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 TRANSE~ERASE
RELATED APPLICATION
The present patent application is a continuation-in-part
application of copending application Serial No. 08/470,690, filed June 6,
1995, which is a continuation-in-part application of copending
application Serial No. 08/412,829, filed March 29, 1995.
BACKGROUND OF THE INVENTION
The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ras)
are part of a ~i~n~lling pathway that links cell surface growth factor
receptors to nuclear ~i~n~l.s initi~ting cellular proliferation. Biological
and biochemical studies of Ras action indicate that Ras functions like a
G-regulatory protein. In the inactive state, Ras is bound to GDP. Upon
growth factor receptor activation Ras is inclllced to exchange GDP for
GTP and undergoes a conformational change. The GTP-bound form of
Ras propagates the growth stimlll~tQry signal until the signal is
termin~te-l by the intrinsic GTPase activity of Ras, which returns the
protein to its inactive GDP bound form (D.R. Lowy and D.M.
Willumsen, Ann. Rev. Biochem. 62:851-891 (1993)). Mutated ras
genes (Ha-ras, Ki4a-ras, Ki4b-ras and N-ras) are found in many human
cancers, including colorectal ca~cinoma, exocrine pancreatic carcinoma,
and myeloid leukemias. The protein products of these genes are
defective in their GTPase activity and constitutively transmit a growth
stimulatory signal.
Ras must be localized to the plasma membrane for both
normal and oncogenic functions. At least 3 post-tr~n~l~tional
modifications are involved with Ras membrane localization, and all 3
modifications occur at the C-terminus of Ras. The Ras C-tenninll~
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 serves as a signal sequence for the
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enzymes farnesyl-protein transferase or geranylgeranyl-protein
transferase, which catalyze the alkylation of the cysteine residue of the .
CAAX motif with a Cls or C20 isoprenoid, respectively. (S. Clarke.,
Ann. Rev. Biochem. 61:355-386 (1992); W.R. Schafer and J. Rine,
5 Ann. Rev. Genetics 30:209-237 (1992)). The Ras protein is one of
several proteins that are known to undergo post-tr~n~l~tional
farnesylation. Other farnesylated proteins include the Ras-related GTP-
binding proteins such as Rho, fungal mating factors, the nuclear l~mins,
andthe ~ mm~ subunit of transducin. James, et al., J. Biol. Chem. 269,
14182 (1994) have identified a peroxisome associated protein Pxf which
is also farnesylated. James, et al., have also suggested that there are
farnesylated proteins of unknown structure and function in addition to
those listed above.
Inhibition of farnesyl-protein transferase has been shown to
15 block the glo~vlh of Ras-transformed cells in soft agar and to modify
other aspects of their transformed phenotype. It has also been
demonstrated that certain inhibitors of farnesyl-protein transferase
selectively block the processing of the Ras oncoprotein intracellularly
(N.E. Kohl et al., Science, 260: 1934-1937 (1993) and G.L. James et al.,
20 Science, 260:1937-1942 (1993). Recently, it has been shown that an
inhibitor of farnesyl-protein transferase blocks the growth of ras-
dependent tumors in nude mice (N.E. Kohl et al., Proc. Natl. Acad. Sci
U.S.A., 91:9141-9145 (1994) and induces regression of m~mm~ry and
salivary carcinomas in ras transgenic mice (N.E. Kohl et al., Nature
25 Medicine, 1:792-797 (1995).
Indirect inhibition of farnesyl-protein transferase in vivo
has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) and
compactin (Hancock et al., ibid; Casey et al., ibid; Schafer et al.,
Science 245:379 (1989)). These drugs inhibit HMG-CoA reductase, the
30 rate limiting enzyme for the production of polyisoprenoids including
farnesyl pyrophosphate. Farnesyl-protein transferase lltili7es farnesyl
pyrophosphate to covalently modify the Cys thiol group of the Ras
CAAX box with a farnesyl group (Reiss et al., Cell, 62:81-88 (1990);
Schaber et al., J. Biol. Chem., 265: 14701-14704 (1990); Schafer et al.,
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Science, 249:1133-1139 (1990); Marlne et al., Proc. Natl. Acad. Sci
USA, 87:7541-7545 (1990)). Inhibition of farnesyl pyrophosphate
biosynthesis by inhibiting HMG-CoA reductase blocks Ras membrane
loc~li7.~tion in cultured cells. However, direct inhibition of farnesyl-
5 protein transferase would be more specific and attended by fewer sideeffects than would occur with the required dose of a general inhibitor of
isoprene biosynthesis.
Inhibitors of farnesyl-protein transferase (FPTase) have
been described in two general classes. The first are analogs of farnesyl
10 diphosphate (FPP), while the second class of inhibitors is related to the
protein substrates (e.g., Ras) for the enzyme. The peptide derived
inhibitors that have been described are generally cysteine cont~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.,
15 PNAS, 88:732-736 (1991)). Such inhibitors may inhibit ~loteh
prenylation while serving as alternate substrates for the farnesyl-L,lolt;i
transferase enzyme, or may be purely competitive inhibitors (U.S.
Patent 5,141,851, University of Texas; N.E. Kohl e~ al., Science,
260:1934-1937 (1993); Graham, et al., J. Med. Chem., 37, 725 (1994)).
20 In general, deletion of the thiol from a CAAX derivative has been
shown to dramatically reduce the inhibitory potency of the compound.
However, the thiol group potentially places limit~tions on the
therapeutic application of FPTase-inhibitors with respect to
ph~rm~rokinetics, ph~rm~codynamics and toxicity. Therefore, a
25 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 and are therefore useful in the prevention and therapy of
arteriosclerosis and diabetic disturbance of blood vessels (JP H7-
30 112930).
It has recently been disclosed that certain tricycliccompounds which optionally incorporate a piperidine moiety are
inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516).
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Tmicl~7.ole-cont~inin~ inhibitors of farnesyl protein transferase have also
been disclosed (WO 95/09001 and EP 0 675 112 A1).
It is, therefore, an object of this invention to develop
peptidomimetic compounds that do not have a thiol moiety, and that will
5 inhibit farnesyl-protein transferase and thus, the post-tr~n.cl~tional
farnesylation of proteins. It is a further object of this invention to
develop chemotherapeutic compositions COI~ lg the compounds of
this invention and methods for producing the compounds of this
invention.
SUMMARY OF THE INVENTION
The present invention comprises peptidomimetic
piperazine-co-~t~ compounds which inhibit the farnesyl-~loteill
transferase. The in~t~nt compounds lack a thiol moiety and thus offer
15 unique advantages in terms of improved ph~ cokinetic behavior in
~nim~l~, prevention of thiol-dependent chemical reactions, such as rapid
autoxidation and disulfide formation with endogenous thiols, and
reduced systemic toxicity. Purther contained in this invention are
chemotherapeutic compositions cont~ininp these farnesyl transferase
20 inhibitors and methods for their production.
The compounds of this invention are illustrated by the
formulae A, B and C:
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V-A1(CR1a2)nA2(cR1a2)n~ (cR1b2)p\ ~N\ /N~
R4 R5
( l ~8)r ~R9)\ R\~
V - A1(CR1a2)nA2(CR1a2)n -\W/- (CR1b2)p~ ~N N--Z
B R3 R4
( lR8)r ~R9~; R2 R3
V ~ A1(CRla2)nA2(CR1a2)n ~W~!- (CR1b2)p~N\ /N Z
C R4
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DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention are useful in the inhibition
of farnesyl-protein transferase and the farnesylation of the oncogene
protein Ras. In a first embodiment of this invention, the inhibitors of
5 farnesyl-protein transferase are illustrated by the formula A:
(R8)r ~9~ R j R3
V ~ Al(CRla2)nA2(CRla2)n ~W~!- (CRlb2)p~ ~N~ /N~
R4 R5
.
whereln:
R1a and R1b are independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-clo cycloalkyl, C2-c6 alkenyl, C2-
C6 alkynyl, R100-, Rl lS(O)m-, RlOC(O)NR10-,
(R10)2N-c(o)-~ CN, NO2, (R10)2N-C(NR10)-, R10C(O)-,
R10OC(O)-, N3, -N(R10)2, or Rl 1OC(O)NR10-,
c) unsubstituted or substituted Cl-C6 aLkyl wherein the
sub~LiLulellt on the substituted Cl-C6 aLkyl is selected from
unsubstituted or substituted aryl, heterocyclic, C3-C10
cycloalkyl, C2-C6 alkenyl, C2-C6 aLkynyl, R100-,
Rl lS(o)m, R10c(o)NRlo-~ (R10)2N-c(o)-~ CN,
(Rlo)2N-c(NRlo)-~ Rloc(o)-~ R10Oc(o)-~ N3,
-N(R10)2, and Rl 1OC(O)-NR10-;
R2 and R3 are independently selected from: H; unsubstituted or
substituted C1 8 aLkyl, unsubstituted or substituted C2 8 aLkenyl,
25 unsubstituted or substituted C2 g alkynyl, unsubstituted or substituted aryl, ~NR6R7 or ~oR6
unsubstituted or substituted heterocycle, ~ ~
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wherein the substituted group is substituted with one or more of:
,, 1) aryl or heterocycle, unsubstituted or substituted with:
a) Cl 4 aLkyl,
b) (CH2)pOR6,
c) (CH2)pNR6R7,
d) halogen,
e) CN,
2) C3-6 cycloaLkyl,
3) oR6,
4) SR6a, s(o)R6a~ So2R6a,
5) --NR6R7
~6
6) --N~ R7
o
8) - O ~ N R6R7
o
9) --o~OR6
10) NR6R7
..
11 ) --SO2--NR6R7
H6
12) --N--SO2--R6a
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13) ~ R6
14) ~ o R6
15) N3, or
16) F; or
R2 and R3 are attached to the same C atom and are combined to form
5 - (CH2)U - wherein one of the carbon atoms is optionally replaced by a
moiety selected from: O, S(O)m, -NC(O)-, and -N(COR10)-;
R4 and R5 are independently selected from H and CH3;
and any two of R2, R3, R4 and RS are optionally attached to the
same carbon atom;
R6, R7 and R7a are independently selected from: H; Cl 4 aL~yl, C3-6
cycloaL~yl, heterocycle, aryl, aroyl, heteroaroyl, arylsulfonyl,
lS heteroarylsulfonyl, unsubstituted or substituted with:
a) Cl 4 aLkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
e) ~R
O "
f) SO2R , or
g) N(R10)2; or
R6 and R7 may be joined in a ring;
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R7 and R7a may be joined in a ring;
R6a is selected from: Cl 4 aL~yl, C3-6 cycloaL~yl, heterocycle, aryl,
unsubstituted or substituted with:
S a) Cl 4 alkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
e) ~R11
fl --SO2R 1 , or
g) N(R10)2;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-
C6 alkynyl, perfluoroalkyl, F, Cl, Br, RlOO-, RllS(O)m-,
RlOC(O)NR10-, (RlO)2Nc(o)-7 RlO2N-c(NRlo)-7 CN,
NO2, RlOC(O)-, RlOOC(O)-, N3, -N(Rl0)2, or
RllOC(O)NR10-, and
c) Cl-C6 aLkyl unsubstituted or substituted by aryl,
cyanophenyl, heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, RlOO-,
RllS(O)m-, RlOC(O)NH-, (RlO)2NC(O)-, R102N-
C(NR10)-, CN, RlOC(O)-, RlOOC(O)-, N3, -N(Rl0)2, or
RlOOC(O)NH-;
R9 is selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br,
R100-, RllS(O)m-, RlOC(O)NR10-, (RlO)2NC(O)-,
RlO2N-c(NRlo)-7 CN, NO2, RlOC(O)-, RlOOC(O)-, N3,
-N(Rl0)2, or Rl lOC(O)NR10-, and
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- 10 -
c) Cl-C6 alkyl unsubstituted or substituted by perfluoroalkyl,
F, Cl, Br, RlOO-, RllS(O)m-, RlOC(O)NR10-,
(RlO)2Nc(o)-~ R102N-C(NR10)-, CN, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or Rl loC(ojNR10-;
R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and
aryl;
Rl 1 is independently selected from Cl-C6 alkyl 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;
15 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 heteroatom selected from O, 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;
25 W is a heterocycle;
X is -cH2-~ -C(=O)-, or -S(=O)m-;
Y is unsubstituted or substituted aryl or unsubstituted or
substituted heterocycle, wherein the substituted aryl or
substituted heterocycle is substituted with one or
more of:
1) Cl 4 alkyl, unsubstituted or substituted with:
a) Cl 4 alkoxy,
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b) NR6R7,
c) C3-6 cycloaLkyl,
d) aryl or heterocycle,
e) HO,
f) -S(O)mR6a, or
g) -c(o)NR6R7~
2) aryl or heterocycle,
3) halogen,
4) oR6~
5) NR6R7,
6) CN,
7) N02,
8) CF3;
9) -S(O)mR6a
10) -C(o)NR6R7, or
11) C3-C6 cycloaLkyl;
m is 0, 1 or 2;
nis 0, 1, 2, 3 or4;
pis 0, 1, 2, 3 or4;
q is 1 or 2;
r is O to 5, provided that r is O when V is hydrogen;
s is O or 1 ;
tis Oorl;and
uis 4 or 5;
or the plh~ reutically acceptable salts thereof.
.
In a second embodiment of this invention, the inhibitors of
30 farnesyl-protein transferase are illustrated by the formula B:
=
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(18)r ~ 9)~ R j ~
V ~ A1(CRla2)nA2(CR1a2)n tW~- (CRlb2)p~ ~N~ ~N--Z
R3 R4
B
.
wherem:
Rla and Rlb are independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-clo cycloaL~yl, C2-c6 alkenyl, C2-
C6 alkynyl, RlOO-, RllS(O)m-, RlOC(O)NR10-,
CN(RlO)2Nc(o)-~ R102N-C(NR10)-, CN, NO2,
RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
Rl 10C(O)NR10
c) unsubstituted or substituted Cl-C6 alkyl wherein the
sub~liLulellt on the substituted Cl-C6 alkyl is selected from
unsubstituted or substituted aryl, heterocyclic, C3-Clo
cycloaL~yl, C2-C6 alkenyl, C2-C6 aLkynyl, R100-,
RllS(O)m-, RlOC(O)NR10-, (RlO)2Nc(o)-~ R102N-
C(NR10)-, CN, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2,
and Rl lOC(o)-NR10;
R2 and R3 are independently selected from: H; unsubstituted or
substituted C1 8 alkyl, unsubstituted or substituted C2 8 alkenyl,
unsubstituted or substituted C2 8 aL~ynyl, unsubstituted or substituted aryl,
~NR6R7 or ~l~~R6
unsubstituted or substitutedheterocycle, ~ O
wherein the substituted group is substituted with one or more of:
1) aryl or heterocycle, unsubstituted or substituted with:
a) Cl 4 alkyl,
b) (CH2)pOR6,
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c) (CH2)pNR6R7,
r d) halogen,
e) CN,
2) C3-6 cycloaLkyl,
3) 0 R6,
4) SR6a,S(o)R6a, S 02R6a
5) - NR6R~
6) n6
o
R6
--N~ NR7R7a
8) --~ NR6R7
O
o
10) ~ N R6R7
1 1 ) --S02--NR6R7
F~6
r 12) --N SO2--R
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- 14 -
13) ~ R6
o
15) N3, or
16) F; or
R2 and R3 are attached to the same C atom and are combined to form
- (CH2)u - wherein one of the carbon atoms is optionally replaced by a
S moiety selected from: O, S(O)m, -NC(O)-, and -N(COR10)-;
R4 is selected from H and CH3;
and any two of R2, R3 and R4 are optionally attached to the same
carbon atom;
R6, R7 and R7a are independently selected from: H; Cl 4 alkyl, C3-6
cycloaL~yl, heterocycle, aryl, aroyl, heteroaroyl, arylsulfonyl,
heteroarylsulfonyl, unsubstituted or substituted with:
a) Cl 4 aL~oxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
e) ~Rl1
f) --SO2Rl 1 , or
g) N(Rlo)2; or
R6 and R7 may be joined in a ring;
R7 and R7a may be joined in a ring;
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R6a is selected from: Cl 4 aL~yl, C3-6 cycloalkyl, heterocycle, aryl,
unsubstituted or substituted with:
a) Cl 4 alkoxy,
S b) aryl or heterocycle,
c~ halogen,
d) HO,
e) ~R
o
f) --SO2R1 1 , or
g) N(R10)2;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-clo cycloaL~yl, C2-c6 alkenyl, C2-
C6 aL~ynyl, perfluoroaL~yl, F, Cl, Br, R100-, RllS(O)m-,
RlOC(O)NR10-, (RlO)2Nc(o)-~ R102N-C(NR10)-, CN,
NO2, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aL~yl unsubstituted or substituted by aryl,
cyanophenyl, heterocycle, C3-clo cycloalkyl, C2-C6
alkenyl, C2-C6 aL~ynyl, perfluoroaL~yl, F, Cl, Br, R100-,
RllS(O)m-, RlOC(O)NH-, (RlO)2Nc(o)-~ R102N-
C(NR10)-, CN, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
RlOOC(O)NH-;
R9 is selected from:
a) hydrogen,
b) aL~enyl, aL~ynyl, perfluoroalkyl, F, Cl, Br, R100-,
RllS(O)m-, RlOC(O)NR10-, (RlO)2Nc(o)-~ R102N-
C(NR10)-, CN, NO2, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, or Rl 1 OC(O)NR10-, and
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c) Cl-C6 aL~yl unsubstituted or substituted by perfluoroalkyl,
F, Cl, Br, RlOO-, RllS(O)m-, RlOC(O)NR10-,
(RlO)2Nc(o)-~ R102N-C(NR10)-, CN, RlOC(O)-,
RlOOC(O)-, 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 selectedfrom: abond, -CH=CH-, -C--C-,
-C(O)-, -C(O)NR10-,-NRlOC(O)-, O, -N(R10)-,
-S(O)2N(R10)-, -N(RlO)S(O)2-~ or S(O)m;
15 G is H2 or O;
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,
25 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;
30 X is -cH2-~ -C(=O)-, or -S(=O)m-;
Z is a unsubstituted or substituted group selected from aryl,
heteroaryl, arylmethyl, heteroarylmethyl, arylsulfonyl,
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heteroarylsulfonyl, wherein the substituted group is
substituted with one or more of the following:
1) Cl 4 aL~yl, unsubstituted or substituted with:
a) Cl 4 aL~oxy,
S b) NR6R7
c) C3-6 cycloaL~yl,
d) aryl or heterocycle,
e) HO,
f) -S(O)mR6a, or
g) -c(o)NR6R7~
2) aryl or heterocycle,
3) halogen,
4) oR6,
S) NR6R7,
6) CN,
7) NO2,
8) CF3;
9) -S(O)mR6a
10) -C(o)NR6R7, or
11) C3-C6 cycloaL~yl;
mis 0,lor2;
nis 0, 1, 2, 3 or4;
pis 0, 1, 2, 3 or4;
q is 1 or 2;
r is 0 to S, provided that r is 0 when V is hydrogen;
s is 0 or 1 ;
t is 0 or 1 ; and
u is 4 or S;
or the pharmaceutically acceptable salts thereof.
