Note: Descriptions are shown in the official language in which they were submitted.
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TITLE OF THE INVENTION
INHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
RELATED APPLICATION
The present patent application is a continuation-in-part
application of copending application Serial No. 08/412,828, filed March
29, 1995.
BACKGROUND OF THE INVENTION
The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ras)
are part of a si~n~llinp pathway that links cell surface growth factor
receptors to nuclear signals initiating cellular proliferation. Biological
and biochernical studies of Ras action indicate that Ras functions like a
G-regulatory ~r~teill. In the inactive state, Ras is bound to GDP. Upon
growth factor receptor activation Ras is induced to exchange GDP for
GTP and undergoes a conformational change. The GTP-bound form of
Ras propagates the growth stim~ tory signal until the signal is
termin~te-l by the intrinsic GTPase activity of Ras, which returns the
protein to its inactive GDP bound forrn (D.R. Lowy and D.M. Willumsen,
Ann. Rev. Biochem. 62:851-891 (1993)). Mutated ras genes (Ha-ras,
Ki4a-ras, Ki4b-ras and N-ras) are found in many hllm~n cancers,
including colorectal carcinoma, exocrine pancreatic carcinoma, and
myeloid leukemias. The protein products of these genes are defective in
their GTPase activity and constitutively transrnit a growth stimlll~tory
signal.
Ras must be localized to the plasma membrane for both
normal and oncogenic functions. At least 3 post-translational
modifications are involved with Ras membrane localization, and all 3
modifications occur at the C-terminus of Ras. The Ras C-terminus
contains a sequence motif termed a "CAAX" or "Cys-Aaal-Aaa2-Xaa"
box (Cys is cysteine, Aaa is an aliphatic amino acid, the Xaa is any amino
~ acid) (Willumsen et al.. Nature 310:583-586 (1984)). Depending on the
specific sequence, this motif ser~ es as a signal sequence for the enzymes
farnesyl-protein transferase or geranylgeran~ l-protein transferase, ~~ hich
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catalyze the aL~ylation of the cysteine residue of ~e CAAX motif with a
C1s or C20 isoprenoid, respec*ively. (S. Clarke., Ann. Rev. Biochem.
61:355-386 (1992); W.R. Schafer and J. Rine, Ann. Rev. Genetics 30:209-
237 (1992)). The Ras protein is one of several proteins that are known to
5 undergo post-tr~n~l~tional farnesyla*ion. Other farnesylated ~ teills
include the Ras-related GTP-binding ~ teil~s such as Rho, fungal m~*n~
factors, the nuclear l~min~, and the ~s~mm~ subunit of transducin. James,
et al., J. BioL Chem. 269, 14182 (1994) have iden*fied a peroxisome
associated protein Pxf which is also farnesylated. James, et al., have also
10 suggested that there are farnesylated proteins of unknown structure and
function in addi*ion to those listed above.
Inhibi*on of farnesyl-~r~teill transferase has been shown to
block the growth of Ras-transformed cells in soft agar and to modify
other aspects of their transformed phenotype. It has also been
15 demonstrated that certain inhibitors of farnesyl-protein transferase
selectively block the processing of the Ras onc~ t~in 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-
20 dependent tumors in nude mice (N.E. Kohl et al., Proc. Natl. Acad. SciU.S.A., 91:9141-9145 (1994) and induces regression of m~mm~ry and
salivary carcinomas in ras transgenic mice (N.E. Kohl et al., Nature
Medicine, 1:792-797 (1995).
Indirect inhibition of farnesyl-protein transferase in vivo has
25 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 including farnesyl
pyrophosphate. Farnesyl-protein transferase utilizes farnesyl
30 pyrophosphate to covalently modify the Cys thiol group of the Ras
CAAX box with a farnesyl, roup (Réiss et al.. Cell, 62:81-88 (1990);
Schaber et al., J. Biol. Chem. . 265: l 4701 - l 4704 (1990); Schafer et al.,
Science, 249: l 133-1139 (1990); Manne et al.. Proc. Natl. Acad. Sci USA,
87:7541-7545 (1990)) Inhibition of famesyl pyrophosphate biosynthesis
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by inhibiting HMG-CoA reductase blc~cks Ras membrane loc~li7~tion in
cultured cells. However, direct inhibition of farnesyl-protein transferase
would be more specific and attended by fewer side effects than would
occur with the required dose of a general inhibitor of isoprene
5 biosynthesis.
Tnhibitors of farnesyl-protein transferase (FPTase) have been
described in two general classes. The first are analogs of farnesyl
diphosphate (FPP), while the second class of inhibitors is related to the
protein substrates (e.g., Ras) for the enzyme. The peptide derived
10 inhibitors that have been described are generally cysteine con~inin~
molecules that are related to the CAAX motif that is the signal for protein
prenylation. (Schaber et al., ibid; Reiss et. al., ibid; Reiss et al., PNAS,
88:732-736 (1991)). Such inhibitors may inhibit ~l~)t~ prenylation
while serving as alternate substrates for the farnesyl-proteill transferase
15 enzyme, or may be purely competitive inhibitors (U.S. Patent 5,141,851,
University of Texas; N.E. Kohl et al., Science, 260: 1934-1937 (1993);
Graham, et al., J. Med. Chem., 37, 725 (1994)). In general, deletion of the
thiol from a CAAX derivative has been shown to dramatically reduce the
inhibitory potency of the compound. However, the thiol group
20 potentially places limit~tions on the therapeutic application of FPTase
inhibitors with respect to pharmacokinetics, ph~ codynamics and
toxicity. Therefore, a functional replacement for the thiol is desirable.
It has recently been reported that farnesyl-protein transferase
inhibitors are inhibitors of proliferation of vascular smooth muscle cells
25 and are therefore useful in the prevention and therapy of arteriosclerosis
and diabetic disturbance of blood vessels (JP H7-112930).
It has recently been disclosed that certain tricyclic
compounds which optionally incorporate a piperidine moiety are
inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516).
30 Imidazole-cont~ining inhibitors of farnesyl protein transferase have also
been disclosed (WO 95/09OO l and EP 0 675 112 Al ).
It is, therefore, an object of this invention to develop
peptidomimetic compounds that do not have a thiol moiety, and that will
inhibit farnesyl-protein transferase and thus. the post-translational
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W O96134010 ' PCTrUS9610397
farnesylation of proteins. It is a further object of this invention to develop
chemotherapeutic compositions cont~inin~ the compounds of this
invention and methods for producing the compounds of this invention.
S SUMMARY OF THE INVENTION
The present invention comprises analogs of the CAlA2X
motif of the protein Ras that is modified by farnesylation in vivo. These
CAlA2X analogs inhibit the farnesylprotein transferase. Furthermore,
these CAlA2X analogs differ from those previously described as
10 inhibitors of farnesyl-protein transferase in that they do not have a thiol
moiety. The lack of the thiol offers unique advantages in terms of
improved ph~rm~t~okinetic behavior in ~nim~l.c, prevention of thiol-
dependent chemical reactions, such as rapid autoxidation and disulfide
formation with endogenous thiols, and re~ re~ systemic toxicity. The
15 compounds of the instant invention also incorporate a cyclic amine
moiety in the A2 position of the motif. Further cont~in~.-l in this
invention are chemotherapeutic compositions cont~ining these farnesyl
transferase inhibitors and methods for their production.
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l~e compounds of this invention are illustrated by the
forrn~
V- A1(CR1a2)nA2(CR1~2)n ~- (CR1b2)p~N~OH
R4a R4b
V A1 (CR1 a2)nA2(CR 1a2)n ~W)- (CR1b2)p~
R4a R4b
HOCH2(CH2)q
V-A1(cR1~2)nA2(cR182)n-(w (CR1b2)p~N~
V - A1 (CR1 a2)nA2(cR 1 a2)n (W)- (CRl bz)p~f N ~H~
X R4b
IV R4a/
DETAILED D~;S(~RIPTION OF THE INVENTION
S The compounds of this invention inhibit the farnesyl-protein
transferase. In a first embodiment of this invention, the farnesyl-protein
transferase inhibitors are illustrated by the forrnula I:
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(R~), ( ), R6 Y ~,
V-A (CR1a2),~AZ(CR1a2)n (CR1b2)p~fN~R4bo
wherem:
Rla and Rlb are independently selected from:
a) hydrogen,
S b) aryl, heterocycle, cycloaLkyl, aL~enyl, aL~cynyl, R100-,
Rl lS(O)m-, RlOC(O)NR10-, CN, NO2, (R10)2N-
C(NR10)-, RlOC(O)-, RlOoc(o)-~ N3, -N(R10)2, or
Rl lOC(o)NRlO
c) Cl-C6 aL~yl unsubstituted or substituted by aryl,
heterocyclic, cycloaLkyl, aL~enyl, alkynyl, R100-,
RllS(O)m-, RlOC(O)NR10-, CN, (R10)2N-C(NR10)-,
RlOc(o)-~ RlOoc(o)-7 N3, -N(R10)2, or Rl loC(O)
NRlO;
15 R2 and R3 are independently selected from:
a) a side chain of a naturally occurring arnino acid,
b) an oxidized form of a side chain of a naturally occurring
amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substituted or unsubstituted C 1 -C20 alkyl, C2-C20 alkenyl,
C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and
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d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group s~ cte-l from aryl, heterocycle and C3-
Clo cycloalkyl; or
S R2 and R3 are combined to forrn - (CH2)S -; or
R2 or R3 are combined with R6 to forrn a ring such that
R6 ~
~;N2~' R7a~H2),
- R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubsti~lte~l or subctitlltç-l by alkenyl, R100-,
RllS(O)m-, RlOC(O)NR10-, CN, N3, (R10)2N-C(NR10)-,
RlOC(O)-, RlOOC(O)-, -N(R10)2, or Rl lOC(O)NR10-,
c) aryl, heterocycle, cycloaLkyl, alkenyl, R100-,
Rl lS(O)m-, RlOC(O)NR10-, CN, NO2, (R10)2N-
C(NRlo)-~ RlOc(o)-~ Rlooc(o)-~ N3, -N(R10)2,
or Rl lOC(O)NR10-, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and
- C3-Clo cycloalkyl;
R5a and Rsb 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 C 1 -C20 alkyl, C2-c2o alkenyl,
C3-C l o cycloalkyl, aryl or heterocycle group,
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W O96/34010 PCTrUS96/03975
wherein the substituent is selected from F, Cl, Br,
(RlO)2Nc(o)-~ N02, R10O-, RllS(o)m
R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-,
R10OC(O)-, N3, -N(R10)2, Rl 1OC(O)NR10- and
S Cl-C20 aL~yl,
d) Cl-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloallyl; or
10 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)-;
R6 is independently selected from hydrogen or C l-C6 aLkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, aL~cenyl, aLkynyl,
perfluoroalkyl, F, Cl, Br, R100-, Rl lS(O)m-,
R10c(o)NRlo-~ CN, NO2, R102N-C(NR10)-, R10C(O)-,
R10oc(o)-~ N3, -N(R10)2, or R1 1OC(O)NR10-, and
c) C 1-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, cycloaL~yl, alkenyl, aLkynyl, perfluoroaL~yl, F,
Cl, Br, R10O-, Rl lS(O)m-, R10C(O)NH-, CN, H2N-
C(NH)-, R10C(O)-, Rl0oc(o)-~ N3, -N(R10)2, or
RlOOC(O)NH-;
R9 is selected from:
a) hydrogen,
30 b) alkenyl, alkynyl, perfluoroaLkyl, F. Cl, Br, R100-,
Rl l~S(O)m-, RlOC(O)NR10-, CN. N02, (R10)2N-C-
(NR 10), R 1 ~C(O)-, R 1 ~OC(O)-. N3, -N(R 1~)2, or
R 1 1 OC(O)NR 10 and
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c) Cl-C6 aL~yl unsubstituted or substituted by perfluoroalkyl,
F, Cl, Br, R100-, Rl lS(O)m-, RlOC(O)NR10-, CN,
(Rlo)2N-c(NRlo)-7 Rloc(o)-7 RlOoc(o)-7 N3, -N(R10)2,
or Rl lOC(O)NR10;
s
R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and
aryl;
Rl 1 is independently selected from Cl-C6 aL~yl and aryl;
Al and A2 are independently selected from: a bond, -CH=CH-, -C--C-,
-C(0)-, -C(O)NR 10 , -NR 1 ~C(0)-, 0, -N(R 10) ,
-S(0)2N(R10)-, -N(RlO)s(o)2-7 or S(O)m;
15 Q is a substituted or unsubstituted nitrogen-cont~ining C4-Cg mono or
bicyclic ring system, wherein the non-nitrogen cont~ining ring may be a
C5-C7 saturated ring;
V is selected from:
a) hydrogen,
b) heterocycle,
c) aryl,
d) Cl-C20 aLkyl wherein from 0 to 4 carbon atoms are replaced
with a a heteroatom selected from 0, S, and N, and
e) C2-C20 aLkenyl,
provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if
Al is a bond, n is 0 and A2 is S(O)m;
W is a heterocycle;
X, Y and Z are independently H2 or 0;
.,
m is 0 1 or ';
nis 0, 1. '.30r4;
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- 10-
pis 0, 1,2, 3 or4;
r is 0 to S, provided that r is 0 when V is hydrogen;
sis 40rS;
t is 3, 4 or S; and
S u is 0 or 1;
or the ph~ ceutically acceptable salts thereof.
In a second embodiment of this invention the prodrugs of
compounds of formula I are illustrated by the formula II:
V - A (cR1a2)nA2(cR1~2)n -(W)- (CR1b2)p~ J ~ ~ J
R4a
wherein:
R1a and Rlb are independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, R100-,
lS R1 lS(O)m, RlOC(O)NR10-~ CN, N02,
(R10)2N-C(NR10)-, R1OC(0)-~ RlOoc(o)-~ N3, -N(R10)2,
or Rl 1OC(O)NR10
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-,
RllS(O)m-, RlOC(O)NR10-, CN, (R10)2N-C(NR10)-,
RlOC(0)-, RlOOC(0)-, N3, -N(Rl0)2, or Rl loC(0)
NRlO;
R2 and R3 are independently selected from:
a) a side chain of a naturally occurrin~ amino acid,
b) an oxidized form of a side chain of a naturally occurrin~
amino acid which is:
i) methionine sulfoxide~ or
ii) methionine sulfone. and
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c) substituted or unsubstituted Cl-C20 aL~yl, C2-C20 aL~enyl,
C3-Clo cycloaL~yl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
S CN, (R10)2N-C(NR10)-, R10C(O)-, R10OC(O)-, N3,
-N(R10)2, Rl 10C(O)NR10- and Cl-C20 aL~yl, and
d) Cl-C6 aL~yl substitllte~1 with an unsubstituted or
sub~titlltç-1 group selected from aryl, heterocycle and C3-
Clo cycloaL~yl; or
R2 and R3 are combined to form - (CH2)S -; or
R2 or R3 are combined with R6 to form a ring such that
R6 R7a ~R7b
R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubstituted or substituted by aL~enyl, R100-,R1 1S(O)m-, R10C(O)NRl0-~ CN, N3, (R10)2N-C(NR10)-,
R10C(O)-, R10OC(O)-, -N(R10)2, or Rl lOC(O)NR10-,
c) aryl, heterocycle, cycloaLkyl, alkenyl, R100-,
Rl1S(O)m-, R10C(O)NRl0-~ CN, NO2, (R10)2N-
C(NRlo)-~ R10C(o), R10oc(o)-~ N3, -N(R10)2,
or R 1 1 OC(O)NR 10 , and
d) C 1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and
C3-C l o cycloalkyl;
c~
30 RSa and R5b are independently selected from:
a) a side chain of a naturallv occurring amino acid~
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- 12 -
b) an oxitli~ecl form of a side chain of a naturally occurring
amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
S c) substituted or unsubstituted Cl-C20 aIkyl, C2-C20 alkenyl,
C3-Clo cycloaL~yl, aryl or heterocycle group,
wherein ~e substituent is selected from F, Cl, Br,
(RlO)2Nc(o)-~ N02, R10O-, Rl lS(o)m
RlOC(O)NR10, CN, (R10)2N-c(NRlo)-~ RlOC(O)-~
RlOOC(O)-, N3, -N(R10)2, RllOC(O)NR10- and
Cl-C20 alkyl,
d) Cl-C6 aL~yl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaIkyl; or
RSa and Rsb 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)-;
R6 is independently selected from hydrogen or Cl-C6 aLkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl,
perfluoroaIkyl, F, Cl, Br, RlOO-, Rl lS(O)m-,
R 1 OC(O)NR 10 , CN, NO2, R 1 02N-C(NR 10) , Rl ~C(O)-,
RlOOC(O)-, N3, -N(R10)2, or R1 lOC(O)NR10-, and
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, cycloalkyl, alkenyl. alkynyl, perfluoroalkyl, F,
3Q Cl, Br, R100-, Rl lS(O)m-, RlOC(O)NH-, CN, H2N-
C(NH)-, R1OC(O)-, RlOOC(O)-. N3, -N(Rl0)2, or
R 1 OOC(O)NH-;
R9 is selected from:
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a) hydrogen,
b) aLkenyl, alkynyl, perfluoroaL~yl, F, Cl, Br, R100-,
Rl1S(O)m-, RlOC(O)NR10-, CN, NO2, (R10)2N-C-
(NR10)-, R10c(o)-~ R10oc(o)-~ N3, -N(R10)2, or
S RllOC(O)NR10-, and
c) Cl-C6 aL~yl unsubstituted or substituted by perfluoroaL~yl,
F, Cl, Br, R1OO-, R11S(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, RlOC(O)-~ RlOOC(O)-, N3, -N(R10)2,
or Rl 1OC(O)NR10;
R10 is independently selected from hydrogen, Cl-C6 aL~yl, benzyl and
aryl;
R11 is independently selected from Cl-C6 aLkyl and aryl;
R12 is
a) substituted or unsubstitllteA Cl-Cg aL~yl, substituted or
unsubstituted C5-Cg cycloaL~yl, or substituted or
unsubstituted cyclic amine, wherein the substituted aLkyl,
cycloaLkyl or cyclic amine is substituted with 1 or 2
substituents independently selected from:
1 ) C 1 -C6 alkyl,
2) aryl,
3) heterocycle,
4) -N(R 1 1)2,
5) -OR10, or
b)
R13 o
0~ R14
30 R13 is independently selected from hydrogen and Cl-C6 alkyl;
R14 is independently selected from Cl-C6 ~l};yl;
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- 14-
Al and A2 are independently selected from: a bond, -CH=CH-, -C-C-,
-C(O)-, -C(O)NR 1 0, -NR 1 ~C(O)- , O, -N(R 1 0),
-S(0)2N(R10)-, -N(RlO)s(o)2-~ or S(O)m;
S Q is a substihlte-l or unsubstituted nitrogen-cont~ining C4-Cg mono or bicyclic ring ~y~ , wherein the non-nitrogen cont~inin~ ring may be a
C5-C7 saturated ring;
V is selected from:
a) hydrogen,
b) heterocycle,
c) aryl,
d) C l-C20 aL~yl wherein from O to 4 carbon atoms are replaced
with a a heteroatom selected from 0, S, and N, and
e) C2-C20 aL~enyl,
provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if
Al is a bond, n is O and A2 is S(O)m;
W is a heterocycle;
X, Y and Z are independently H2 or 0;
m is 0, 1 or 2;
n is 0, 1, 2, 3 or4;
pis 0,1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
sis 40rS;
tis 3,40rS;and
uis Oorl;
30 or the pharrnaceutically acceptable salts thereof.
