2(S)-t-Butoxycarbonylamino)-3-
methylpentyl]-N-( l -naphthylmethyl)glycine-methionine
methyl elster
N-~2(S)-t-Butoxycarbonylamino)-3-methylpentyl]-N-(l -
naphthylmethyl)glycine (2.29 g, 5.53 mmol), dissolved in DMF (20
mL), was treated with HOBT (0.822 g, 6.08 mmol), EDC (1.17 g,
6.08 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 mix~ure was stirred at ambient temperature for 24 h. The
mixture was concentrated, and the residue was partitioned between
EtOAc (50 mL) and saturated NaHCO3 solution (25 mL). The
W096/09836 ~ 2 n ~ PCT/US9S/12319
- 87 -
aqueous layer was extracted with EtOAc (lx30 mL). The organic
layers were combined, washed with brine (lx25 mL), dried (Na2S04),
filtered, and concentrated to give crude product which was purified by
chromatography (silica gel eluting with 1 :3 to 1 :2 ethyl acetate in
hexane) to provide the title compound. 1H NMR (CD30D); ~ 8.36-
8.29 (d, lH, J=6Hz), 7.93-7.86 (d, lH, J=6Hz), 7.85-7.80 (d, lH,
J=6Hz), 7.61-7.39 (m, 4H), 6.60-6.52 (m, lH), 4.32-4.06 (m, 2H),
3.90-3.69 (m, lH), 3.65 (s, 3H), 3.27-3.14 (m, 2H), 2.93-2.70 (m, 2H),
2.19-1.78 (m, 6H), 1.63-1.30 (m, 13H), 1.19-1.05 (m, lH), 0.95-0.81
0 (m, 6H).
Step E: Preparation of N-~2(S)-amino-3-methylpentyl)-N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
hydrochloride
N-[2(S)-(t-Butoxycarbonylamino)-3-methylpentyl]-N-(1-
naphthylmethyl)-glycyl-methionine methyl ester (2.82 g, 5.04 mmol)
was dissolved in EtOAc (50 mL) and cooled to -25C. HCI was
bubbled through the mixture until TLC (95:5 CH2C12:MeOH)
indicated complete reaction. Nitrogen was bubbled through the
20 mixture to remove excess HCI and the mixture was then concentrated
to provide the title compound. lH NMR (CD30D); ~ 8.34-8.28 (d,
lH, J=6Hz), 8.00-7.92 (d, 2H, J=6Hz), 7.83-7.71 (m, lH), 7.68-7.49
(m, 3H), 4.76-4.55 (m, 4H), 3.~4-3.75 (m, 2H), 3.71 (s, 3H), 3.59-3.70
(m~ lH), 3.21-3.00 (m, 2H), 2.57-2.38 (m, 3H), 2.17-2.04 (m, 4H),
25 1.97-1.81 (m, lH), 1.63-1.50 (m, lH), 1.39-1.20 (m, lH), 1.19 1.00
(m, lH), 0.95-0.79 (m, 6H).
Step F: Preparation of (4-nitrobenzylthio)acetic acid
4-Nitrobenzyl chloride (2.5 g, 15 mmol) was added to a
30 solution of mercaptoacetic acid (80% in H2O, 1 mL, ~ 12 mmol) in
aqueous sodium hydroxide (2.75 N, 10 mL) and THF (10 mL) with
stirring at 50C. After 1 hour, the reaction was cooled to ambient
temperature, diluted with water (30 mL) and extracted with Et2O to
remove excess halide. The aqueous layer was distributed between
W0 96/09836 a ~ 6 PCT/US95/12319
- 88 -
EtOAc and 5% citric acid and the organic extract then washed with
H20, dried (MgSO4) and evaporated to give the title compolmd as a
pale yellow solid, m.p. 101-104C [lit. m.p. 114C]. 1H NMR
(CDCl3); ~ 3.10 (s, 2H), 3.94 (s, 2H), 5.37 (d, 2H, J = 8.7 Hz), 8.21
5 (d, 2H, J = 8.7 Hz).
Step G: Prepara~ion of N-{2~S)-[(4-Nitrobenzyl~io)acetamido]-
3(S)-methylpentyl } -N-(l -naphthylmethyl)-glycyl-
methionine methyl ester
(4-Nitrobenzylthio)acetic acid (68 mg, 300 !lmol),
dissolved in DMF (4 rnL), was treated with HOBT (51 mg, 300
~mol), EDC (65 mg, 300 !lmol), and N-r2(S)-amino-3-methylpentyl)-
N-( l -naphthylmethyl)-glycyl-methionine methyl ester hydrochloride
(133 mg, 250 !lmol). The pH was adjusted to 7.5 with Et3N (110 ,uL,
250 ~lmol) and the mixture was stirred at ambient temperature for 16
h. The mixture was concentrated and the residue was partitioned
between EtOAc (100 mL) and H20 (100 mL). The organic layer was
washed with H2O (2x50 mL), dried (MgSO4), filtered, and
concentrated to give a crude product which was purified by
chromatography (silica gel, eluting with 1 :1 to 1 :2 hexane: EtOAc) to
provide the title compound. lH NMR (CDCl3); ~ 0.89 (t, 3H, J = 7.5
Hz), 0.91 (d, 3H, J = 6.9 Hz), 1.02-1.19 (m, lH), 1.37-1.6 (m, 2H),
1.67-1.79 (m, lH), 1.85-1.98 (m, lH), 1.95 (s, 3H), 2.0-2.1 (m, 3H),
2.74 (dd, lH, J = 3, 12 Hz), 2.86-3.01 (m, 3H), 3.21 (d, lH, J = 14
Hz), 3.29 (d, lH, 14 Hz), 3.69 (s, 3H), 3.74 (d, lH, J = 12 Hz), 3.8 (d,
lH,J=12Hz),3.83-3.94(m,1H),4.05(d,1H,J=9Hz),4.18(d,1H,
J = 9 Hz), 4.41-4.49 (m, lH), 6.7 (d, lH, J = 8 Hz), 7.39-7.53 (m, 7H),
7.~s-7.9 (m, 2H), 8.14-8.22 (m, 2H).