In a third embodiment of this invention, the inhibitors of
farnesyl-protein transferase are illustrated by the formula C:
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- 18 -
(R~)r ~1~ R~ R3
V - A (CR a2)nA2(CR1a2)n - (CR1b2)p\ ,N /N--Z
/~G
C R
.
wherem:
R1a and Rlb are independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-clo cycloalkyl, C2-c6 alkenyl, C2-
C6 alkynyl, R1OO-, R11S(O)m-, R1OC(O)NR10-,
(R10)2Nc(o)-7 RlO2N-c(NRlo)-7 CN, NO2, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or R1 1OC(O)NR10-,
c) unsubstituted or substituted Cl-C6 alkyl wherein the
sub~liLulellt on the substituted Cl-C6 alkyl is selected from
unsubstituted or substituted aryl, heterocyclic, C3-Clo
cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, RlOO-,
RllS(O)m-, RlOC(O)NR10-, (RlO)2NC(O)-, R102N-
C(NR10)-, CN, RlOC(O)-, R1OOC(O)-, N3, -N(R10)2,
and Rl 1Oc(o)-NRlo;
R2 and R3 are independently selected from: H; unsubstituted or
substituted C1 8 alkyl, unsubstituted or substituted C2 8 alkenyl,
unsubstituted or substituted C2 g alkynyl, unsubstituted or substituted aryl,
~NR6R7 or ~oR6
unsubstituted or substituted heterocycle, ~ O
wherein the substituted group is substituted with one or more of:
1) aryl or heterocycle, unsubstituted or substituted with:
a) C1 4 alkyl,
b) (CH2)pOR6,
c) (CH2)pNR6R7,
d) halogen,
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- 19 -
e) CN,
2) C3-6 cycloaLkyl,
3) oR6,
4) SR6a, s(o)R6a~ So2R6a,
5) --NR6R7
6) --N~ R7
o
8) - O ~ N R6R7
o
10) 'f NR6R7
1 1 ) --S02--NR6R7
12) --N--S02--R6a
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- 20 -
13) ~ R6 , ,
o
15) N3, or
16) F; or
R2 and R3 are attached to the same C atom and are combined to form
- (CH2)U - wherein one of the carbon atoms is optionally replaced by a
5 moiety selected from: O, S(O)m, -NC(O)-, and -N(COR10)-;
R4 is selected from H and CH3;
and any two of R2, R3 and R4 are optionally ~tt~c.hed to the same
carbon atom;
R6, R7 and R7a are independently selected from: H; Cl 4 alkyl, C3-6
cycloaLkyl, heterocycle, aryl, aroyl, heteroaroyl, arylsulfonyl,
heteroarylsulfonyl, unsubstihlte~l or substituted with:
a) Cl-4 aLkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
11
O
f) --SO2R1 1 , or
g) N(R10)2; or
R6 and R7 may be joined in a ring;
R7 and R7a may be joined in a ring;
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R6a is selected from: Cl 4 aL~yl, C3-6 cycloalkyl, heterocycle, aryl,
unsubstituted or substituted with:
a) Cl 4 alkoxy,
S b) aryl or heterocycle,
c) halogen,
d) HO,
e) ~ R11
f) --SO2R1 1 , or
g) N(R10)2;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-clo cycloalkyl, C2-C6 alkenyl, C2-
C6 alkynyl, perfluoroalkyl, F, Cl, Br, RlOO-, RllS(O)m-,
RlOC(O)NR10-, (RlO)2Nc(o)-~ R102N-C(NR10)-, CN,
NO2, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aLkyl unsubstituted or substituted by aryl,
cyanophenyl, heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, RlOO-,
RllS(O)m-,RlOC(O)NH-,(RlO)2Nc(o)-~R102N-
C(NR10)-, CN, RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
RlOOC(O)NH-;
~ 25 R9 is seleeted from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br,
R100-, RllS(O)m-, RlOC(O)NR10-, (RlO)2Nc(o)-~
R102N-C(NR10)-, CN, NO2, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, or Rl lOC(O)NR10-, and
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c) Cl-C6 aL~yl unsubstituted or substituted by perfluoroaL~yl,
F, Cl, Br, R1OO-, R11S(O)m-, R1OC(O)NR10-,
(R10)2Nc(o)-~ R102N-C(NR10)-, CN, R1OC(O)-,
R1OOC(O)-, N3, -N(R10)2, or R1 1OC(O)NR10-;
R10 is independently selected from hydrogen, Cl-C6 aL~yl, benzyl and
aryl;
R11 is independently selected from Cl-C6 aL~yl and aryl;
A1 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(R1O)S(O)2-, or S(O)m;
15 G is O;
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,
25 provided that V is not hydrogen if A1 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;
30 X is -CH2-, -C(=O)-, or -S(=O)m-;
Z is a unsubstituted or substituted group selected from aryl,
heteroaryl, arylmethyl, heteroarylmethyl, arylsulfonyl,
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heteroarylsulfonyl, wherein the substituted group is
,~ substituted with one or more of the following:
1) Cl 4 alkyl, unsubstituted or substituted with:
a) Cl 4 alkoxy,
S b) NR6R7,
c) C3-6 cycloalkyl,
d) aryl or heterocycle,
e) H0,
f) -S(O)mR6a, or
g) -C(o)NR6R7,
2) aryl or heterocycle,
3) halogen,
4) oR6~
S) NR6R7,
lS 6) CN,
7) N02,
8) CF3;
g) -S(o)mR6a~
10) -C(o)NR6R7, or
11) C3-C6 cycloalkyl;
m is 0, 1 or 2;
nis 0, 1, 2, 3 or4;
p is 0, 1, 2, 3 or 4;
q is 1 or 2;
r is 0 to S, provided that r is 0 when V is hydrogen;
sis l;
t is 0 or 1; and
uis 40rS;
or the ph~ ceutically acceptable salts thereof.
A preferred embodiment of the compounds of this
invention is illustrated by the following formula:
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- 24 -
V - A (CR 2)nA2(CR 1a2)n -(W)- (CR1 b2)p~X~N\ ~f N~
R4 R5
A
whereln:
S 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 C2-C6
aLkenyl,
c) unsubstituted or substituted Cl-C6 aLkyl wherein the
substitutent on the substituted Cl-C6 aLkyl is selected from
unsubstituted or substituted aryl, heterocycle, cycloalkyl,
alkenyl, RlOO- and N(R10)2;
R3, R4 and R5 are independently selected from H and CH3;
~ NR6R7
R2 is H; O or Cl 5 aLkyl, unbranched or branched,
unsubstituted or substituted with one or more of:
1 ) aryl,
2) heterocycle,
3) oR6,
4) SR6a, SO2R6a, or
5) ~ NR6R7
o
and any two of R2, R3, R4, and R5 are optionally attached to the
same carbon atom;
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R6, R7 and R7a are independently selected from:
H; Cl 4 aL~yl, C3-6 cycloalkyl, aryl, heterocycle,
unsubstituted or substituted with:
a) Cl 4 aL~oxy,
S b) halogen, or
c) aryl or heterocycle;
R6a is selected from:
Cl 4 aL~yl or C3-6 cycloaL~yl,
unsubstituted or substituted with:
a) C 1-4 aL~oxy,
b) halogen, or
c) aryl or heterocycle;
15 R8 is independently selected from:
a) hydrogen,
b) Cl-c6 aL~yl, C2-c6 aL~enyl, C2-c6 alkynyl, Cl-C6
perfluoroaL~yl, F, Cl, R100-, RlOC(O)NR10-, CN, N02,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aL~yl substituted by Cl-C6 perfluoroaL~yl, R100-,
RlOC(o)NR10, (Rlo)2N-c(NRlo)-~ RlO
RlOOC(O)-, -N(R10)2, or Rl lOC(O)NR10-;
25 R9 is selected from:
a) hydrogen,
b) C2-C6 aL~enyl, C2-C6 aL~ynyl, Cl-C6 perfluoroalkyl, F,
Cl, RlOO, Rl lS(o)m, Rloc(o)NRlo-~ CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)- -N(R10)2 or
Rl 1 OC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6
perfluoroaL~yl, F, Cl, R100-, RllS(O)m-, RlOC(O)NR10-,
CN, (Rlo)2N-c(NRlo)-~ RlOC(O)-, RlOOC(O)-,
-N(R10)2, or Rl lOC(O)NR10-;
=
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R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and
aryl;
5 Rl 1 is independently selected from Cl-C6 alkyl and aryl;
Al and A2 are independently selected from: a bond, -CH=CH-, -~C-,
-C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;
~0 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 0 to 4 carbon atoms are
replaced with a a heteroatom selected from O, S, and N,
and
e) C2-C20 aL~enyl, and
20 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 selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or
25 isoquinolinyl;
X is -CH2- or -C(=O)-;
Y is mono- or bicyclic aryl, or mono- or bicyclic heterocycle,
unsubstituted or substituted with one or more of:
a) Cl 4 aLkyl,
b) Cl 4 alkoxy,
c) halogen, or
d) NR6R7;
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m is 0, 1 or 2;
nis 0, 1, 2, 3 or4;
pis 0, 1, 2, 3 or4;
r is O to 5, provided that r is O when V is hydrogen;
s is O or l; and
- tis Oor l;
or the phalmaceutically acceptable salts thereof.
In a second preferred embodiment of this invention, the
inhibitors of farnesyl-protein transferase are illustrated by the formula
B:
( I 8)r ~R9~ R2,~
V - A1 (CR1 a2)nA2(CR 1 a2)n -\W~!- (CR1 b2)p\ ~ N~ JN--Z
R3 R4
B
wherein:
Rla is independently selected from: hydrogen or Cl-C6 alkyl;
20 Rlb is independently selected from:
a) hydrogen,
- b) aryl, heterocycle, cycloalkyl, R100-, -N(R10)2 or C2-C6
alkenyl,
c) unsubstituted or substituted Cl-C6 alkyl wherein the
sub~Lilu~ellt on the substituted Cl-C6 alkyl is selected from
unsubstituted or substituted aryl, heterocycle, cycloalkyl,
alkenyl, R100- and N(R10)2;
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R3 and R4 are independently selected from H and CH3;
~ NR6R7
R2 is H; O or Cl 5 aLkyl, unbranched or branched,
unsubstituted or substituted with one or more of:
1) aryl,
S 2) heterocycle,
3) oR6,
4) SR6a, SO2R6a, or
5) NR6R7
~ ;
and any two of R2, R3, R4, and RS are optionally attached to the
same carbon atom;
R6, R7 and R7a are independently selected from:
H; Cl 4 alkyl, C3-6 cycloalkyl, aryl, heterocycle,
unsubstituted or substituted with:
lS a) Cl 4 alkoxy,
b) halogen, or
c) aryl or heterocycle;
R6a is selected from:
Cl 4 aLkyl or C3-6 cycloalkyl,
unsubstituted or substituted with:
a) Cl-4 aL~oxy,
b) halogen, or
c) aryl or heterocycle;
R8 is independently selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-C6 aL~enyl, C2-C6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, RlOO-, RlOC(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2, or
RllOC(O)NR10-, and
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-
c) Cl-C6 aL~yl substituted by Cl-C6 perfluoroalkyl, R100-,
RlOC(o)NR10, (Rlo)2N-c(NRlo)-~ RlO
RlOOC(O)-, -N(R10)2, or RllOC(O)NR10-;
5 R9 is selected from:
a) hydrogen,
b) C2-C6 aL~enyl, C2-C6 aL~ynyl, Cl-C6 perfluoroaL~yl, F,
Cl, RlOO-, RllS(O)m-, RlOC(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOc(o)-~ RlOOC(O)-, -N(R10)2, or
RllOC(O)NR10-, and
c) Cl-C6 aL~yl unsubstituted or substituted by Cl-C6
perfluoroalkyl, F, Cl, R100-, RllS(O)m-, RlOC(O)NR10-,
CN, (RlO)2N-c(NRlo)-~ 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;
Rll is independently selected from Cl-C6 alkyl and aryl;
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 aL~enyl, and
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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;
G is H2 or O;
s
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or
isoquinolinyl;
10 X is -CH2- or -C(=O)-;
Z is mono- or bicyclic aryl, mono- or bicyclic heteroaryl,
mono- or bicyclic arylmethyl, mono- or bicyclic
heteroarylmethyl, mono- or bicyclic arylsulfonyl,
mono- or bicyclic heteroarylsulfonyl, unsubstituted or
substituted with one or two of the following:
1) Cl 4 alkyl, unsubstituted or substituted with:
a) Cl 4 aLkoxy,
b) NR6R7,
c) C3-6 cycloaL~yl,
d) aryl or heterocycle,
e) HO,
f) -S(O)mR6, or
g) -c(o)NR6R7~
2) aryl or heterocycle,
3) halogen,
4) oR6,
S) NR6R7,
6) CN,
7) NO2,
8) CF3;
9) -S(O)mR6,
10) -C(o)NR6R7, or
11) C3-C6cycloaLkyl;
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m is 0, 1 or 2;
nis 0, 1, 2, 3 or4;
pis 0, 1, 2, 3 or4;
r is 0 to 5, provided that r is 0 when V is hydrogen;
s is 0 or 1;
t is 0 or 1; and
u is 4 or 5;
10 provided that when G is H2 and W is imicl~7.olyl, then the substitutent
(R8)r- V - Al(CRla2)nA2(CRla2)n - is not H and
provided that when X is -C(=O)-, or -S(=O)m-~ then t is 1 and the
sub~Lilu~ellt (R8)r- V - Al(CRla2)nA2(CRla2)n - is not H;
or the ph~rm~eutically acceptable salts thereof.
The preferred compounds of this invention are as follows:
20 2(S)-Butyl-1-(2,3-~ minoprop-1-yl)-4-(1-naphthoyl)-
plperazme
1-(3-Annino-2-(2-naphthylmethylamino)prop- 1-yl)-2(S)-butyl-
4-(1-naphthoyl)piperazine
2(S)-Butyl-1-{5-[1-(2-naphthylmethyl)]-4,5-
dihydroimidazol }methyl-4-( 1 -naphthoyl)piperazine
1-[5-(1-Benzylimidazol)methyl]-2(S)-butyl-4-(1-naphthoyl)piperazine
1-{5-[1-(4-Nitrobenzyl)imidazolyl]methyl}-2(S)-butyl-4-(1-
naphthoyl)piperazine
1 -(3-Acetamidomethylthio-2(R)-aminoprop- l-yl)-2(S)-butyl-4-(1-
35 naphthoyl)piperazine
~ =
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2(S)-Butyl- 1-[2-(1-imidazolyl)ethyl]sulfonyl-4-(1-naphthoyl)piperazine
2(R)-Butyl- 1 -imidazolyl-4-methyl-4-( 1 -naphthoyl)piperazine
2(S)-Butyl-4-( 1 -naphthoyl)- 1 -(3-pyridylmethyl)piperazine
1-2(S)-butyl-(2(R)-(4-nitrobenzyl)amino-3-hydroxypropyl)-4-(1-
naphthoyl)piperazine
1-(2(R)-Amino-3-hydroxyheptadecyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine
2(S)-Benzyl- 1 -imidazolyl-4-methyl-4-( 1 -naphthoyl)piperazine
1-(2(R)-Amino-3-(3-benzylthio)propyl)-2(S)-butyl-4-(1-naphthoyl)-
piperazine
1 -(2(R)-Amino-3-[3-(4-nitrobenzylthio)propyl]))-2(S)-butyl-4-( 1-
20 naphthoyl)piperazine
2(S)-Butyl-1-[(4-imidazolyl)ethyl]-4-(1-naphthoyl)piperazine
2(S)-Butyl-1-[(4-imi(1~7.olyl)methyl]-4-(l-naphthoyl)piperazine
2(S)-Butyl-l-[(l-naphth-2-ylmethyl)-lH-imi(1~7ol-5-yl)acetyl]-4-(l-
naphthoyl)piperazine
2(S)-Butyl- 1- [( 1 -naphth-2-ylmethyl)- 1 H-imidazol-5-yl)ethyl]-4-( 1-
30 naphthoyl)piperazine
1 -(2(R)-Amino-3-hydroypropyl)-2(S)-butyl-4-( 1-
naphthoyl)piperazine
1-(2(R)-Amino-4-hydroxybutyl)-2(S)-butyl-4-(1-naphthoyl)piperazine
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1-(2-Amino-3-(2-benzyloxyphenyl)propyl)-2(S)-butyl-4-(1-
- naphthoyl)piperazine
1-(2-Amino-3-(2-hydroxyphenyl)propyl)-2(S)-butyl-4-(1-
5 naphthoyl)piperazine
1- [3-(4-imidazolyl)propyl] -2(S)-butyl-4-( 1-
naphthoyl)piperazine
10 2(S)-n-Butyl-4-(l-naphthoyl)-l-[l-(l-naphthylmethyl)imi-1~7.ol-5-
ylmethyl] -piperazine
2(S)-n-Butyl-4-(1-naphthoyl)- 1-[1 -(2-naphthylmethyl)imicl~701-5-
ylmethyl] -piperazine
2(S)-n-Butyl-1-[1-(4-cyanobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine
2(S)-n-Butyl-1-[1-(4-methoxybenzyl)imidazol-5-ylmethyl]-4-(1-
20 naphthoyl)piperazine
2(S)-n-Butyl-1-[1-(3-methyl-2-butenyl)imicl~7.ol-5-ylmethyl]-4-(l-
naphthoyl)piperazine
25 2(S)-n-Butyl-1-[1-(4-fluorobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine
2(S)-n-Butyl-1-[1-(4-chlorobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine
1-[1 -(4-Bromobenzyl)imidazol-5-ylmethyl]-2(S)-n-butyl-4-( 1-
naphthoyl)piperazine
1-[1-(4-Bromobenzyl)imidazol-5-ylmethyl]-2(S)-n-butyl-4-(1-
35 naphthoyl)piperazine .
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2(S)-n-Butyl-4-(1-naphthoyl)-1-[1 -(4-trifluoromethylbenzyl)imidazol-5-
ylmethyl] -piperazine
2(S)-n-Butyl- 1- [ 1 -(4-methylbenzyl)imidazol-5-ylmethyl] -4-(1-
5 naphthoyl)-piperazine
2(S)-n-Butyl-1-[1-(3-methylbenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)-piperazine
1-[l-(4-phenylbenzyl)imic1~7ol-5-ylmethyl]-2(s)-n-butyl-4-(
naphthoyl) -piperazine
2(S)-n-Butyl-4-(1-naphthoyl)-1-[1-(2-phenylethyl)irnidazol-5-ylmethyl]-
piperazine
2(S)-n-Butyl-4-(l-naphthoyl)-1-[1-(4-trifluoromethoxy)-imic1~7.ol-5-
ylmethyl]piperazine
1-{ [1-(4-cyanobenzyl)-lH-imicl~7.ol-5-yl]acetyl }-2(S)-n-
20 butyl-4-(1-naphthoyl)piperazine
5(S)-n-Butyl- 1 -(2,3-dimethylphenyl)-4-(4-imidazolylmethyl)-piperazin-
2-one
5(S)-n-Butyl-4-[1-(4-cyanobenzyl)imidazol-5-ylmethyl]-1-(2,3-
dimethylphenyl)piperazin-2-one
4-[1 -(4-Cyanobenzyl)imidazol-5-ylmethyl]- 1 -(2,3-dimethylphenyl)-
5(S)-(2-methoxyethyl)piperazin-2-one
(S)-1-(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-
(methanesulfonyl)ethyl] -2-piperazinone
(S)-1-(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-
(ethanesulfonyl)ethyl]-2-piperazinone
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(S)- 1-(3-Chlorophenyl)-4-[ 1 -(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-
(ethanesulfonyl)methyl] -2-piperazinone
(S)- 1-(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imicl~7.olylmethyl]-5-
5 [N-ethyl-2-acetamido]-2-piperazinone
(+)-5-(X-Butynyl)- 1 -(3-chlorophenyl)-4-[1 -(4-cyanobenzyl)-5-
imidazolylmethyl] -2-piperazinone
10 1-(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imicl~7.olylmethyl]-2-
piperazinone
5(S)-Butyl-4-[1-(4-cyanobenzyl-2-methyl)-5-imidazolylmethyl]-1-(2,3-
dimethylphenyl) -piperazin-2-one
4-[1-(2-(4-Cyanophenyl)-2-propyl)-5-imi~1~7.olylmethyl]- 1-(3-
chlorophenyl)-5(S)-(2-methylsulfonylethyl)piperazin-2-one
5(S)-n-Butyl-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-1-(2-
20 methylphenyl)piperazin-2-one
4-[1-(4-Cyanobenzyl)-5-imidazolylmethyl]-5(S)-(2-fluoroethyl)-1-(3-
chlorophenyl)piperazin-2-one
25 4-[3-(4-Cyanobenzyl)pyridin-4-yl]-1-(3-chlorophenyl)-5(S)-(2-
methylsulfonylethyl)-piperazin-2-one
4-[5-(4 -Cyanobenzyl)- 1 -imidazolylethyl]- 1 -(3-chlorophenyl)piperazin-
2-one.
or the ph~ eutically acceptable salts thereof.