In a third embodiment of this in- ention, the inhibitors of
farnesyl transferase are illustrated by the forrnula III:
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HOCH2(CH2)q
V-A (CR 2)nA2(CR1a2)n-(W) (CR1bz)p~
wherein:
Rla and R1b are independently selected from:
S a) hydrogen,
b) aryl, heterocycle, cycloaLkyl, aLkenyl, aLkynyl, R100-,
Rl 1S(O)m-, RlOc(o)NRlo-~ CN, NO2, (R10)2N-
C(NR10)-, RlOC(O)-, R1OOC(O)-, N3, -N(R10)2, or
RllOC(O)NR10
c) Cl-C6 alkyl unsubstituted or substitllte-l by aryl,
heterocyclic, cycloaLkyl, aLkenyl, alkynyl, R100-,
R1 lS(O)m-, R1OC(O)NR10-, CN, (R10)2N-C(NR10)-,
R1OC(O)-, R10oc(o)-~ N3, -N(R10)2, or Rl loC(O)-
NR10;
R2 and R3 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, and
c) substituted or unsubstituted C 1 -C20 alkyl, C2-C20 aLkenyl,
C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(RlO)2~ NO2, RlOO-, Rl lS(O)m, RlOC(O)NR10-
CN, (R 1 0)2N-C(NR 10), R 1 ~C(O)-, R 1 ~OC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 alk~l, and
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- 16-
d) Cl-C6 aL~yl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
5 R2 and R3 are combined to form - (CH2)S -; or
R2 or R3 are combined with R6 to form a ring such ~at
R6 ~
~-,N2~ R7a ~HR2)b
R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubstituted or substituted by aL~enyl, R100-,
Rl lS(O)m-, R10C(O)NRl0-~ CN, N3, (R10)2N-C(NR10)-,
R10C(O)-, R10OC(O)-, -N(R10)2, or Rl lOC(O)NR10-,
c) aryl, heterocycle, cycloaL~yl, aL~enyl, R100-,
R1 1S(O)m-, R10C(O)NRl0-~ GN, NO2, (R10)2N-
C(NR10)-, R10C(O)-, R10OC(O)-, N3, -N(R10)2,
or Rl lOC(O)NR10-, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and
C3-Clo cycloaLkyl;
R6 is independently selected from hydrogen or Cl-C6 aLkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, a~enyl, alkynyl,
perfluoroalkyl, F, Cl, Br, R10O-. Rl lS(O)m-,
R 1 0C(O)NR 1 0-, CN. NO2, R 10 N-C(NR 10)-, Rl ~C(O)-~
R 1 ~OC(O)-. N3, -N(R 1 ~)2. or R l l OC(O)NR 10-. and
CA 02216~32 1997-09-26
W O 96/34010 PCTrUS96/0397
c) Cl-C6 aL~yl unsubstituted or substituted by aryl,
heterocycle, cycloaL~yl, alkenyl, alkynyl, perfluoroalkyl, F,
Cl, Br, RlOO-, Rl lS(O)m-, RlOC(O)NH-, CN, H2N-
C(NH)-, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
RlOOC(O)NH-;
R9 is selected from:
a) hydrogen,
b) alkenyl, aL~ynyl, per~1uoroaL~yl, F, Cl, Br, R100-,
RllS(O)m-, RlOC(O)NR10-, CN, NO2, (R10)2N-C-
(NR10)-, RlOC(O)-, RlOoc(o)-~ N3, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aL~yl unsubstituted or substit-lte-l by perfluoroallyl,
F, Cl, Br, RlOO-, RllS(O)m-, RlOc(o)NRlo-~ CN,
(R10)2N-C(NR10)-, RlOC(O)-, RlOoc(o)-~ N3, -N(R10)2,
or Rl lOC(O)NR10;
R10 is independently selected from hydrogen, Cl-C6 aL~yl, benzyl and
aryl;
Rl 1 is independently selected from Cl-C6 aL~yl and aryl; -
Al and A2 are independently selected from: a bond, -CH=CH-, -C_C-,
-C(O)-, -C(O)NR 10 , -NR 1 ~C(O)-, O, -N(R 10) ,
-S(O)2N(R10)-, -N(RlO)S(O)2-, or S(O)m;
Q is a substituted or unsubstituted nitrogen-cont~ining C4-Cg mono or
bicyclic ring system, wherein the non-nitrogen cont~inin~: ring may be a
Cs-C7 saturated ring;
r 30
V is selected from:
a) hydrogen,
b) heterocycle,
c) aryl
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- 18-
d) Cl-C20 aLkyl wherein from O to 4 carbon atoms are replaced
with a a heteroatom selected from 0, S, and N, and
e) C2-C20 alkenyl,
provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if
5 Al is a bond, n is O and A2 is S(O)m;
W is a heterocycle;
X, Y and Z are independently H2 or 0;
mis 0,1 or2;
nis 0, 1,2,30r4;
pis 0, 1,2,30r4;
qis 0,1 or2;
lS r is O to S, provided that r is O when V is hydrogen;
sis 40r5;
tis 3,40rS;and
uis Oor l;
20 or the ph~ çeutically acceptable salts thereof.
In a fourth embodiment of this invention the prodrugs of
compounds of formula III are illustrated by the formula IV:
V Al(CR1a2)nA2(CR1a2 n ~w - (CR1b )p~ à
25 wherein:
Rla and Rlb are independently selected from:
a) hydrogen,
CA 02216~32 1997-09-26
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- 19-
b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, R100-,
R11S(o)m, Rloc(o)NRlo-~ CN, NO2, (R10)2~-
C(NR10)-, RlOC(O)-, RlOOC(O)-, N3, -N(Rl0)2, or
RllOC(O)NR10
Sc) Cl-C6 alkyl unsubstitllte-l or substitllterl by aryl,
heterocyclic, cycloaL~yl, aL~enyl, alkynyl, R100-,
R11S(O)m-, R1OC(O)NR10-, CN, (R10)2N-C(NR10)-,
R1OC(O)-, R1OOC(O)-, N3, -N(R10)2, or Rl loC(O)-
NR10,
R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxi~li7e-1 form of a side chain of a naturally occurring
amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substituted or unsubstituted C1-C20 alkyl, C2-C20 alkenyl,
C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, NO2, RlOO-, R11S(O)m-, R1OC(O)NR10-,
CN, (R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, R1 1OC(O)NR10- and C1-C20 alkyl, and
d) C 1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; or
R2 and R3 are combined to form - (CH2)S -; or
R2 or R3 are combined with R6 to form a rin~ such that
R6 r~
;N~ is ~ (~H2)~ ;
R2 R7a R7b
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- 20 -
R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubstituted or substituted by aL~cenyl, R100-,
S RllS(O)m-, RlOC(O)NR10-, CN, N3, (R10)2N-C(NR10)-,
R10C(O)-, R10OC(O)-, -N(R10)2, or Rl lOC(O)NR10-,
c) aryl, heterocycle, cycloaL~yl, aL~enyl, R100-,
Rl1S(O)m-, R10C(O)NRl0-~ CN, NO2, (R10)2N-
C(NR10)-, R10C(O)-, R10OC(O)-, N3, -N(R10)2,
or Rl lOC(O)NR10-, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and
C3-Clo cycloalkyl;
15 R6 is independently selected from hydrogen or Cl-C6 alkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, alkenyl, aLkynyl,
perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(O)m-,
Rl0c(o)NRlo-~ CN, NO2, R102N-C(NR10)-, R10C(O)-~
R10OC(O)-, N3, -N(R10)2, or R1 1OC(O)NR10-, and
c) C l-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, cycloaLkyl, alkenyl, aL~ynyl, perfluoroalkyl, F,
Cl, Br, R10O-, R1 1S(O)m-, R10C(O)NH-, CN,H2N-
C(NH)-, R10C(O)-, R10OC(O)-, N3, -N(R10)2, or
RlOOC(O)NH-;
R9is selected from:
a) hydrogen,
b) aL~cenyl, aL~cynyl, perfluoroaL~cyl, F, Cl, Br, R100-,
Rl ls(o)m-~ RlOC(O)NR10-, CN. N02, (R10)2N-C-
(NR10), Rl0C(o), R10OC(O)-~ N3, -N(R10)2. or
R I 1 OC(O)NR 10-. and
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- 21 -
c) Cl-C6 alkyl unsubstituted or substituted by perfluoroaL~yl,
F, Cl, Br, RlOO-, Rl 1S(O)m-, RlOC(O)NR10-, CN,
(Rlo)2N-c(NRlo)-~ Rloc(o)-~ R10Oc(o)-~ N3, -N(R10)2,
or Rl lOC(O)NR10;
s
R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and
aryl;
R11 is independently selected from Cl-C6 aL~yl and aryl;
A1 and A2 are independently selected from: a bond, -CE~=CH-, -C_C-,
-C(O)-, -C(O)NR 10 , -NR 1 ~C(O)-, O, -N(R 10
-S(O)2N(R10)-, -N(RlO)S(O)2-, or S(O)m;
15 Q is a substi1~lte-1 or unsubstihlte~l nitrogen-cont~ining C4-Cg mono or
bicyclic ring system, wherein the non-nitrogen cont~inin~ ring may be a
C5-C7 saturated ring;
V is selected from:
a) hydrogen,
b) heterocycle,
c) aryl,
d) Cl-C20 aL~yl wherein from 0 to 4 carbon atoms are replaced
with a a heteroatom selected from O, S, and N, and
e) C2-C20 aL~enyl,
provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if
A1 is a bond, n is 0 and A2 is S(O)m;
W is a heterocycle;
X, Y and Z are independently H2 or O;
y
m is 0~ l or'';
n is 0. 1, ', 3 or4:
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- 22 -
pis 0,1,2,30r4;
qis 0,lor2;
r is 0 to S, provided that r is 0 when V is hydrogen;
sis 40rS;
S tis 3,40rS;and
u is 0 or 1;
or the ph~ eutically acceptable salts thereof.
In a more preferred embodiment of this invention, the Ras
farnesyl transferase inhibitors are illustrated by the Formula I:
V-A1(CR1a2)nAZ(cR1l2)n-(w) (CR1b2)p~N? ~
wherein:
Rla is independently selected from: hydrogen or Cl-C6 aLkyl;
R1b is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, R100-, -N(R10)2 or alkenyl,
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, cycloalkyl, alkenyl, RlOO-, or-N(R10)2;
R2 and R3 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~
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- 23 -
c) substihlte~l or unsubstituted Cl-Clo aL~yl, C2-Clo aL~enyl,
C3-Clo cycloaL~yl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R10O-, R11S(O)m-, R10C(O)NR10-, CN,
(R10)2N-c(NRlo)-~ R10C(O)-~ R10OC(O)-~ N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 aL~yl, and
d) Cl-C6 aL~yl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaL~yl, or
R2 and R3 are combined to form - (CH2)S -; or
R2 or R3 are combined with R6 to form a ring such that
~,; N~, R7a ~R7b
R4a and R7a are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubstituted or substituted by alkenyl, R100-,
R11S(O)m-, R10C(O)NRl0-~ CN, N3, (R10)2N-C(NR10)-,
R10C(O)-, R10OC(O)-, -N(R10)2, or R1 lOC(O)NR10-,
c) aryl, heterocycle, cycloaLkyl, alkenyl, R100-,
R1 lS(O)m-, R10C(O)NRl0-~ CN, NO2, (R10)2N-
C(NR10), RlOC(O)-~ RlOOC(O)-, N3,-N(R10)2,
or R 1 1 OC(O)NR 10, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and C3-
C l o cycloalkyl;
~0 R4b and R7b are hydrogen;
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- 24 -
RSa is selected from:
a) a side chain of a naturally occurring ~mino acid, wherein the
aII~ino acid is selected from methionine and glllt~mine,
b) an oxidized form of a side chain of a naturally occurring
S amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substihlte~l or unsubstituted Cl-Clo aL~yl, C2-Clo aL~enyl,
C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02, R100-, RllS(O)m-, RlOC(O)NR10-,
(R10)2Nc(o)-~ CN, (R10)2N-C(NR10)-, RlOC(O)-,
R10OC(O)-, N3, -N(R10)2, R11OC(O)NR10- and
Cl-C20 alkyl, and
15 d) Cl-C6 aL~yl substihlte~l with an unsubstituted or
sub~titllte.-1 group selected from aryl, heterocycle and C3-
Clo cycloaLkyl;
RSb is selected from:
a) hydrogen, and
b) Cl-C3 alkyl;
R6 is independently selected from hydrogen or C 1-C6 alkyl;
2~ 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)-, R10C(O)-, R10OC(O)-~ -N(R10)2, or
R 1 1 OC(O)NR 10 , and
c) C l ~C6 alkyl substituted by C l -C6 perfluoroaLkyl, R l ~O-~
R10C(O)NR10-, (R10)2N-C(NR10)-, RlOC(O)-,
R10OC(O)-, -N(R10)2, or Rl lOC(03NR10-;
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- 25 -
R9 is selected from:
a) hydrogen,
b) C2-C6 aL~enyl, C2-C6 alkynyl, C l-C6 perfluoroaL~yl, F, Cl,
R1OO-, Rl 1S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
S C(NR10)-, R1OC(O)-, R1OOC(O)-, -N(R10)2, or
R1 1OC(O)NR10-, and
c) Cl-C6 aLkyl unsubstitllte~l or substituted by Cl-C6
perfluoroaLkyl, F, Cl, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, R10c(o)-~ R10oc(o)-~ -N(R10)2,
or R1 1OC(O)NR10;
R10 is independently selected from hydrogen, Cl-C6 aLkyl, benzyl and
aryl;
15 Rl 1 is independently selected from Cl-C6 aLkyl and aryl;
Q is selected from:
,~
~~N~ N~ and -~- N~
Al and A2 are independently selected from: a bond, -CH=CH-, -C_C-,
-C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;
25 V is selected from:
a) hydrogen,
b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl,
isoquinolinyl, and thienyl,
c) aryl,
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- 26 -
d) Cl-C20 alkyl wherein from O to 4 carbon atoms are replaced
with a a heteroatom selected from 0, S, and N, and
e) C2-C20 aLkenyl, and
provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if
5 A1 is a bond, n is O and A2 is S(O)m;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or
isoquinolinyl;
X, Y and Z are independently H2 or 0;
mis 0,1 or2;
nis 0,1,2,30r4;
pis 0,1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
tis 3,40r5;and
uis Oor l;
20 or the pharmaceutically acceptable salts thereof.