Anal. Calcd for C34H44N4O6S2-0.15 EtOAc
C, 60.92; H, 6.68; N, 8.21
C, 60.64; H, 6.65, N, 8.23
W096/09836 2 ~ e PCT/US95/12319
- 89 -
1. .
EXAMPLE 2
N- { 2(S)-[(4-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(l -
naphthylmethyl)-glycvl-methionine
N- { 2(S)-[(4-Nitrobenzylthio)acetamido]-3(S)-
methylpentyl)-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
(from Example 1; 100 mg, 150 !lmol) was dissolved in MeOH (1 mL)
and 1.0N NaOH (300 ~L, 300,umol) was added. The mixture was
stirred at 45C under argon for 45 minutes then the solution was
0 partitioned between EtOAc (100 mL) and 5% citric acid (50 mL).
The organic layer was washed with H2O (2x50 mL), dried (MgSO4),
filtered and evaporated to give the title compound. lH NMR; o 0.84
(t, 3H, J = 7.5 Hz), 0.8$8 (d, 3H, J = 7 Hz), 0.91-1.0 (m, lH), 1.0-1.2
(m, lH), 1.2-2.1 (m, 5H), 1.90 (s, 3H), 2.8-2.86 (m, 2H), 3.0-3.3 (m,
15 4H), 3.90 (s, 2H), 3.91-4.22 (m, 4H), 7.37-7.56 (m, 6H), 7.78-7.88 (m,
2H), 8.15 (d, 2H, J = 8Hz), 8.27 (d, lH, J = 8.3 Hz).
Anal. Calcd for C33H42N406S2-0.25 CHCl3-1.0 CH30H
C, 57.39; H, 6.50; N, 7.82
C, 57.71; H, 6.42; N, 7.46
EXAMPLE 3
The following acids were prepared according to the
procedure described for Example 1, Step F or were obtained
25 commercially:
(Benzylthio)acetic acid
(3-Nitrobenzylthio)acetic acid
(2-Nitrobenzylthio)acetic acid
(4-Cyanobenzylthio)acetic acid
(4-Trifluoromethylbenzylthio)acetic acid
(4-Methoxybenzylthio)acetic acid
(4-Methylsulfonylbenzylthio)acetic acid
(4-Phenylbenzylthio)acetic acid
W096/09836 22 ~ ~ 3 ~ ~ PCTIUS95/12319
- 90 -
(4-Methylbenzylthio)acetic acid
(8-Chloronaphth-l-ylthio)acetic acid
(2-Methylindol-3-ylthio)acetic acid
3-(Benzylthio)propionic acid
(4-Picolinylthio)acetic acid
(4-Pyridylthio)acetic acid
EXAMPLE 4
The following compounds were prepared using the
procedure described for Example 1, Step G, but substituting the
appropriate acid from Example 3 for (4-nitrobenzylthio)acetic acid,
and, in some cases, subsli~ulillg N-t-butoxycarbonylleucinal for N-t-
butoxycarbonyl-isoleucinal in Step A in the preparation of the
intermediate.
N- { 2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethvl)-~lycyl-methionine methyl ester
FAB mass spectrum m/z = 624 (M+1).
Anal. Calcd for C34H45N304S2-0.30 TFA-0-95 H2O
C, 55.70; H, 6.16; N, 5.32
C,55.72;H,6.17;N,5.30
N- { 2(S)-[(3-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)- lycyl-methionine methyl ester
Anal. Calcd for C34H44N4O6S2-0.2 EtOAc
C, 60.88; H, 6.70, N, 8.16
C, 60.52; H, 6.63; N, 8.31
N- { 2(S)-[(2-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)- Iycyl-methionine methyl ester
W096/09836 ~2n~46 PCTIUS95/12319
- 91 -
Anal. Calcd for C34H44N4O6S2-0.2 EtOAc
C,60.88;H,6.70;N,8.16
C, 60.44; H, 6.67; N, 8.19
5 N-{2(S)-[(4-Cyanobenzylthio)acetamido]-3(S)-methylpentyl}-N-(1-
naphthvlmethyl)-glycvl-methionine methvl ester
Anal. Calcd for C35H44N404S2-0.5 EtOAc
C, 64.13; H, 6.98; N, 8.09
C, 63.85; H, 6.91; N, 8.17
N- ~ 2(S)-~(4-Trifluoromethylbenzylthio)acetamido]-3(S)-methylpentyl } -
N-(l-naphthylmethyl)-glycyl-methionine methyl ester
Anal. Calcd for C35H44F3N3O4S2-0.4 EtOAc-1.0 H2O
C, 58.99; H, 6.66; N, 5.64
C,58.97;H,6.37;N,5.61
N- { 2(S)-[(4-Methoxybenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glycvl-methionine methyl ester
Anal. Calcd for C35H47N305S2-0.6 EtOAc 0.35 H2O
C, 62.99; H, 7.42; N, 5.89
C, 62.64; H, 7.17; N, 6.29
N- { 2(S)-~(4-Methylsulfonylbenzylthio)acetamido]-3(S)-methylpentyl } -
N-(l-naphthylmethyl)- Iycyl-methionine methyl e.ster
Anal. Calcd for C35H47N306S3
C, 59.~s9; H, 6.75; N, 5.99
C, 60.14; H, 6.90; N, 5.96
N- { 2(S)-[(4-Phenylbenzylthio)acetamido] -3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glvcvl-methionine methyl ester
W 0 96/09836 ~ PC~rrUS95/12319
- 92 -
FAB mass spectrum m/z = 700 (M+1).