Specific examples of the compounds of the invention are:
1-{5-[1-(4-Nitrobenzyl)imidazolyl]methyl}-2(S)-butyl-4-(1-
naphthoyl)piperazine
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PCTrUS96/04019
- 36 -
<'N~
1-[5-(1-Benzylimidazol)methyl]-2(S)-butyl-4-(1-naphthoyl)piperazine
~0
S N~
1-(2(R)-Amino-3-(3-benzylthio)propyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine
NH2 S
1-(2(R)-Amino-3-[3-(4-nitrobenzylthio)propyl])-2(S)-butyl-4-(1-
naphthoyl)piperazine
- CA 022l6707 l997-09-29
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- 37 -
02N ~S ,N~NJ~
2(S)-n-Butyl-1-[1-(4-cyanobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine
O
N~
2(S)-n-Butyl-1-[1-(4-cyanobenzyl)imil1~7.ol-5-ylmethyl]-4-(2,3-
dimethylphenyl)piperazin-5-one
~ H3C CH3
NC~ ~N~N~
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2(S)-n-Butyl- 1- [ 1 -(4-chlorobenzyl)imidazol-5-ylmethyl]-4-( 1-
naphthoyl)~i~erazine
Nf
1-{ [1-(4-Cyanobenzyl)-lH-imidazol-5-yl]acetyl}-2(S)-n-butyl-4-(1-
naphthoyl)piperazine
~\N~
1-[ 1 -(4-Cyanobenzyl)imidazol-5-ylmethyl]-4-(2,3-dimethylphenyl)-
2(S)-(2-methoxyethyl)piperazin-5-one
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,CH3
O~
NC ~ ~N~N~
5(S)-n-Butyl-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-1-(2-
methylphenyl)piperazin-2-one
CH3
NC
N~
(S)-1-(3-Chlorophenyl)-4-tl-(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-
10 (methanesulfonyl)ethyl]-2-piperazinone
CH3
o2
NC ~
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(S)- 1 -(3-Chlorophenyl)-4-[ 1 -(4-cyanobenzyl)-5-imicl~7.olylmethyl]-5-[2-
(ethanesulfonyl)ethyl] -2-piperazinone
,CH2CH3
o2
NC ~N~N--
<' O
(S)-1-(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imicl~zolylmethyl]-5-[2-
(ethanesulfonyl)methyl]-2-piperazinone
<CH3
SO2
NC~ ~N N~
N
1 -(3-Chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imicl~zolyl-methyl]-2-
piperazinone
NC ~
N
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or the ph~ r.eutically acceptable salts thereof.
The compounds of the present invention may have
asymmetric centers and occur as racemates, racemic rnixtures, and as
5 individual diastereomers, with all possible isomers, including optical
isomers, being included in the present invention. When any variable
(e.g. aryl, heterocycle, Rl, R2 etc.) occurs more than one time in any
constituent, its definition on'each occurence is independent at every
other occurence. Also, combinations of substituents/or variables are
10 permis~ible only if such combinations result in stable compounds.
As used herein, "aL~yl" is intended to include both branched
and straight-chain saturated aliphatic hydrocarbon groups having the
specified number of carbon atoms; "aL~oxy" represents an alkyl group
of indicated number of carbon atoms ~tt~rhed through an oxygen
15 bridge. "Halogen" or "halo" as used herein means fluoro, chloro,
bromo and iodo.
As used herein, "aryl" is intended to mean any stable
monocyclic or bicyclic carbon ring of up to 7 members in each ring,
wherein at least one ring is aromatic. Fx~mples of such aryl elements
20 include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl,
phenanthryl, anthryl or acenaphthyl.
The term heterocycle or heterocyclic, as used herein,
represents a stable S- to 7-membered monocyclic or stable 8- to 11-
membered bicyclic heterocyclic ring which is either saturated or
25 Im.~tllrated, 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
30 creation of a stable structure. Examples of such heterocyclic elements
include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl,
benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl,
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dihydrobenzothiopyranyl sulfone, furyl, imidazolidinyl, imidazolinyl,
imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl,
isothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl,
naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-oxopiperazinyl,
S 2-oxopiperdinyl, 2-oxopyrrolidinyl, piperidyl, piperazinyl, pyridyl,
pyrazinyl, pyrazolidinyl, pyrazolyl, pyridazinyl, pyrimidinyl,
pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl,
tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl,
thienofuryl, thienothienyl, and thienyl.
As used herein, "heteroaryl" is intended to mean any stable
monocyclic or bicyclic carbon ring of up to 7 members in each ring,
wherein at least one ring is aromatic and wherein from one to four
carbon atoms are replaced by heteroatoms selected from the group
consisting of N, O, and S. Examples of such heterocyclic elements
include, but are not limited to, ben7imidazolyl, benzisoxazolyl,
benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl,
isochromanyl, isoindolinyl, isoquinolinyl, isothiazolyl, naphthyridinyl,
oxadiazolyl, pyridyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl,
pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl,
tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiazolyl, thienofuryl,
2~ thienothienyl, and thienyl.
As used herein in the definition of R2 and R3, the term "the
substituted group" intended to mean a substituted Cl ~ alkyl, substituted
C2 ~ alkenyl, substituted C2 ~, alkynyl, .substituted aryl or substituted
heterocycle from which the substitutent(s) R2 and R3 are selected.
As used herein in the definition of R6, R7 and R7a, the
substituted Cl ~ alkyl, substituted C3-6 alkenyl, substituted aroyl,
sub.stituted aryl, substituted heteroaroyl, substituted arylsulfonyl,
substituted heteroarylsulfonyl and substituted heterocycle include
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moieties cont~ining from 1 to 3 substitutents in addition to the point of
attachment to the rest of the compound.
As used herein, the term "substituted aryl"is intended to
include the aryl group which is substituted with 1 or 2 substitutents
S selected from the group which includes but is not limited to F, Cl, Br,
CF3, NH2, N(cl-c6 aLkyl)2, NO2, CN, (cl-c6 alkyl)O-, -OH, (Cl-C6
alkyl)s(o)m-~ (cl-c~6 alkyl)C(O)NH-, H2N-c(NHk (Cl-C6
aLkyl)C(O)-, (Cl-C6 alkyl)OC(O)-, N3,(C1-C6 aLkyl)OC(O)NH- and
Cl-C20 aLkyl.
When R2 and R3 are combined to form - (CH2)U -, cyclic
moieties are formed. Examples of such cyclic moieties include, but are
not limited to:
In addition, such cyclic moieties may optionally include a
heteroatom(s). Examples of such heteroatom-cont~ining cyclic moietie~s
include, but are not limited to:
OJ ~S~
~"~ ~ ~ ~ ~10
Lines drawn into the ring systems from substituents (such
as from R2, R3, R4 etc.) indicate that the indicated bond may be
attached to any of the substitutable ring carbon atoms.
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Preferably, R1a and Rlb are independently selected from:
hydrogen, -N(R 1~)2, RlOC(O)NR 10 or unsubstituted or substituted
Cl-C6 alkyl wherein the substituent on the substituted C1-C6 alkyl is
selected from unsubstituted or substituted phenyl, -N(R10)2, R100- and
5 R 1 OC(O)NR 10
Preferably, R2 is selected from: H,
~NR6R7 ~oR6
~ ~ and an unsubstituted or substituted group, the
group selected from Cl ~S aL~yl, C2 ~ aL~enyl and C2 ~s alkynyl;
wherein the substituted group is substituted with one or more of:
1) aryl or heterocycle, unsubstituted or substituted with:
a) C 1-4 alkyl,
b) (CH2)pOR6,
c) (CH2)pNR6R7,
d) halogen,
2) C3-6 cycloalkyl,
3) oR6,
4) SR6a, s(o)R6a~ So2R6a,
5) --N R6R7
R6
I
6) --N~ R7
o
R6
--N~ N R7R7a
-
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8) --O~ NR6R7
O
9) --o~OR6
10) ~NR6R7
11) - S02- NR6R7
R6
12) N SO2 R
o
1 4) oR6
~r
15) N3, or
16) F.
Preferably, R3 is selected from: hydrogen and Cl-C6 aIkyl.
Preferably, R4 and R5 are hydrogen.
Preferably, R6, R7 and R7a is selected from: hydrogen,
- unsubstituted or substituted Cl-C6 alkyl, unsubstituted or substituted
aryl and unsubstituted or substituted cycloaLkyl.
- 10 Preferably, R6a is unsubstituted or substituted Cl-C6 aLkyl,
unsubstituted or substituted aryl and unsubstituted or substituted
cycloalkyl.
Preferably, R9 is hydrogen or methyl. Most preferably,
Ra is hydrogen.
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Preferably, R10 is selected from H, Cl-C6 alkyl and
benzyl.
Preferably, A 1 and A2 are independently selected from: a
bond, -C(O)NR 10, -NR I ~C(O)-, O, -N(R 10), -S(O)2N(R 10)- and-
N(R l ~)S(O)2 .
Preferably, V is selected from hydrogen, heterocycle and '
aryl. More preferably, V is phenyl.
Preferably, Y is selected from unsubstituted or substituted
phenyl, unsubstituted or substituted naphthyl, unsubstituted or
substituted pyridyl, unsubstituted or substituted furanyl and
unsubstituted or substituted thienyl. More preferably, Y is unsubstituted
or substituted phenyl.
Preferably, Z is selected from unsubstituted or substituted
phenyl, unsubstituted or substituted naphthyl, unsubstituted or
substituted pyridyl, unsubstituted or substituted furanyl and
unsubstituted or substituted thienyl. More preferably, Z is unsubstituted
or substituted phenyl.
Preferably, W is selected from imidazolinyl, imidazolyl,
oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl. More
preferably, W is selected from imidazolyl and pyridyl.
Preferably, n and r are independently 0, l, or 2.
Preferably p is 1, 2 or 3.
Preferably s is 0.
Preferably t is 1.
Preferably, in compounds of the formula B, when G is H2
and W is imidazolyl, then the substitutent (R~)r- V -
A l (CR 1 a2)nA2(CR l a2)n - is not H.
Preferably, in compounds of the formula B, when X is
-C(=O)-, or -S(=O)m-, then t is 1 and the substitutent (R~)r- V -
A 1 (CR 1 a2)nA2(CR 1 a2)n - is not H;
It is intended that the definition of any substituent or
variable (e.g., Rla, R9, n, etc.) at a particular location in a molecule be
independent of its definitions elsewhere in that molecule. Thus,
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-N(R10)2 represents -NHH, -NHCH3, -NHC2Hs, etc. It is understood
that substituents and substitution patterns on the compounds of the
instant invention can be selected by one of ordinary skill in the ~rt to
provide compounds that are chemically stable and that can be readily
S .synthesized by techniques known in the art, as well as those methods set
forth below, from readily available starting materials.
The ph~ ceutically acceptable salts of the compounds of
this i~vention include the conventional non-toxic salts of the compounds
of this invention as formed, e.g., from non-toxic inorganic or organic
10 acids. For example, such conventional non-toxic salts include those
derived from inorganic acids such as hydrochloric, hydrobromic,
sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared
from organic acids such as acetic, propionic, succinic, glycolic, stearic,
lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,
15 phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic,
fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic,
isethionic, trifluoroacetic and the like.
The phz~ eeutically acceptable salts of the compounds of
this invention can be synthesized from the compounds of this invention
20 which contain a basic moiety by conventional chemical methods.
Generally, the salts are prepared either by ion exchange
chromatography or by reacting the free base with stoichiometric
amoumts or with an excess of the desired salt-forming inorganic or
organic acid in a suitable solvent or various combinations of solvents.
Reactions used to generate the compounds of this invention
are prepared by employing reactions as shown in the Schemes 1-22, in
addition to other standard manipulations such as ester hydrolysis,
cleavage of protecting groups, etc., a,s may be known in the literature or
exemplified in the experimental procedure~. Substituents R, Ra and Rb,
30 as shown in the Schemes, represent the substituents R2, R3, R4, and R5;
however their point of attachment to the ring is illustrative only and i~s
not meant to be limiting.
These reactions may be employed in a linear sequence to
provide the compounds of the invention or they may be used to
=
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- 4~ -
synthesize fragments which are subsequently joined by the alkylation
reactions described in the Schemes.
Synop.sis of Schemes 1-22:
The requisite interrnediates are in some cases commercially
available, or can be prepared according to literature procedures, for the
most part. In Scheme 1, for example, the synthesis of 2-alkyl
substituted piperazines is outlined, and is essentially that described by J.
S. Kiely and S. R. Priebe in Or~anic Preparation,s and Proceedings Int.,
1990, 22, 761-76~. Boc-protected amino acids I, available
commercially or by procedures known to those skilled in the art, can be
coupled to N-benzyl amino acid esters using a variety of dehydrating
agents such as DCC (dicyclohexycarbodiimide) or EDC-HCl (l-ethyl-3-
(3-dimethylaminopropyl)carbodiimide hydrochloride) in a solvent such
as methylene chloride, chloroform, dichloroethane, or in
dimethylformamide. The product II is then deprotected with acid, for
example hydrogen chloride in chloroform or ethyl acetate, or
trifluoroacetic acid in methylene chloride, and cyclized under weakly
basic conditions to give the diketopiperazine III. Reduction of III with
lithium aluminum hydride in re~uxing ether gives the piperazine IV,
which is protected as the Boc derivative V. The N-benzyl group can be
cleaved under standard conditions of hydrogenation, e.g., 10%
palladium on carbon at 60 psi hydrogen on a Parr apparatus for
24-48 h. The product VI can be treated with an acid chloride, or a
carboxylic acid under standard dehydrating conditions to furnish the
carboxamides VII; a final acid deprotection as previously described
gives the intermediate VIII (Scheme 2). The intermediate VIII can be
reductively alkylated with a variety of aldehydes, such as IX. The
aldehydes can be prepared by standard procedures, such as that
described by O. P. Goel, U. Krolls, M. Stier and S. Kesten in Or~anic
Syntheses. 1988, 67, 69-75, from the appropriate amino acid (Scheme
3). The reductive alkylation can be accomplished at pH 5-7 with a
variety of reducing agents, such as sodium triacetoxyborohydride or
sodium cyanoborohydride in a solvent such as dichloroethane, methanol
-
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-~ or dimethylformamide. The product X can be deprotected to give the
final compounds XI with trifluoroacetic acid in methylene chloride.
The final product XI is isolated in the salt form, for example, as a
trifluoroacetate, hydrochloride or acetate salt, among others. The
S product ~ mine XI can further be selectively protected to obtain XII,
which can ~ubsequently be reductively alkylated with a second aldehyde
to obtain XIII. Removal of the protecting group, and conversion to
cyclized products such as the dihydroimidazole XV can be accomplished
by literature procedures.
Alternatively, the protected piperazine intermediate VII can
be reductively alkylated with other aldehydes such as l-trityl-4-
imidazolyl-carboxaldehyde or l-trityl-4-imidazolylacetaldehyde, to give
products such as XVI (Scheme 4). The trityl protecting group can be
removed from XVI to give XVII, or alternatively, XVI can first be
15 treated with an alkyl halide then subsequently deprotected to give the
alkylated imidazole XVIII. Alternatively, the intermediate VIII can be
acylated or sulfonylated by standard techniques. The imidazole acetic
acid XIX can be converted to the acetate XXI by standard procedures,
and XXI can be first reacted with an alkyl halide, then treated with
20 refluxing methanol to provide the regiospecifically alkylated imidazole
acetic acid ester XXII. Hydrolysis and reaction with piperazine VIII in
the presence of condensing reagents such as 1-(3-dimethylaminopropyl)-
3-ethylcarbodiimide (EDC) leads to acylated products such as XXIV.
If the piperazine VIII is reductively alkylated with an
25 aldehyde which also has a protected hydroxyl group, such as XXV in
Scheme 6, the protecting groups can be subsequently removed to
unmaslc the hydroxyl group (Schemes 6, 7). The alcohol can be
oxidized under standard conditions to e.~. an aldehyde, which can then
be reacted with a variety of organometallic reagents such as Grignard
30 reagents, to obtain secondary alcohols such as XXIX. In addition, the
fully deprotected amino alcohol XXX can be reductively alkylated
(under conditions described previously) with a variety of aldehydes to
obtain secondary amines, such as XXXI (Scheme 7), or tertiary amines.
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The Boc protected amino alcohol XXVII can also be
utilized to synthesize 2-aziridinylmethylpiperazines such as XXXII
(Scheme 8). Treating XXVII with l,l'-sulfonyl~liimi(1~7ole and sodium
hydride in a solvent such as dimethyl~ormamide led to the formation of
S ~iricline XXXII. The aziridine reacted in the presence of a
nucleophile, such as a thiol, in the presence of base to yield the ring-
opened product XXXIII.
In addition, the piperazine VIII can be reacted with
aldehydes derived from amino acids such as O-alkylated tyrosines,
10 according to standard procedures, to obtain compounds such as XXXIX.
When R' is an aryl group, XXXIX can first be hydrogenated to llnm~k
the phenol, and the amine group deprotected with acid to produce XL.
~ltern~tively, the amine protectin~ group in XXXIX can be removed,
and O-alkylated phenolic amines such as XLI produced.
Depending on the identity of the amino acid I, various side
chains can be incorporated into the piperazine. For example when I is
the Boc-protected ~-benzyl ester of aspartic acid, the intermediate
diketopiperazine XLII where n=l and R=benzyl is obtained, as shown in
Scheme 10. Subsequent lithium aluminum hydride reduction reduces
the ester to the alcohol XLIII, which can then be reacted with a variety
of alkylating agents such as an aLkyl iodide, under basic conditions, for
example, sodium hydride in dimethylformamide or tetrahydrofuran.
The resulting ether XLIV can then be carried on to final products as
described in Schemes 3-9.
N-Aryl piperazines can be prepared as described in Scheme
11. An aryl amine XLV is reacted with bis -chloroethyl amine
hydrochloride (XLVI) in refluxing n -butanol to furnish compounds
XLVII. The resulting piperazines XLVII can then be carried on to ~mal
products as described in Schemes 3-9.
Piperazin-5-ones can be prepared as shown in Scheme 12.
Reductive ~min~tion of Boc-protected amino aldehydes XLIX
(prepared from I as described previously) gives rise to compound L.
This is then reacted with bromoacetyl bromide under Schotten-B~llm~nn
conditions; ring closure is effected with a base such as sodium hydride
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..
in a polar aprotic solvent such as dimethylform~micle to give LI. The
carbamate protecting group is removed under acidic conditions such as
trifluoroacetic acid in methylene chloride, or hydrogen chloride gas in
methanol or ethyl acetate, and the resulting piperazine can then be
5 carried on to final products as described in Schemes 3-9.
The isomeric piperazin-3-ones can be prepared as described
in Schelme 13. The imine formed from arylcarboxamides LII and 2-
aminoglycinal diethyl acetal (LIII) can be reduced under a variety of
conditions, including sodium triacetoxyborohydride in dichloroethane,
10 to give the amine LIV. Amino acids I can be coupled to amines LIV
under st~nc~rd conditions, and the resulting amide LV when treated
with aqueous acid in tetrahydrofuran can cyclize to the lln~ lrated
LVI. Catalytic hydrogenation under standard conditions gives the
requisite intçrmediate LVII, which is elaborated to final products as
15 described in Schemes 3-9.
Access to alternatively substituted piperazines is described
in Scheme 14. Following deprotection with trifluoroacetic acid, the N-
benzyl piperazine V can be acylated with an aryl carboxylic acid. The
resulting N-benzyl aryl carbox~ e LIX can be hydrogenated in the
20 presence of a catalyst to give the piperazine carbox~mi~le LX which can
then be carried on to final products as described in Schemes 3-9.
Reaction Scheme 15 provides an illustrative example the
synthesis of compounds of the instant invention wherein the substituents
R2 and R3 are combined to form - (CH2)U -. For example, 1-
25 aminocyclohexane-l-carboxylic acid LXI can be converted to the
spiropiperazine LXVI essentially according to the procedures outlined
in Schemes 1 and 2. The piperazine interrnediate LXIX can be
deprotected as before, and carried on to final products as described in
Schemes 3-9. It is understood that reagents utilized to provide the
30 substituent Y which is 2-(naphthyl) and the imidazolylalkyl substituent
may be readily replaced by other reagents well known in the art and
readily available to provide other N-substituents on the piperazine.