In a second more preferred embodiment of this invention,
the prodrugs of the preferred compounds of Formula I are illustrated by
the Formula II:
V ~ A1(CR1a2)nA2(CRla2)n (W)- (CR1b2)p~oRl2
X R4b
11 R4a
wherein:
Rla is independently selected from: hvdro~en or Cl-C6 alkyl;
CA 02216~32 1997-09-26
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R1b is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloaL~yl, R100-, -N(R10)2 or aL~enyl,
S c) Cl-C6 aL~yl unsubstituted or substituted by aryl,
heterocycle, cycloalkyl, aL~enyl, R100-, or-N(R10)2;
R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally occurring
amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-Clo alkyl, C2-Clo alkenyl,
C3-Clo cycloaLkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, R11S(O)m-, R10c(o)NRlo-~ CN,
(R10)2N-C(NR10)-, R10c(o)-~ R10oc(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
R2 and R3 are combined to form - (CH2)s -; or
R2 or R3 are combined with R6 to form a ring such that
R6 ~
R7a ~HR2)b
30 R4a and R7a are independently selected from:
a) hvdrogen,
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- 28 -
b) Cl-C6 aL~yl unsubstituted or substituted by alkenyl, R100-,
Rl lS(O)m-, R1OC(O)NR10-, CN, N3, (R10)2N-C(NR10)-,
R1OC(O)-, R1OOC(O)-, -N(R10)2, or R1 1OC(O)NR10-,
c) aryl, heterocycle, cycloaL~yl, aL~enyl, R100-,
R1 1S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
C(NR10)-, R1OC(O)-, R1OOC(O)-, N3, -N(R10)2,
orR11OC(O)NR10-, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and C3-
Clo cycloaL~yl;
R4b and R7b are hydrogen;
RSa is selected from:
a) a side chain of a naturally occl-rnn~ amino acid, wherein the
amino acid is selected from methionine and gl~
b) an o~itli7e~1 form of a side chain of a naturally occurring
amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substituted or unsubstituted Cl-Clo aL~yl, C2-Clo alkenyl,
C3-Clo cycloaL~yl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02,R100-,RllS(O)m-,RlOC(O)NR10-,
(R10)2Nc(o)-~ CN, (R10)2N-C(NR10)-, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, Rl lOC(O)NR10- and
Cl-C20 aL~yl, and
d) Cl-C6 aL~cyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl;
R~b is selected from:
a) hydro~en. and
b) C1-C~ al~yl;
- ==
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- 29 -
.,
R6 is independently selected from hydrogen or Cl-C6 aL~yl;
R8 is independently selected from:
5a) hydrogen,
b) Cl-c6 aL~yl, C2-c6 aL~enyl, C2-c6 alkynyl, Cl-c6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2, or
R11OC(O)NR10-, and
10c) Cl-C6 aL~yl substituted by Cl-C6 perfluoroaL~yl, R100-,
RlOC(O)NR10-, (RlO)2N-C(NR10)-, RlOC(O)-,
R1OOC(O)-, -N(R10)2, or Rl 1OC(O)NR10-;
R9 is selected from:
15a) hydrogen,
b) C2-C6 aL~enyl, C2-C6 aL~ynyl, C l-C6 perfluoroaL~yl, F, Cl,
RlOO-, Rl 1S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
C(NR10)-, RlOC(O)-, R1OOC(O)-, -N(R10)2, or
R1 lOC(O)NR10-, and
20c) Cl-C6 aL~yl unsubstituted or substituted by Cl-C6
perfluoroaL~yl, F, Cl, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOc(o)-~ RlOoc(o)-~ -N(R10)2,
or Rl lOC(O)NR10;
25 R10 is independently selected from hydrogen, Cl-C6 aLkyl, benzyl and
aryl;
R l 1 is independently selected from C l -C6 aLkyl and aryl;
30 R 1 2 is
a) substituted or unsubstitutéd C I -C8 aL~cyl or substituted or
unsubstituted C5-Cg cycloalkyl. wherein the substituent on
the aLkyl or cycloalkvl is selected from:
1 ) aryl,
-
CA 02216532 1997-09-26
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- 30 -
2) heterocycle,
3) -N(R1 1)2,
4) -OR10, or
b)
R13 0
~,loJ~ Rl4
R13 is independently selected from hydrogen and Cl-C6 allyl;
R14 is independently selected from Cl-C6 alkyl;
Q is selected from:
~ ~\
N~> and -~- N 2~>
Al and A2 are independently selected from: a bond, -CH=CH-, -~C-,
-C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;
V is selected from:
a) hydrogen,
b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl,
isoquinolinyl, and thienyl,
c) aryl,
d) Cl-C20 aL~cyl wherein from 0 to 4 carbon atoms are replaced
with a a heteroatom selected from O, S, and N, and
e) C2-C20 aLkenyl, and
provided that V is not hydro~en if Al is S(O)m and V is not hydro~en if
A1 is a bond, n is 0 and A2 is S(O)m;
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- 31 -
W is a heterocycle selected from pyrrolidinyl, imi~ 7.olyl, pyridinyl,
thiaaolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or
isoquinolinyl;
X, Y and Z are independently H2 or 0;
m is 0, 1 or 2;
nis 0, 1,2,30r4;
pis 0,1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
tis 3,40rS;and
uis Oor l;
15 or the ph~ ceutically acceptable salts thereof.
In a third more preferred embotliment of this invention, the
inhibitors of farnesyl transferase are illustrated by the formula m
HOCH2(CH2)q
V ~ A1(CR1a2)nA2(CR1~z)n (W)~ (CR1bZ)p~N~OH
Il 4a
R
wherein:
Rla is independently selected from: hydro~en or Cl-C6 aL~cyl;
Rlb is independently selected from:
a) hydro~en,
b) aryl. heterocycle. cycloalkyl. R l Oo, -N(R l ~)~ or alkenyl.
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- 32 -
c) Cl-C6 aL~cyl unsubstituted or substituted by aryl,
heterocycle, cycloaL~yl, alkenyl, R100-, or-N(R10)2;
R2 and R3 are independendy selected from:
a) a side chain of a naturally occurring ~mino acid,
b) an oxidized form of a side chain of a naturally occurring
amino acid which is:
i) me~ionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-Clo aL~yl, C2-Clo alkenyl,
C3-Clo cycloaL~yl, 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)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and
d) C l-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaLkyl; or
20 R2 and R3 are combined to forrn - (CH2)S -; or
R2 or R3 are combined with R6 to form a ring such that
R6 ~
~N~, is ~ (CH2)t;
R2 R3 R7a ~ R7b
R4a and R7a are independently selected from:
a) hydrogen,
b) C l-C6 alkyl unsubstituted or substituted by alkenyl, R l Oo,
R 1 1 S(O)m-~ R 1 OC(O)NR 10, CN. N3, (R 1 0)2N-C(NR 10),
RlOC(O)-, RlOoc(o)-~ -N(R10)~. or Rl lOC(O)NR10-,
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c) aryl, heterocycle, cycloaL~yl, alkenyl, R100-,
Rl lS(O)m-, RlOC(O)NR10-, CN, N02, (R10)2N-
C(NR10)-, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2,
or Rl lOC(O)NR10-, and
S d) Cl-c6 aL~yl substituted with an unsubstituted or
substituted group selected from aryl, heterocyclic and C3-
Clo cycloaL~yl;
R4b and R7b are hydrogen;
R6 is independently selected from hydrogen or C l-C6 alkyl;
R8 is independently selected from:
a) hydrogen,
15 b) Cl-C6 aL~yl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-c6
perfluoroaIkyl, F, Cl, R10O-, R10C(O)NRl0-~ CN, NO2,
(R10)2N-C(NR10)-, R10C(O)-, Rl0oc(o)-~ -N(R10)2, or
Rl lOC(O)NR10-, and
c) Cl-C6 aL~yl substit~lteA by Cl-C6 perfluoroalkyl, R100-,
Rl0c(o)NRlo, (Rlo)2N-c(NRlo)-~ Rl0C(o),
Rl0oc(o)-~ -N(R10)2, or Rl lOC(O)NR10-;
R9 is selected from:
a) hydrogen,
25 b) C2-C6 alkenyl, C2-C6 alkynyl, C l-C6 perfluoroalkyl, F, Cl,
R10O-, Rl lS(O)m, Rl0c(o)NRlo-~ CN, NO2, (R10)2N-
C(NR 10) , R 1 ~C(O)-, R 1 ~OC(O)-, -N(R 1~)2, or
R 1 1 OC(O)NR 10 , and
c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6
perfluoroalkyl, F, Cl, R10O-, Rl lS(O)m-. R10C(O)NR10-,
CN, (Rl0)2N-c(NRlo)-~ Rl0C(O)-, RlOOC(O)-, -N(Rl0)2
or R l l OC(O)NR 10 ;
R10 is independently selected from hydro~en. Cl-C6 alkyl, benzyl and
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- 34-
aryl;
Rl 1 is independently selected from Cl -C6 aL~yl and aryl;
5 Q is selected from:
s~-N~ N~ and -~-N =>
~0 Al and A2 are independently selected from: a bond, -CH=CH-, -C_C-,
-C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;
V is selected from:
a) hydrogen,
b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl,
isoquinolinyl, and thienyl,
c) aryl,
d) Cl-C20 aLkyl wherein from 0 to 4 carbon atoms are replaced
with a a heteroatom selected from O, S, and N, and
e) C2-C20 aLkenyl, and
provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if
Al is a bond, n is 0 and A2 is S(O)m;
25 W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl. quinolinyl, or
isoquinolinyl;
X, Y and Z are independently H2 or O;
m is 0, 1 or'';
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nis 0, 1,2,30r4;
pis 0,1,2,30r4;
q is 0, l or 2;
r is O to 5, provided that r is O when V is hydrogen;
t is 3, 4 or 5; and
uis Oorl;
or the ph~ eutically acceptable salts ~ereof.
In a fourth more ~lefcl,ed embodiment of this invention, the
prodrugs of the preferred compounds of Follllula III are illustrated by the
Formula IV:
V A1 (CR182)nA2(CR 1 ~2)n 4~~ (CR1 b2)p~N~
IV R4a R
wherein:
Rla is independently selected from: hydrogen or Cl-C6 aLkyl;
Rlb is independently selected from:
a) hydrogen,
b) aryl, heterocycle, cycloalkyl, R100-, -N(R10)2 or alkenyl,
c) C 1 -C6 alkyl unsubstituted or substituted by aryl,
heterocycle, cycloaLkyl, alkenyl, R100-, or-N(R10)2;
- S R2 and R3 are independently selected from:
a) a side chain of a naturally occurrinC amino acid,
b) an oxidized form of a side chain of a naturally occurrin~
amino acid which is:
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i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstit~lte~l Cl-Clo alkyl, C2-Clo alkenyl,
C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R10O-, R11S(O)m-, R10C(O)NR10-, CN,
(Rlo)2N-c(NRlo)-~ Rloc(o)-~ R10Oc(o)-~ N3,
-N(R10)2, Rl 10C(O)NR10- and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsul~s~i~uled or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; or
R2 and R3 are combined to form - (CH2)S -; or
15 R2 or R3 are combined with R6 to form a ring such that
R6 ~_~
~;N~; is ~ H2)t;
R2 -R3 R7a ~ R7b
R4a and R7a are independently selected from:
a) hydrogen,
b) Cl-C6 alkyl unsubstituted or substituted by alkenyl, R100-,
R11S(O)m-, R10C(O)NRl0-~ CN, N3, (R10)2N-C(NR10)-,
R10C(O)-, R10OC(O)-, -N(R10)2, or R1 lOC(O)NR10-,
c) aryl, heterocycle, cycloalkyl, aLkenyl, R100-,
Rl lS(O)m-, R10C(O)NRl0-~ CN. NO2, (R10)2N-
C(NR10)-~ Rl0c(o), R10OC(O)-~ N3, -N(R10)2,
or R 1 1 OC(O)NR 10, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted ~roup selected from arvl. heterocyclic and C3-
Clo cycloalkyl;
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R4b and R7b are hydrogen;
R6 iS independently selected from hydrogen or Cl-C6 alkyl;
5 R8 is independently selected from:
a) hydrogen,
b) Cl-c6 alkyl, C2-c6 aL~enyl, C2-c6 aL~ynyl, Cl-C6
perfluoroaLkyl, F, Cl, R100-, RlOC(O)NR10-, CN, N02,
(R10)2N-C(NR10)-, RlOc(o)-7 R10oc(o)-7 -N(R10)2, or
R11OC(O)NR10-, and
c) Cl-C6 aL~yl substituted by Cl-C6 perfluoroaL~yl, R100-,
RlOC(O)NR10-, (R10)2N-C(NR10)-, RlOC(O)-,
RlOOC(O)-, -N(R10)2, or Rl lOC(O)NR10-;
15 R9 is selected from:
a) hydrogen,
b) C2-C6 aL~enyl, C2-C6 aL~ynyl, C l-C6 perfluoroaL~cyl, F, Cl,
RlOO-, R11S(O)m-, RlOc(o)NRlo-~ CN, NO2, (R10)2N-
C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aL~cyl unsubstituted or substituted by Cl-C6
perfluoroaL~yl, F, Cl, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2,
or Rl lOC(O)NR10;
R10 is independently selected from hydrogen, Cl-C6 aL~yl, benzyl and
aryl;
Rl 1 is independently selected from Cl-C6 aL~yl and aryl;
Q is selected from:
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- 38 -
~ N~ -~ N~ and -~- N~>
A1 and A2 are independently selected from: a bond, -CH=CH-, -C_C-,
S -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;
V is selected from:
a) hydrogen,
b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl,
isoquinolinyl, and thienyl,
c) aryl,
d) Cl-C20 aL~yl wherein from O to 4 carbon atoms are replaced
with a a heteroatom selected from 0, S, and N, and
e) C2-C20 aL~enyl, and
provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if
Al is a bond, n is O and A2 is S(O)m;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
20 thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or
isoquinolinyl;
X, Y and Z are independently H2 or 0;
mis 0, 1 or2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or4;
q is 0, 1 or 2;
r is O to 5, provided that r is O when ~' is hydrogen;
tis 3,40r5:and
uis Oor l;
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- 39 -
or the ph~ ceutically acceptable salts thereof.
The ~refell~d compounds of this invention are as follows:
s
N-[1-(lH-Tmicl~7.ol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-
prolyl-methionine methyl ester
N-[1-(lH-Tmi~1~7.ol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-
10 prolyl-methionine
N-[l-tl-(4-Cyanobenzyl)-lH-imidazol-S-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine isopropyl ester
15 N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine sulfone isopropyl ester
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine sulfoxide
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine sulfoxide isopropyl ester
N-[ 1- [ 1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-
25 ylmethyl]-3(S)-ethyl-prolyl-methionine sulfone
N- [ 1-[1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[ l -(1 H-Imidazol-4-ylacetyl)-pyrrolidin-2(S )-ylmethyl]-3 (S )-ethyl-
prolyl-methionine methyl ester
N-[ l-(lH-Imidazol-4-ylacetyl)-pyrrolidin-r'(S)-ylmethyl]-3(S)-ethyl-
prolyl-methionine
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- 40 -
N-[1-Glycyl-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine
methyl ester
5 N-[1-Glycyl-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[ 1 -(3-[lH-Tmi~1~7.ol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine methyl ester
10 N-[1-(3-[lH-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine
N-[1-[3-(1-(4-Cyanobenzyl)- lH-imidazol-5-yl)propionyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
N-[1-[3-(1-(4-Cyanobenzyl)-lH-imidazol-5-yl)propionyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[2(S)-(lH-Tmi-1~7.ol-4-ylacetyl-amino)-3(S)-methylpentyl]-prolyl-
20 methionine methyl ester
N-[2(S)-(lH-Imidazol-4-ylacetyl-amino)-3(S)-methylpentyl]-prolyl-
methionine
25 N-[1-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S-)ylmethyl]-prolyl-
methionine methyl ester
N-~ 1-(1 H-Imidazol-4-ylacetyl)-pyrrolidin-2(S-)ylmethyl] -prolyl-
methionine
N-[ 1-[1 -(4-Cyanobenzyl)- lH-imidazol-5-ylacetyl~pyrrolidin-2(S)-
ylmethyl]-prolyl-methionine methyl ester
N-[ 1-[1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-
35 ylmethyl]-prolyl-methionine
N- [ l -( l H-Imidazol-4-ylacetyl )-3 ( S )-ethylpyrrol idin-2(S )-ylmethyl ]- 3 ( S )
ethyl-prolyl-methionine methvl ester
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- 41 -
., .
N-[1-(lH-Imidazol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine
5 N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
N-[1-[1-(4-Cyanobenzyl)-lH-imicl~7.ol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine isopropyl ester
N-[1-(3-[lH-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-
15 ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
N-[1-(3-[lH-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine
20 N-[1-Glycyl-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine methyl ester
N-[ 1 -Glycyl-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine
N-[1-[1-(4-Nitrobenzyl)-lH-imidazol-4-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[ 1-[1 -(4-Nitrobenzyl)- 1 H-imidazol-5-ylacetyl] pyrrolidin-2(S)-
30 ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[ 1-(1-(1 -Farnesyl)- 1 H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
3(S)-ethyl-prolyl methionine
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N-[l-(l-(1-Geranyl)-lH-imida_ol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
3(S)-ethyl-prolyl-methionine
N-[1-[1-(4-Methoxyben_yl)-lH-imida_ol-5-ylacetyl]pyrrolidin-2(S)-
5 ylmethyl]-3(S)-ethyl-prolyl-methionine
N-[1-[1-(2-Naphthylmethyl)-lH-imi~1~7.ol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine and
10 N-[l-(lH-Tmi~l~7.ol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-
prolyl-(~-acetylamino)~l~nine
or the ph~rm~eeutically acceptable salts thereof.