N- { 2(S)-[(4-Methylbenzylthio)acetamido]-3(S)-methylpentyl } -N-( l -
naphthylmethyl)-glycyl-methionine methyl ester
s
FAB mass spectrum m/z = 638 (M+1).
N- { 2(S)-[(8-Chloronaphth- 1 -ylthio)acetamido] -3(S)-methylpentyl } -
N-(l-naphthylmethyl)-glvcyl-methionine methyl ester
Anal. Calcd for C37H44ClN3O4S2-0.5 EtOAc
C, 63.43; H, 6.55; N, 5.69
C,63.33;H,6.35;N,6.07
N- ~ 2(S)-[((2-Methylindol-3-yl)thio)acetamido]-4-methylpentyl } -N-
(1-naphthylmethyl)-glycyl-methionine methyl ester
Anal. Calcd for C36H46N404S2-0.35 EtOAc
C, 64.74; H, 7.09; N, 8.08
C, 64.73; H, 6.93; N, 8.32
N- { 2(S)-[(4-Pyridylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethvl)- lycvl-methioninemethyl e.ster
25 Anal. Calcd for C32H42N4O4S2-2.0 TFA-0.75 H2O
C, 50.72; H, 5.3~s; N, 6.57
C, 50.~4; H, 5.2~s; N, 6.56
N- ~ 2(S)-[(4-Picolinylthio)acetamidol-3(S)-methylpentyl } -N-(l -
30 naphthvlmethyl)-~lvcvl-methionine methvl e,ster
Anal. Calcd for C33H44N404S2-2.0 TFA-0.75 H20
C, 51.29; H, 5.53; N, 6.47
C, 51.27; H, 5.35; N, 6.70
I W096t09836 22 a ~ ~ 4 ~ PCT/US95/12319
- 93 -
,
N- { 2(S)-[3-(Benzylthio)propionamido] -3 (S)-methylpentyl } -N-(1 -
naphthylmethvl)-glycvl-methionine methvl ester
lH NMR (CDCl3); o 0.~4 (t, 3H), 0.88 (d, 3H), 1.08 (m, lH), 1.40
(m, 2H~, 1.66-2.0 (m,7H), 2.33 (m, 2H), 2.7 (m, 3H), 2.99 (dd, lH),
3.21 (s, 2H), 3.68 (s, 3H), 3.7 (m, 3H), 4.4 (m, lH), 6.33 (d, lH), 7.2-
7.4 (m, 10H), 7.7-7.9 (m, 4H), 8.19 (m, lH).
0 EXAMPLE 5
N- { 2(S)-[(Phenylthio)acetamido]-3(S)-methylpentyl } -N-(l -
naphthylmethyl)-~lyGyl-m.ethionine m.ethyl ~st~r
To solution of N-(2(S)-amino-3(S)-methylpentyl)-N-(1-
naphthylmethyl)-glycyl-methionine methyl ester hydrochloride (125
mg, 0.23 mmol) and Et3N (65.5 ,uL, 0.47 mmol) in THF (2.5 mL) at
room temperature was added bromo acetylbromide (24 ~L, 0.28
mmol). After 10 minutes, thiophenol (29 ,uL, 0.28 mmol) and Et3N
(49 IlL, 0.35 mmol) were added and the mixture was stirred for 4
20 hours. The solution was poured into saturated aqueous NaHCO3,
extracted with EtOAc (x3), washed with brine, dried (MgSO4) and
evaporated. Purification of the residue by column chromatography
(silica gel; hexane/EtOAc 3:2) gave the title compound as an oil.
25 FAB mass spectrum m/z = 610 (M+l).
Anal. Calcd for C33H43N304S2 0.5 TFA-1.15 H2O
C, 53.94; H, 5.88; N, 5.24
C, 53.93; H, 5.91; N, 5.11
EXAMPLE 6
N- { 2(S)-[(4-(1 -H-Tetrazol-5-yl)benzylthio)acetamido] -3(S)-
methylpentyl ~ -N-( l -naphthylmethyl)-glycyl-methionine methvl ester
W 0 96/09836 ~7 ~ ~ ~ 3 ~ ~ PC~rrUS95/12319 ~
Step A: Preparation of ~4-(lH-tetrazol-5-yl)benzylthiolacetic acid
(4-Cyanobenzylthio)acetic acid (from Fx~mple 3; 207
mg, 1 mmol) was added to a stirred solution of trimethylsilylazide
(400 ,uL, 3 mmol) and dibutyltin oxide (25 mg, 100 !lmol) in toluene
(0.5 mL) at 110C. After 6 hours, the reaction was quenched with
MeOH (1 mL) and evaporated. The residue was purified by flash
chromatography (silica gel, 1000:100:3 to 10Q0:100:13
CHC13/MeOH/HOAc) to give the title compound. Rf (silica;
1000:100:6 CHC13/MeOH/HOAc) = 0.2 compared to the nitrile
10 starting material of Rf = 0.67. lH NMR (d6 DMSO), ~ 3.15 (s, 2H),
3.88(s,2H),7.51 (d,2H,J=8.1 Hz),7.99(d,2H,J=8.1 Hz). FAB
mass spectrum m/z = 251 (M+1).
Step B: N- ~ 2(S)-[~4-(1 -H-Tetrazol-5-yl)benzylthio)acetamido]-
3(S)-methylpentyl } -N-(1 -naphthylmethyl)-glycyl-
methionine methyl e~ter
Following the procedure described for Example 1, Step
G, [4-(lH-tetrazol-5-yl)benzylthio]acetic acid was coupled with N-
(2(S)-amino-3(S)-methylpentyl)-N-( l -naphthylmethyl)-glycyl-
20 methionine methyl ester hydrochloride to give the title compound.