The aldehyde XLIX from Scheme 12 can also be
reductively aLkylated with an aniline as shown in Scheme 16. The
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product LXXI can be converted to a piperazinone by acylation with '~
chloroacetyl chloride to give LXXII, followed by base-intlllce-1
cyclization to LXXIII. Deprotection, followed by reductive alkylation
with a protected imicl~7.ole carboxaldehyde leads to LXXV, which can
S be aLkylation with an arylmethylh~licle to give the imit1~7olium salt
LXXVI. Final removal of protecting groups by either solvolysis with a
lower alkyl alcohol, such as methanol, or treatment with triethylsilane
in methylene chloride in the presence of trifluoroacetic acid gives the
final product LXXVII.
Scheme 17 illustrates the use of an optionally substituted
homoserine lactone LXXIX to prepare a Boc-protected piperazinone
LXXXII. Intermediate LXXXII may be deprotected and reductively
aLkylated or acylated as illustrated in the previous Schemes.
~ltern~tively, the hydroxyl moiety of intermediate LXXXII may be
15 mesylated and displaced by a suitable nucleophile, such as the sodium
salt of ethane thiol, to provide an intermediate LXXXIII. Tntermediate
LXXXII may also be oxidized to provide the carboxylic acid on
intermediate LXXXIV, which can be lltili7e-1 form an ester or amide
moiety.
Amino acids of the general formula LXXXVI which have a
sidechain not found in natural amino acids may be prepared by the
reactions illustrated in Scheme 18 starting with the readily prepared
imine LXXXV.
Schemes 19-22 illustrate syntheses of suitably substituted
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 moieties for
variable W are also well known in the art.
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SCHEME 1
O Ra Rb
~OJ~ N ~OH PhCH2NHCHCO2C2H5
H o DCC, CH2CI2
O N ~N~,COzc2Hs
H I ~ Rb
Ra
1) HCI, CH2CI2 R)~o LAH ~ HN N
~ HN N \ THF, reflux \
2) N~HCO3 O Rb ~ \Rb
111 lV
Ra Ra
Boc20 0,\ ) \ 10% Pd/C ~~ )
V ;~ Vl
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SCHEME 2
a EDC-HCI, HOBT
R ~ ~ DMF
BocN NH
Y, Rb ~CO2H
) ~ ~ HCI, EtOAc
Y O
Rb
Vll
Vlll
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SCHEME 3
Boc NHl IX
Ra~ ~ ~ Boc NH CHO
HCI N N NaBH(OAc)3
YRb Et3N, CICH2CH2CI
Vlll
Ra Q; 3 CF3CO2H
Boc NH~NyN~ CH2C12
NHBoc R
Ra ~ Boc20
NH~ N N CH2C12
NH2 Rb
Xl
Ra ~Q3 ~CHO
BocNH~ NyN NaBH(OAc)3
NH2 Rb Et3N, CICH2CH2CI
Xll
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SCHEME 3 (continllerl)
BocNH~N N~ CF3CO2H, CH2CI2;
y O NaHCO3
=\ NH R~b
~/ Xlll
NH~ N N $~ ~ NC
NH R AgCN
~ XIV
Ra
~ NyN
N~N~ R
~3 XV
-
-
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SCHEME 4
a /= NaBH(OAc)3
R~ Et3N, CICH2CH2CI
HCI ~ NyN~ (CH2)nCHO
Rb o N
Vlll Tr
RN~ ~3
(CH2)n+1 y o
N~ Rb
N, XVI
Tr 1 ) Ar CH2X, CH3CN
CF3CO2H, CH2CI2 (c2H5)3siH
Ra N~3
9~H2)n+1 y O
N;~ Rb
H XVII
.
Ra~
Ar (/CH2)n+1 y o
N~ Rb
N~ XVIII
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SCHEME S
N~ 2 02H CH N~CH2C02CH3
H HCI H HCI
XIX XX
CH2C02CH3 1) ArCH2X CH3CN
(C6H5)3CBr ~ reflux
(C2H )3FN Tr 2) CH30H, reflux
XXI
Ar~\N~cH2c02cH3 2.5N HCla
N 55~C
XXII
Ar~\ CH2C02H
N~
N
XXIII
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SCHEME 5 (continued)
.
Ar~\N~cH2c02H Ra ~3
+ HCI H N~N
XXIII ~ . Rb
Vlll
EDC- HCI
HOBt
DMF
~ HCI
~3~N ~N~3
N Rb
XXIV
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SCHEME 6
NaBH(OAc)3
~ Et3N, CICH2CH2CI
HCI N N~ ~=' BnO~
y 0
Rb BocNH CHO
Vlll
XXV
BnO ~NyN~ zoyO Pd(OH)2 H2
NHBoc Rb CH3CO2H
XXVI
HO N N ~ CICOCOCI
DMSO CH2CI2
NHBoc Rb (C2Hs)3N
XXVII
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SCHEME 6 (CONTINUED)
- R~ ~ ~ 1. R'MgX
0~N N ~ \=/ (C2H5)20
HNHBoc Rb CH2Cl2
XXVIII
Ra ~ 9
HO~N N
R' NH2 Rb
XXIX
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SCHEME 7
Ra /~
~ ~ CF3CO2H
HO~ NyN~ CH2CI2
NHBoc Rb
XXVII
Ra /~
~ ~ R'CHO
HO,~N N~ NaBH(OAc)3
~ b ~ CIC H2C H2CI
XXX
HO ~N <N~
NH Rb
R'CH2
XXXI
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F~a ~3 8
HO ~N N
y o NaH, DMF0~C
NHBoc Rb
XXVII
R~ ~ ~ R'SH
<~ y o CH30H
H Rb
XXXII
Ra
R'S ~N N~
NH2 Rb
XXXIII
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SCHEME 9
HO~ 1) Boc20, K2C~3 HO~
~ THF-H20
J~ 2) CH2N2, EtOAc J~
H2NCO2H BocNH CO2CH3
XXXIV
XXXV
HO,~
LiAlH4 ~ R'CH2X
THF l Cs2CO3
0-20~C BocNH CH2OH DMF
XXXVI
R'CH ~ pyrldine SO
BocNH CH2OH 20~C BocNH CHO
XXXVII XXXVIII
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SCHEME 9 (continued)
R'CH~3 + HCI N N~
BocNH CHO \~Rb
XXXVIII Vlll
NaBH(OAc)3
CICH2CH2CI
R'CH20~ ~\
NHBoc Rb
XXXIX \ HCI
ETOAc
1 ) 20% Pd(OH)2
CH30H, CH3CO2H /
2) HCI, EtOAc / R'CH20~ y
HO~; \N~ XLI
NH2 Rb
XL
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SCHEME 10
CO2R '
n( S~~ 1) LAH, Et20
HN N 2) Boc20
0~ ~
XLII
R60
HO
R61 . n ( < )
O n( S~, aH, DMF ~N~\N~
XLIV
XLIII
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SCHEME 1 1
Rb
ArNH2 + Cl~)2NH ~ HCI
XLV Ra
XLVI
Ra Rb
n- butanol ArN NH ~ HCI
reflux >~
Ra Rb
XLVII
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SCHEME 12
OH CH3NHOCH3 ~ HCI
O N ~EDC . HCI, HOBT
~ DMF, Et3N, pH 7
O Ra
>~oJ~ N ~ N(CH3)0CH3 LAH, Et20
XLVIII
>~ ~ Ra ArCH2NH2
O N ~ NaBH(OAc)3
H O CICH2CH2CI
XLIX pH 6
1, C 1) BrCH2COBr
H 2) NaH, THF, DMF
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SCHEME 12 (CONT'D)
r
Ra
o ) ~ 1) TFA, CH2CI2
_~ ~N N~
o
Ra~
HN N~
~~ Ar
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S C ~I]~n~ 13
NaBH(OAc)3
ArCHO +NH2CH2CH(Oc2H5)2
Lll Llll ~ Ra
Ar CH2NHCH2CH(Oc2Hs)2 >~o~ N~
LIV EDC. HCI, HOBT
DMF, Et3N, pH 7
O Ra ~Ar 6N HCI
>~0~ N~N~CH(OC2Hs)2 THF
LV
Ra O H2 1 0%Pd/C
N~=~N~ CH30H
t~ Ar
LVI
Ra o
~N N~
~0 \~ Ar
LVII
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.
Rb
GF CO H CH Cl /~ ArCO2H
V 1) 3 2 ~ 2 2 rN NH
2) NaHCO3 /=~, RaY EDC HCI
HOBT, DMF
LVIII
Rb Rb
N N~ 10% Pd/C , HN N~
LIX Ar H2 CH3oH RLY Ar
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SCHEME 15
r ul PhcH2NHcH2co2c2H5
~ DCC, CH2C12
BocNH C02H
LXI
a) TFA, CH2CI2
BocNH~N~C02c2Hs b) NaHCO3
~ LXII
H N~N~,C02C2H5 CHC13
LXIII
~/<~
0~ ~ LiAlH4 HN \N~
LXIV ~ LXV
Boc20 BocN~ N H2 Pd/C
CH2CI2 ~ CH30H "
LXVI
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SCHEME 15 (continued)
COCI
~\ ~
BocN~NH NaHCO3 EtOAc
LXVII
~u~
~ \ a) TFA, CH2C12
BocN N~ /=\ b) NaBH(OAc)3
~ ~C(=O)H
LXVIII ,N
~ CPh3
~ u~ .
~N ~~ TFA CH2CI2
(c2H5)3siH
N,
CPh3LXIX
/ \ ,0
~ H 2 TFA
~- LXX
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SC~nE~nE 16
R R H
BocNH CHO ArNH2 ~ BocNH~N~Ar
NaBH(OAc)3
CICH2CH2CI
XLIX LXXI
CI~J~cI BocNH N-Ar
EtOAc / H20 ~
NaHCO3 Cl O
LXXII
BocN N-Ar HCI
DMF ~ EtOAc
o
LXXIII
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SCHEME 16 (continued)
CHO
~\ ' N N--Ar
HCIHN~N--Ar C(Ph)3 N~ \~O
O NaBH(OAc)3 N
LXXIV pH 5-6 (Ph)3C
LXXV
R
~,
Ar~ ~N N--Ar
ArCH2X N~ ~~
CH3CN ~ O
,N ~
(Ph)3c X
LXXVI
MeOH ~
or Ar~ ~ N N--Ar
TFA, CH2CI ~N~ ~
(C2H5)3SiH
LXXVII
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SCHEME 17
~o 1. Boc20, i-Pr2EtN \-\0
H2N O 2. DIBAL BocHN~
LXXIX OH
~ sub
ArNH2 /~ N
NaBH(OAc)3 ' BocNH Ar
CICH2CH2CI LXXX
HO sub
O ~/
C~ CI BocNH N-Ar
EtOAc / H20 /~
NaHCO3 Cl O
LXXXI
HO sub
,'1~
Cs2CO3 ~
BocN N-Ar
DMF \~
o
LXXXII
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SCHEME 17 (continued)
HO /sub
BocN~N--Ar
~ 1. (COCI)2, Et3N
1. MsCI, iPr2NEt LXXXII \
2. NaCI02, t-BuOH
2. NaSEt,~ ~ 2-Me-2-butene
EtS /sub HO /sub
BocN N--Ar BocN~N--Ar
O O
LXXXIII LXXXIV
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SCHEME 18
1. KOtBu, THF R2
r CO2Et R2x ~ CO2Et
r N ~ H2N
Ph 2. 5% aqueous HCI HCI
LXXXV
1. Boc20, NaHCO3 R2
~ CO2H
BocHN
2. LiAlH4, Et20 LXXXVI
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- REACIION SCHEME 19
CH3 1 ) HNo2~Br2 CO2CH3
~ 2) KMnO
H2N N 3) MeOH,H+ Br~N~
~\~\ MgCI R6
~ ~,CO2CH3
zncl2~Nicl2(ph3p)2 N
NaBH4 (excess) ~CH20H
DMSO ~CHO
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REACTION SCHEME 20
1. EtO(CO)CI R6
Br 2 R6 MgCI [~
~C02CH3 Zn CuCN ~CO2CH3
N 3. S, xylene, heat N
R6 R6
NaBH4 ~j~ SO3Py, Et3N ~
(excess) ~CH20H DMSO ,~CHO
Br~ ~c02cH3 [~\MgCI ¢~j
~,CO2CH3
N ZnC12, NiC12(Ph3P)2 N~
R6 R6
NaBH4 ¦ S03Py, Et3N
~CH20H ~ ~CHO
(excess) N DMSO N ~I
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- REACTION SCHEME 21
co2CH3
Br~ 1. LDA, C02 Br~
N 2. MeOH, H~ N
ZnCI2, Nici2(ph3p)2 N
R6
NaE3H4(excess) ¢~OH SO3Py, Et3N
DMSO
R6
CHO
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REACTION SCHEME 22
cO2CH3 r
1. LDA, CO2 [~Br
2. (CH3)3SiCHN2
R6 ~\ Br R6 ~q
Zn, NiCI2(Ph3P)2 N~CO2CH3
R6 1~
excess NaBH4 1~ SO3 Py, Et3N
~CH20H DMSO
R6 ~
N ~,CHO
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J'
The instant compounds are useful as ph~rm~reutical agents
for m~mm~l.c, especially for hllm~n~. These compounds may be
~t1mini~tered to patients for use in the treatment of cancer. Examples of
the type of cancer which may be treated with the compounds of this
5 invention include, but are not limited to, colorectal carcinoma, exocrine
pancreatic carcinoma, myeloid leukemias and neurological tumors.
Such tumors may arise by mutations in the ras genes themselves,
mutations in the proteins that can regulate Ras activity (i.e.,
neurofibromin (NF-l), neu, scr, abl, lck, fyn) or by other mech~ni.~m~.
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 growth of tumors (J. Rak et al. Cancer Research, 55:4575-
4580 (1995)). Such anti-angiogenesis properties of the instant
compounds may also be useful in the treatment of certain forms of
blindness related to retinal vasc~ ri7~tion.
The compounds of this invention are also useful for
inhibiting other proliferative diseases, both benign and m~li n~nt
wherein Ras ~loteills 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 ~1mini~tration of an effective amount of the compounds of the
inventiom to a m~mm~l in need of such tre~tment 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 translumin~l coronary
angioplasty by inhibiting neointim~l 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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American Journal of Pathology, 142:1051-1060 (1993) and B. Cowley,
Jr. et al.FASEB Journal, 2:A3160 (1988)). ..
The instant compounds may also be useful for the treatment
of fungal infections.
The compounds of this invention may be ~lmini.~tered to
m~mm~l~, preferably hnm~n~, either alone or, preferably, in
combination with ph~nn~ceutically acceptable ca~Tiers or diluents,
optionally with known adjuvants, such as alum, in a ph~rTn~ceutical
composition, according to standard ph~rtn~eutical practice. The
compounds can be ~-lmini~tered orally or parenterally, including the
intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and
topical routes of ~-lmini~tration.
For oral use of a chemotherapeutic compound according to
this invention, the selected compound may be ~tlmini~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
commonly used include lactose and corn starch, and lubricating agents,
such as magnesium stearate, are commonly ~-lclecl For oral
~1mini.stration in capsule form, useful diluents include lactose and dried
corn starch. When aqueous suspensions are required for oral use, the
active ingredient is combined with emulsifying and suspending agents.
If desired, certain sweetening and/or flavoring agents may be ~ e~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 the preparation isotonic.
The compounds of the instant invention may also be co-
~tlmini.~tered with other well known therapeutic agents that are selected
for their particular usefulness against the condition that is being treated.
For example, the instant compounds may be useful in combination with
known anti-cancer and cytotoxic agents. Simil~rly, the instant
compounds may be useful in combination with agents that are effective
in the treatment and prevention of NF-l, restinosis, polycystic kidney
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o
~ disease, infections of hepatitis delta and related viruses and fungal infections.
If formulated as a fixed dose, such combination products
employ the compounds of this invention within the dosage range
5 described below and the other ph~rm~ceutically active agent(s) within
its approved dosage range. Compounds of the instant invention may
alternatively be used sequentially with known ph~rm~ceutically
acceptable agent(s) when a combination formulation is inaL~ iate.
The present invention also encompasses a ph~ reutical
10 composition useful in the treatment of cancer, comprising the
~lmini.~tration of a therapeutically effective amount of the compounds
of this invention, with or without pharmaceutically acceptable carriers
or diluents. Suitable compositions of this invention include aqueous
solutions comprising compounds of this invention and ph~rm~colo-
15 gically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4. Thesolutions may be introduced into a patient's blood-stream by local bolus
injection.
When a compound according to this invention is
~tlmini.~tered into a human subject, the daily dosage will normally be
20 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 sy~ toms.
In one exemplary application, a suitable amount of
compound is ~1mini~tered to a m~mm~l undergoing treatment for
25 cancer. ~Clmini.~tration occurs in an amount between about 0.1 mg/kg
of body weight to about 60 mg/kg of body weight per day, preferably
of between 0.5 mg/kg of body weight to about 40 mg/kg of body weight
per day.
The compounds of the instant invention are also useful
30 as a component in an assay to rapidly determine the presence and
quantity of farnesyl-protein transferase (FPTase) in a composition.
Thus the composition to be tested may be divided and the two
portions contacted with mixtures which comprise a known substrate
of FPTase (for example a tetrapeptide having a cysteine at the amine
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terminus) and farnesyl pyrophosphate and, in one of the mixtures, a
compound of the instant invention. After the assay mi~ules 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
5 the assay mixhlres may be deter~nined by well known
immunological, radiochemical or chromatographic techniques.
Because the compounds of the instant invention are selective
inhibitors of FPTase, absence or qll~ntit~tive reduction of the amount
of substrate in the assay mixture without the compound of the instant
10 invention relative to the presence of the unchanged substrate in the
assay cont~inin~ the instant compound is indicative of the presence of
FPTase in the composition to be tested.
It would be readily apparent to one of ordinary skill in the
art that such an assay as described above would be useful in identifying
15 tissue s~mples which contain farnesyl-protein transferase and
q~l~ntit~tin~ the enzyme. Thus, potent inhibitor compounds of the
instant invention may be used in an active site titration assay to
determine the quantity of enzyme in the sample. A series of samples
composed of aliquots of a tissue extract cont~inin~ an unknown amount
20 of farnesyl-protein transferase, an excess amount of a known substrate
of FPTase (for example a tetrapeptide having a cysteine at the amine
terminus) and farnesyl pyrophosphate are incubated for an a~pro~liate
period of time in the presence of varying concentrations of a compound
of the instant invention. The concentration of a sufficiently potent
25 inhibitor (i.e., one that has a~Ki subst~nti~lly smaller than the
concentration of enzyme in the assay vessel) required to inhibit the
enzymatic activity of the sample by 50% is approximately equal to half
of the concentration of the enzyme in that particular sample.
EXAMPLES
Examples provided are intended to assist in a further
understanding of the invention. Particular materials employed, species
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and conditions are intended to be further illustrative of the invention
and not limit~tive of the reasonable scope thereof. Purification by
HPLC was accomplished with a 40 X 100 mm Waters PrepPak(~
reverse phase HPLC column (Delta-PakTM C18 15 ,um, 100 A).
5 Gradient elution employed 0.1% trifluoroacetic acid in water (Solvent
A) and 0.1~o trifluoroacetic acid in acetonitrile (Solvent B). Chloride
salts were obtained by passing an aqueous solution of the trifluoroacetic
acid salt through a Biorad A~@~ 3X4 ion exchange resin column (100-
200 mesh, C1-form). Purification by HPLC was utilized for each of the
10 Fx~mples 1-23, 27, 48 and 49 as set forth below.
EXAMPLE 1
2(S)-Butyl- 1 -(2,3-~ minoprop- 1 -yl)-4-(1 -naphthoyl)piperazine
15 trihydrochloride
Step A: 1-Benzyl-3(S)-n -butylpiperazine-2~5-dione
The title compound was prepared according to the
procedure described by John S. Kiely and Stephen R. Priebe in Organic
20 Preparations and Procedures Int.. 22 (6), 761-768 (1990). Thus
dicyclohexylcarbodiimide (9.33 g, 45.2 mmol) in methylene chloride
(0.5 M) was added to methylene chloride (250 mL). This solution was
cooled to 0 C under nitrogen and Boc-L-norleucine (10.5 g, 45.2
mmol) was added. The resulting slurry was stirred for 5 min, and then
ethyl N-benzylglycinate (8.72 g, 45.2 mmol) was added. The reaction
was stirred for 2 h at 0 C, then at 20 C overnight. The precipitate was
removed by filtration, and hydrogen chloride gas bubbled through the
methylene chloride solution for 2-4 h, until the reaction was shown to
be complete by tlc. The solvent was removed in vacuo~ and the residue
partitioned between ethyl acetate (150 mL) and saturated sodium
" bicarbonate solution (42 mL). The organic phase was washed with
saturated sodium chloride, dryed over magnesium sulfate, filtered and
evaporated. The crude diketopiperazine was triturated with hexane to
give the title compound as a white powder. 1HNMR (300 MHz, CDCl3)
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o 7.24-7.40 (5H, m), 6.22 (lH, br s), 4.07 (lH, dt, J=3, 6 Hz), 3.87 (lH,
d, J=17 Hz), 3.80 (lH, d, J=17 Hz), 1.88 (2H, m), 1.35 (4H, m), 0.91
(3H, t, J=7 Hz).