Specific examples of compounds of the invention are:
N-[l-(lH-Imida_ol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-
prolyl-methionine
O
HN~ H ' N ~OH
N-[ 1-(1 H-Imidazol-4-ylacetyl)-pyrrolidin-2(S )-ylmethyl]-3(S)-ethyl-
prolyl-methionine methyl ester
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SCH3
H~ H OCH3
N-[1-(3-[lH-~mi~l~7.ol-4-yl]propionyl)-pyrrolidin-2(S)-ylme~yl]-3(S)-
ethyl-prolyl-methionine
SCH3
O
H ~ ~ ~OH
N-t 1-(3-~ lH-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine methyl ester
SCH3
O
~ H ~ N ~OCH3
N~ N~N~
- 10 H
.
N- [ 1-(3-[ l H-Imidazol-4-yl]propionyl )-3 (S )-ethylpyrrolidin-2(S )-
ylmethyl]-3(S)-ethyl-prolyl-methionine
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SCH3
<N ~ ~ ~
N-tl-(3-[lH-~mi-1~7.ol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
O
~ H \~N~fOCH3
<N~ ~\
N-~2(S)-(lH-Imida_ol-4-ylacetyl-amino)-3(S)-methylpentyl]-prolyl-
methionine methyl ester
~SCH3
O
H ~ N--~OCH3
HN/~N O
\=N O
N-[2(S)-( 1 H-Imida_ol-4-ylacetyl-amino)-3(S )-methylpentyl]-prolyl-
methionine
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SCH3
H ~ N OH
HN/~ I - ~
\= N O ~ -
N- [ 1- [ 1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine
NC
SCH3
r~ 1~
~,
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-S-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester
NC
SCH3
N ~ H ~ NH ~OCH3
\~N~N
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- 46 -
N-[1-[1-(4-Cyanobenzyl)-lH-imi-l~7.ol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine isopropyl ester
NC
Q~ ~ CH3
r~ H ~ N ~ ~
5 or the ph~n~eutically acceptable salts thereof.
In the present invention, the amino acids which are disclosed
are identified both by conventional 3 letter and single letter abbreviations
as indicated below:
Alanine Ala A
Arginine Arg R
Asparagine Asn N
Aspartic acid Asp D
Asparagine or
Aspartic acid Asx B
Cysteine Cys C
Glutamine Gln Q
Glutamic acid Glu E
Ghlt~mine or
Glutamic acid Glx Z
Glycine Gly G
Histidine His H
Isoleucine Ile
Leucine Leu L
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Lysine Lys K
Methionine Met M
Phenyl~l~nint- Phe F
Proline Pro P
Serine Ser S
Threonine Thr T
Tryptophan Trp W
Tyrosine Tyr Y
Valine Val V
The compounds of the present invention may have
asymmetric centers and occur as racemates, racemic mixtures, and as
individual diastereomers, with all possible isomers, including optical
15 isomers, being included in the present invention.
As used herein, "aL~yl" is int~qnc1e~1 to include both branched
and straight-chain saturated aliphatic hydrocarbon groups having the
specified number of carbon atoms.
As used herein, "cycloaLkyl" is intended to include non-
20 aromatic cyclic hydrocarbon groups having the specified number of
carbon atoms. Examples of cycloaLkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl and the like.
"Alkenyl" groups include those groups having the specified
number of carbon atoms and having one or several double bonds.
25 Examples of alkenyl groups include vinyl, allyl, isopropenyl, pentenyl,
hexenyl, heptenyl, cyclopropenyl, cyclobutenyl, cyclopentenyl,
cyclohexenyl, l-propenyl, 2-butenyl, 2-methyl-2-butenyl, isoprenyl,
farnesyl, geranyl, geranylgeranyl and the like.
As used herein, "aryl" is intended to include any stable
30 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, biphen~ l~ tetrahydronaphthyl,
indanyl, phenanthrenyl and the like.
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- 48 -
The term heterocycle or heterocyclic, as used herein,
represents a stable 5- to 7-membered monocyclic or stable 8- to 11-
membered bicyclic or stable 11-15 membered tricyclic heterocycle ring
which is either saturated or lm~ ated, 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 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 limite~l to, azepinyl,
benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl,
benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl,
benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydro-
benzothienyl, dihydrobenzothiopyranyl, dihydrobenzothio-pyranyl
sulfone, furyl, imidazolidinyl, imi-l~701inyl, imidazolyl, indolinyl,
indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolidinyl,
isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl,
2-oxoazepinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
piperidyl, piperazinyl, pyridyl, pyridyl N-oxide, pyridonyl, pyrazinyl,
pyrazolidinyl, pyrazolyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl,
quinolinyl, quinolinyl N-oxide, quinoxalinyl, tetrahydrofuryl,
tetrahydroisoquinolinyl, tetrahydro-quinolinyl, thiamorpholinyl,
thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl, thienofuryl,
thienothienyl, and thienyl.
As used herein, the terms "substituted aryl", "substituted
heterocycle" and "substituted cycloalkyl" are intended to include the
cyclic group which is substituted with 1 or 2 substitutents selected from
the group which includes but is not limited to F, Cl, Br, CF3, NH2, N(Cl-
C6 alkyl)2, N02, CN, (cl-c6 alkyl)O-, -OH, (Cl-C6 alkyl)s(o)m-~ (Cl-
C6 alkyl)C(O)NH-, H2N-c(NH)-~ (cl-c6 alkyl)C(O)-, (Cl-C6 >
alkyl)OC(O)-. N3 ,(C 1 -C6 alkyl)OC(O)NH- and C 1 -C20 alkyl.
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- 49 -
The following structure:
,N~
( ~H2)t
represents a cyclic amine moiety having 5 or 6 members in the ring, such
5 a cyclic amine which may be optionally fused to a phenyl or cyclohexyl
ring. Examples of such a cyclic amine moiety include, but are not limite-l
to, the following specific structures:
It is also understood that substitution on the cyclic amine moiety by R8a
and R8b may be on different carbon atoms or on the same carbon atom.
When R3 and R4 are combined to form - (CH2)S -, cyclic
moieties are formed. Examples of such cyclic moieties include, but are
15 not limited to:
When Rsa and R5b are combined to form - (CH2)S -, cyclic
moieties as described hereinabove for R3 and R4 are formed. In addition,
20 such cyclic moieties may optionally include a heteroatom(s). Examples
of such heteroatom-containing cyclic moieties include. but are not limited
to:
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~0~ SJ '
'~ ~0 ~S NJ
~ CORl~
As used herein, the phrase "nitrogen co,.t~i..inp C4-Cg mono
S or bicyclic ring system wherein the non-nitrogen cont~inin.~ ring may be
a C5-C7 saturated ring" which defines moiety "Q" of the instant invention
includes but is not limited to the following ring systems:
~- N~ N~
N~>
~,
The pharmaceuticallv acceptable salts of the compounds of
this invention include the conventional non-toxic salts of the compounds
of this invention as formed. e.~.. from non-toxic inor~anic or or~anic
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acids. For example, such conventional non-toxic salts include those
derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric,
slllf~mic, phosphoric, nitric and the like: and the salts prepared from
organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic,
malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenyl-
acetic, glllt~mic, benzoic, salicylic, slllf~nilic, 2-acetoxy-benzoic,
fumaric, toluenesulfonic, methanesuLfonic, ethane disulfonic, oxalic,
isethionic, trifluoroacetic and the like.
It is inte.ncled that the definition of any substituent or variable
(e.g., R10, Z, n, etc.) at a particular location in a molecule be independent
of its definitions elsewhere in that molecule. Thus, -N(R10)2 represents
-NHH, -NHCH3, -NHC2Hs, etc. It is understood that substituents and
substitution p~ .rn~ on the compounds of the instant invention can be
selected by one of ordinary skill in the art to provide compounds that are
chemically stable and that can be readily synthesized by techniques
known in ~e art as well as those methods set forth below.
Preferably, R1a and R1b are independently selected from:
hydrogen, -N(R8)2, R8C(O)NR8- or Cl-C6 alkyl unsub~ti~lte~l or
substituted by -N(R8)2, R8O- or R8C(O)NR8-.
Preferably, R2 is the sidechain of glycine (hydrogen).
Preferably, R3 is selected from:
a) a side chain of a naturally occurring amino acid,
b) substituted or unsubstituted C1-C20 alkyl,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, NO2, RlOO-, R1 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
c) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
- 30 Clo cycloalkyl; or
R3 is combined~with R6 to form pyrrolidinyl ring.
Preferably, R4a, R4b~ R7a and R7b are independently
selected from: hydro~en, Cl-C6 alkyl. aryl and benzyl.
Preferably, R5~ and RSb are independentlv selected from:
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a side chain of a naturally occurring amino acid, methionine sulfoxide,
methionine sulfone and unsubstituted or substituted C l-C6 alkyl .
Preferably, R6 is: hydrogen or is combined with R3 to forIn
pyrrolidinyl ring.
Preferably, R8 is selected from: hydrogen, per~luoroaL~yl, F,
Cl, Br, R100-, RllS(O)m-, CN, NO2, Rlo2N-c(NRlo)-~ RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or Rl lOC(O)NR10- and Cl-C6 aL~yl.
Preferably, R9 is hyd~ogen.
Preferably, R10 is selected from H, Cl-C6 alkyl and benzyl.
Preferably, R12 is selected from Cl-C6 alkyl and benzyl.
Preferably, Al and A2 are independently selected from: a
bond, -C(O)NR10-, -NR10C(O)-, O, -N(R10)-, -S(O)2N(R10)- and-
N(R10)S(0)2-.
Preferably, Q is a pyrrolidinyl ring.
Preferably, V is selected from hydrogen, heterocycle and
aryl.
Preferably, n, p and r are independently 0, 1, or 2.
Preferably t is 3.
The pharmaceutically acceptable salts of the compounds of
this invention can be synthesized from the compounds of this invention
which contain a basic moiety conventional chemical methods. Generally,
the salts are prepared by ion exchange chromatography or 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
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 following
works: Schroeder et al., "The Peplides", Vol. I~ Academic Press 1965, or
Bo~l~n~7ky et al~, "Peptide Svnthesis", Interscience Publishers, 1966, or
McOmie (ed.) "Protective Groups in Organic Chemistr~ ", Plenum Press,
1973, or Barany et al., "The Peptides: Anal~sis. S~nthesis, Biolog~" 2,
Chapter 1, Academic Press, 1980 or Stewart et al., "Solid Phase Peptide
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Synthesis", Second Edition, Pierce Chernical Company, 1984. The
te~cllin~.~ of these works are hereby incorporated by reference.
J 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 Dimethylform~mi~le;
EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbo~liimicl.-
hydrochloride;
HOBT l-Hydroxybenzotriazole hydrate;
Et3N Triethyl~rninP;
EtOAc Ethyl ~et~t~;
FAB Fast atom bombar~lm~.nt;
HOOBT 3-Hydroxy- 1 ,2,2-benzotriazin-4(3H)-one;
HPLC High-performance liquid chromatography;
MCPBA m-Chloroperoxybenzoic acid;
MsCl Methanesulfonyl chloride;
NaHMDS Sodium bis(trimethylsilyl)amide;
Py Pyridine;
TFA Trifluoroacetic acid;
THF Tetrahydrofuran.
Compounds of this invention are prepared by employing the
reactions shown in the following Reaction Schemes A-J, in addition to
other standard manipulations such as ester hydrolysis, cleavage of
30 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:
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Reaction A Amide bond formation and protecting group cleavage using
~t~n~l~rd solution or solid phase methodologies.
Reaction B Preparation of a reduced peptide subunit by reductive
S aL~ylation of an amine by an aldehyde using sodium
cyanoborohydride or other reducing agents.
Reaction C Deprotection of the rerlllce-l peptide subunit
Reaction D Peptide bond formation and protecting group cleavage using
standard solution or solid phase methodologies.
Reaction E ~ al~tion of a re~l~lce~l subunit by borane reduction of the
amide moiety.
Reaction Schemes A-E illustrate bond-forming and peptide
modifying reactions incorporating acyclic peptide units. It is well
understood that such reactions are equally useful when the - NHC(RA) -
moiety of the reagents and compounds illustrated is replaced with the
following moiety:
(~H2)t
~R7b
R7a
These reactions may be employed in a linear sequence to provide the
compounds of the invention or they may be used to synthesize fragments
2~ which are subsequently joined by the reactions described in the Reaction
Schemes.
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REACTION SCHEME A
Reaction A. Coupling of residues to form an amide bond
O RA CO2R
>~O~N I OH + ~R4b
EDC, HOBT H ~ CO2R
or HOOBT >~O~NJI~ <
Et3N, DMF ( l
R4a />~/~ R4b
TFA H N~J~ CO2R
~ R4b
R4a
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REACTION SCHEME B
Reaction B. Preparation of reduced peptide subunits by reductive
aLkylation
>~oJ~N~H +
R4a
NaCNBH3 >~0~ N ~ <02R
~R4b
R4a
REACTION SCHEME C
Reaction C. Deprotection of reduced peptide subunits
H CO2R
>~ ~ ~~,~~~ TFA or
O RA ~ Q
~ R4b HCI
R4a
~ CO2R
RA ~Q ~
~R4b
R4a/
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REACTION SCHEME D
Reaction D. Coupling of residues to form an amide bond
EDC, HOBT
>~oJI, < + H2N
fl ~) o Et3N, DMF
R4a~ R4b
O ~ O
R4a R4b
H _~--H o
R4a ~ R4b
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REACTION SCHEME E
Reaction E. Plc;pal~tion of reduced dipeptides from peptides
BH3 THF
O RA ~Q ~
>~ R4b
R a
H CO2R
>~ O ~ Q
4a ~ R4b
R
s
where RA is R2, R3, RSa or Rsb as previously defined; R4a and R4b are
as previously defined; and R is an a~lo~fiate protecting group for the
carboxylic acid.
Reaction Schemes F - M illustrate reactions wherein the non-
sulfhydryl-cont~ining moiety at the N-terminus of the compounds of the
instant invention is attached to an acyclic peptide unit which may be
further elaborated to provide the instant compounds. It is well understood
that such reactions are equally useful when the - NHC(RA) - moiety of
1~ the reagents and compounds illustrated is replaced with the following
moiety:
(Cj H2)t
>~ R7b
R7a
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_ 59 _
These reactions may be employed in a linear sequence to provide the
compounds of the invention or they may be used to synthesize fragments
which are subsequently joined by the reactions described in Reaction
Schemes A - E.
S The intermediates whose synthesis are illustrated in Reaction
Schemes A and C can be reductively aL~ylated with a variety of
aldehydes, such as I, as shown in Reaction Scheme F. The aldehydes can
be prepared by t~n~l~rd procedures, such as that described by O. P. Goel,
U. Krolls, M. Stier and S. Kesten in Organic Syntheses. 1988, 67, 69-75,
from the a~rol~liate amino acid (Reaction Scheme F). The reductive
aL~ylation can be accomplished at pH 5-7 with a variety of reducing
agents, such as sodium triacetoxyborohydride or sodium
cyanoborohydride in a solvent such as dichloroethane, meth~nol or
dimethylform~mide. The product II can be deprotected to give the final
compounds m with trifluoroacetic acid in methylene chloride. The final
product m is isolated in the salt form, for example, as a trifluoroacetate,
hydrochloride or acetate salt, among others. The product ~ minlo m can
further be selectively protected to obtain IV, which can subsequently be
reductively aL~ylated with a second aldehyde to obtain V. Removal of
the protecting group, and conversion to cyclized products such as the
dihydroimidazole VII can be accomplished by literature procedures.
Alternatively, the protected dipeptidyl analog intermediate
can be reductively aIkylated with other aldehydes such as 1-trityl-4-
carboxaldehyde or l-trityl-4-imidazolylacetaldehyde, to give products
2~ such as vm (Reaction Scheme G). The trityl protecting group can be
removed from VIII to give IX, or alternatively, VIII can first be treated
with an alkyl halide then subsequently deprotected to give the alkylated
imidazole X. Alternatively, the dipeptidyl analog interrnediate can be
acylated or sulfonylated by standard techniques.
The imidazole acetic acid XI can be converted to the acetate
XIII by standard procedures, and XIII can be first reacted with an alkyl
halide, then treated with refluxine methanol to provide the
reoiospecifically alkylated imidazole acetic acid ester XI~7. Hydrolysis
and reaction with the protected dipeptidyl analocr intermediate in the
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- 60 -
presence of condensing reagents such as 1-(3-dimethyl~minopropyl)-3-
ethylcarbo-liimi~le (EDC) leads to acylated products such as XV.
If the protected dipeptidyl analog intermediate is reductively
aL~ylated with an aldehyde which also has a protected hydroxyl group,
S such as XVI in Reaction Scheme I, the protçctin~: groups can be
subsequently removed to llnm~k the hydroxyl group (Reaction Schemes
I, J). The alcohol can be oxicli7~1 under standard conditions to e.g. an
aldehyde, which can then be reacted with a variety of organometallic
reagents such as Grignard reagents, to obtain secondary alcohols such as
10 XX. In addition, the fully deprotected amino alcohol XXI can be
reductively aL~ylated (under conditions described previously) with a
variety of aldehydes to obtain secondary amines, such as XXII (Reaction
Scheme K), or tertiary ~mines.