Anal. Calcd for C35H45N7O4S2-0.8 TFA
C, 51.67; H, 5.26; N, 10.93
C, 51.66; H, 5.26; N, 10.84
2s
EXAMPLE 7
N- (2(S)-[(Benzyloxy)acetamido]-3(S)-methylpentyl } -N-(l -
naphthylmethvl)-glycyl-methionine methyl e~ter
The title compound was prepared from N-[2(S)-amino-3-
methylpentyl)-N-( l -naphthylmethyl)-glycyl-methionine methyl ester
hydrochloride (Example 1, Step E) and benzyloxyacetyl chloride
(Aldrich) under standard conditions.
FAB mass spectrum m/z = 608 (M+l).
WO 96/09836 PCT/US95/12319
~ ~2 ~ ~ 3 4 6
EXAMPLE 8
N- { 2(S)-~(N'-Benzyl-N'-t-butoxycarbonylglycyl)amino]-3(S)-
methylpentyl ~-N-(l -naphthylmethyl)-glvcyl-methionine methyl ester
Step A: Preparation of N-benzyl-N-t-butoxycarbonylglycine
N-Benzylglycine ethyl ester hydrochloride was converted
to N-benzyl-N-t-butoxycarbonylglycine ethyl ester using di-t-butyl
o dicarbonate under standard conditions. The ethyl ester was saponified
using sodium hydroxide in methanol to give the title compound.
Step B: Preparation of N-{ 2(S)-[(N'-benzyl-N'-t-
butoxycarbonylglycyl)amino]-3(S)-methylpentyl }-N-(1 -
naphthylmethyl)-~lycvl-methionine methvl ester
The glycine derivative prepared in Step A was converted
to the title compound using the method of Example 1, Step G.
Anal. Calcd for C39H54N4O6S-0.5 H20
C, 65.42; H, 7.74; N, 7.83
C,65.31;H,7.33;N,8.01
EXAMPLE 9
N- { 2(S)-[(N'-Benzylglycyl)amino]-3(S)-methylpentyl } -N-(1-
naphthvlmethyl)-~lvcyl-methionine methyl ester
The t-butoxycarbonyl group of N-{2(S)-[(N'-benzyl-N'-t-
butoxycarbonylglycyl)amino] -3 (S)-methylpentyl } -N-( 1 -naphthyl-
methyl)-glycyl-methionine methyl ester, Example 8, was cleaved
3 0 using the method of Example 1, Step E to provide the title compound.
Anal. Calcd for C34H46N4O4S-2.0 HC1-0.75 H2O
C, 58.90; H, 7.20; N, 8.08
C, 59.07; H, 6.88; N, 7.87
W096/09836 22 ~ ~ ~ 4 ~ P~l/U~g51l23l9
- 96 -
EXAMPLE 10
N- { 2(S)-[(N'-Acetyl-N'-benzylglycyl)amino]-3(S)-methylpentyl } -N-
(1-naphthylmethyl)-~lvcyl-methionine methvl ester
A solution of N- { 2(S)-[(N'-benzylglycyl)amino]-3(S)-
methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine methyl ester
from Example 9 (200 mg, 0.3 mmol) was dissolved in CH2C12 (4
mL). Acetic anhydride (0.1 1 mL) and N,N-diisopropylethylamine
(0.31 mL) were added and the mixture was stirred for 48 hours. The
o mixture was concentrated and partitioned between ethyl acetate and
saturated sodium bicarbonate solution. The organic layer was dried
and evaporated to give a residue which was purified by silica gel
chromatography (1-3% MeOH in CH2C12) to give the title compound.
Anal. Calcd for C36H4~N405S-0.5 H20
C,65.72,H,7.51;N,8.52
C, 65.87, H, 7.23, N, 8.69
EXAMPLE 1 1
N- { 2(S)-[(Benzylthio-S-oxide)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
Step A: Preparation of (benzvlthio)acetic acid S-oxide
Benzylthioacetic acid was converted to the known title
compound (Lindstrom and Mark, US Patent 4,637,833) by oxidation
with sodium periodate in methanol.
Step B: N- { 2(S)-~(Benzylthio-S-oxide)acetamido]-3(S)-
3 0 methylpentyl } -N-( 1 -naphthylmethyl)-glycyl-methionine
methvl ester
The sulfoxide derivative from Step A was converted to
the title compound using the method of Example 1, Step G.
FAB mass spectrum m/z = 640 (M+l).
-- WO 96/09836 PCI/US95112319
2a~34~ `
- 97 -
EXAMPLE 12
N- { 2(S)-[(Benzylthio-S,S-dioxide)acetamido]-3(S)-methylpentyl } -N-
5 ( l -naphthylmethyl)-~lycyl-methionine methvl ester
Step A: Preparation of (benzylthio)acetic acid S,S-dioxide
Benzylthioacetic acid was converted to the known title
compound (Lindstrom and Mark, US Patent 4,637,833) by oxidation
with Oxone in methanol.
Step B: N- { 2(S)-[(Benzylthio-S,S-dioxide)acetamido]-3(S)-
methylpentyl }-N-(1-naphthylrnethyl)-glycyl-methionine
methyl ester
The sulfone derivative from Step A was converted to the
title compound using the method of Example 1, Step G.
FAB mass spectrum m/z = 656 (M+l).
EXAMPLE 13
N- ~ 2(S)-[(Benzylthio)acetamido] -3(S)-methylpentyl } -N-(1-
naphthylmethyl)-~lycyl-methionine .sulfone methyl ester
Using the method of Example 1, but substituting
25 methionine sulfone and benzylthioacetic acid as appropriate, the title
compound was prepared.
FAB mass spectrum m/z = 6~6 (M+l).