Step B: 4-Benzyl-l-tert -butoxycarbonyl-2(S)-n -buLyl~i~elazine
The product from Step A (4.95 g, 0.019 mol) was dissolved
in THF (200 mL) and cooled under nitrogen to 0 C with mechanical
stirrin3~. Lithium al~lminllm hydride (2.60 g, 0.0685 mol) was added
slowly. The reaction was refluxed for 18 h, cooled to 0 C, and
quenched by the sequential slow addition o~ 5 rnL H20, 5 mL 10%
sodium hydroxide solution and 5 mL H2O. The reaction was stirred for
30 min and filtered. The solvent was removed in vacuo, the crude
product taken up in methylene chloride and dried over magnesium
sulfate. The drying agent was removed by filtration, and the filtrate
treated with di-tert -butyl dicarbonate ((4.35 g, 0.020 mol). After 2 h
at 20 C, saturated sodium bicarbonate was added. The layers were
separated, and the organic phase washed with saturated sodium chloride
solution, then dried over magnesium sulfate. Filtration and evaporation
gave the crude product which was purified by column chromatography
on silica gel, eluting with 5% ethyl acetate in hexane. The title
compound was obtained as a foam. lHNMR (300 MHz, DMSO-d6)
7.25 (5H, m), 3.90 (lH, br s,), 3.73 (lH, d, J=13 Hz), 3.51 (lH, d, J=13
Hz), 3.34 (lH, d, J=13 Hz), 2.93 (lH, m), 2.75 (lH, d, J=ll Hz), 2.62
(lH, d, J=ll Hz), 1.90 (2H, m), 1.60 (2H, m), 1.38 (9H,s), 1.26 (2H,
m), 1.04 (2H, m), 0.84 (3H, t, J=7 Hz).
Step C: l-tert -Butoxycarbonyl-2(S)-n -butylpiperazine
The product from Step B (3.75 g, 11.3 mmol) was
dissolved in methanol (75 mL) in a Parr bottle, and the vessel purged
with argon. To this was added 10 % palladium on carbon (0.80 g) and
the reaction hydrogenated under 60 psi hydrogen for 24 h. The catalyst O
was removed by filtration through Celite, and the filtrate evaporated in
vacuo to give the title compound as an oil. lHNMR (300 MHz, CDC13)
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t!
o 4.08 (lH, br s), 3.90 (lH, d, J=12 Hz), 2.5-3.8 (6H, m), 1.80 (lH,
m), 1.60 (lH, m), 1.46 (9H, s), 1.30 (4H, m),0.90 (3H, t, J=7 Hz).
Step D: l-tert -Butoxycarbonyl-2(S)-n -butyl-4-(1-naphthoyl)-
piperazine
l-tert-butoxycarbonyl-2(S)-n -butylpiperazine (0.325 g,
1.34 mrnol), l-hydroxybenzotriazole (HOBT) (0.203 g, 1.34 mmol) and
l-ethyl-3-(3-dimethylamino-propyl)carbodiimide hydrochloride (EDC -
HCl) (0.254 g, 1.34 mmol) were added to dry, degassed
10 dimethylform~mi~le (7 mL). The pH of the reaction was adjusted to 7
with triethyl~mine, and the reaction stirred for 2 h. The
dimethylform~mide (DMF) was distilled in vacuo. and the residue
partitioned between ethyl ~cet~te and water. The organic phase was
washed with 2% aqueous potassium hydrogen sulfate, saturated sodium
15 bicarbonate solution, saturated sodium chloride solution, and dried over
m~gnesium sulfate. The title compound was obtained as a thick oil.
Step E: 2(S)-n -Butyl-4-(1-naphthoyl)piperazine hydrochloride
The product from Step D was dissolved in ethyl ~cet~te,
20 cooled to -40~C under nitrogen, and the solution saturated with HCl(g).
The solution was warmed to 0~C for 30 min, and then purged with
nitrogen. The solvent was removed in vacuo. The product was
evaporated from ethyl acetate three times. The title compound was
obtained as a white solid.
Step F: 2.3-(bis-tert-Butoxycarbonylamino)propanoic acid
Diaminopropanoic acid monohydrochloride (2.86 g, 0.0204
mol) was suspended in 1:1 water-dioxane (100 mL) cont~ining
triethyl~mine (5.97 mL, 0.0204 mol). BOC-ON (11.0 g, 0.0448 mol)
30 was added along with additional triethylamine to adjust the pH to 9.5.
The reaction was stirred under nitrogen overnight at 20~C. The clear
solution was diluted with water and extracted with diethyl ether (5 x
100 mL). The aqueous solution was adjusted to pH 1 with cold 5%
aqueous hydrochloric acid and extracted with ethyl acetate. The organic
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layer was washed with water, saturated sodium chloride, then dried
over m~gnesium sulfate. The resulting foam (5.46 g) was cryst~lli7e-1
from ethyl acetate to give the title compound as a white solid.
5 Step G: N-Methoxy-N-methyl-2,3-(bis-tert-butoxycarbonyl-
amino)proprionamide
2,3-bis-(ter~-Butoxycarbonylamino)propanoic acid (1.80 g,
5.92 mmol) in dimethylfonn~mide (25 mL) was stirred overnight with
N-methoxy-N-methylamine hydrochloride (0.635 g, 6.51 mmol), EDC
10 HCl (1.24 g, 6.51 mmol), N-hydroxybenzotriazole (0.80 g, 5.92 mmol)
and triethylamine (0.825 mL, 5.92 mmol). The dimethylform~mi~1e
was removed in vacuo and the residue partitioned between ethyl acetate
and water. The organic phase was washed with 10% hydrochloric acid,
s~Lulat~;d sodium bicarbonate solution, saturated brine, and dried over
15 m~gnesium sulfate. The crude product was chromatographed on silica
gel with 30% ethyl acetate in hexane. The title compound was obtained
as a foam. lHNMR (CDC13, 300 MHz) d 5.51 (lH, br d), 4.87 (lH, br
s), 4.72 (lH, br s), 3.77 (3H, s), 3.50 (lH, m), 3.40 (lH, dt, J=12, 6
Hz), 3.20 (3H, s), 1.44 (9H, s), 1.42 (9H, s).
Step H: 2~3-(bis-tert-Butoxycarbonylamino)propanal
Lithium aluminum hydride (0.384 g, 10.14 mmol) was
suspended in diethyl ether (20 mL) and cooled to -45~C under nitrogen.
N-Methoxy-N-methyl-2,3-(bis-tert-butoxycarbonylamino)-
25 proprionamide (2.07 g, 5.96 mrnol) in 1:1 ether-tetrahydrofuran (60
mL) was added at such a rate so as to keep the reaction temperature less
than -35~C. The reaction was allowed to warm to 5~C, then cooled to
-45~C and quenched with a solution of potassium hydrogen sulfate (3.08
g, 22.6 mmol) in water (20 mL). The reaction was stirred at 20~C lh,
30 then ~lltered through celite. The organic phase was washed with 10%
citric acid and saturated brine, then dried over magnesium sulfate. The
title compound was obtained as a foam. NMR (CDC13, 300 MHz) d
9.61 (lH,s),5.60(1H,brs),4.88(1H,brs),4.24(1H,brd,J=6Hz),
3.68 (lH, m), 3.50 (lH, m), 1.40 (9H, s), 1.39 (9H, s).
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Step I: 1-[(2,3-bis-tert-Butoxycarbonylamino)prop-l-yl]-2(S)-
butvl-4-(1-naphthoyl)piperazine
A solution of 3(S)-butyl-l-(l-naphthoyl)piperazine (1.83 g,
6.20 mmol) (free base of the product from Step E), acetic acid (0.17
mL, 2.9 mmol) in dichloroethane (20 mL) was adjusted to pH 5.5 with
triethyl~nnine. Sodium triacetoxyborohydride (1.87 g, 8.79 mmol) and
crushed molecular sieves (1 g) were added, and the reaction cooled to
0~C under nitrogen. A solution of 2,3-(bis-tert-butoxycarbonyl-
amino)propanal (1.69 g, 5.86 mmol) in dichloroethane (10 mL) was
added dropwise, and the reaction stirred at 20~C overnight The
reaction was cooled to 0~C, quenched with saturated sodium bicarbonate
and stirred for lh. The layers were separated and the organic layer
washed with saturated brine, then dried over m~n~sium sulfate. The
crude product was chromatographed on silica gel with 40-50% ethyl
acetate in hexane, and the title compound was isolated as a foam (Rf
0.30, 50% ethyl acetate in hexane).
Step J: 1 (2,3 Diaminoprop-l-yl)-2(S)-butyl-4-(1-
naphthoyl)piperazine trihydrochloride
Trifluoroacetic acid (25 mL) was added to a solution of 1-
[(2,3-bis-tert-butoxycarbonylamino)prop- 1 -yl]-2(S)-butyl-4-( 1-
naphthoyl)piperazine (2.13 g, 3.75 mmol) in dichloromethane (75 mL).
After 25 min at 20~C, the solvent was evaporated and the residue
partitioned between chlorform and 20% aqueous sodium hydroxide.
The organic layer was washed with saturated brine and dried over
m~gnesium sulfate. The free base of the title compound was obtained as
a yellow gum (1.72 g). A portion of this material (52 mg) was pllrifie-1
by preparative HPLC using a 100 mm Waters PrepPak@~ reverse phase
column (DeltaPakTM Clg, 50 mM, 100 A), and pure product isolated by
gradient elution using 100% 0.1% trifluoroacetic acid in water (Solvent
A) to 50% Solvent A and 50% 0.1% trifluoracetic acid in acetonitrile
(Solvent B). The pure fractions were combined and the solvent
evaporated, and the crude product dissolved in water and passed
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through an ion exchange column (Biorad AG(~) 3X4 ion exchange resin,
chloride form). A~ter lyophili7~tion, the title compound was obtained
as a white solid. Anal. Calc. for C22H32N4O ~ 4.45 HCl C, 49.78; H,
6.92; N, 10.56. Found: C, 49.75, H, 6.72; N, 10.36.
EXAMPLE 2
1-(3-Amino-2-(2-naphthylmethylamino)prop-1-yl)-2(S)-butyl-4-(1-
naphthoyl)piperazine trihydrochloride
Step A: 1-[(2-Amino-3-tert-butoxycarbonylaminoprop-1-yl]-2(S)-
butyl-4-(1 -naphthoyl)piperazine
Di-tert-butyl dicarbonate (0.282 g, 1.29 mmol) was added
to a solution of 4-(2,3-~ minoprop-1-yl)-2(S)-butyl-1-(1-
naphthoyl)piperazine (0.476 g, 1.29 mmol) in dichloromethane (10
mL). The reaction was stirred a total of 2h at 20~C, then quenched and
extracted with water. The organic phase was washed with sa~ul~ted
brine and dried over magnesium sulfate. The crude product was
chromatographed on silica gel with 5% methanol in chloroform
followed by 5% (9:1 methanol-ammonium hydroxide) in chloroform.
The title compound was isolated as the major product. FAB ms (m+1)
469.
Step B: 1-(3-tert-Butoxycarbonylamino-2-(2-
naphthylmethylamino)prop- 1 -yl)-2(S)-butyl-4-(1 -
naphthoyl)piperazine
The title compound was prepared according to the
procedure described in Fx~mple 1, Step D except using 1-[(2-amino-3-
tert-butoxycarbonylaminoprop- 1 -yl] -2(S)-butyl-4-(1 -
naphthoyl)piperazine (0.287 g, 0.613 mmol), naphthalene-2-
carboxaldehyde (0.95 g, 0.613 mmol), sodium triacetoxyborohydride
(0.194 g, 0.919 mmol), in dichloroethane (15 mL) at pH 6. The crude
product was chromatographed on silica gel with 5% methanol in
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chloroform (Rf 0.30), and the title compound isolated as a foam. FAB
ms (m+l) 609.
Step C: 1-(3-Amino-2-(2-naphthylmethylamino)prop-1-yl)-2(S)-
butvl-4-(1-naphthovl)piperazine trihvdrochloride
A solution of 1-(3-tert-butoxycarbonylamino-2-(2-
naphthylmethylamino)prop-l-yl)-2(S)-butyl-4-(1-naphthoyl)piperazine
(0.313 g, 0.514 mmol) in methylene chloride (10 mL) was deprotected
with trifluoroacetic acid (5 mL) and converted to the free base (255
mg) according to the procedure described in F,x~mple 1, Step E.
Purification of 40 mg by preparative HPLC used gradient elution with
solvents A and B (from Fx~mple 1; 95% to 5% solvent A). Ion
exchange and lyophili7~tion as described fi~ hed the title compound as
a white solid. FAB ms (m+l) 509. Anal. Calc. for C33H40N4O 0.05
H2O 4.45 HCl: C, 59.00; H, 6.68; N, 8.34. Found: C, 59.00; H, 6.51;
N, 8.44.
EXAMPLE 3
2(S)-Butyl-1-{5-[1-(2-naphthylmethyl)]-4,5-dihydroimidazol}methyl-4-
(l-naphthoyl)piperazine ditrifluoroacetate
1 -(3-Amino-2-(2-naphthylmethylamino)prop- 1 -yl)-2(S)-
butyl-4-(1-naphthoyl)piperazine (0.105 g, 0.207 mmol), tert-
butylisocyanate (1.5 mL) and silver cyanide (0.023 g, 0.207 mmol)
were h~,~te,-l in a sealed tube under nitrogen at 90~C overni~ht The
volatiles were removed in vacuo, and the residue chromatographed on
silica gel with 5-10% methanol in chloroform to give the free base (73
,., mg). This was purified by preparative HPLC as described in Example 1
by gradient elution with solvents A and B (from Example 1; 95% to 5%
solvent A ). Lyophili7~tion furnished the title compound as a white
solid. FAB ms (m+l) 519. Anal. Calc. for C34H38N4O ~ 0.85 H2O
3.75 TFA: C, 51.84; H, 4.55; N, 5.83. Found: C, 51.83; H, 4.56; N,
6.32.
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EXAMPLE 4
1-[5-(1-Benzylimi~l~7.ol)methyl]-2(S)-butyl-4-(1-naphthoyl)piperazine
dihydrochloride
Benzyl bromide (0.012 mL, 0.103 mmol) was added to a
solution of 2(S)-butyl-1-[5-(3-triphenylmethylimidazol)]methyl-4-(1-
naphthoyl)piperazine (63 mg, 0.103 mmol) in acetonitrile (0.5 mT ) at
20~C under nitrogen. The rèaction was stirred overni~ht concentrated
in vacuo, and taken up in dichlormethane (2 mL) cont~inin~?
triethylsilane (0.100 mL). Trifluoroacetic acid was added and the
reaction stirred for lh at 20~C. The solvent was evaporated and the
residue purified by HPLC (95% to 5% solvent A). Pure fractions were
combined and converted to the HCl salt as described in Fx~mrle 1, Step
E. After lyophili7~tion, the title compound was isolated as a white
solid. FAB ms (m+1) 467. Anal. Calc. for C30H34N4O ~ 0.05 H20 -
3.70 HCl: C, 59.81; H, 6.32; N, 9.30. Found: C, 59.78; H, 6.33; N,
9.30.
EXAMPLE 5
1 - { 5-[1 -(4-nitrobenzyl)]imidazolylmethyl }-2(S)-butyl-4-(1 -
naphthoyl)piperazine ditrilluoroacetate
The title compound was prepared according to the
procedure described in Example 4, except using p-nitrobenzylbromide
(0.043 g, 0.199 mmol) and 2(S)-butyl-1-[5-(3-
triphenylmethylimidazol)]methyl-4-(1-naphthoyl)piperazine (123 mg,
0.199 mmol) in acetonitrile (2 mL). The crude product was treated
with triethylsilane (0.127 mL, 0.80 mmol) and trifluoroacetic acid (2
mL) in dichloroethane (4 mL). Preparative HPLC (95-5% solvent A)
provided the title compound as a white solid. FAB ms (m+1) 512. "
Anal. Calc. for C30H33N5O3 2 CF2CO2H 0.03 H2O: C, 41.42; H,
3.98; N, 10.18. Found: C, 41.43; H, 3.96; N, 10.51.
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- EXAMPLE 6
1 -(3-Acetamidomethylthio-2(R)-aminoprop- 1 -yl)-2(S)-butyl-4-(1 -
naphthoyl)piperazine ditrifluoroacetate
1-(3-Acetamidomethylthio-2(R)-aminoprop-1-yl)-2(S)-
butyl-4-(1-naphthoyl)piperazine dihydrochloride and N-
hydroxymethyl~cet~mide (0.010 g, 0.105 mmol) (prepared as described
in M. Bodansky, A. Bodansky, "The Practice of Peptide Synthesis",
Springer-Verlag, 1984, p.82) were dissolved in trifluoroacetic acid for
0.5h. The solvent was removed in vacuo and the residue purified by
preparative HPLC (85-60% solvent A). After lyophili7~tion, the title
compound was isolated as a solid. FAB ms (m+1) 457. Anal. Calc. for
C25H36N4O2 S ~-2 CF2CO2H ~ 1.9 H2O . C, 44.38 H, 5.12; N, 6.64.
Found: C, 44.35; H, 5.11, N, 6.97.
EXAMPLE 7
2(S)-Butyl-1-[2-(1-imidazolyl)ethyl]sulfonyl-4-(1-naphthoyl)piperazine
ditrifluoroacetate
Step A: 2(S)-Butyl-4-(1 -naphthoyl)- 1 -vinylsulfonylpiperazine
Chloroethylsulfonylchloride (0.038 mL, 0.314 mmol) was
added to a solution of 3(S)-butyl-1-(1-naphthoyl)piperazine (0.095 g,
0.285 mmol) and diisopropylethyl~mine (0.119 mT~, 0.685 mmol) in
dichloromethane (3 mL). The reaction was stirred overnight under
nitrogen, quenched with saturated sodium bicarbonate and extracted into
ethyl acetate. After drying with magnesium sulfate, the title compound
was isolated.
Step B: 2(S)-Butyl-1-[2-(1-imidazolyl)ethyl]sulfonyl-4-(1-
naphthoyl)piperazine ditrifluoroacetate
Imidazole (0.043 g, 0.627 mmol) was added to sodium
hydride (60% dispersion in oil, 0.024 g, 0.598 mmol) suspended in
dimethylform~mide (2 mL). The reaction was cooled to 0~C under
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nitrogen, and 2(S)-butyl-4-(1 -naphthoyl)- 1 -vinylsulfonylpiperazine
(0.011 g, 0.29 mmol) in dimethylform~micle (5 mL) was ~cldecl The
reaction was stirred at 20~C overnight. The dimethylforrn~ e was
removed in vacuo and the residue dissolved in ethyl acetate. This was
S extracted with saturated sodium bicarbonate solution, satu.ated brine
and dried over m~gnesium sulfate. The crude product was first purified
by silica gel chromatography using 8% methanol in chloroform, then by
preparative HPLC (80 to 40% solvent A). The trifluoroacetate salt was
dissolved in water and partitioned between saturated sodium bicarbonate
10 and ethyl acetate. The organic phase was washed with saturated brine
and dried over m~gnesium sulfate. The title compound was obtained as
a foam. FAB ms (m+1) 455. Anal. Calc. for C25H36N4O2 S ~0-8 H20
. C, 61.46; H, 6.79; N, 11.95. Found: C, 61.44; H, 6.97; N, 10.72.