The Boc protected amino alcohol XVIII can also be lltili7e-1
15 to synthesi_e 2-a_iridinylmethylpipera_ines such as xxlll (Reaction
Scheme L). Treating XVIII with l,1'-sulfonyl-liimi-1~7.ole and sodium
hydride in a solvent such as dimethylform~mide led to the formation of
z~7~ inf~ X X l l l . The aziridine reacted in the presence of a nucleophile,
such as a thiol, in the presence of base to yield the ring-opened product
20 XXIV.
In addition, the protected dipeptidyl analog intermediate can
be reacted with aldehydes derived from amino acids such as O-aL~cylated
tyrosines, according to standard procedures, to obtain compounds such as
XXX, as shown in Reaction Scheme M. When R' is an aryl group, XXX
25 can first be hydrogenated to llnm~.~k the phenol, and the amine group
deprotected with acid to produce XXXI. Alternatively, the amine
protecting group in XXX can be removed, and O-aL~ylated phenolic
amines such as XXXll produced.
Similar procedures as are illustrated in Reaction Schemes F-
30 M may be employed using other peptidyl analog intermediates such asthose whose synthesis is illustrated in Reaction Schemes B - E.
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Reaction Schemes N-R illustrate syntheses of suitably
substihltç~l aldehydes useful in the syntheses of the instant compounds
wherein the variable W is present as a pyridyl moiety. Similar synthetic
strategies for preparing aLkanols that incorporate other heterocyclic
S moieties for variable W are also well known in the art.
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REACTION SCHEME F
Boc NH ~ I
2 J~\ CO2R Boc NH CHO
R (~J Q ~ NaBH(OAc)3
~ R4b Et3N, CICH2CH2CI
R4a
NHBoc
Boc NH/~2R CF3CO2H
R4a R4b
NH2
NH/~HN~JI~ CO2R Boc20
RA ~ Q ~ CH2CI2
R4a~ R4b
BocNH/~N~ ~ CHO
,~ NaBH(OAc)3
Et3N, CICH2CH2CI
IV ~ R4b
R4a
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REACTION SCHEME F (conhnued)
/=\
~3,
NH CF3CO2H, CH2C12;
BocNH/~ _<02R NaHCO3
RA ~ Q ~
/= R4a>~ R4b
NH
NH/~N~JI~ < ~NC
RA ~ R4b AgCN
Vl R4a
~N~ <
RA ~Q ~
~1 R4a R4b
Vll
W
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-- 64 --
REACTION SCHEME G
H2N ~ CO2R NaBH(OAc)3
RA 1/~ Q Et3N, CICH2CH2CI
R4a ~ R4b ~(CH2)nCHO
Tr
H2)n+1 \,~
~C RA ~_ R4b
D4a
Tr ' ' 1 ) Ar CH2X, CH3CN
Vlll 2) CF3C02H, CH2C12
H CH Cl (C2H5)3siH
CF3C ~2 ~ 2 2
(c2H5)3siH
H Y CO2R
(CH2)n+1
N RA ~--R4b
A (CH2)~ ~ 4b
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REACTION SCHEME H
~N~ CH30H ~N~
Xl Xll
N CH2C02CH31 ) ArCH2X CH3CN
(c6H5)3cBr ~ reflux
(C2DH )3FN NTr 2) CH30H, reflux
Xlll
Ar~\N~cH2c02cH3 2.5N HClaq,
55~C
XIV
Ar~\N j~CH2C~2H
N
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REACTION SCHEME I
,~5 H2N ~JJ~
RA ~ Q
>~ R4b
R4a
EDC- HCI
HOBt
DMF
Ar~~~2R
XV R4a R4b
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REACTION SCHEME J
NaBH(OAc)3
H2N ~ ~02R BnO 1
,~ R4b BocNH CHO
R4a
XVI
NHBoc
~N~L~ CO2R 20% Pd(OH)2 H2
BnO H - J~~< CH30H
R ~ Q ~ CH3C02H
\/ ~ R4b
XVII R4a~
NHBoc
N I I CO2R CICOCOCI
HO H~ ~ DMSO CH2C12
-A ~ (C2H5)3N
XVIII R4a
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REACTION SCHEME J (CONTINUED)
H NHBoc y
O~HN~ < (c2H5)2o
R ~ Q ~ 2. TFA, CH2C12
~ R4b
XIX R4a
R' NH2
HOX--HN~JI~ <
RA 1/~ Q
XX >~ R4b
R4a
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REACTION SCHEME K
NHBoc y CF3C02H
HO~--H ~02R CH2CI2
RA ~Q ~
XVIII ~ R4b
R4a
NH2 y R'CHO
HO/~H CO2R NaBH(OAc)3
RA (PI Q ) CICH2CH2CI
~ ~' ~4b
XXI R4a~
R'CH2
NH y
HO/~H .J~ <02R
RA ~Q ~
XXII >~ R4b
R4a
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REACTION SCHEME L
NHBoc y H H
HO/~--N~ll <02R ~\N N
XVIII ~ R4b NaH, DMF 0~C
N y
H ~ ~OzR (C2Hs)3N
NH2 y
R'S/~H \JI ~02R
RA ~Q ~
R4a R4b
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REACTION SCHEME M
HO~ 1) Boc2O, K2CO3 HO~
~ THF-H20 .~/
J~ 2) CH2N2, EtOAc ~1~
H2NC02H BocNH CO2CH3
XXV XXVI
HO~
LiAlH4 ~bJ R'CH2X
THF ,1~ Cs2CO3
0-20~C BocNH CH2OH DMF
XXVII
R'CH20 R'CH20
~ pyndine so3"~
BocNH CH2OH 20~C BocNH CHO
XXVIII XXIX
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REACTION SCHEME M (continued)
R'CH20~ ~ ~ C02R
+ RA ~ Q ~
BocNH CHO >~ R4b
R a
XXIX NaBH(OAc)3
CICH2CH2CI
NHBoc
R'CH20~ ~2R
R4a R4b
HC~ EtOAC
1) 20% Pd(OH)2
CH30H, CH3CO2H /
NH2
2) HCI, EtOAc / ~ y CO2R
XXXI I ~"
NH2
XXXI ~ 4b
R4a
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REACTION SCHEME N
CH3 1) HN~2~Br2 ,~CO2CH3
1' ~ 2) KMnO4 . 1' JJ
H2N N 3) MeOH,H+ Br N
R6
~MgCI R~l C02CH3
Zncl2~Nicl2(ph3p)2
NaBH4 (excess) ~CH20H
S03 Py, Et3N ~CHO
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REACTION SCHEME P
R6
R6 [~i~
[~CO2CH3 ~ MgCI ~CO2CH3
N Zn, CuCN R6
NaBH4 [~j~ SO3Py, Et3N [~
(excess) [~,CH20H DMSO ~CHO
Br CO2CH3 ~\ MgCI
~ ZnC12, Nicl2(ph3p)2 ~CO2CH3
R6 R6
NaBH4 ~ SO3 Py, Et3N CHO
(excess) N DMSO N
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REACTION SCHEME Q
co2CH3
Br~1. LDA, CO2 Br~
N 2. MeOH, H+ N
R6
ZnCI2, NiCI2(Ph3P)2 N
~/'1
NaBH4 (excess) \~2OH SO3 Py, Et3N
~ ,~ DMSo
N
R6
CHO
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REACTION SCHEME R
co2CH3
~3~ 1. LDA, C02 ~Br
2. (CH3)3SiCHN2
R6 ~\ Br R6 3~
~1~ co2CH3
Zn, NiC12(Ph3P)2 N~
R6 ~
excess NaBH4 ~ S03Py, Et3N
N~CH20H DMSO
R6 ~
N~,CHO
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The instant compounds are useful as pharmaceutical agents
for m~mm~l~, especially for hllm~n~. These compounds may be
~lmini~tered to p~ti~nt~ for use in the treatment of cancer. Fx~mples of
the type of cancer which may be treated with the compounds of this
S invention include, but are not limitetl to, colorectal carcinoma, exocrine
pancreatic carcinoma, myeloid lellk~omi~ and neurological tumors. Such
tumors may arise by mutations in the ras genes themselves, mllt~tions in
the ~lott;ills that can regulate Ras formation (i.e., neurofibromin (NF-l),
neu, scr, abl, lck, fyn) or by other mech~nicmc.
The compounds of the instant invention inhibit farnesyl-
protein transferase and the farnesylation of the oncogene protein Ras.
The instant compounds may also inhibit tumor angiogenesis, thereby
affecting the ~,lOWlll of tumors (J. Rak et al. Cancer Research, 55:4575-
4580 (1995)). Such anti-angiogenesis properties of the in~t~nt
compounds may also be useful in the tre~tm~nt of certain forms of
blindness related to retinal vasclll~ri7~tion.
The compounds of this invention are also useful for
inhibiting other proliferative diseases, both benign and malignant,
wherein Ras proteins are aberrantly activated as a result of oncogenic
mutation in other genes (i.e., the Ras gene itself is not activated by
mutation to an oncogenic form) with said inhibition being accomplished
by the ~-lmini~tration of an effective amount of the compounds of the
invention to a m~mm~l in need of such treatment. For example, a
component of NF-l is a benign proliferative disorder.
The instant compounds may also be useful in the treatment
of certain viral infections, in particular in the treatment of hepatitis delta
and related viruses (J.S. Glenn et al. Science, 256:1331-1333 (1992).
The compounds of the instant invention are also useful in the
prevention of restenosis after percutaneous translllmin~l coronary
3Q angioplasty by inhibiting neointimal formation (C. Indolfi et al. Nature
medicine, 1:541-545(1995).
The instant compounds may also be useful in the treatment
and prevention of polycystic kidney disease (D.L. Schaffner et al.
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- 78 -
American Journal of Pathology, 142:1051-1060 (1993) and B. Cowley,
Jr. et al.FASEB Journal, 2:A3160 (1988)).
The compounds of this invention may be ~1mini.ctered to
m~mm~l~, preferably hllm~n~, either alone or, preferably, in combination
S with pharmaceutically acceptable carriers or diluents, optionally with
known adjuvants, such as alum, in a ph~rm~relltical composition,
according to standard ph~rm~eutical practice. The compounds can be
~1mini.~tered orally or parenterally, including the intravenous,
intramuscular, i~ ilolleal, subcutaneous, rectal and topical routes of
10 ~lmini.~tration.
For oral use of a chemotherapeutic compound according to
this invention, the selected compound may be ~rlmini~tered, for example,
in the form of tablets or capsules, or as an aqueous solution or
suspension. In the case of tablets for oral use, carriers which are
15 commonly used include lactose and corn starch, and lubricating agents,
such as m~nesium stearate, are commonly added. For oral
~tlmini~tration in capsule form, useful diluents include lactose and dried
corn starch. When aqueous suspensions are required for oral use, the
active ingredient is combined with emulsifying and suspending agents. If
20 desired, certain sweetening and/or flavoring agents may be ~ l For
intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile
solutions of the active ingredient are usually prepared, and the pH of the
solutions should be suitably adjusted and buffered. For intravenous use,
the total concentration of solutes should be controlled in order to render
25 the preparation isotonic.
The present invention also encompasses a pharmaceutical
composition useful in the treatment of cancer, comprising the
~rlminictration of a therapeutically effective amount of the compounds of
this invention, with or without pharmaceutically acceptable carriers or
30 diluents. Suitable compositions of this invention include aqueous
solutions comprising compounds of this invention and pharrnacologically
acceptable carriers, e.g., saline, at a pH level, e.~., 7.4. The solutions may
be introduced into a patient's intramuscular blood-stream by local bolus
injection.
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Y When a compound according to this invention is
~lmini~tered into a hllm~n 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 S~ tOlllS.
In one exemplary application, a suitable amount of
compound is ~rlmini~tered to a m~mm~l undergoing tre~tm~nt for cancer.
~clmini.ctration occurs in an amount between about 0.1 mgfkg 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 10 mg/kg of body weight per day.
The compounds of the instant invention are also useful as
a component in an assay to rapidly ~leterminp the presence and
quantity of fa~nesyl-protein transferase (E~PTase) in a composition.
Thus the composition to be tested may be divided and the two portions
contacted with mixtures which comprise a known substrate of l;PTase
(for example a tetrapeptide having a cysteine at the ~mine tel...;..l.s)
and farnesyl pyrophosphate and, in one of the mixtures, a compound
of the instant invention. After the assay mixtures are incubated for an
sufficient period of time, well known in the art, to allow the FPTase to
farnesylate the substrate, the chemical content of the assay mixtures
may be determined by well known immunological, radiochemical or
chromatographic techniques. Because the compounds of the instant
invention are selective inhibitors of ~Tase, absence or quantitative
reduction of the amount of substrate in the assay mixture without the
compound of the instant invention relative to the presence of the
unchanged substrate in the assay containing 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
~ 30 identifying tissue samples which contain farnesyl-protein transferase and
quantitating the enzyme. Thus, potent inhibitor compounds of the instant
invention may be used in an active site titration assay to determine the
quantity of enzyme in the sample. A series of samples composed of
aliquots of a tissue extract containing an unknown amount of farnesyl-
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protein transferase, an excess amount of a known substrate of FPTase (for
example a tetrapeptide having a cysteine at the amine terminll~) and
farnesyl pyrophosphate are incubated for an ayylopliate period of time in
the presence of varying concentrations of a compound of the instant
5 invention. The concentration of a sufficiently potent inhibitor (i.e., one
that has a Ki substantially smaller than the concentration of enzyrne in the
assay vessel) required to inhibit the enzymatic activity of the sample by
50% is approximately equal to half of the concentration of the enzyme in
that particular sample.
EXAMPLES
Examples provided are intended to assist in a further
understanding of the invention. Particular materials employed, species
15 and conditions are intended to be further illustrative of the invention and
not limit~tive of the reasonable scope thereof.
The standard workup referred to in the examples refers to
solvent extraction and washing the organic solution with 10% citric acid,
lO~o sodium bicarbonate and brine as appropriate. Solutions were dried
20 over sodium sulfate and evaporated in vacuo on a rotary evaporator.
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EXAMPLE 1
Preparationof N-[l-(lH-Tmicl~7ol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-prolyl-methionine methyl ester and
S N-[l-(lH-Tmi~l~7ol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-
prolyl-methionine trifluoroacetate
Step A: P~ al~lionofDiethyl l-acetyl-5-hydroxy-3-
ethylpyrrolidine-2.2-dicarboxylate
Sodium (4.02 g, 0.175 mol) was dissolved in a stirred
solution of diethyl acetamidomalonate (235.4 g, 1.19 mol) in abs EtOH
(1.4 L) at ambient te~ el~ule under argon. The reaction mixture was
cooled to 0~C, and trans-2-pentenal (100 g, 1.08 mol) was added
d~ wise m~int~inin~ the reaction tempel~Lule at <5~C. After the
addition, the reaction was allowed to warm to room telll~el~Lule, stirred
for 4 h, then quenched with acetic acid (28 mL). The solution was
concentrated in vacuo, and the residue dissolved in EtOAc (1.5 L),
washed with 10% NaHCO3 solution (2 x 300 mL), brine, and dried
(MgSO4). The solution was filtered and concentrated to 700 rnL, then
heated to reflux and treated with hexane (1 L). On cooling, the title
compound precipitated and was collected, mp 106 - 109~C. lH NMR
(CD30D) ~ 5.65 (d, lH, J= 5 Hz), 4.1 - 4.25 (m, 4H), 2.7-2.8 (m, lH),
2.21 (s, 3H), 2.10 (dd, lH, J = 6, 13, Hz),1.86- 1.97 (m, 2H), 1.27 (t, 3H,
J= 7 Hz), 1.23 (t, 3H, J= 7 Hz), 1.1- 1.25 (m, lH), 0.97 (t, 3H, J= 7 Hz).
~tep B: Preparation of Diethyl l-acetyl-3-ethylpyrrolidine-2,2-
dicarboxvlate
To a solution of diethyl l-acetyl-5-hydroxy-3-
- 30 ethylpyrrolidine-2,2-dicarboxylate (287 g, 0.95 mol) and triethylsilane
(228 mL, 1.43 mol) in CH2Cl2 (3 L) under argon was added
trifluoroacetic acid (735 mL, 9.53 mol) dropwise with stirring while
maintainin~ the internal temperature at 25 ~C b~ means of an ice bath.
After stirring for 3 h at 23~C, the solution was concentrated in vacuo. ~
the residue diluted with CH2CI~ (1.5 L). then treated ~ith H2O (l L) and
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solid Na2CO3 with vigorous stirring until the solution was basic. The
organic layer was separated, dried (Na2SO4), filtered, then concentrated
to give the title compound as a yellow oil which was used without further
puri~lcation.
s
Step C: P~ al~lion of 3-Ethylproline hydrochloride (Cis:Trans
Mixture)
Diethyl l-Acetyl-3-ethylpyrrolidine-2,2-dicarboxylate (373
g, 0.95 mol) was suspended in 6N HCl (2 L) and HOAc (500 mL) and
heated at reflux for 20 h. The reaction mixture was cooled, washed with
EtOAc (lL), then concentrated in vacuo to give an oil which crystallized
upon l~ilul~tion with ether to give the title compound. lH NMR (D20) o
4.23(d, lH,J=8Hz),3.84(d, lH,J=8Hz),3.15-3.4(m,4H),2.33-2.44
(m, lH), 2.19-2.4 (m, lH), 2.02- 2.15 (m, 2H), 1.53- 1.72 (m, 3H), 1.23-
1.43 (m, 2H), 1.0- 1.15 (m, lH), 0.75 - 0.83 (m, 6H).