EXAMPLE 14
N- { 2(S)-[(N'-4-Nitrobenzoylglycyl)amino]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)- Iycyl-methionine methyl ester
Following the procedure described for Example 1, Step
G, but substituting (4-nitrobenzylthio)acetic acid with N-(4-
nitrobenzoyl)glycine (Aldrich) the title compound was obtained.
W096/09836 2~ ~ ~ 3 4 6 PCI/US95/12319
- 98 -
Anal. Calcd for C34H43N507S-0.8 H2O
C, 60.03; H, 6.61; N, 10.30
C, 59.99; H, 6.32; N, 10.28
EXAMPLE 15
N- ~ 2(S)-[(N'-Methyl-N'-4-nitrobenzoylglycyl)amino]-4-methyl-
pentyl ~-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
0 Following the procedure described for Example 1, but
substituting (4-nitrobenzylthio)acetic acid with N-(4-
nitrobenzoyl)sarcosine (A. Morgan et al, J. Med. Chem., 34:2126
(1991)) and N-t-butoxycarbonylleucinal for N-t-
butoxycarbonylisoleucinal the title compound was obtained.
Anal. Calcd for C35H45N507S-0.35 EtOAc
C,61.51;H,6.78;N,9.86
C, 61.13; H, 6.69; N, 10.11
EXAMPLE 16
N- ~ 2(S)-[S-Benzyl-L-cysteinyl]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-~lycvl-methionine methvl ester
S-Benzyl-N-t-butoxycarbonyl-L-cysteine (Bachem) was
2s coupled with N-[2(S)-amino-3-methylpentyl)-N-(1-naphthylmethyl)-
glycyl-methionine methyl ester hydrochloride according to the
procedure described for Example 1, Step G. The product thus
obtained was deprotected using TFA in CH2C12 to give the title
compound.
FAB mass spectrum m/z = 653 (M+1).
Anal. Calcd for C35H4~N404S2-2.~ TFA-0.1 H2O
C, 50.07; H, 5.28; N, 5.75
C, 50.07; H, 5.32; N, 5.9
WO 96/09836 PCT/US95/12319
a2 0 ~ 3~ 6
_ 99 _
EXAMPLE 17
.
N- { 2(S)-[S-benzyl-D-cysteinyl]-3(S)-methylpentyl } -N-(1-
5 naphthylmethyl)- lycyl-methionine methyl ester
S-Benzyl-N-t-butoxycarbonyl-D-cysteine (Bachem) was
coupled with N-[2(S)-amino-3-me~ylpentyl)-N-(1-naph~ylmethyl)-
glycyl-methionine methyl ester hydrochloride according to the
procedure described for Example 1, Step G. The product thus
obtained was deprotected using TFA in CH2Cl2 to give the title
compound.
FAB mass spectrum m/z = 653 (M+l).
Anal. Calcd for C35H48N404S2-2.3 TFA-0-5 H2O
C, 51.47; H, 5.60; N, 6.06
C, 51.49; H, 5.57; N, 6.33
EXAMPLE 1~
The following compounds were prepared from the
corresponding esters (Examples 3-17) using the procedure described
for Example 2.
N- ( 2(S)-[(Benzylthio)acetamido] -3(S)-methylpentyl } -N-( 1-
naphthylmethyl)-glycyl-rnethionine
FAB mass spectrum m/z = 610 (M+1).
Anal. Calcd for C33H43N304S2-0.05 TFA- 1.05 H2O
C, 56.32; H, 6.22; N, 5.61
C, 56.32; H, 5.99; N, 6.01
N- { 2(S)-[(Phenylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)- lycyl-methionine
FAB mass spectrum m/z = 596 (M+1).
W096/09836 ~a Q ~ 3 ~ ~ PcTluS95/12319`
- 100-
Anal. Calcd for C32H41N3O4S2-0.25 TFA-0.75 H20
C, 55.11; H, 5.87; N, 5.59
C, 55.14; H, 5.86; N, 5.46
N- { 2(S)-[(3-Nitrobenzylthio)acetamido] -3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-~lycvl-methionine
Anal. Calcd for C33H42N4O6S2-1.2 H2O-0.5 EtOAc
C, 58.34; H, 6.77; N, 7.78
C, 58.34; H, 6.54; N, 7.51
N- { 2(S)-[(2-Nitrobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glvcvl-methionine
Anal. Calcd for C33H42N4O6S2-1.2 H2O-0.5 EtOAc
C, 58.34; H, 6.77; N, 7.78
C,58.34;H,6.51;N,7.63
N- { 2(S)-[(4-Cyanobenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glycyl-methionine
Anal. Calcd for C34H42N4O4S2-0.3 CHCl3
C, 61.42; H, 6.36; N, 8.35
C,61.73;H,6.31;N,8.41
N- { 2(S)-[(4-Trifluoromethylbenzylthio)acetamido]-3(S)-
methylpentyl ~ -N-( l -naphthylmethyl)-~lycyl-methionine
Anal. Calcd for C34H42F3N3O4S2-0.5 EtOAc
C, 59.89; H, 6.42; N, 5.82
C, 59.60; H, 6.25; N, 5.82
N- { 2(S)-[(4-Methoxybenzylthio)acetamido]-3(S)-methylpentyl } -N-
( l -naphthvlmethyl)- lycyl-methionine
W096/09836 ` a~n~34~ PCT/US9S/12319
- 101 -
Anal. Calcd for C34H45N305S2-0.5 EtOAc
C,63.22;H,7.22,N,6.14
C, 63.22; H, 7.03; N, 6.19
5 N- { 2(S)-[(4-Methylsulfonylbenzylthio)acetamido]-3(S)-
methvlpentyl ~ -N-( l -naphthylmethyl)-glycyl-methionine
Anal. Calcd for C34H45N306S3-0.3 H2O
C, 58.~9; H, 6.63; N, 6.06
C, 58.85; H, 6.64; N, 6.00
N- ~ 2(S)-[(4-Phenylbenzylthio)acetamido] -3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glycyl-methionine
Anal. Calcd for C3gH47N3O4S2-0.5 TFA-0.5 H2O
C, 58.25; H, 5.76; N, 4.85
C, 58.26; H, 5.67; N, 4.87
N- (2(S)-[(4-Methylbenzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)- lvcyl-methionine
Anal. Calcd for C34H4sN3O4S2-0.25 H20
C, 64.98; H, 7.30; N, 6.69
C, 64.95; H, 7.17; N, 6.66
N- ~ 2(S)-[(~s-Chloronaphth- 1 -ylthio)acetamido]-3(S)-methylpentyl } -
N-( l -naphthylmethyl)-glvcyl-methionine
Anal. Calcd for C36H42ClN3O4S2-1.0 EtOAc 1.0 H2O
C, 61.08; H, 6.66; N, 5.34
C, 61.09; H, 6.94; N, 5.38
N- { 2(S)-[((2-Methylindol-3-yl)thio)acetamido]-4-methylpentyl } -N-
(I-naphthylmethyl)- Iycvl-methionine
W096/09836 2~ 0 1 3 4 6 PCT/US95112319
- 102-
Anal. Calcd for C35H44N404S2-0.5 EtOAc-0.5 H2O
C, 63.30; H, 7.04; N, 7.98
C,63.17;H,6.77;N,7.98
5 N- { 2(S)-[(4-Pyridylthio)acetamido]-3(S)-methylpentyl } -N-(l -
naphthylmethyl)-glycyl-methionine
FAB mass spectrum m/z = 579 (M+1).