EXAMPLE 8
2(R)-Butyl- 1 -imidazolyl-4-methyl-4-(1 -naphthoyl)piperazine
Step A: 2(R)-Butyl-4-(1-naphthoyl)-1-[4-(1-
triphenylmethylimidazolyllmethyl-piperazine
3(R)-Butyl-l-(1-naphthoyl)piperazine (0.202 g, 0.607
mmol) (prepared as described for the (S) enantiomer in Example 1) was
reacted with 1-triphenylmethylimidazole-4-carboxaldehye (0.226 g,
0.667 mmol), sodium triacetoxyborohydride (0.321 g, 1.52 mmol), in
dichloroethane (7 mL) in the presence of crushed molecular sieves. The
pH was adjusted to 5-6 with triethylamine/acetic acid. The reaction was
stirred overnight, then quenched with saturated sodium bicarbonate.
The organic layer was washed with saturated brine, and dried over
m~gnesium sulfate. The crude product was purified by chromatography
on silica gel with 30% ethyl acetate in hexane followed by 5% methanol
in chloroform, to obtain the title compound.
Step B: 2(R)-Butyl-1-imidazolyl-4-methyl-4-(1-
naphthoyl)piperazine ditrifluoroacetate
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~,
Triethylsilane (1.0 mL, 11.80 mmol) was added to a
solution of 2(R)-butyl-4-(1-naphthoyl)-1-[4-(1-triphenylmethyl-
imitl~zolyl]methyl-piperazine (0.381 g, 0.616 mmol) in dichloro-
methane, followed by trifluoroacetic acid (8 mL). After lh, the
5 solvents were evaporated and the residue partitioned between water and
hexaIle. The aqueous phase was injected directly onto a preparative
HPLC column (100-40% solvent A) and the title compound isolated
after lyophili7~tion. FAB ms'(m+l) 377. Anal. Calc. for
C25H36N4O2 ~ 2.35 CF3co2H ~0.32 H2O . C, 48.93; H, 4.52; N, 7.98.
Found: C, 48.93; H, 4.55; N, 8.26.
EXAMPLE 9
2(S)-Butyl-4-(1 -naphthoyl)- 1 -(3-pyridylmethyl)piperazine
dihydrochloride
3 (S)-Butyl- 1 -(1 -naphthoyl)piperazine hydrochloride (0.200
g, 0.601 mmol) was reacted with pyridine-3-carboxaldehyde (0.062
mL, 0.661 mmol), sodium triacetoxyborohydride (0.321 g, 1.52 mmol),
in dichloroethane (7 mL) at pH 5-6 in the presence of crushed
molecular sieves as described in F.x~mple 8, Step A. The crude product
was puri~led by silica gel chromatography with 30% acetone in hexane,
followed by preparative HPLC (80-75% solvent A). After ion
exchange, the title compound was isolated. FAB ms (m+l) 388. Anal.
Calc. for C25H29N3O ~ 2.3 HCl ~0.95 H2O . C, 61.49; H, 6.85; N,
8.60. Found: C, 61.49; H, 7.01; N, 8.76.
EXAMPLE 10
1 -2(S)-Butyl-(2(R)-(4-nitrobenzyl)amino-3-hydroxypropyl)-4-(1-
30 naphthoyl)piperazine dihydrochloride
1 -(2(R)-amino-3-hydroxypropyl)-2(S)-butyl-4-(1 -
naphthoyl)piperazine (120 mg, 0.326 mmol) was converted to the title
compound according to the procedure described in Example 8, Step A
using 4-nitrobenzaldehyde (0.493 g, 0.327 mmol), sodium
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triacetoxyborohydride (0.173 g, 0.817 mmol), in dichloroethane. The
crude product was purified by preparative HPLC (100-75% solvent A),
and after ion exchange to the HCl salt and lyophili7z~tion, the title
compound was obtained. FAB ms (m+l) 505. Anal. Calc. for
5 C25H29N3O ~ 3.6 HCl -0.10 H2O . C, 54.69; H, 6.30; N, 8.80. Found:
C, 54.66; H, 5.85; N, 8.31.
EXAMPLE 11
1-(2(R)-Amino-3-hydroxyheptadecyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine di~ luoroacetate
Step A: 1-(2(R)-t-Butyoxycarbonylamino-2-formylethyl)-2(S)-
butyl-4-(1-naphthoyl)piperazine
A solution of oxalyl chloride (1.36 mL, 14.9 mmol) in
dichloromethane (35 mL) was cooled to -65~C under nitrogen, and
dimethylsulfoxide (2.30 mL, 32.4 mmol) in methylene chloride (7 mL)
added, and the reaction stirred 2 min. A solution of 1-(2(R)-amino-3-
hydroxypropyl)-2(S)-butyl-4-(1-naphthoyl)piperazine (3.19 g, 6.79
mmol) was added to this solution at -10~C and the reaction stirred at this
temperature for 15 min. The reaction was cooled to -55~C and
triethylamine (4.76 mL, 34 mmol) ~ 1e-1 The reaction was stirred for
5 min then warmed to room temperature. Additional methylene
chloride is added and the reaction is extracted with water. The organic
phase was washed with 2% potassium hydrogen sulfate, water, dilute
sodium bicarbonate solution, and saturated brine. After drying with
magnesium sulfate, the title compound was obtained.
Step B: 1-(2(R)-Amino-3-hydroxyheptadecyl)-2(S)-butyl-4-(l-
naphthoyl)piperazine ditrifluoroacetate
A solution of 0.25 g, 0.53 mmol)l-(2(R)-t-
butoxycarbonylamino-2-formylethyl)-2(S)-butyl-4-(1 -
naphthoyl)piperazine in dry tetrahydrofuran (5 rnL) was cooled to 0~C
under nitrogen in a flame-dried three necked flask. A solution of
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heptadecy1m~gnesium chloride (1.01 mL of a lM solution in ether, 1.01
mmol) ~as added via syringe, and the reaction allowed to warm to
room temperature. The reaction was quenched with saLulated sodium
bicarbonate solution, and then extracted with ethyl acetate. After
5 drying over magnesium sulfate, the crude product was
chromatographed on silica gel with 25% ethyl acetate in hexane
followed by 5% methanol in chloroform. The puri~led product was
dissolved in methylene chloride (7 rnL) and treated with trifluoroacetic
acid (3.5 mL). After 45 min, the solvents were removed in vacuo and
10 the residue purified by preparative HPLC (95-40% solvent A). Two
isomers were separated. After lyophili7~tion, the title compound was
isolated as diastereomer A (retention time 8.405 min, gradient 100-50%
solvent A overlS min), FAB ms (m+l) 566, Anal. Calc. for
C36H59N302 ~ 2.35 CF3C02H ~ 0.35 H20. C, 58.19; H, 7.44; N,
15 5.00. Found: C, 58.21; H, 7.46; N, 5.36. After lyophili7~tion, the title
compound was also isolated as diastereomer B (retention time 9.269
min, gradient 100-50% solvent A overl5 min), FAB ms (m+l) 566,
Anal. Calc. for C36H59N302 ~ 2.35 CF3C02H ~ 0.05 H20. C, 58.56;
H, 7.42; N, 5.03. Found: C, 58.53; H, 7.41; N, 5.17.
EXAMPLE 12
2(S)-Benzyl-l-imidazolyl-4-methyl-4-(1-naphthoyl)piperazine
25 Step A: 1~(3S)-Dibenzylpiperazine-2.5-dione
The title compound was prepared according to the
procedure described in Example 1, Step A, except using Boc-L-
phenyl~l~nine (12.8 g, 48.2 mmol), ethyl N-benzylglycinate (9.32 g,
48.2 mmol) and dicyclohexylcarbodiimide (96.5 mL 0.5 M in
30 dichloromethane, 48.2 mmol). The crude diketopiperazine was
triturated with hexane to give the title compound as a white powder.
lHNMR (300 MHz, CD30D) o 7.0-7.4 (lOH, m), 4.61 (lH, d, J=16
Hz), 4.37 (lH, t, J=5 Hz), 4.24 (lH, d, J=16 Hz), 3.42 (lH, d, J=18 Hz),
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3.28 (lH, dd, J=4, 16 Hz), 2.96 (lH, dd, J=6, 16 Hz), 2.55 (lH, d, J=18
Hz).
Step B: l-tert-Butoxycarbonyl-2(S)-~(S)~4-dibenzylpiperazine
The title compound was prepared according to the
procedure described in Example 1, Step B, except using 1,3(S)-
dibenzylpiperazine-2,5-dione (5.01 g, 17.1 mmol) and lithium
alllmimlm hydride (2.33 g, 61.4 mmol), followed by di-tert -butyl
dicarbonate (4.02 g, 18.4 mmol). The crude product was purified by
column chromatography on silica gel, eluting with 7.5% ethyl acetate in
hexane. The title compound was obtained as a white solid. lHNMR
(300 MHz, CD30D) ~ 7.2-7.4 (5H, m), 7.0-7.2 (5H, m), 4.15 (lH, m),
3.90 (lH, d, J=15 Hz), 3.60 (lH, d, J=15 Hz), 3.15 (lH, m), 2.95 (3H,
m), 2.7 (lH, d, J=13 Hz), 2.02 (lH, dt, J=6, 13 Hz), 1.95 (lH, br d),
1.35 (9H, s).
Step C: 2(S)-Benzyl-l-tert -butoxycarbonylpiperazine
The title compound was prepared according to the
procedure described in Fx~mple 1, Step C, except using l-tert -
20 butoxycarbonyl-2(S),4-dibenzylpiperazine (4.78 g, 11.3 mmol) and
10% palladium on carbon (1.04 g). The title compound was obtained as
an oil. lHNMR (300 MHz, CD30D) o 7.25 (SH, m), 4.35 (lH, m),
4.00 (lH, d, J=12 Hz), 2.7-3.3 (7H, m), 1.25 (9H, s).
25 Step D: 2(S)-Benzyl-l-tert-butoxycarbonyl-4-(1-
naphthoyl)piperazine
The title compound was prepared according to ~e
procedure described for Example 1, Step A except using 2(S)-benzyl-l-
tert-butoxycarbonylpiperazine (0.292 g, 1.06 mmol), 2,3-dimethyl-
30 benzoic acid (0.159 g, 1.06 mmol), HOBT (0.157 g, 1.02 mmol),EDC-HCl (0.213 g, 1.11 mmol) and triethylamine to adjust the pH to 7.
The title compound was obtained as a thick oil. lHNMR (DMSO-d6,
300 MHz) o 7.15 (2H, m), 6.06 (lH,m), 4.42 (lH,m), 3.6-4.2 (2H, m),
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- 2.7-3.24 (4H, m), 2.24 (3H, s), 2.03-2.20 (3H, 4s), 1.10-1.6 (lSH, m),
0.72-1.00 (3H, m).
Step E: 2(S)-Benzyl-4-(1-naphthoyl)-1-[4-(1-
triphenylmethylimidazolyllmethyl-piperazine
3(S)-Benzyl-l-(l-naphthoyl)piperazine (0.173 g, 0.472
mmol) was reacted with l-triphenylmethylimitl~701e-4-carboxaldehyde
(0.160 g, 0.472 rnmol), sodium triacetoxyborohydride (0.300 g, 1.42
mmol), in dichloroethane (7 mL) in the presence of crushed molecular
sieves as described in Example 8, Step A. The title compound was
obtained as an oil.
Step F: 2(S)-Benzyl-l-imidazolyl-4-methyl-4-(1-
naphthoyl)piperazine ditrifluoroacetate
Triethylsilane (0.300 mL, 1.89 mmol) was added to a
solution of 2(S)-benzyl-4-(1-naphthoyl)-1-[4-(1-
triphenylmethylimi~ olyl]methyl-piperazine (0.310 g, 0.472 mmol) in
dichloromethane (5 mL), followed by trifluoroacetic acid (5 mL).
After lh, the solvents were evaporated and the residue partitioned
between water and hexane. The aqueous phase was injected directly
onto a preparative HPLC column (85-45% solvent A) and the title
compound isolated after lyophili7~tion. FAB ms (m+l) 411. Anal.
Calc. for C26H26N4O2 ~ 2.75 CF3CO2H ~0.05 H2O. C, 52.11, H, 4.14;
N, 7.72. Found: C, 52.10, H, 4.03, N, 8.16.
EXAMPLE 13
1-(2(R)-Amino-3-(3-benzylthio)propyl)-2(S)-butyl-4-(1-naphthoyl)-
piperazine ditrifluoroacetate
Step A: l-[(l-Aziridinyl)methyl]-2(S)-butyl-4-(1-naphthoyl)-
piperazine
A solution of 1-(2(R)-butoxycarbonylamino-3-
hydroxypropyl)-2(S)-butyl-4-(1-naphthoyl)piperazine (1.67 g, 3.56
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mmol) in dimethylform~rnide (10 mL) was cooled to 0~C under
nitrogen. Sodium hydride (0.427 g, 10.6 mmol, 60% dispersion in oil)
was ~(l(le~l, followed by l,l'-sulfonyldiimidazole (0.704 g, 3.56 mol).
The reaction was warmed to 20~C for lh, cooled to 0~C and quenched
5 with water. Dimethylform~mide was distilled in vacuo. and the residue
partitioned between ethyl acetate and water. The organic phase was
washed with saturated brine and dried over magnesium sulfate. The
crude product was chromatographed on silica gel with 70% ethyl acetate
in hexane, followed by 5~o methanol in chloroform. The title
10 compound was obtained as the major product, FAB ms (m+l) 352. A
lesser amount of l-[(l-butoxycarbonylaziridinyl)methyl]-2(S)-butyl-4-
(l-naphthoyl)piperazine was also isolated.
Step B: 1-(2(R)-Amino-3-(3-benzylthio)propyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine ditrifluoroacetate
l-[(l-Aziridinyl)methyl]-2(S)-butyl-4-(1-
naphthoyl)piperazine (0.050 g, 0.142 mmol) was refluxed for 18h with
benzyl mercaptan (0.100 mL, 0.852 mmol) and triethylamine (0.200
mL) in methanol (4 mL). The crude product was first
chromatographed on silica gel with 3% methanol in chloroform, and
then purified by preparative HPLC (85% to 10% solvent A). The title
compound was obtained after lyophili7~tion. FAB ms (m+l) 476.
Anal. Calc. for C29H37N3OS ~ 2.6 CF3CO2H ~0.3 H2O. C, 52.83; H,
5.21; N, 5.40. Found: C, 52.78; H, 5.17; N, 5.66.
EXAMPLE 14
1 -(2(R)-Amino-3-[3-(4-nitrobenzylthio)propyl]))-2(S)-butyl-4-(1 -
naphthoyl)piperazine ditrifluoroacetate
A solution ofl-[(l-butoxycarbonylaziridinyl)methyl]-2(S)-
butyl-4-(1-naphthoyl)piperazine (0.050 g, 0.111 mmol) in methanol (4
mL) was reluxed with p-nitrobenzylthioacetate (0.070 g, 0.333 mmol)
and triethylamine (0.200 mL) for 2h. The crude product was
chromatographed on silica gel with 3~o methanol in chloroform. The
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.,
purified product was treated with 33% trifluoroacetic acid in methylene
chloride for 20 min. The solvents were evaporated and the product
puri~led by preparative HPLC (85%-10~o solvent A). The title
compound was obtained after lyophili7~tion. FAB ms (m+l) 529.
Anal. Calc. for C29H36N4O3 ~ 2 CF3CO2H ~0.08 H2O. C, 51.94; H,
5.23; N, 7.34. Found. C, 51.87; H, 5.06; N, 7.47.
EXAMPLE 15
2(S)-Butyl-1-[(4-imidazolyl)ethyl]-4-(1-naphthoyl)piperazine
dihydrochloride
Step A: N-Methyl-N-methoxy-2-(1 -triphenylmethyl- 1 H-imidazol-4-
yl)acetamide
To a solution of 4-imill~7oleacetic acid (1.04g, 6.40 mmol)
and triphenylmethyl bromide (2.48 g, 7.68 mmol) in
dimethylform~micle (40 ml) was added triethyl~mine (4.46 ml, 32
mmol) and the suspension allowed to stir for 18 hours at room
temperature. After this time, the mixture was treated with 3-hydroxy-
1,2,3-benzotriazin-4(3)-one (HOOBT) (1.31g, 8 mmol), N, O-
dimethylhydroxyamine hydrochloride (1.56g, 16 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (1.53
g, 8 mmol) and stirred for 24 hours at room temperature. After this
time, sat. aq. sodium bicarbonate (50 ml) and water (50 ml) were
added and the mixtllre was extracted with ethyl acetate ( 2 x 100 ml ).
The combined extracts were washed with sat. aq. NaHCO3 (100 ml) and
then brime (50 ml) and the solvent evaporated in vacuo. The residue
was suspended in ether (20 ml) and the white solid filtered to give the
title compound as a white solid.
1NMR(CD3OD, 300 MHz) ~ 7.37(10H, m), 7.16(6H, m), 6.84(1H, s),
3.73(2H, s), 3.68(3H, s) and 3.18(3H, s)ppm.
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Step B: 2-(1-triphenylmethyl-lH-imidazol-4-yl)acetaldehyde
To a solution of the product from Step A (300 mg, 0.73
mmol) in freshly distilled THF (15 ml) cooled to -40~C over dry
ice/acetone was added lithium aluminum hydride (33.2 mg, 0.874
5 mmol). The resulting suspension was allowed to warm to +5~C and
then m~int~in~.d at 0~C for 30 min. After this time, the reaction mixture
was recooled to -40~C and quenched sequentially with water (33 ,ul),
1.0N NaOH (33 ,ul) and water (100 ,ul). The resulting suspension was
stirred for 30 min, filtered and the solvent removed in vacuo. The
10 residue was dissolved in methylene chloride (5 ml) and washed with
10% aq. citric acid (5 ml) and then water (Sml). The organic layer was
dried(MgSO4) and the solvent evaporated in vacuo to give the title
compound.
lHNMR (CDCl3, 300 MHz) ~ 9.79(1H, t, J=3Hz), 7.44(1H, s), 7.4-
15 7.1(15H, m), 6.76(1H, s) and 3.63(2H, d, J=3Hz)ppm.
Step C: 2(S)-Butyl-1-[(4-imi~1~7olyl)ethyl]-4-(1-
naphthoyl)piperazine di~ luoroacetate
To a solution of the product from Step B (89 mg, 0.263
20 mmol) and 3(S)-butyl-1-naphthoylpiperazine hydrochloride (62.6 mg,
0.188 mmol) in 1,2-dichloroethane (4 ml) was added 3A molecular
sieves (400 mg) and sodium triacetoxyborohydride (200 mg, 0.94
mmol). This mixture was stirred at room temperature for 4 days. After
this time, filtered the mixture through sintered glass. The filtrate was
25 diluted with methylene chloride (50 ml) and washed with water (25 ml).
The solvent was evaporated in vacuo and the residue was purified by
flash chromatography eluting with 2-5% methanol/methylene chloride
to provide the trityl protected title compound as an oil. This oil was
dissolved in methylene chloride (2 ml) and trifluoroacetic acid (1 ml)
30 and treated with triethylsilane (2 drops) to give a colorless solution.
This solution was stirred at room temperature for 3 hours and then the
solvent was evaporated in vacuo. The resulting residue was dissolved in
water (20 ml) and washed with hexanes (20 ml). The aqueous layer was
lyophilized to give the title compound.
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.,
lHNMR (CD30D, 300 MHz) o 8.83(1H, s), 8.08-7.75(3H, m), 7.67-
7.49(4H, m), 7.45(1H, s), 4.5-3.8(2H, m), 3.65-2.95(9H, m), 2.0-
- 1.3(4H, m) and 2.2-0.2(5H, m)ppm.