Step D: Plc~ tion of N-[(tert-Butyloxy)carbonyl]-cis:trans-3-
ethylproline methyl ester
3-Ethylproline hydrochloride (Cis:Trans Mixture) (20 g,
0.11 mol) was dissolved in CH30H (200 mL), and the solution was
saturated with HCl gas, then stirred at 23~C for 24 h. Argon was bubbled
through the solution to remove excess HCl. The solution was treated
with NaHCO3 (>84 g) to a pH of 8, then di-tert-butyl dicarbonate (25.1 g,
0.115 mol) dissolved in CH30H (20 mL) was added slowly. After
stirring for 18 h at 23~C, the mixture was filtered, the filtrate
concentrated, and the residue triturated with EtOAc, filtered again, and
concentrated to give the title compound as an oil.
Step E: Preparation of N-[(tert-Butyloxy)carbonyl]-trans-3-
ethylproline and N-[(tert-Butyloxy)carbonyl]-cis-3-
ethylproline methyl ester
N-[(tert-Butyloxy)carbonyl]-cis.Irans-3-ethylproline methyl
ester ('~9.1 g, 0.113 mol) was dissolved in CH~OH (114 mL) with cooling
to 0~C, then treated with 1 N NaOH (114 mL). After stirring for '~0 h at
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23~C, the solution was conce~ ted to remove the CH30H and then
extracted with EtOAc (3 x). The organic layers were combined, dried
(MgSO4), filtered, and conc~ ated to give 12.8 g of N-[(tert-
Butyloxy)carbonyl]-cis-3-ethylproline methyl ester as an oil. The
5 aqueous layer was acidified with solid citric acid and extracted with
EtOAc (2 x), the organic layers combined, dried (MgSO4), filtered, and
concentrated to give N-[(tert-Butyloxy)carbonyl]-trans-3-ethylproline as
an oil. lH NMR (CD30D) o 3.86 and 3.78 (2 d, lH, J = 6 Hz), 3.33 -
3.58 (m, 2H), 2.01 - 2.22 (m, 2H), 1.5 - 1.74 (m, 2H), 1.33 - 1.5 (m, lH),
1.45 and 1.42 (2 s, 9H), 0.98 (t, 3H, J= 8 Hz).
Step F: Preparation of 3(S)-Ethvl-2(S)-proline hydrochloride
N-[(tert-Butyloxy)carbonyl]-trans-3-ethylproline (15.5 g,
0.064 mol), S-a-methylbenzyl~mine (9.03 mL, 0.070 mol), HOBT (10.73
15 g, 0.70 mol), and N-methylmorpholine (8 mL, 0.076 mol) were dissolved
in CH2C12 (150 mL) with sitrring in an ice-H2O bath, treated with EDC
(13.4 g, 0.070 mol) stirred at 23~C for 48 h. The reaction mixture was
partitioned between EtOAc and 10% citric acid solution, the organic layer
washed with satd NaHCO3 solution, brine and dried (MgSO4), filtered,
20 and concentrated to give an oil. This oil was dissolved in a minimum
amount of ether (10 mL) to crystallize the desired S,S,S diastereomer (4.2
g), mp 118-121~C. A solution of this product in 8N HCl (87 mL) and
glacial acetic acid (22 mL) was heated at reflux overnight. The solution
was concentrated on a rotary evaporator, and the residue taken up in H20
25 and extracted with ether. The aqueous layer was concentrated to dryness
to give a 1: 1 mixture of 3(S)-ethyl-2(S)-proline hydrochloride and a-
methylbenzylamine.
3(S)-Ethyl-2(S)-proline containing a-methylbenzylamine
(2.0 g, 0.0128 mol) was dissolved in dioxane (10 mL) and H2O (10 mL)
30 with stirring and cooling to 0~C. N,N-diisopropylethylamine (2.2 mL,
0.0128 mol) and di-tert-butyl-dicarbonate (2.79 g, 0.0128 mol) were
added and stirring was continued at '3~C for 48 h. The reaction mixture
was partitioned between EtOAc (60 mL) and H~O (30 mL), the organic
layer washed with 0.5N NaOH (2 x 40 mL). the aqueous layers
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combined and washed with EtOAc ( 30 mL) and this layer back-extracted
with 0.5 N NaOH (30 mL). The aqueous layers were combined and
carefully acidified at 0~C with lN HCl to pH 3. This mixhlre was
extracted with EtOAc (3 x 40 mL), the organics combined, dried
S (MgSO4), filtered and conc~ ted to give N-[(tert-Butyloxy)carbonyl-
3(S)-ethyl-2(S)-proline as a colorless oil. N-t(tert-Butyloxy)carbonyl-
3(S)-ethyl-2(S)-proline was dissolved in EtOAc (50 mL) and the solution
was saturated with HCl gas with cooling in an ice-H2O bath. The
solution was stoppered and stirred at 0~C. for 3 hr. Argon was bubbled
through the solution to remove excess HCl, and the solution was
concentrated to dryness to give 3(S)-ethyl-2(S)-proline hydrochloride.
Step G: N-r(tert-Butyloxy)carbonyll-3(S)-ethyl-2(S)-prolinol
3(S)-Ethyl-2(S)-proline hydrochloride cont~inin~ a-
methylbenzyl~mine (2.0 g, 0.0128 mol) was dissolved in dioxane (10
mL) and H2O (10 mL) with stirring and cooling to 0~C. N,N-
diisopropylethyl~min~ (2.2 mL, 0.0128 mol) and di-tert-butyl-
dicarbonate (2.79 g, 0.0128 mol) were added and stirring was continued
at 23~C for 48 h. The reaction rnixture was partitioned between EtOAc
(60 mL) and H20 (30 mL), the organic layer washed with 0.5N NaOH (2
x 40 mL), the aqueous layers combined and washed with EtOAc ( 30 mL)
and this layer back-extracted with 0.5 N NaOH (30 mL). The aqueous
layers were combined and carefully acidified at 0~C with lN HCl to pH
2. T-his mixture was extracted with EtOAc (3 x 40 mL), the organics
combined, dried (MgSO4), filtered and concentrated to give N-[(tert-
Butyloxy)carbonyl--3(S)-ethyl-2(S)-proline as a colorless oil which was
used without punfication.
N-~(lert-Butyloxy)carbonyl]-3(S)-ethyl-2(S)-proline (1.6 g, 6.58 mmol)
was dissolved in dry THF (10 mL) and treated with borane (lM in THF~
12.5 mL, 12.5 mmol) with stirring at 0 ~C for ~ h, then 23~C for l h. The
solution was cooled to 0~C, treated with H2O (20 mL). and extracted
with EtOAc (2 x 30 mL). The organics were u ashed u ith brine, satd
NaHCO3. H2O dried (MgSO4). filtered and concentrated to give a
3~ viscous oil. The oil uas dissolved in CH~CI~. filtered throu~h dr~ SiO~.
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and the filtrate concentrated to give the title compound as an oil. 1H
NMR (CDC13) ~ 4.97 (d, lH, J= 7 Hz), 3.71 (t, lH, J = 8 Hz), 3.51-3.62
(m, 3H), 3.18 - 3.26 (m, lH), 1.9 - 2.0 (m, lH), 1.53-1.7 (m, 2H), 1.47 (s,
9H), 1.26 - 1.43 (m, 2H), 0.95 (t, 3H, J = 7 Hz).
s
Step H: N-r(tert-Butvloxv)carbonyll-3(S)-ethyl-2(S)-prolinal
N-[(ter~-Butyloxy)carbonyl-3(S)-ethyl-2(S)-prolinol (0.638
g, 2.78 mmol) and Et3N (1.4 mT ., 9.74 mmol) were dissolved in dry
CH2Cl2 (10 mL) with stirring and cooling to -10~C and treated dropwise
with a solution of S03.pyr (1.33 g, 8.35 mmol) in dry DMSO (5 mL)
keeping the reaction mixture temperature at < 0~C. The mixture was
stirred at 0~C. for 20 min then at 5~C for 20 min, and at 15~C for 1 h, then
poured into ice-cold 0.5 N HCl and the layers separated. The aqueous
layer was extracted with CH2C12 (3 x 20 mL), organics combined,
washed with H2O, aq satd NaHCO3 solution, brine, and dried (Na2SO4).
Filtration and concentration to dryness gave the title compound which
was used without purification.
Step I: N-t(tert-Butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-
ylmethyll-proline methyl ester
N-[(tert-Butyloxy)carbonyl]-3(S)-ethyl-2(S)-prolinal (0.315
g, 0.0014 mol) and proline methyl ester hydrochloride (0.233 g, 0.0014
mol) were dissolved in MeOH (5 mL) at ambient temperature under
argon with cooling in an ice-H2O bath, and treated with sodium
cyanoborohydride (0.131 g, 0.002 mol) with stirring. After 18 h the
mixture was poured into 5% NaHCO3 solution (20 mL), the CH30H
removved and the aq layer washed with EtOAc (3 x 30 mL), the organics
combined, washed with brine, and dried (MgSO4). Filtration and
concentration to dryness gave the title compound as a colorless oil after
chromatography (sio2~ hexane: EtOAc, 6:1). lH NMR (CDCl3) ~ 3.70
(s, 3H), 3.1 - 3.7'(m, SH), 2.2 - 2.65 (m, 4H), 1.7 - 2.15 (m, 5H), 1.5 -
1.65 (m, lH), 1.46 (s, 9H) 1.2 - 1.5 (m, 2H), 0.93 (t, 3H, J = 7 Hz).
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Step J: N-[(tert-Butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-
ylmethyll -proline
N-[(tert-Butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-proline methyl ester 0.081 g, 0.238 mmol) was dissolved in
CH30H (2 mT .), cooled to 0~C and treated with lN NaOH solution
(0.952 mL, 0.952 mmol). After stirrin~ at 23~C for 3h, the solution was
neutralized with lN HCl (0.952 mL, 0.952 mmol), concentrated to
remove the CH30H, then lyophilized and the residue used as is.
Step K: N-[(tert-Butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-
ylmethyll-prolyl-methionine methyl ester
N-[(tert-Butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-proline (0.238 mrnol), HOBT (0.048 g, 0.262 mmol), EDC
(0.068 g, 0.0357 mmol), and methionine methyl ester hydrochloride
(0.048 g, 0.238 mmol) were dissolved in CH2CL2 (10 mL) and stirred at
23~C for 18 h. EtOAc (100 mL) was ~lrl~rl, and the mixture washed with
satd NaHCO3 solution, H20, bnne, and dried (MgSO4). Filtration and
concentration to dryness gave 0.085 g of title compound. lH NMR
(CD30D) ~i (major rotamer) 4.63 (t, lH, J = 7 Hz), 3.73 (s, 3H), 3.55 -
3.7 (m, lH), 3.0-3.5 (m, 4H), 2.3 - 2.7 (m, 5H?, 1.9 - 2.2 (m, 3H), 2.08 (s,
3H), 1.6 - 1.9 (m, 4H), 1.46 (s, 9H), 1.3 - 1.45 (m, 2H), 0.92 - 1.02 (m,
3H).
Step L: N-[3(S)-Ethylpyrrolidin-2(S)-ylmethyl]-prolyl-methionine
methyl ester hydrochloride
HCl gas was bubbled into a solution of N-[(tert-
butyloxy)carbonyl-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-prolyl
methioninine methyl ester (0.085 g, 0.18 mmol) in EtOAc (5 mL) with
stirring and cooling in an ice-H20 bath until saturation. The solution was
stoppered and stirred at 0~C for 2 h, then purged with Ar and
concentrated to give the title compound as a yellow foam.
Step M: N-[ l -( lH-Imidazol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyll-prc-lyl-methionine methyl ester
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N-[3(S)-Ethylpyrrolidin-2(S)-ylmethyl]-prolyl methionine
methyl ester hydrochloride (0.075 g, 0.169 mmol), lH-imida_ol-4-
ylacetic acid (0.052 g, 0.253 mmol), HOBT (0.34 g, 0.253 mmol), EDC
(0.49 g, 0.253 mmol) and Et3N ( 0.176 mL, 1.27 mmol) were dissolved
in DMF (4 mL) and stirred at 23~C for 18 h. The solvent was removed in
vacuo, EtOAc (60 mL) was ~.1.1~,.17 and the solution was washed
seqllPnti~lly with satd NaHCO3 solution, H2O, brine, and dried
(MgSO4). Filtration and concentration to dryness gave dle title
compound after chromatography (SiO2, 5 - 10% CH30H/CH2C12). lH
NMR (CD30D) o (major rotamer) 7.62 (s, lH), 6.93 (s, lH), 4.6 - 4.67
(m, lH), 4.1-4.16 (m, lH), 3.74 (s, 3H), 3.65 (s, 2H), 3.5 - 3.68 (m, 2H),
3.2 - 3.25 (m, lH), 3.04 - 3.1 (m, lH), 2.44 - 2.7 (m, 5H), 2.05 - 2.26 (m,
4H), 2.08 (s, 3H), 1.68 - 1.87 (m, 4H), 1.26 - 1.5 (m, 3H), 0.99 (t, 3H, J =
7 H_). FAB MS 480 (M + 1).
Step N: N-tl-(lH-Tmicl~7ol-4-ylacetyl)-3(S)-ethylpyrrolidin-
2(S)- ylmethyll-prolyl-methionine trifluoroacetate
N-[l-(lH-Imidazol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyl]-prolyl-methionine methyl ester (0.020 g, 0.042 mmol) was
dissolved in CH30H (2 mL) at O~C and treated with lN NaOH solution
(0.167 mL, 0.167 mmol) with stirring. After S h at 23~C lN HCl (0.167
mL, 0.167 mmol) was added and the mixture was purified by preparative
RP HPLC on a Vydac column eluting with 0.1% TFA/CH3CN: 0.1%
TFA/ H20 gradient to give the title compound.
lH NMR (CD30D) ~ 8.88 (d, lH, J = lHz)), 7.43 (d, lH, J = lHz), 4.53
- 4.58 (m, lH), 4.25-4.31 (m, lH), 3.96 (ABq, 2H), 3.7 - 3.85 (m, 3H),
3.58-3.66(m, lH),3.50(dd, lH,J=3, 14Hz),3.39(dd, lH,J=3, 14
Hz), 3.23 - 3.42 (m, lH), 2.45 - 2.67 (m, 3H), 2.12 - 2.28 (m, 4H), 2.08
(s, 3H), 1.98 - 2.05 (m, 3H), 1.54- 1.68 (m, 2H), 1.26 - 1.4 (m, lH),
1.03 (t, 3H, J = 7 Hz). FAB MS 466 (M + 1).
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EXAMPLE 2
r~ lion of N-[1-[1-(4-Cyanobenzyl)-lH-imitl~7.ol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine
isopropyl ester
Step A: N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-proline
3(S)-Ethyl-2(S)-proline hydrochloride (from Example 1,
Step F) (2.33 g, 0.013 mol) was dissolved in CH30H (20 mL), treated
with 3A molecular sieves (2 g) and KOAc (1.27 g, 0.013 mol) to adjust
the pH of the reaction mixture to 4.5-5, then N-[(tert-
Butyloxy)carbonyl-prolinal (Pettit et al., J. Org. Chem. (1994) S9, [21]
6287-95) (3.36 g, 0.017 mol) was ~tlde~l, and the mixture was stirred for
16 hrs at room temperature. The reaction mixture was filtered, quenched
with aq satd NaHCO3 (5 mL) and concentrated to dryness. The residue
was extracted with CHC13. The extract was dried (MgSO4), filtered, and
conce~ t~d to give the title compound and inorganic salts.
Step B: N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine isopropyl ester
N-t(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-proline (2.4 g, 0.008 mol), methionine isopropyl ester hydrochloride
(2.21 g, 0.0097 mol), HOBT (1.49 g, 0.0097 mol) and EDC (1.86 g,
0.0097 mol) were dissolved in DMF (15 mL) at room temperature and
treated with N-methylmorpholine (3 mL, 0.024 mol). The reaction
mixture was stirred overnight at room temperature, then concentrated and
partitioned between EtOAc and H2O. The organic layer was washed
with aq satd NaHCO3 solution, brine, and dried (MgSO4). The crude
product was chromatographed on a flash silica gel column eluting with
hexane: EtOAc, 7:3 to give N-(t-butyloxycarbonyl)-pyrrolidin-2(S)-
ylmethyl]-3(S)-ethyl-prolyl-methionine isopropyl ester.
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Step C: N-(Pyrrolidin-2(S)-ylmethyl)-3(S)-ethyl-prolyl methionine
isopropyl ester hydrochloride
N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine isopropyl ester (1.38 g, 0.0028 mol) was
dissolved in EtOAc (40 mL), cooled to -20~C, saturated with HCl gas,
and stirred at 0~C. for 1.25 hr, and room tempe~alul~ for 0.25 hr.
Concentration to dryness gave the title compound.