10 N- { 2(S)-[(4-Picolinylthio)acetamido] -3 (S)-methylpentyl } -N-( l -
naphthylmethyl)- Iycvl-methionine
Anal. Calcd for C32H42N404S2-2.0 TFA-1.7 H2O
C, 49.72; H, 5.49; N, 6.44
C, 49.51; H, 5.10; N, 6.57
N- { 2(S)-[3-(Benzylthio)propionamido]-3 (S)-methylpentyl } -N-( l -
naphthylmethyl)-~lycyl-methionine
20 FAB mass spectrum m/z = 624 (M+l).
Anal. Calcd for C34H4sN3O4S2-1.0TFA
C, 58.60; H, 6.28; N, 5.69
C, 58.63; H, 6.29; N, 5.92
25 N- { 2(S)-~(Benzyloxy)acetamido]-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-~lvcyl-methionine
FAB mass spectrum m/z = 594 (M+1).
30 N- { 2(S)-[(N'-Benzylglycyl)amino] -3(S)-methylpentyl } -N-(1 -
naphthylmethvl~- Iycyl-methionine
Anal. Calcd for C33H44N404S-2.0 TFA-1.0 H2O
C, 52.97; H, 5.77; N, 6.68
C, 52.91; H, 5.61; N, 6.63
W096/09836 ~ 4 6 PCI/US95/12319
- 103-
., .
N- { 2(S)-[(N'-Acetyl-N'-benzylglycyl)amino]-3(S)-methylpentyl } -N-
( l -naphthylmethyl)-~lycyl-methionine
Anal. Calcd for C35H46N405S-1.0 TFA-0.75 H2O
C, 58.29; H, 6.41; N, 7.35
C,58.13;H,6.19;N,7.68
N- { 2(S)-[(Benzylthio-S-oxide)acetamido]-3(S)-methylpentyl } -N-(1-
o naphthylmethvl)-glvcvl-methionine
FAB mass spectrum m/z = 626 (M+1).
N- { 2(S)-[(Benzylthio-S,S-dioxide)acetamido]-3(S)-methylpentyl } -N-
( 1 -naphthylmethyl)- lycyl-methionine
FAB mass spectr~m m/z = 642 (M+1).
N- { 2(S)-[(Benzylthio)acetamido]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)- lycyl-methionine ~ulfone
The 1 H-NMR wa~ consistent with the titled structure.
N- { 2(S)-[(N'-4-Nitrobenzoylglycyl)amino]-3(S)-methylpentyl } -N-(1-
naphthylmethyl)-~lycyl-methionine
Anal. Calcd for C33H41N507S-0.5 H2O
C, 59.9g; H, 6.41; N, 10.60
C, 59.~4; H, 6.31; N, 10.22
N- { 2(S)-[(N'-Methyl-N'-4-nitrobenzoylglycyl)amino]-4-
methylpentyl ~ -N-( I -naphthylmethyl)-glycyl-methionine
Anal. Calcd for C34H43N507S 0.6 EtOAc-0.5 H2O
W096t09836 22 ~ ~ ~ 4 6 PCItUS95/12319 ~
- 104-
.
C, 60.08; H, 6.76; N, 9.62
C,60.06;H,6.45;N,9.60
N- { 2(S)-[(S-Benzyl-L-cysteinyl] -3 (S)-methylpentyl } -N-(1 -
naphthvlmethyl)- lycyl-methionine
FAB mass spectrum m/z = 639 (M+1).
Anal. Calcd for C34H46N4O4S2-2.4 TFA-0-7 H20
C, 50.37; H, 5.43; N, 6.06
o C, 50.34; H, 5.39; N, ~.14
N-~2(S)-[S-Benzyl-D-cysteinyl]-3(S)-methylpentyl}-N-(1 -
naphthylmethyl)-~lycyl-methionine
FAB mass spectrum m/z = 639 (M+1).
Anal. Calcd for C34H46N4O4S2-2.9 TFA-0.5 H2O
C, 48.85; H, 5.14; N, 5.73
C, 48.82; H, 5.12; N, 5.84
EXAMPLE 19
3-Benzylthiopropionyl-valyl-isoleucyl-methionine
This compound wa.s synthesized using standard techniques
of solution phase peptide synthesis starting from methionine methyl ester
25 and other commercially available materials.