Anal. calc'd for C24H30N4O 3.45 TFA 0.75H20: C, 46.54; H, 4.42; N,
7.03. Found: C, 46.54; H, 4.41; N, 7.35.
FAB H~MS exact mass calc'd for C23H31N40 391.249787 (MH+),
found 391.249028
EXAMPLE 16
2(S)-Butyl-1-[(4-imidazolyl)methyl]-4-(1-naphthoyl)piperazine
dil~ uoroacetate
To a solution of the product from Step C (102 mg, 0.30
mmol) and 3(S)-butyl-l-naphthoylpiperazine hydrochloride (62.6 mg,
15 0.188 mmol) in 1,2-dichloroethane (5 ml) was added sodium
triacetoxyborohydride (200 mg, 0.94 mmol) and triethyl~minP to pH =
5.5 and stirred at room temp. for 18 hours. After this time, the
mixtllre was filtered. The filtrate was concentrated in vacuo and
dissolved in ethyl acetate (25 ml) and washed with sat. aq. NaEICO3 (10
20 ml) and then brine (10 ml). The solvent was evaporated in vacuo and
the residue was purified by ilash chromatography eluting with 3-8%
methanol/methylene chloride to provide the trityl protected title
compound. This oil was dissolved in methylene chloride (4 ml) and
trifluoroacetic acid (2 ml) and treated with triethylsilane until the
25 yellow color disappeared to give a colorless solution. Stirring
continued at room temp. for 10 min. and the solvent evaporated in
vacuo. The resulting white solid was partially dissolved in water (7 ml)
and filtered. The filtrate was purified by Prep HPLC using a Nova
Prep 5000 Semi Preparative HPLC system and a Waters PrepPak
30 cartridge (47 X 300mm, C18, 15 ,um, lOOA) eluting with 5 - 95%
acetonitrile/water ( 0.1% TFA) at 100 ml/min ( Chromatography A ) to
give the title compound after lyophili7~tion
lHNMR (DMSO-d6, 400 MHz, 150~C) o 8.53(1H, s), 7.94(2H, m),
7.79(1H, m), 7.53(3H, m), 7.41(1H, m), 7.40(1H, s), 4.12(1H, d,
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,.
J=4.8Hz), 3.95(1H, d, J=4.8Hz), 3.70(1H, s), 3.63(1H, s), 3.48(1H, s),
3.40(1H, s), 3.01(1H, s), 2.82(1H, s), 2.74(1H, s), 1.70(1H, m),
1.49(1H, m). 1.18(2H, s), 1.08(2H, s) and 0.77(3H, t, J=5.5Hz)ppm.
Anal. calc'd for C23H2gN4o 2 TFA 0.70H20: C, 52.55; H, 5.13; N,
9.08. Found: C, 52.54; H, 5.11; N, 9.35.
FAB MS mass calc'd for C23H30N4o 377 (MH+), found 377.
EXAMPLE 17
2(S)-Butyl-l-[(l-naphth-2-ylmethyl)-lH-imiclz~7.ol-5-yl)acetyl]-4-(l-
naphthoyl~piperazine dihydrochloride
Step A: Preparation of lH-Tmicl~7ole-4- acetic acid methyl ester
hydrochloride
To a solution of lH-imidazole-4-acetic acid hydrochloride
(4.00g, 24.6 mmol) in methanol (100 ml) was bubbled hydrogen
chloride gas until satu~ted. This solution was allowed to stand for 18h
at room temperature and the solvent evaporated in vacuo to give the
title compound as a white solid.
lH NMR(CDC13, 400 MHz) ~ 8.85(1H, s),7.45(1H, s), 3.89(2H, s) and
3.75(3H, s) ppm.
Step B: Preparation of l-(Triphenylmethyl)-lH-imi~1~7ol-4-ylacetic
acid methyl ester
To a suspension of the product from Step A (7.48g, 42.4
mmol) in methylene chloride (200ml) was added triethyl~tnine (17.7
ml, 127 mmol) and triphenylmethyl bromide(l6.4g, 50.8 mmol) and
stirred for 72h. After this time, the reaction mixture was washed with
sat. aq. NaHCO3 (100 ml) and water ( 100 ml). The organic layer was
evaporated in vacuo and the residue was purified by flash
chromatography( 30-100% ethyl acetate in hexanes gradient elution) to
provide the title compound as a white solid. lH NMR(CDC13, 400
MHz) ~ 7.35(1H, s), 7.31(9H, m), 7.22(6H, m), 6.76(1H, s), 3.68(3H,
s) and 3.60(2H, s) ppm.
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;
Step C: Preparation of 2- [1-(Naphth-2-ylmethyl)-lH-imidazol-S-
yl)lacetic acid methyl ester
To a solution of the product of Step B (4.36g, 11.4 mmol)
S in acetonitrile (70 ml) was added 2-(bromomethyl)naphthalene and
heated to 55~C for 4h. After this time, the reaction was cooled to room
temperature and the resulting white precipitate was collected by
filtration. The filtrate was concentrated to 30 ml and heated to 55~C for
18h. After this time, the reaction was cooled to room teperature and
10 filtered the resulting white precipitate collected by filtration. The
ltrate was concellLl~ted to 10 ml volume and heated to 55~C for lh.
After this time, the reaction was again cooled to room tempel~lur~ and
diluted with ethyl acetate (25 ml). The resulting preci~ te was
collecte~ by filtration and combined with the previous 2 precipitates in
15 methanol (100 ml) and heated to reflux for 30m. After this time, the
solvent was removed in vacuo and the resulting residue was partioned
between methylene chloride(200 ml) and sodium bicarbonate (100 ml).
The organic layer was evaporated in vacuo to dryness and the residue
was puri~led by flash chromatography (0-6% methanol/methylene
20 chloride gradient) to provide the title compound as an off white solid:
lHNMR (CDC13, 400 MHz) ~ 7.82(2H, m), 7.75(1H, m), 7.70(1H, s),
7.49(3H, m), 7.20(1H, d, J=8.4Hz), 7.06(1H, s), 5.32(2H, s), 3.57(3H, s)
and 3.49(2H, s) ppm.
25 Step D: Preparation of 2-[1-(Naphth-2-ylmethyl)-lH-imidazol-5-
yllacetic acid hydrochloride
2-[1-(Naphth-2-ylmethyl)-lH-imidazol-5-yl]acetic acid
methyl ester (0.92g, 3.28mmol ) was dissolved in 2.5N hydrochloric
acid ( 50ml ) and heated to 55~C for 3h. After this time, the solution
30 was concentrated to dryness in vacuo to give the title compound as a
white solid.
lHNMR (CD30D, 400 MHz) ~ 8.92(1H, s),7.94(1H, d, J=8.6Hz),
7.88(2H, m), 7.83(1H, s), 7.54(3H, m), 7.43(1H, d, J=14Hz), 5.60(2H,
s) amd 3.82(2H, s) ppm.
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Step E: Preparation of 2(S)-Butyl-l-[(l-(naphth-2-ylmethyl)-lH-
imidazol-5-yl)acetyl]-4-(1 -naphthoyl)piperazine
dihydrochloride
To a solution of the product from Step D (100 mg,
0.330mmol), 3(S)-butyl-l-naphthoylpiperazine hydrochloride (100 mg,
0.300 mmol) and 3-hydroxy-1,2,3-benzotriazin-4(3H)-one (HOOBT)
(54 mg, 0.33mmol) in dimethylform~mide (2 ml) was added 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)
10 (63mg, 0.33 mmol) and triethylamine (161ul, 1.16mmol) and ~e
resulting suspension stirred for 18 hours. After this time, sat. aq.
NaHCO3 (7 ml) was added and the resulting precipitate filtered. The
solid was redissolved in methylene chloride and washed with water (25
ml) and brine (20 ml). The solvent was evaporated in vacuo and the
15 residue puri~led by Preparative HPLC (Chromatography A ) to give
after lyophilli7~tion the title compound.
lHNMR (CDC13, 400 MHz) o 8.98(1H, m), 8.03-7.27(15H, m), 5.69-
5.50(2H, m), 4.79-4.34(2H, m), 4.15-2.50(7H, m), 1.8-0.2(9H, m)ppm.
Anal. calc'd for C35H35N402 HCl 0.65H20: C, 67.30; H, 6.16; N,
20 8.97. Found: C, 67.25; H, 6.16; N, 9.15.
FAB HRMS exact mass calc'd for C35H36N4O2 545.291652 (MH~),
found 545.292050.
EXAMPLE 18
2(S)-Butyl-l-[(l-naphth-2-ylmethyl)-lH-imidazol-5-yl)ethyl]-4-(1-
naphthoyl)piperazine ditrifluoroacetate
Step A: Preparation of N-Methyl-N-methoxy-2-[1-(naphth-2-
ylmethyl)-lH-imidazol-5-yl)lacetamide
To a solution of 2-[1-(naphth-2-ylmethyl)-lH-imidazol-5-
yl]acetic acid hydrochloride (0.819 mg, 2.70 mmol) in
dimethylform~rnide (15 ml) was added sequentially N, O-
dimethylhydroxylamine hydrochloride (293 mg, 3.0 mmol ), 3-
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hydroxy-1,2,3-benzotriazin-4(3H)-one (HOOBT) (489 mg, 3.0 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)
(575 mg, 3.0 mmol) and triethylamine (1.67 ml, 12.0 mmol). This
mixture was stirred at room temperature for 18h. After this time, sat.
aq. NaHCO3 (30ml) and water (30ml) were added and the mixture was
extracted with methylene chloride ( 2 x 50rnl ). The combined organic
extracts were washed with brine (50 ml) and the solvent evaporated in
vacuo. The residue was purified by flash chromatography eluting with
2-4% methanol/methylene chloride gradient to provide the title
compound as an oil.
lHNMR (CDC13, 400 MHz) o 7.80(2H, m), 7.74(1H, m), 7.56(1H, s),
7.47(3H, m), 7.22(1H, d, J=8.6Hz), 6.97(1H, s), 5.37(2H, s), 3.58(2H,
s), 3.51(3H, s) and 3.12(3H, s)ppm.
Step B: Preparation of 2-[1-(Naphth-2-ylmethyl)(lH-imi~ ol-5-
yl)lacetaldehyde
To a suspension of lithillm ~ hydride (40.8mg,
1.07mmol) in tetrahydrofuran (Sml) cooled to -45~C over a dry
ice/acetone bath was added a solution of the product from Step A (243
mg, 0.895 mmol) in tetrahydrofuran (S ml) via canula at such a rate to
m~int~in tempel~lure the temperaLulc; at <-35~C. After the addition was
complete, the solution was allowed to warm to +5~C and then recooled
to -35~C. To this solution was added a solution of potassium bisulfate
(272 mg) in water (1 ml). This mix1~1re was stirred for 30 min at room
temperature and filtered through celite. The celite pad was washed with
ethyl acetate (25 ml). The combined filtrates were washed with sat.
sodium bicarbonate (10 ml) and then water (lOml). The organic layer
was dried over m~p~nesium sulfate, filtered and evaporated to give the
title compound as a clear oil.
lHNMR (CDC13, 400 MHz) ~ 9.50(1H, t, 2Hz), 7.85-7.70(3H, m),
7.64(1H, s), 7.53-7.40(3H, m), 7.16(1H, d, J=12Hz), 7.06(1H, s),
5.20(2H, s) and 3.53(2H, m)ppm.
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Step C: Preparation of 2(S)-Butyl-l-[(l-naphth-2-ylmethyl)-lH-
imidazol-5-yl)ethyl]-4-(1-naphthoyl)piperazine
ditrifluoroacetate
To a solution of the product from Step B (58.4 mg, 0.232
5 mmol) and 3(S)-butyl-l-naphthoylpiperazine hydrochloride (96.5 mg,
0.279 mmol) in 1,2-dichloroethane (lOml) and dimethylform~mide
(Sml) was added 3A molecular sieves (250mg) and sodium
triacetoxyborohydride (236.5 mg, 1.12 mmol). This mixture was
stirred at room temperature for 18 h. After this time, the mixture was
10 filtered . The filtrate was diluted with methylene chloride (lOOml) and
washed with sat sodium bicarbonate (SOml). The organic layer was
dried(MgS04), filtered and evaporated in vacuo. The residue was
purified first by flash chromatography eluting with 2-5~o methanol /
methylene chloride and then by Prep HPLC (Chromatography A) to
15 provide the title compound.
lH NMR(CD30D, 400 MHz) ~i 9.04(1H, s), 8.17-7.30(15H, m),
5.65(2H, s), 4.6-2.2(11H, m) and 1.6-0.2(9H, m)ppm.
FAB HRMS exact mass calc'd for C35H39N40 531.312387 (MH+),
found 531.313011.
EXAMPLE 19
1 -(2(R)-Amino-3-hydroypropyl)-2(S)-butyl-4-(1 -naphthoyl)piperazine
bis trifluoracetate salt
Step A: N-Boc-O-Benzylserine-(N'-Methoxy) methyl amide
N-Boc-O-Benzylserine (Bachem; 5.0g, 16.9 mmol) and
HOBT (2.29g, 16.9 mmol) were dissolved in dry DMF (100 mL) under
argon. To this solution was added N,O-dimethylhydroxyl amine
30 hydrochloride (1.98 g, 20.3 mmol) and then, at 0~C, EDC
hydrochloride (3.56 g, 18.6 mmol). 4-Methylmorpholine was added to
bring the pH to ~7 (4.5 mL) and the mixture was stirred at room
temperature for 3h. The solution was diluted with EtoAc and poured
into 0.5N HCl. After extraction with EtoAc (twice), the organic layers
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were washed with water then brine, dried (MgSO4) and evaporated in
vacuo to give a pale yellow oil. Column chromatograph (silica gel;
hexanel/EtoAc 2:1) gave the title compound as an oil Rf (silica;
hexare/EtoAc 1:1) = 0.45.
Step B: N-Boc-O-Benzyl-serine aldehyde
The amide from Step A (5.7 g, 16.9 mmol) in dry ether
(20 mL) was added L~)~wise to a suspension of LAH (0.705 g, 18.5
mmol) in 80 mL of ether at -50~C. After the addition was complete, the
solution was stirred at 0~C for 45 min. then cooled back to -50~C and a
solution of KHSO4 (4 g in 11 mL H2O) was added slowly. This
mixhlre was then stirred at room temperature for 1 h, filtered through
celite, washed successively wit~h 10% citric acid solution, sa~ ted
NaHCO3 solution and brine, dried (MgSO4) and evaporated to give an
oil which was used as such in the next step. Rf (silica; hexane/EtoAc
2:1) = 0.58.
Step C: 1-(2(R)-N-Boc-Amino-3-benzyloxy propyl)-2(S)-butyl-4-
(l-naphythoyl) piperazine
A solution of the piperazine hydrochloride from Example
1, Step E (1.7 g, 574 mmol) in CH2C12 (25 mL) was adjusted to pH 6
using Et3N then freshly ground and activated 4 A sieves were added
followed by sodium triacetoxyborohydride (4.85 g, 22.9 mmol). The
aldehyde from Step B (2.08g, 747 mmol) dissolved in 20 mT . CH2C12
was added dro~wise at 0~C over 20 rnin. then the mixture was sitrred at
room temperature for 16 h. After this time, the mixture was filtered
through celite, diluted with EtoAc and washed successively with H2O,
KHSO4, solution, NaHCO3 solution and then brine. The dried
(MgSO4) solution was evaporated in vacuo to give a pale yellow oil
~, 30 which was subjected to column chromatography (silica gel;
hexane/EtOAc 1:1) to give the title compound as a foam. Rf (silica;
hexane/EtoAc 2:1) = 0.15.
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Step D: 1-(2(R)-N-Boc-Amino-3-hydroxy propyl)-2-(S)-butyl-4-
(l-naphythoyl) piperazine
The benzyl ether from Step C (700 mg, 1.25 mmol) was
dissolved in 20 mL MeOH with 150 ,~LL of acetic acid and then 20%
5 Pd(OH)2 on carbon (500 mg) was added and the mixture hydrogenated
at 50 psi for 16 hr. After filtration through celite, the solvent was
removed and the residue was chromatographed on silica gel
(EtoAc/hex~ne 1:1 then 5% hIeOH/EtoAc) to give the title compound as
an oil.
Step E: 1-(2(R)-Amino-3-hydroxy propyl)-2(S)-butyl-4-(1-
naphythoyl)piperazine bis trifluoroacetate salt
A solution of the N-Boc amine from Step D (140 mg) in 10
mT~ EtoAc was treated with HCl (gas) until saturated. After 5 min, the
15 solution was purged with argon then the solvent was removed to give a
solid which was pllrifi~tl by preparative HPLC (C-18 column;
H2O/CH3CN with 0.1% TGA; gradient). The aqueous solution was
frozen and lyophilized to give the title compound as a hygroscopic
powder. FAB mass spectrum m/z = 370 (M+l).
Analysis calculated for C22H32N3O2-2.35 TFA
C, 50.31; H, 5.27; N, 6.59
Found: C, 50.28; H, 5.49; N, 6.70
EXAMPLE 20
1-(2(R)-Amino-4-hydroxybutyl)-2(S)-butyl-4-(1-naphthoyl)piperazine
bishydrochloride salt
Following the procedure of Example 19, Steps A to E but
using N-Boc-O-benzyl homoserine (Bachem) as starting material, the
30 title compound was obtained as the bis hydrochloride salt. FAB Mass
spectrum, m/z = 384 (M+l).
Analysis calculated for C23H33N3O2-1.5 HCl
C, 54.04; H, 7.20; N, 8.22
Found: C, 53.95; H, 7.23; N, 8.50
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EXAMPLE 21
..
1-(2-Amino-3-(2-benzyloxyphenyl)propyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine bistrifluoroacetate salt
s
Step A: D.L-N-Boc-ortho-tyrosine methyl ester
The title compound was prepared as a crystalline solid
from D,L-orthotyrosine (Sigma) in two steps ((Boc)20/K2C03 in
THF/H20 followed by diazomethane in EtOAc).
Step B: 3-(2-Hydroxyphenyl)-2-(N-Boc-Amino)propanol
To a solution of the ester from Step A (1.34 g, 4.54 mmol)
in THF (20 mL) at 0~C was added LAH (400 mg, 10.5 mmol) in
portions. After 4 hs at room temperatrue, 0.4 mL H2O was added
15 d,o~wise followed by 0.4 mL lN NaOH and then 1.2 mL H20. The
slurry was stirred for 1 h, filtered through celite rin~inp with THF and
the solvent was removed chromatography of the residue (silica gel;
hexane/EtoAc 1:1) gave the title compound as a solid. Rf (silica;
hexane/EtoAc 1:1) = 0.45.
Step C: 3-(2-Benzyloxyphenvl)-2-(N-Boc-amino)propanol
A mixture of the alcohol from Step B (280 mg, 1.05 mmol)
benzyl bromide (150 ,uL, 1.26 mmol) and CS2CO3 (513 mg, 1.57
mmol) in DMF (10 mL) was stirred at room tempe-atul~ under argon
25 for 16 h. The mixtllre was poured into H2O, extracted twice with
EtOAc, washed with water then brine, dried (MgSO4) and evaporated
to give an oil. Purification by chromatography (silica gel;
hexane/EtoAc 2: 1) gave the title compound as an oil. Rf (silica;
hexane/EtoAc 2:1) = 0.36.
Step D: 3-(2-Benzyloxyphenyl)-2-(N-Boc-amino)propanol
To a solution of the alcohol from Step C (280 mg, 0.78
mmol) in 3 mL DMSO, 3 mL CH2C12 and 0.55 mL Et3N at room
temperature under argon was added pyridine SO3 complex (500 mg,
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3.14 mmol) and the mixture stirred for lh. The solution was poured
into saturated NaHCO3 solution, extracted twice with EtoAc, washed
with water then brine, dried (MgSO4) and concentrated in vacuo. The
title compound was thus obtained as an oil and was used as such in the
following step. Rf (silica; hexane/EtoAc 4:1) = 0.41.
Step E: 1-(2-N-Boc-Amino-3-(2-benzyloxyphenyl)propyl)-2(S)-
butyl-4-(1-naphthoyl)piperazine
The aldehyde from Step D and the piperazine
hydrochloride from Example 1, Step E were coupled via reductive
aL~yltion using the procedure described for Example 19, Step C.
Purification by column chromatgraphy (silica gel; hexane/EtoAc 3:2)
gave the title compound as a 1:1 mixture of diastereomers. Rf (silica;
hexane/EtOAc 1:1) = 0.51 and 0.45.
Step F: 1-(2-Amino-3-(2-benzyloxyphenyl)propyl)-2(S)-butyl-4-
(l-naphthoyl)piperazine bis trifluoroacetate salt
A solution of the N-Boc amine from Step E (70 mg) in
EtOAc (15 mL) was treated with HCl (g) until saturated. After 15
min., the solution was purged with argon and then the solvent removed
to yield a solid. Purification by preparative HPLC (C-18 column,
H2O/CH3CN with 0.1% TFA; gradient) afforded (after lyophili~tion)
the title compound as a powder. FAB mass spectrum, m/z = 536
(M+ 1).