Step D: lH-Imidazole-4- acetic acid methyl ester hydrochloride
A solution of lH-imidazole-4-acetic acid hydrochloride
(4.00g, 24.6 mmol) in methanol (100 ml) was saturated with gaseous
hydrogen chloride. The resulting solution was allowed to stand at room
temperature (RT) for 18hr. The solvent was evaporated in vacuo to afford
the title compound as a white solid.
lH NMR(CDC13, 400 MHz) o 8.85(1H, s),7.45(1H, s), 3.89(2H, s) and
3.75(3H, s) ppm.
Step E: l-(Triphenylmethyl)-lH-imidazol-4-ylacetic acid methyl
ester
To a solution of lH-Imidazole-4- acetic acid methyl ester
hydrochloride (24.85g, 0.141mol) in dimethyl formamide (DMF)
(115ml) was added triethylamine (57.2 ml, 0.412mol) and tribenzyl
bromide(55.3g, 0.171mol) and the suspension was stirred for 24hr. After
this time, the reaction mixture was diluted with ethyl acetate (EtOAc) (1
1) and water (350 ml). The organic phase was washed with sat. aq.
NaHCO3 (350 ml), dried (Na2SO4) and evaporated in vacuo. The
residue was purified by flash chromatography (sio2~ 0-100% ethyl
acetate in hexanes; gradient elution) to provide the title compound as a
white solid.
1 H NMR (CDC13, 400 MHz) o 7.35(1 H, s), 7.31 (9H, m), 7.22(6H, m),
6.76(1H, s), 3.68(3H, s) and 3.60('H, s) ppm.
Step F: [1-(4-Cyanobenzyl)-lH-imidazol-:--yl]acetic acid methyl
ester
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To a solution of l-(Triphenylmethyl)-lH-imidazol-4-
ylacetic acid methyl ester (8.00g, 20.9mmol) in acetonitrile (70 ml) was
added bromo-p-toluonitrile (4.10g, 20.92 mmol) and heated at 55~C for 3
hr. After this time, the reaction was cooled to room tempel~lul~ and the
S resulting imicl~7olium salt (white precipitate) was collected by filtration.
The filtrate was heated at 55~C for 18hr. The reaction mixt~lre was
cooled to room tempel~lul~ and evaporated in vacuo. To the residue was
added EtOAc (70 ml) and the resulting white preci~ te collected by
filtration. The precipitated imicl~7.olium salts were combined, suspended
10 in methanol (100 ml) and heated to reflux for 30min. After this time, the
solvent was removed in vacuo, the resulting residue was suspended in
EtOAc (75ml) and the solid isolated by filtration and washed (EtOAc).
The solid was treated with sat aq NaHCO3 (300ml) and CH2C12 (300ml)
and stirred at room temp~l~lul~ for 2 hr. The organic layer was
15 separated, dried (MgSO4) and evaporated in vacuo to afford the title
compound as a white solid:
lHNMR(CDCl3, 400 MHz) o7.65(1H, d, J=8Hz), 7.53(1H, s), 7.15(1H,
d, J=8Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.
~0 Step G: rl-(4-Cyanobenzyl)-lH-imidazol-5-yllacetic acid
A solution of [1-(4-cyanobenzyl)-lH-imid~7ol-5-yl]acetic
acid methyl ester (4.44g, 17.4mmol ) in THF (lOOml) and 1 M lithium
hydroxide (17.4 ml, 17.4 mmol) was stirred at RT for 18 hr. 1 M HCl
(17.4 ml) was added and the THF was removed by evaporation in vacuo.
25 The aqueous solution was lyophili7e-1 to afford the title compound
containing lithium chloride as a white solid.
lH NMR(CD30D, 400 MHz) o 8.22(1H, s), 7.74(1H, d, J=8.4Hz),
7.36(1H, d, J=8.4Hz), 7.15(1H, s), 5.43(2H, s) and 3.49(2H, s) ppm.
~0 Step H: N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-
pyr~olidin-2(S )-ylmethyl] -3 (S )-ethyl-prolyl-methionine
isopropyl ester
[1-(4-Cyanobenzyl)-lH-imidazol-5-yl]acetic acid ~ LiCl
(0.416 c. 1.47 mmol). N-(pyrrolidin-2(S)-ylmethyl)-3(S)-ethyl-prolyl-
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methionine isopropyl ester hydrochloride (Step I) ( 0.63 g, 1.33 rnmol),
HOOBT (0.239 g, 1.47 mmol), and EDC (0.281 g, 1.47 rnmol) were
dissolved in degassed DMF (20 mL) with stirnng at room tempelalul~,
N-methylmorpholine (0.8 mL, 5.32 rnmol) was added to achieve a pH of
5 7, and stilTing was continued overni~ht The reaction mix~lre was
concentrated to remove most of the DMF, and the residue was partitioned
between EtOAc and aq satd NaHCO3 solution. The aq layer was washed
with EtOAc, the organics combined, washed with brine and dried
(MgSO4). Filtration and concentration to dryness gave the title
10 compound after chromatography on silica gel eluting with
CH2C12:CH30H, 95:5.
Anal. calcd for C33H46N6O4S ~ 0.7 H2O: C, 62.38; H, 7.52; N, 13.23;
found: C, 62.40; H, 7.17; N, 13.11.
FAB MS 623 (M+1)
Following the procedures outlined above, but substit~ltin~ methionine
sulfone isopropyl ester for methionine isopropyl ester, the following
compound was prepared:
N-tl-~1-(4-Cyanobenzyl)-lH-irnidazol-S-ylacetyl]pyrrolidin-2(S)-
20 ylmethyll-3(S)-ethyl-prolyl-methionine sulfone isopropyl ester
Anal. calcd for C33H46N6O6S ~ 0.9 H2O: C, 59.07; H, 7.18; N, 12.52,
found: C, 58.99; H, 6.87; N, 12.86.
FAB MS 655 (M+1)
EXAMPLE 3
Preparation of N-[ l -[1 -(4-Cyanobenzyl)- 1 H-imidazol-5-
ylacetyl] pyrrolidin-2(S )-ylmethyl] -3 (S )-ethyl-prolyl-methionine
sulfoxide
To a solution of N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine (0.15 g,
0.186 mM) in 3 mL of MeOH:H ~O 1: 1 was added sodium periodate
(0 048 c~ 0.~23 mM). The mixture was stirred for 1 h. diluted with 3 mL
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of H2O and purified by prep HPLC (Delta-pak, C-18). The pure fractions
were pooled and lyophilli7e~1 to yield the title compound.
Anal. calcd for C30H40N6oss ~ 4.2 CF3C02H ~ 0.5 H20:
C, 42.52; H, 4.20; N, 7.75;
found: C, 42.51; H, 4.21; N, 8.11.
FAB MS 597 (M+l)
Following the procedure above the following compound was prepared:
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
ylmethyll-3(S)-ethyl-prolyl-methionine sulfoxide isopropvl ester
Anal. calcd for C33H46N605S ~ 1.0 H2O:
C, 60.34; H, 7.37; N, 12.80;
found: C, 60.32; H, 7.19; N, 12.42.
EXAMPLE 4
Pl~al~ion of N-[1-[1-(4-Cyanobenzyl)-lH-imi~1~7ol 5
ylacetyl~pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine sulfone
20 To a solution of N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine (0.15 g,
0.186 mM) in 5 mL of MeOH:H2O 1:1 was added Oxone (1.1 g, 0.372
mM). After stirring for 0.5 h, the mixture was partially evaporated and
diluted with 5 mL of H2O and purified by prep HPLC (Vydac, C-18).
25 The pure fractions were pooled and lyophilized to yield the title
compound.
Anal. calcd for C30H4oN6o6s ~ 3.2 CF3C02H ~ 1.2 H20:
C, 43.75; H, 4.60; N, 8.41;
found: C, 43.75; H, 4.59; N, 8.45.
30 FAB MS 613 (M+l)
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E~AMPLE 5
Preparation of N-[1-[1-(4-Cyanobenzyl)-lH-imi~1~7.ol 5
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine methyl
5 esterand N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine
Step A: N- [ 1- [ 1 -(4-Cyanobenzyl)- 1 H-iII~idazol-5-ylacetyl]-
pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine
methvl ester
Following the procedures described for Fx~mrle 2, but
substihltin~ methionine methyl ester hydrochloride for methionine
isopropyl ester hydrochloride in Step B, the title compound was prepared.
Anal. calcd for C31H42N6O4S ~ 3.7 CF3C02H ~ 0.3 H20:
C, 45.13; H, 4.57; N, 8.22;
found: C, 45.10; H, 4.53; N, 8.39.
Step B: N-[l-tl-(4-Cyanobenzyl)-lH-imidazol-5-l)acetyl]pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine trifluoroacetate
N- [ 1- [ 1 -(4-Cyanobenzyl)- 1 H-imidazol-5 -
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine methyl
ester (0.016 g, 0.02 mmol) was dissolved in CH30H (1 mL) and H2O (1
mL) at ambient temperature and treated with 1 N NaOH (0.3 mL, 0.3
mmol) with stirring. After 1 hr the reaction mixture was neutralized with
lN HCl (0.3 mL) and purified on a VYDAC preparative RP HPLC
column and lyophilized to give the title compound.
Anal. calcd for C30H40N604S ~ 3.9 CF3C02H ~ 0.6 H20:
C, 43.81; H, 4.39; N, 8.11;
found: C, 43.79; H, 4.39; N, 8.27.
Followin~ the procedures outlined in Examples 2 and 3, but substituting
the appropriate carboxylic acid in Example 2. Step H; the followin~
compounds were prepared:
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N-rl-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S-)vlmethyll-3(S)-ethyl-
prolyl-methionine methyl ester
lH NMR (CD30D) o 8.88 (s, lH), 7.43 (s, lH), 4.64 - 4.71 (m, lH),
4.19-4.27 (m, lH), 3.94 (s, 2H), 3.75 - 3.88 (m, 2H), 3.74 (s, 3H), 3.57 -
3.61 (m, 2H), 3.34 - 3.5 (m, 3H), 3.15 - 3.25 (m, lH), 2.45 - 2.67 (m, 2H),
1.98 - 2.37 (m, 6H), 2.08 (s, 3H), 1.83 - 1.98 (m, 3H), 1.4 - 1.56 (m, lH),
1.01 (t, 3H, J = 7 Hz).
Anal. calcd for C23H37N504S ~ 2.8 CF3C02H ~ 1.3 H20:
C, 41.88; H, 4.96; N, 8.54;
Found: C, 41.85; H, 4.95; N, 8.54.
FAB MS 480 (M + 1).
N-rl-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-
prolyl-methionine
lH NMR (CD30D) ~ 8.87 (s, lH), 7.43 (s, lH), 4.61 - 4.71 (m, lH),
4.2-4.3 (m, lH), 3.94 (brs, 2H), 3.75 - 3.88 (m, 2H), 3.6 - 3.73 (m, lH),
3.16 - 3.48 (m, SH), 2.5 - 2.7 (m, 2H), 2.0 - 2.38 (m, 6H), 2.10 (s, 3H),
1.83 - 1.98 (m, 3H), 1.4- 1.55 (m, lH), 1.01 (t, 3H, J = 7 Hz).
Anal. calcd for C22H35N504S ~ 2.8 CF3C02H:
C, 42.24; H, 4.85; N, 8.92;
Found: C, 42.18; H, 4.86; N, 8.95.
FAB MS 466 (M + 1).
N-r 1 -Glycyl-pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine
methyl ester
FAB MS 429 (M + 1).
N-r l -Glycyl-pyrrolidin-2(S)-ylmethyl~-3(S)-ethyl-prolyl-methionine
Anal. calcd for ClgH34N404S ~ 3.0 CF3C02H ~ 0.5 H20:
C, 39.72; H, 5.00; N, 7.3 ';
Found: C. 39.21; H. 5.02: N, 7.68.
FAB MS 415 (M + l ).
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N-rl-(3-rlH-Imidazol-4-yllpropionyl)-pyrrolidin-2(S-)vlmethyll-3(S)-
ethyl-prolyl-methionine methyl ester
Anal. calcd for C24H3gN5O4S ~ 0.75 H20:
C, 56.84; H, 8.05; N, 13.81,
Found: C, 56.79; H, 7.95; N, 13.90.
FAB MS 494 (M + 1).
N-rl -(3-rlH-Imidazol-4-yllpropionyl)-pvrrolidin-2(S-)ylmethyll-3(S)-
ethyl-prolvl-methionine
FAB MS 480(M+ 1).
N-rl-r3-(1-(4-Cyanobenzyl)-lH-imidazol-S-vl)propionyllpyrrolidin-2(S)-
vlmethyll-3(S)-e~yl-prolyl-methionine methyl ester
Anal. calcd for C32H44N604S ~ 2.0 HCl ~ 0.4 H20:
C, 55.79; H, 6.85; N, 12.20;
Found: C, 55.86; H, 6.85; N, 11.95.
FAB MS 609 (M + 1).
N-r 1 -r3-(1 -(4-Cyanobenzyl)- 1 H-imidazol-5-yl)propionyllpyrrolidin-2(S)-
20 ylmethyll-3(S)-ethyl-prolyl-methionine
Anal. calcd for C31H42N604S ~ 2.9 CF3C02H ~ 0.8 H20:
C, 47.03; H, 4.99; N, 8.94;
Found: C, 47.05; H, 4.96; N, 9.31.
FAB MS 595 (M + 1).
EXAMPLE 6
Preparation of N-[2(S)-(lH-Imidazol-4-ylacetyl-amino)-3(S)-
methylpentyl]-prolyl-methionine methyl ester and N-[2(S)-(lH-
30 Imidazol-4-ylacetyl-amino)-3(S)-methylpentvll-prolvl-methionine
Step A: N-~2(S)-( l H-Imidazol-4-ylacetyl-amino)-3(S)-
methylpentyll-prolyl-methionine methyl ester
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Following the methods outlined in Example 1, but
sub~ N-(t-Butyloxycarbonyl)-isoleucinal for N-t(t-
Butyloxy)carbonyl]-3(S)-ethyl-2(S)-prolinal in Step I, the title compound
was prepared.
5 Anal. calcd for C22H37N504S ~ 0.5 H20: C, 55.43; H, 8.04; N, 14.69;
Found: C, 55.75; H, 7.82; N, 14.36.
FAB MS 468 (M + 1).
Step B: N-[2(S)-(lH-Imidazol-4-ylacetyl-amino)-3(S)-
methvlpentyll-prolyl-methionine
The title compound was prepared following the method
dscribed in Example 1, Step N.
Anal. calcd for C21H35N504S ~ 2.5 CF3C02H:
C, 42.27; H, 5.12; N, 9.48;
Found: C,41.91;H,5.17;N,9.51.
FAB MS 454 (M + 1).
EXAMPLE 7
Preparationof N-[l-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
prolyl-methionine methyl ester and N-[l-(lH-Imidazol-4-ylacetyl)-
pyrrolidin-2(S)-ylmethyll -prolyl-methionine
Step A: N-[l-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
prolyl-methionine methyl ester
Following the methods outlined in Example 1, but
substituting N-(t-Butyloxycarbonyl)-prolinal for N-[(t-
Butyloxy)carbonyl]-3(S)-ethyl-2(S)-prolinal in Step I, the title compound
was prepared.
Anal. calcd for C21H33N504S ~ l.9 CF3C02H ~ 2 HCl:
C, 39.80; H, 5.00; N, 9.36;
Found: C. 39.82; H, 5.01; N, 9.33.
FAB MS 452 (M + l).
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Step B: N-[1-(lH-Tmi~1~7O1-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
prolyl-methionine
The title compound was prepared following the method
dscribed in Example 1, Step N.
Anal. calcd for C20H31N504S ~ 2.6 CF3C02H ~ 1.1 HCl:
C,40.15,H,4.79;N,9.29;
Found: C, 40.15; H, 4.85; N, 9.02.
FAB MS 438 (M + 1).
10 Following the procedures outlined in Examples 1, 2, and 7, the following
compounds were prepared:
N-[1-[1-(4-Cyanoberlzyl)-lH-imidazol-S-ylacetyl]pyrrolidin-2(S)-
ylmethyll-prolyl-methionine methyl ester
Anal. calcd for C29H3gN6O4S ~ 1.2 H20: C, 59.20; H, 6.92; N, 14.28;
Found: C,59.25;H,6.81;N, 14.14.
FAB MS 567 (M+ 1).
N-[ l -[1 -(4-Cyanobenzyl)- lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-
20 ylmethvll-prolyl-methionine
Anal. calcd for C28H36N6O4S ~ 3.4 CF3C02H ~ 1.0 H20:
C, 43.61; H, 4.35; N, 8.77;
Found: C, 43.59; H, 4.35; N, 8.91.
FAB MS 553 (M + 1).
EXAMPLE 8
Using the procedures described in Example l, but substituting 3(S)-Ethyl-
2(S)-proline hydrochloride for proline methyl ester in Step I, the
- 30 following compounds were prepared:
N-[1 -( lH-Imidazol-4-ylacetyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine methyl ester
Anal. calcd for Cr~5H4lNsO4S ~ 0.5 H2O: C. 50.93: H, 7.52; N l 1.88;
Found: C. 50.. 90; H. 7.38; 1~,'. l l.87.