Anal. Calcd for C26H4 l N3O5S2
C, 57.86; H, 7.66; N, 7.79
C, 57.54; H, 7.58; N, 7.81
EXAMPLE 20
N- ~ 2(S)-[(4-(1 -H-Tetrazol-5-yl)benzylthio)acetamido]-3(S)-
methylpentyl~-N-(I-naphthvlmethyl)- Iycyl-methionine sodium salt
W096/09836 ~a~346 PCT/US95/12319
- 105-
The sodium salt of N-{2(S)-[(4-(1-H-tetrazol-5-yl)-
benzylthio)acetamido]-3(S)-methylpentyl } -N-(1 -naphthylmethyl)-
glycyl-methionine was prepared from the corresponding methyl ester
(2.1 mg) in 200 ~L of MeOH treated with 150 ,uL of 0.1N NaOH and
the solution diluted to 1 mL for assay.
EXAMPLE 21
N- { 2(S)-[(N'-Benzyl-N'-t-butoxycarbonylglycyl)amino]-3(S)-
methylpentyl~-N-(l-naphthylmethyl)-glycvl-methionine sodium salt
The sodium salt of N-{2(S)-[(N'-benzyl-N'-t-butoxy-
carbonylglycyl)amino]-3(S)-methylpentyl } -N-( 1 -naphthylmethyl)-
glycyl-methionine was prepared in situ by hydrolysis of the
corresponding methyl ester (2.2 mg) in 0.3 mL of MeOH with 6.2 ~L
of lN NaOH to give a 10 mM solution for assay.
EXAMPLE 22
N- { [ 1 -(4-Nitrobenzylthio)acetylamino]cyclopent- 1 -ylmethyl } -N-(1-
naphthylmethyl)-glycyl-methionine methvl ester
Step A: Preparation of 1-tert-(butyloxy)carbonylaminocyclopentane-
1 -carboxaldehyde
The title compound was prepared by a Swern oxidation
(3 equivalents of pyridine-sulfurtrioxide complex and excess triethyl
arnine in DMSO/CH2C12) of BOC protected 1-amino-1-
hydroxymethyl-cyclopentane (Aldrich).
Step B: Preparation of N-{ [1-(4-Nitrobenzylthio)acetylamino]-
cyclopent- 1 -ylmethyl } -N-( 1 -naphthylmethyl)-glycyl-
methionine methyl ester
Following the procedure described in Example 1, but
substituting the 1-tert-(butyloxy)carbonylaminocyclopentane-
1-carboxaldehyde from Step A for N-t-butoxycarbonylisoleucinal
provided the title compound.
W0 96/09836 ~2 2 01~ 4 6 PCT/US95/12319
- 106-
C34H42N406S2 FAB mass spectrum, m/e 667 (M+1).
EXAMPLE 23
5 N- { [ 1 -(4-Nitrobenzylthio)acetylamino]cyclopent- 1 -ylmethyl } -N-( 1-
naphthylmethvl)-glycyl-methionine
Following the procedure described in Example 2, but
substituting the ester from Example 22 provided the title compound.
C33H40N4o6s2 FAB mass spectrum, m/e 653 (M+1).
EXAMPLE 24
N- { 2(S)-r2-(4-Nitrophenyl)ethylcarbamoylamino]-4-methylpentyl } -N-
(l-naphthylmethvl)-glycyl-methionine methyl e~ster
2-(4-Nitrophenyl)ethylamine (0.5 mmol) and p-
nitrophenylchloroformate (0.5 mmol) were allowed to react in the
presence of triethylamine ( 1 mmol) in 5 ml of DMF to provide 2-(4-
nitrophenyl)ethyl isocyanate. Without isolation, the isocyanate was
mixed with N-(2(S)-amino-4-methylpentyl)-N-(1-naphthylmethyl)-
20 glycyl-methionine methyl ester dihydrochloride (0.4 mmol) and
triethylamine (1 mmol) in 4 mL of DMF to provide the title compound.
C34H45N5O6S-0.15 hexane
Anal. Calcd for C26H4lN3oss2
C, 63.05; H, 7.14; N, 10.54
C, 63.10; H, 7.Q9; N, 10.16
EXAMPLE 25
N- { 2(S)-[2-(4-Nitrophenyl)ethylcarbamoylamino]-4-methylpentyl } -N-
3 o ( I -naphthylmethyl)- Iycyl-methionine
Following the procedure described in Example 2, but
substituting the ester from Example 24 provided the title compound.
C33H43NsO6S EtOAc
-
W096/09836 ~ 3 ~ 6 PCT/US95112319
- 107-
C, 61.22; H, 7.08; N, 9.65
C, 61.32; H, 6.72; N, 9.73
~.
EXAMPLE 26
N- { 2(S)-[(3(S)-tetrahydrofuryloxy)carbonylamino]-3(S)-methylpentyl}-
N-(l-naphthylmethyl)-~lycvl-methionine methyl ester
The succinimidyl carbonate of 3(S)-hydroxytetrahydrofuran,
was prepared as described by Ghosh, et al., (J. Med. Chem. 1993,
o 36:292). The carbonate (46 mg, 0.20 mmol) and N-(2(S)-amino-3(S)-
methylpentyl)-N-(l-naphthylmethyl)-glycyl-methionine methyl ester
dihydrochloride (106 mg, 0.20 mmol) were dissolved in 3 mL of
methylene chloride and 56 ,uL (0.40 mmol) of triethylamine was added.
After stirring overnight the mixture was diluted with ethyl acetate and
washed successively with 10% citric acid, 10% sodium bicarbonate and
brine. After drying, the solvent was evaporated and the residue was
chromatographed on silica (1.5% methanol in methylene chloride) to
provide N-{2(S)-[(3(S)-tetrahydrofuryloxy)-carbonylamino]-3(S)-
methylpentyl}-N-(1-naphthylmethyl)-glycyl-methionine methyl ester.
EXAMPLE 27
N- ~ 2(S)-[(3(S)-tetrahydrofuryloxy)carbonylamino~-3(S)-methylpentyl } -
N-( l -naphthylmethyl)-~lycyl-methionine
Following the procedure described in Example 2, but
substituting the ester from Example 26 provided the title compound.
C29H4 1 N306S-CF3C02H-2. 1 H20
C, 52.32; H, 6.54; N, 5.91
C,51.85;H,6.01;N,6.34
EXAMPLE 28
N- { 2(S)-[(cis-4-benzyloxytetrahydrofur-3-yloxy)carbonylamino]-3(S)-
methylpentyl ~ -N-t 1 -naphthylmethvl)-~lycyl-methionine methyl ester
W096109836 ~2 ~ 1 3 4 6 PCT/US95/12319
- 108-
Following the procedure described in Example 26, but
substituting the succinimidyl carbonate of racemic cis-4-benzyloxy-3-
hydroxytetrahydrofuran (prepared as described by Vacca, et al., (Can.
Pat. 2,084,800, June 17, 1993)) provided a 1:1 mixture of diastereomers
5 of N- { 2(S)-[(cis-4-benzyloxytetrahydrofur-3-yloxy)carbonylamino]-3(S)-
methylpentyl } -N-( l -naphthylmethyl)-glycyl-methionine methyl ester
which were inseparable by HPLC chromatography.
EXAMPLE 29
N- { 2(S)-[(cis-4-benzyloxytetrahydrofur-3-yloxy)carbonylamino]-3(S)-
methylpentyl ~ -N-( l -naphthylmethyl)-glycyl-methionine
Following the procedure described in Example 2, but
substituting the ester from Example 28 provided the title compound as a
mixture of diastereomers that were separated by reverse phase HPLC
chromatography .
Isomers A and B, C36H47N3O7S gave FAB MS m/e = 666 (M+l).
EXAMPLE 30
N- { 2(S)-(t-butoxycarbonylamino)-3(S)-methylpentyl } -N-(1 -
naphthylmethyl)-glycvl -methionine
Using the procedure described in Example 2, N-[2(S)-(t-
butoxycarbonylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)-glycyl-
25 methionine methyl ester (prepared in Example 1, Step B, was convertedto the title compound.
C29H43N3O5S-Q.4 H20
C, 62.99; H, 7.98; N, 7.60
C, 62.98; H, 7.83; N, 7.62
W096109836 ~2 ~ PCT/US95/12319
- 109- -
EXAMPLE 31
In vitro inhibition of ras farnesyl tran.sferase
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). Bovine FPTase was assayed in a volume of
o 100 ~l containing 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 ,ug/ml FPTase at 31 C
for 60 min. Reactions were initiated 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 ,~-plate counter. The 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 radioactivity in the presence of 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 ~lM ZnCl2 and 100 nM Ras-CVIM were added to 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 human FPTase by the assay described above
and were found to have ICso of < 10 ~M.
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W096/09836 ~2 0 13 4 6 PCT/US95/12319
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EXAMPLE 32
In vivo ras farnesvlation assay
The cell line used in this assay is a v-ras line derived from
5 either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21. The
assay is perfo~ned 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 (~1nal concentration of
solvent, methanol or dirnethyl sulfoxide, is 0.1%). After 4 hours at 37C,
o the cells are labelled in 3 ml methionine-free DMEM supple-meted with
10% regular DMEM, ~% fetal bovine serum and 400
mCil35S]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 PMS~) and the lysates cleared by centrifugation at
100,000 x g for 45 min. Aliquots of lysates cont~ining equal numbers of
acid-precipitable counts are bought to 1 ml with IP buffer (lysis buffer
lacking DTT) and immunoprecipitated with the ras-specific monoclonal
antibody Y13-259 (Furth, M.E. et al., J. Virol. 43:294-304, (1982)).
20 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
added for 45 min. The immunoprecipitates are washed four times with IP
buffer (20 nM HEPES, pH 7.5/1 mM EDTA/1% Triton X-100Ø5%
deoxycholate/0.1 %/SDS/0.1 M NaCl) boiled in SDS-PAGE sample
25 buffer and loaded on 13% acrylamide gels. When the dye front reached
the bottom, the gel is fixed, soaked in Enlightening, dried and
autoradiographed. The intensities of the bands corresponding to
farnesylated and nonfarnesylated ras proteins are compared to determine
the percent inhibition of farnesyl transfer to protein.
>
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W096/09836 ~ Q ~ ~ ~ 6 PCT/US95/12319
EXAMPLE 33
In vivo growth inhibition assay
To determine the biological consequences of FPTase
inhibition, the effect of the compounds of the instant invention on the
anchorage-independent growth of Ratl cells transformed with either a v-
ras, v-raf, or v-mos oncogene is tested. Cells transformed by v-Raf and
v-Mos maybe included in the analysis to evaluate the specificity of
instant compounds for Ras-induced cell transformation.
o Rat 1 cells transformed with either v-ras, v-raf, or v-mos are
seeded at a density of 1 x 104 cells per plate (35 mm in diameter) in a
0.3% top agarose layer in medium A (Dulbecco's modified Eagle's
medium supplemented with 10% fetal bovine serum) over a bottom
agarose layer (0.6%). Both layers contain 0.1% methanol or an
appropriate concentration of the instant compound (dissolved in methanol
at 1000 times the final concentration used in the assay). The cells are fed
twice weekly with 0.5 ml of medium A cont~ining 0.1% methanol or the
concentration of the instant compound. Photomicrographs are taken 16
days after the cultures are seeded and comparisons are made.