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N-[1-(lH-Imidazol-4-ylacety1)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-
ethyl-prolyl-methionine trifluoroacetate
Anal. calcd for C24H39N504S ~ 2.95 CF3C~2H:
C, 43.27; H, 5.09; N, 8.44;
Found: C,43.17;H,5.16;N,8.54.
N-[1-[1-(4-Cyanobenzyl)-lH-imicl~7.ol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine methyl ester
Anal. calcd for C33H46N6O4S: C, 63.63; H, 7.45; N, 13.50;
Found: C, 63.53; H, 7.36; N, 13.39.
N-[1-[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine trifluoroacetate
Anal. calcd for C32H44N6O4S ~ 3.2 CF3C02H ~ 0.6 H20:
C, 46.85; H, 4.96; N, 8.54;
Found: C, 46.86; H, 4.96; N, 8.78.
N-[1 -[1 -(4-Cyanobenzyl)- lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine isopropyl ester
Anal. calcd for C35H50N6o4s ~ 0.25 H2O: C, 64.14; H, 7.77; N, 12.82;
Found: C, 64.16; H, 7.73; N, 12.82.
N-[1-(3-[lH-IIr~idazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyll-3(S)-ethyl-prolyl-methionine methyl ester
FAB MS 522 (M + 1).
N-[1-(3-[lH-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-
ylmethyll-3(S)-ethyl-prolyl-methionine trifluoroacetate
FAB MS 508 (M + 1).
N-[ l -Glycyl-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine methvl ester
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N-[ l -Glycyl-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine trifluoro~et~te
EXAMPLE 9
s
Preparation of N-~1-(1-(4-Nitrobenzyl)-lH-imidazol4-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
trifluoroacetate and N-[1-(1-(4-Nitrobenzyl)-lH-imi(1~7ol-5-ylacetyl]
pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
trifluoroacetate.
Step A: 1-(4-Nitrobenzyl)-lH-imidazol-4-ylacetic acid methyl ester
and 1-(4-Nitrobenzyl)-lH-imic1~7ol-5-ylacetic acid methyl
ester (3: 1 rnixture)
To a solution of sodium hydride (60% in mineral oil, 99 mg,
2.5 mmol) in dimethylform~mide (2 ml) cooled to 0~C was ~ cl via
c~nn~ , a solution of lH-imidazole-4-acetic acid methyl ester
hydrochloride (200 mg, 1.13 mmol) in dimethylform~n~ide (3 ml). This
suspension was allowed to stir at 0~C for 15 min. To this suspension was
added 4-nitrobenzyl bromide (244 mg, 1.13 mmol) and stirred at room
temperature for 2 h. After this time, the mixture was quenched with sat.
aq. sodium bicarbonate (15 ml) and water (20 ml) and extracted with
methylene chloride (2 x 50 ml). The combined organic extracts were
washed with brine (20 rnl), dried (MgSO4), filtered and the solvent was
evaporated in vacuo. The residue was purified by flash chromatography
using acetonitrile as eluent to give the title compounds as a yellow oil.
1H NMR (CDC13, 400 MHz) o 8.20 (2H, d, J=8.5 Hz), 7.49 (lH, s), 7.27
(2H, d, J=8.5 Hz), 7.03 (0.25H, s), 6.87 (0.75H, s), 5.28 (O.SH, s), 5.18
(1.5H, s), 3.70 (2.25H, s), 3.65 (l.SH, s), 3.61 (0.75H, s) and 3.44 (O.SH,
- 30 s) ppm.
Step B: 1-(4-Nitrobenzyl)-1H-imidazol-~-ylacetic acid
hydrochloride and 1 -(4-Nitrobenzyl)- 1 H-imidazol-5-ylacetic
acid (3: I mixture)
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To a solution of a mixture of 1-(4-Nitrobenzyl)-lH-
imi~l~7.ol-4-ylacetic acid methyl ester and 1-(4-Nitrobenzyl)-lH-
imidazol-5-ylacetic acid methyl ester (3:1mixture, 216 mg, 0.785 mmol)
in methanol (3 ml) and tetrahydrofuran (3 ml) under argon was added 1.0
5 M sodium hydroxide (1.18 ml, 1.18 mmol) and stirred for 18 h. After this
time, 1.0 N hydrochloric acid (2.36 ml, 2.36 mmol) was added and the
.~ixL..~ evaporated in vacuo to give the title compounds.
lH NMR (CDC13, 400 MHz) o 9.04 (0.75H, s), 8.83 (0.25H, s), 8.28
(2H, d, J=8.8 Hz), 7.61 (2H, d, J=8.8 Hz), 7.54 (0.75H, s), 7.43 (0.25H,
10 s), 5.61 (0.SH, s), 5.58 (l.SH, s), 3.84 (0.SH, s) and 3.82 (l.SH, s) ppm.
Step C: N-[1-(1-(4-Nitrobenzyl)-lH-imidazol-4-ylacetyl]pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester bis
trifluoroacetate and N-[1-(1-(4-Nitrobenzyl)-lH-imidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl
methionine methyl ester bis trifluoro~et~te
To a solution of 1-(4-nitrobenzyl)-lH-imidazol-4-ylacetic
acid hydrochloride and 1-(4-nitrobenzyl)-lH-imidazol-5-ylacetic acid
hydrochloride (3:1 mixtllre, 0.392 mmol), N-[pyrrolidin-2(S)-ylmethyl]-
3(S)-ethyl-prolyl methionine methyl ester hydrochloride (0.392 mmol),
prepared as described in Example 1, Steps A-L (but lltili7ing the
substitutions described in Example 2, Step A), and 3-hydroxy-1,2,3-
benzotriazin-4(3H)-one (HOOBT, 0.39 mmol) in methylene chloride (10
ml) are added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (EDC, 0.392 mmol) and triethyl~mine (1.57 mmol) and the
mixture is stirred overnight at room temperature. After this time, sat. aq.
sodium bicarbonate (10 ml) is added and the mixture is extracted with
methylene chloride. The combined extracts are washed with sat. aq.
sodium bicarbonate (10 ml) and the solvent evaporated in vacuo. The
regioisomers are separated by preparative HPLC using a Nova Prep 5000
Semi preparative HPLC system and a Waters PrepPak cartridge
(47X300mm, C18, l5,um, 100 A) eluting with 5-95% acetonitrile/water s
(0.1% TFA) at 100 ml/min (chromatography method A) to give the title
compounds after lyophilization.
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..
Step D: N-[1-(1-(4-Nitrobenzyl)-lH-imi~1~7ol-4-ylacetyl]pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
trifluoroacetate
To a solution of N-[1-(1-(4-nitrobenzyl)-lH-imidazol4-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl methioninemethyl
ester bis trifluoroacetate (0.023 mmol) in methanol (1 ml ) at room
tempel~ul~ is added l.ON lithillm hydroxide (135 ~Ll, 0.135 mmol). This
solution is stirred for 4 h and treated with trifluoroacetic acid (100 ~Ll).
10 This mixture is pllrifi~.rl by preparative HPLC using chromatography
method A to give the title compound.
Step E: N-[1-(1-(4-Nitrobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
trifluoroacetate
Toasolutionof N-[1-(1-(4-nitrobenzyl)-lH-imidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl methioninemethyl
ester bis trifluoroacetate (0.031 mmol) in methanol (1 ml ) is added l.ON
20 lithium hydroxide (187 ~Ll, 0.187 mmol ). This solution is stirred for 4 h
and treated with trifluoroacetic acid (100 ,ul). This mixture is purified by
preparative HPLC using chromatography method A to give the title
compound.
EXAMPLE 10
Preparation of N-[ 1-(1-(1 -Farnesyl)- 1 H-imidazol-5-ylacetyl)-pyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl methionine bis trifluoroacetate
- 30 Step A: 1-(1-Farnesvl)-lH-imidazol-5-ylacetic acid methyl ester
To a solution of l-(tribenzyl)-lH-imidazol~-ylacetic acid
methyl ester (200 mg, 0.523 mmol) in acetonitrile (5 ml) was added trans,
trans-farnesyl bromide (156 ~I, 0.575 mmol) and heated at 55~C for 16 h.
After this time. the reaction was heated at 80~C for 3 h and then the
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reaction mixture was evaporated in vacuo. The residue was dissolved in
me~anol (5 ml ) and heated to reflux for 30 min and dlen evaporated in
vacuo. The residue was purified by flash chromatography (2-4%
me~anolJmethylene chloride gradient elution) to provide the title
5 compound.
lH NMR (CDC13, 400 MHz) ~i 7.50 (lH, s), 6.92 (lH, s), 5.24 (lH, t,
J=5.9 Hz), 5.09 (2H, m), 4.49 (2H, d, J=6.9 Hz), 3.69 (3H, s), 3.60 (2H,
s), 1.91-2.15 (8H, m), 1.72 (3H, s), 1.65 (3H, s), 1.59 (3H, s) and 1.57
(3H, s) ppm.
Step B: N-t 1 -(1 -(1 -Farnesyl)- 1 H-imidazol-5-ylacetyl)pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl methioninemethylester
bis trifluoroacetate
Following the procedure described in Example 9, Steps C-D,
but using l-farnesyl-lH-imic1~7ol-5-ylacetic acid me~yl ester described
in Step A in place of 1-(4-nitrobenzyl)-lH-imic1~7ol-5-ylacetic acid
me~yl ester provides the title compound.
Step C: N-~l-(l-(l-Farnesyl)-lH-imicl~701-5-ylacetyl)pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl methionine bis
trifluoroacetate
Following the procedure described in Example 1, Step N,
but using the methyl ester prepared as described in Step B provides the
title compound.
EXAMPLE 11
Preparation of N-[ l -(1 -(1 -Geranyl)- 1 H-imidazol-5-ylacetyl)-pyrrolidin-
2(S)-ylmethyll-3(S)-ethyl-prolyl-methionine bis trifluoroacetate
Step A: N-~ 1 -(1 -(1 -Geranyl)- 1 H-imidazol-5-ylacetyl)pyrrolidin-
2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine methyl ester bis .
trifluoroacetate
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Following the procedure described in Example 10, Steps A-
B, but using trans-geranyl bromide in place of farnesyl bromide provides
the title compound.
S Step B: N-[1-(1-(1-Geranyl)-lH-imi~1~7.ol-5-
ylacetyl)pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine bis trifluoroacetate
Following the procedure described in Example 1, Step N,
but using the methyl ester prepared as described in Step A provides the
title compound.
EXAMPLE 12
P,~alation of N-[1-(1-(4-Methoxybenzyl)-lH-imidazol-5-
ylacetyl)pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
trifluoroacetate
Step A: N-[1-(1-(4-Methoxybenzyl)-lH-imidazol-5-
yl)acetyl)pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine methyl ester bis trifluoroacetate
Following the procedure described in Example 9, Steps B-D,
but using 4-methoxybenzyl chloride in place of 4-nitrobenzylbromide
provides the title compound.
Step B: N-[ 1-(1 -(4-Methoxybenzyl)- 1 H-imidazol-5-
ylacetyl)pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine bis trifluoroacetate
Following the procedure described in Example 1, Step N, but
substituting the methyl ester from Step A provides the title compound.
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EXAMPLE 13
Preparation of N-[1-(1-(2-Naphthylmethyl)-lH-irnidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-methionine bis
5 trifluoroacetate
Step A: N-[1-(1-(2-Naph~ylmethyl)-lH-imi~1~7O1-5-ylacetyl]3(S)-
ethylpyrrolidin-2(S)-ylmethyl]-prolyl-methionine methyl
ester bis trifluoroacetate
Following the procedure described in Example 9, Steps B-D,
but using 2-(bromomethyl)naphthylene in place of 4-nitrobenzylbromide
provided the title compound.
Step B: N-[1-(1-(2-Naphthylmethyl)-lH-imidazol-5-
ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-prolyl-
methionine bis trifluoroacetate
Following the procedure described in Example 1, Step N,
but using the methyl ester prepared as described in Step A provided the
title compound.
EXAMPLE 14
Preparation of N-[ l -(1 H-Irnidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl] -
3(S)-ethyl-prolyl-(~-acetylamino)alanine trifluoroacetate
Step A: Methyl 2(S)-benzyloxycarbonylamino-3-amino propionate
A solution of 2(S)-benzyloxycarbonylamino-3-
aminopropionic acid (2.4 g) in methanol at 0~ C was saturated with HCl
gas. After stirring for 2 h at 20~ C the solution was evaporated to obtain
the title compound. 1HNMR (300 MHz, CD30D ) o 7.35 (5H, m), 5.13
(2H, s), 4.50 (lH, m), 3.77 (3H, s), 3.45 (lH, m), 3.22 (lH, m).
Step B: Methyl 2(S)-benzyloxycarbonylamino-3-acetylamino-
propionate
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To a solution of methyl 2(S)-benzyloxycarbonyl~mino-3-
amino propionate (2.5 g) in methylene chloride was added pyridine (20
mL) and acetic anhydride (5 mL). After ~tirrin~ for 2 h the solution was
conce~ ted in vacuo. The residue was partitioned between ethyl ~ et~te
and water. The ethyl acetate layer was extracted w/ 50 mL each of 2%
potassium hydrogen slllf~te, saturated sodium bicarbonate, saturated
sodium chloride, dried over m~gnesium sulfate and concel~ ted in
vacuo. Upon evaporation pyridine hydrochloride precipitated and was
removed by filtration. The filtrate was evaporated to obtain the title
compound. 1HNMR (300 MHz, CDC13 ) ~ 7.28 (5H, s), 6.14 (lH, s),
5.97 (lH, d), 5.10 (2H, s), 4.41 (lH, m), 3.78 (3H, s), 1.93 (3H, s).
Step C: Methyl 2(S)-amino-3-acetylaminopropionate
To a solution of methyl 2(S)-benzyloxycarbonylamino-3-
acetylamino-propionate (2.2 g ) in ethanolic HCl was added 10% Pd/C (
0.3 g) under nitrogen atmosphere. Hydrogen was applied to the mixt lre
at 60 psi for 16 h. The mixture was filtered and concellLI~t~d in vacuo.
The residue was ~ ul~ted with diethyl ether to obtain the product.
lHNMR (300 MHz, CD30D) ~ 4.20 (lH, m), 3.88 (3H, s), 3.82 (lH, m),
3.60 (lH, m), 1.99 (3H, s).
Step D: N-t 1 -(1 H-Imidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
3(S)-ethyl-prolyl-(,3-acetylamino)alanine methyl ester
trifluoroacetate
Following the procedures outlined in Examples 1 and 2, but
substituting the methyl 2(S)-amino-3-acetylaminopropionate of Step C
for methionine methyl ester hydrochloride in Example 1, Step K, the title
compound is prepared.
- 30 Step E: N-[l-(lH-Imidazol-4-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-
3(S)-ethvl-prolyl-(~-acetylamino)alanine trifluoroacetate
The title compound is prepared following the method
described in Example 1, Step N.
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EXAMPLE 15
In vitro inhibition of ras farnesyl transferase
Assays offarnesyl-protein transferase. Partially purified
bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and RAS-
CAIL) were ~lG~aled 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.A. 86:6630-6634 (1989), respectively.
Bovine FpTase was assayed in a volume of 100,ul cont~inin~ 100 mM N-
(2-hydroxy ethyl) piperazine-N'-(2-ethane sulfonic acid) (HEPES), pH
7.4, 5 mM MgC12, 5 mM dithiothreitol (DTT), 100 mM [3H]-farnesyl
diphosphate ([3H]-FPP; 740 CBq/mmol, New Fn~l~ncl Nuclear), 650 nM
Ras-CVLS and 10 ~g/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.
Preci~i~ates 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 lltili7ell
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 arnount 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 rnixture. Reactions were performed for 30 min., stopped with
100 ~l 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 a~ainst human FPTase by the assay described above
and were found to have IC50 of < l 0 ~LM.
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EXAMPLE 16
In vivo ras farnesylation assav
The cell line used in this assay is a v-ras line derived from
S either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21. The
assay is ~lrol,lled essentially as described in DeClue, J.E. et al., Cancer
Research 51:712-717, (1991). Cells in 10 cm dishes at 50-75%
confluency are treated with the test compound (final concentration of
solvent, methanol or ~lim~.thyl sulfoxide, is 0.1 %). After 4 hours at 37~C,
10 the cells are labelled in 3 ml methionine-free DMEM supple-meted with
10% regular DMEM, 2% fetal bovine serum and 400
mCit35S]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
5 ~ntip~in/O.S mM PMSF) 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 immllnoprecipitated 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 4~C, 200 ml of a 25%
suspension of protein A-Sepharose coated with rabbit anti rat IgG is
added for 45 min. The immllnoprecipitates 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% acryl~micle 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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EXAMPLE 17
In vivo ~rowth inhibition assay
To dete~ le the biological consequences of FPTase
5 inhibition, the effect of the compounds of the in~t~nt 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
in~t~nt compounds for Ras-in-lllce~l cell transformation.
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
15 a~ro~liate concentration of the instant compound (dissolved in methanol
at 1000 times the final concentration used in the assay). The cells are fed
twice weeldy with 0.5 ml of medium A cont~inin~ 0.1% methanol or the
concentration of the instant compound. Photomicrographs are taken 16
days after the cultures are seeded and comparisons are made.