Analysis calculated for C~35H41N302-2.2 TFA-0.35 H20
C, 59.68; H, 5.58; N, 5.30
Found: C, 59.70; H, 5.60; N, 5.56
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EXAMPLE 22
..
1 -(2-AmLino-3 -(2-hydroxyphenyl)propyl)-2(S)-butyl-4-(1 -
naphthoyl)piperazine bis trifluoroacetate salt~ diastereomer A
s
Step A: 1-(2-N-Boc-Amino-3-(2-hydroxyphenyl)propyl)-2(S)-
butyl-4-(1-naphthoyl)piperazine
A mix~lre of 1-~2-N-Boc-Amino-3-(2-benzyloxy-
phenyl)propyl)-2(S)-butyl-4-(1-naphthoyl)piperazine (from Fx~mple
19, Step E; 290 mg, 0.5 mmol), acetic acid (60 ,UL, 1.0 mmol) and 20%
Pd(OH)2 on carbon (100 mg) in MeOH (20 mL) was hydrogenated at
67 psi for 5 h. The solution was filtered through celite and the solvent
evaporated to give an oil which showed 2 well resolved spots on silica
tlc. Column chromatography (silica gel; hexane:EtoAc 1:1) gave:
a) diastereomer A of the title compound RF (silica;
hexane/EtoAc 1:1) = 0.49.
b) diastereomer B of the title compound RF (silica;
hexane/EtoAc 1:1) = 0.35.
Step B: 1-(2-Amino-3-(2-hydroxyphenyl)propyl)-2(S)-butyl-4-(1-
naphthoyl)piperazine bis trifluoroacetate salt, diastereomer
A
The N-Boc amine, diastereomer A from Step A, was
deprotected using HCl (gas) in EtoAc. Removal of the solvent followed
25 by preparative HPLC (C-18 column; H2O/CH3CN with 0.1% TFA;
gradient) gave (after lyophili~tion) the title compound as a powder.
FAB mass spectrum, m/z = 446 (M+l).
Analysis calculated for C2gH35N3O2-2 TFA
C, 57.05; H, 5.54; N, 6.24
30 Found: C, 57.08; H, 5.64; N, 6.32
EXAMPLE 23
1-[3-(4-imidazolyl)propyl]-2(S)-butyl-4-(1-naphthoyl)piperazine bis
35 trifluoroacetate salt
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Step A: Ethyl 3-(4-imidazolyl)propionate
The title compound was prepared from urocanic acid
(Aldrich) in 2 steps using standard chemical procedures (esterification
using HCl in EtOH followed by hydrogenation with 10% Pd-C in
EtOH).
lH NMR (CDC13): o 1.23 (3H, t), 2.65 (2H, t), 2.94 (2H, t), 4.15 (2H,
q), 6.81 (lH, S), 7.56 (lH, s).
Step B: 3-(4-imidazolyl) propanol
To a solution of the ester from Step A (120 g, 71.4 mmol)
in dry THF (230 mL) at 0~C was added LAH (2.99 g, 78.6 mmol)
portionwise over 30 min. The mixtllre was stirred for 2 h at room
temperatrue then cooled back to 0~C and H2O (4.2 mT~) was added
(care!) ~l~wise. This was followed by the dro~wise addition of 10.6
mE of lN NaOH then the resulting slurry was stirred at room
temperature for 1 h. After filtration through celite w~hing with
EtOAc, the solvent was removed to give the title compound as a clear
oil (8.16 g).
lH NMR (CD30D): ~ 1.84 (2H, pentet), 2.68 (2H, t), 3.58 (2H, t), 6.88
(lH, s), 7.80 (lH, s).
Step C: 3-(4-N-BOC-imidazolyl)propanol
The imidazole from Step B (166 mg, 1.32 mmol), (BOC)2o
(302 mg, 1.38 mmol) and K2CO3 (190 mg, 1.38 mmol) were stirred in
THF (10 mL) for 2 h. After filtration, the solvent was removed in
vacuo to give the desired compound as an oil. Rf (silica; 5%
MeOH/CHCl3) = 0.17. This was used as such in the next step.
Step D: 1-[3-(4-imidazolyl)propyl]-2(S)-butyl-4-(1-naphthoyl)-
piperazine bis trifluoroacetate salt
The alcohol from Step C (~1.32 mmol) was dissolved in 4
mL DMSO, 4 mL CH2cl2 and 0.92 mL Et3N and then pyridine SO3
complex (600 mg, 5.28 mmol) was added in portions. After 3 h, the
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mixture was poured into EtOAc, extracted with saturated NaHCO3
solution then brine, dried and evaporated to afford the corresponding
- aldehyde which was used without further purification.
This aldehyde was coupled with the piperazine from
5 Fx~ le 1, Step E by reductive alkylation following the procedure
described for F.x~mrle 19, Step C and the product was obtained by
column chromatography (silica gel; 5% MeOH/CHCl3). This product
was dissolved in 10 mL EtOAc saturated with HCl (gas). The solvent
was removed and the residue purified by preparative HPLC (C-18;
10 H2O/CH3CN with 0.1% TFA; gradient). Lyophili7~tion of the frozen
aqueous solution gave the title compound as a hygroscopic powder.
FAB mass spectrum, m/z = 405 (M+l).
Analysis calculated for C25H32N40-2.35 TFA 0.4 H2O
C, 52.48; H, 5.21; N, 8.24
15 Found: C, 52.45; H, 5.22; N, 8.27
EXAMPLE 24
2(S)-n-Butyl-4-(1 -naphthoyl)- 1 -[1 -(1 -naphthylmethyl)imidazol-5-
20 ylmethyll-piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1 -naphthoyl)piperazine (0.124
g, 0.200 mmol) and l-bromomethylnaphthalene (0.046 g, 0.21 mmol)
according to the procedure described in Example 4. The title
25 compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 30%-75% acetonitrile/0.1% TFA; 70%-
25% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
(m+l) 517. Anal. Calc. for C34H36N4O ~ 0.05 H2O ~ 2.0 TFA: C,
60.55; H, 5.22; N, 7.43. Found: C, 60.48; H, 5.12; N, 7.42.
.,
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EXAMPLE 25
2(S)-n-Butyl-4-(1-naphthoyl)-1-[1-(2-naphthylmethyl)imidazol-5-
ylmethyll-piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimill~7.olyl)methyl]-4-( l -naphthoyl)piperazine (0.124
g, 0.200 mmol) and 2-bromomethylnaphthalene (0.046 g, 0.21 mmol)
according to the procedure described in Fx~mI)le 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 30%-75% acetonitrile/0.1% TFA; 70%-
25% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
(m+l) 517. Anal. Calc. for C34H36N4O ~ 1.7 H2O ~ 2.0 TFA: C,
58.87; H, 5.38; N, 7.23. Found: C, 58.90; H, 4.93; N, 7.13.
EXAMPLE 26
2(S)-n-Butyl-1-[1-(4-cyanobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine dihilluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1-naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-cyanobenzylbromide (0.041 g, 0.21 mmol)
according to the procedure described in Example 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 25%-65% acetonitrile/0.1% TFA; 75%-
35% 0.1% aqueous TFA over 50 rnin.) and lyophili~tion. FAB ms
(m+l) 492. Anal. Calc. for C31H33N5O ~ 0.35 H20 2.0 TFA: C,
57.91; H, 4.96; N, 9.65. Found: C, 57.93; H, 4.91; N, 9.55.
EXAMPLE 27
2(S)-n-Butyl- 1 -[1 -(4-methoxybenzyl)imidazol-5-ylmethyl]-4-(1 -
naphthoyl)piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1 -naphthoyl)piperazine (0.124
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g, 0.200 mmol) and 4-methyoxybenzylchloride (0.041 mL, 0.21 mmol)
according to the procedure described in Example 4, with the addition of
potassium iodide (100 mg) to the reaction mixtnre. The title compound
was obtained after purification by reverse phase ~lepal~tive HPLC
5 (gradient elution with 25%-65% acetonitrile/0.1% TFA; 75%-35%
0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms (m+1)
497. Anal. Calc. for C31H36N402 ~ 1.7 H20 ~ 2.0 TFA: C, 53.40; H,
5.30; N, 7.12. Found: C, 53.37; H, 4.78; N, 7.00.
EXAMPLE 28
2(S)-n-Butyl-1-[1-(3-methyl-2-butenyl)imi~1~7ol-5-ylmethyl]-4-(1-
naphthoyl)piperazine dillinuoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
15 (3-triphenylmethylimi~l~7Olyl)methyl]-4-(l-n~phtlloyl)piperazine (0.124
g, 0.200 mmol) and 4-bromo-2-methyl-2-butene (0.024 mT~, 0.21
mmol) a.ccording to the procedure described in Example 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 5%-95% acetonitrile/0.1% TFA, 95%-5%
20 0.1% aqueous TFA over 50 min.) and lyophili7~tiom. FAB ms (m+1)
445. Anal. Calc. for C2gH36N4O ~ 1.8 H2O ~ 2.0 TFA: C, 54.51; H,
5.95; N, 7.95. Found: C, 54.54; H, 5.39; N, 7.73.
EXAMPLE 29
2(S)-n-Butyl-1-[1-(4-fluorobenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1-naphthoyl)piperazine (0.124
30 g, 0.200 mmol) and 4-fluorobenzylbromide (0.026 mL, 0.21 mmol)
according to the procedure described in Example 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 25%-65% acetonitrile/0.1% TFA; 75%-
35% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
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(m+l) 485. Anal. Calc. for C30H33FN4o ~ 3.0 H2O ~ 2.0 TFA: C,
53.26, H, 5.39; N, 7.31. Found: C, 53.21; H, 4.56; N, 7.08.
EXAMPLE 30
s
2(S)-n-Butyl-1-[1-(4-chlorobenzyl)imi-1~7.ol-5-ylmethyl]-4-(l-
naphthoyl)piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl] -4-(1 -naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-chlorobenzylchloride (0.034 mg, 0.21 mmol)
according to the procedure described in Fx~mple 4, with the addition of
sodium iodide (100 mg) to the reaction mixtllre. The title compound
was obtained after purification by reverse phase preparative HPLC
(gradient elution with 25%-65% acetonitrile/0.1% TFA; 75%-35%
0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms (m+l)
501. Anal. Calc. for C30H33ClN4O ~ 4.8 H2O ~ 2.0 TFA: C, 50.07; H,
5.51; N, 6.87. Found: C, 50.10; H, 4.25; N, 6.48.
EXAMPLE 31
1 - [1 -(4-Bromobenzyl)imidazol-5-ylmethyl] -2(S)-n-butyl-4-(1 -
naphthoyl)piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1 -naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-bromobenzylbromide (0.053 mg, 0.21 mrnol)
according to the procedure described in Example 4, with the addition of
sodium iodide (100 mg) to the reaction mixture. The title compound
was obtained after purification by reverse phase preparative HPLC
(gradient elution with 30%-65% acetonitrile/0.1% TFA; 70%-35%
0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms (m+l)
545. Anal. Calc. for C30H33BrN4O ~ 1.7 H2O ~ 2.0 TFA: C, 50.78; H,
4.81; N, 6.97. Found: C, 50.81; H, 4.39; N, 6.88.
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EXAMPLE 32
-
2(S)-n-Butyl-4-(1-naphthoyl)-1-[1-(4-trifluoromethylbenzyl)imidazol-5-
ylmethyll-piperazine dillinuoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-l-[S-
(3-triphenylmethylimi~ olyl)methyl]-4-(1-naphthoyl)pipera_ine (0.124
g, 0.200 mmol) and 4-trifluoromethylbenzylbromide (0.053 mg, 0.21
mmol) according to the procedure described in Fx~mple 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 30%-65% acetonillilelo.l% TFA; 70%-
35% 0.1% aqueous TFA over 50 min.) and lyophili~tion. FAB ms
(m+l) 535. Anal. Calc. for C30H33BrN4O ~ 2.0 TFA: C, 55.12; H,
4.63; N, 7.35. Found: C, 57.46; H, 4.98; N, 7.84.
EXAMPLE 33
2(S)-n-Butyl-1-[1-(4-methylbenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)-piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimirl~7.olyl)methyl]-4-(l-naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-methylbenzylbromide (0.029 mL, 0.21 mmol)
according to the procedure described in Fx~mple 4, with the addition of
sodium iodide (100 mg) to the reaction mixture. The title compound
was obtained after purification by reverse phase preparative HPLC
(gradient elution with 30%-65% acetonitrile/0.1% TFA; 70%-35%
0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms (m+l)
481. Anal. Calc. for C31H36N4O ~ 2.6 H2O ~ 2.0 TFA: C, 55.64; H,
5.76; N, 7.42. Found: C, 55.61,- H, 5.09; N, 7.43.
.
-
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EXAMPLE 34
2(S)-n-Butyl-1-[1-(3-methylbenzyl)imidazol-5-ylmethyl]-4-(1-
naphthoyl)-piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1-naphthoyl)piperazine (0.124
g, 0.200 mmol) and 3-methylbenzylbromide (0.029 mL, 0.21 mmol)
according to the procedure described in Example 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 25%-65% acetonitrile/0.1% TFA; 75%-
35% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
(m+l) 481. Anal. Calc. for C31H36N4O ~ 2.0 TFA: C, 67.26; H, 6.92;
N, 10.12. Found: C, 69.60; H, 6.98; N, 10.51.
EXAMPLE 35
1-[1-(4-Phenylbenzyl)imicl~7.ol-5-ylmethyl]-2(S)-n-butyl-4-(l-
naphthoyl)-piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimidazolyl)methyl]-4-(1-naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-phenylbenzylbromide (0.029 mL, 0.21 mmol)
according to the procedure described in F.x~mple 4. The title
compound was obtained after purification by reverse phase preparative
HPLC (gradient elution with 30%-65% acetonitrile/0.1% TFA; 70%-
35% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
(m+l) 543. Anal. Calc. for C36H3gN4o ~ 4.95 H2O ~ 2.0 TFA: C,
55.87; H, 5.85; N, 6.52. Found: C, 55.55; H, 4.58; N, 6.23.
EXAMPLE 36
2(S)-n-Butyl-4-(1-naphthoyl)-1-[1-(2-phenylethyl)imidazol-5-ylmethyl]-
piperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-l-[S-
(3-triphenylmethylimidazolyl)methyl]-4-(1 -naphthoyl)piperazine (0.124
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g, 0.200 mmol) and 2-phenylethylbromide (0.029 mL, 0.21 mmol)
according to the procedure described in Example 4, except with the
addition of sodium iodide (120 mg) and refluxing for 12 h. The title
compound was obtained after purification by reverse phase preparative
5 HPLC (gradient elution with 30%-65% acetonitrile/0.1% TFA; 70%-
35% 0.1% aqueous TFA over 50 min.) and lyophili7~tion. FAB ms
(m+l) 481. Anal. Calc. for C36H3gN4O ~ 4.20 H2O ~ 2.0 TFA: C,
53.60; H, 5.96; N, 7.14. Found: C, 53.54; H, 4.86; N, 6.86.
EXAMPLE 37
2(S)-n-Butyl-4-(l-naphthoyl)-1-[1-(4-trifluoromethoxy)imitl~7.ol-5-
ylmethyllpiperazine ditrifluoroacetic acid salt
The title compound was prepared from 2(S)-n-butyl-1-[5-
(3-triphenylmethylimi~ olyl)methyl]-4-(1-naphthoyl)piperazine (0.124
g, 0.200 mmol) and 4-trifluoromethoxybenzylbromide (0.032 mT~, 0.21
mmol) according to the procedure described in F.x~mple 4, except with
the addition of sodium iodide (120 mg) and refluxing for 12 h. The
title compound was obtained after puri~lcation by reverse phase
preparative HPLC (gradient elution with 35%-70% acetonitrile/0.1%
TFA; 65%-30% 0.1% aqueous TFA over 50 min.) and lyophili7~tion.
FAB ms (m+l) 551. Anal. Calc. for C31H33F3N4O2 ~ 4.00 H2O ~ 2.0
TFA: C, 49.42; H, 5.09; N, 6.59. Found: C, 48.95; H, 4.06; N, 6.26.
EXAMPLE 38
Preparation of 1 - { [1 -(4-cyanobenzyl)- lH-imidazol-5-yl] acetyl }-2(S)-n-
butyl-4-(1-naphthoyl)piperazine trifluoroacetate
30 Step A: Preparation of lH-Imidazole-4- acetic acid methyl ester
hydrochloride.
A solution of lH-imidazole-4-acetic acid hydrochloride
(4.00g, 24.6 mmol) in methanol (100 ml) was saturated with gaseous
hydrogen chloride. The resulting solution was allowed to stand at room
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temperature (RT) for 18hr. The solvent was evaporated in vacuo to
afford the title compound as a white solid.
lH NMR(CDCl3, 400 MHz) o 8.85(1H, s), 7.45(1H, s), 3.89(2H, s) and
3.75(3H, s) ppm.
s
Step B: Preparation of l-(Triphenylmethyl)-lH-imidazol-4-ylacetic
acid methyl ester.
To a solution of the product from Step A (24.85g,
0.141mol) in dimethyl form~micle (DMF) (115m1) was added
triethylamine (57.2 ml, 0.412mol) and triphenylmethyl bromide(SS.3g,
0.171mol) and the suspension was stirred for 24hr. After this time, the
reaction mixtllre was diluted with ethyl acetate (EtOAc) (1 L) 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.
lH NMR (CDC13, 400 MHz) o 7.35(1H, s), 7.31(9H, m), 7.22(6H, m),
6.76(1H, s), 3.68(3H, s) and 3.60(2H, s) ppm.
Step C: Preparation of [1-(4-cyanobenzyl)-lH-imi~1~7ol-5-yl]acetic
acid methyl ester.
To a solution of the product from Step B (8.00g,
20.9mmol) in acetonitrile (70 ml) was added bromo-p-toluonitrile
(4.10g, 20.92 mmol) and heated at 55~C for 3 hr. After this time, the
reaction was cooled to room temperature and the resulting imidazolium
salt (white precipitate) was collected by filtration. The filtrate was
heated at 55~C for 18hr. The reaction mixture was cooled to room
temperature and evaporated in vacuo. To the residue was added EtOAc
(70 ml) and the resulting white precipitate collected by filtration. The
precipitated imidazolium salts were combined, suspended in methanol
(100 ml) and heated to re~ux 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
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solid was treated with sat aq NaHCO3 (300ml) and CH2C12 (300ml)
and stirred at room temperature for 2 hr. The organic layer was
separated, dried (MgSO4) and evaporated in vacuo to afford the title
compound as a white solid:
1HNMR(CDC13, 400 MHz) ~ 7.65(1H, d, J=8Hz), 7.53(1H, s), 7.15(1H,
d, J=8Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.
Step D: Preparation of [I-(4-cyanobenzyl)-lH-imi-1~7ol-S-yl]acetic
acid
A solution of [1-(4-cyanobenzyl)-lH-imic1~ol-5-yl]acetic
acid methyl ester (4.44g, 17.4mmol ) in THF (100ml) and 1 M lithium
hydoxide (17.4 ml, 17.4 mmol) was stirred at RT for 18 hr. 1 M HCl
(17.4 ml) was added and the THF was removed by evaporation in
vacuo. The aqueous solution was lyophilised to afford the title
compound cont~ining lithium chloride as a white solid.
lH NMR(CD30D, 400 MHz) ~i 8.22(1H, s), 7.74(1H, d, J=8.4Hz),
7.36(1H, d, J=8.4Hz), 7.15(1H, s), 5.43(2H, s) and 3.49(2H, s) ppm.
Step E: Preparation of 1-{ [1-(4-cyanobenzyl)-lH-imidazol-5-
yl]acetyl}-2(S)-n-butyl-4-(1-naphthoyl)piperazine
trifluoroacetate
To a solution of the acid from step D (100 mg, 0.35
mmol), the amine hydrochloride salt from example step (117 mg,
0.35mmol), HOOBT (58 mg, 0.35 mmol), and triethyl~mine (0.123ml,
0.88 mmol) in DMF (2ml) was added EDC (75 mg, 0.38mmol). The
reaction was stirred at room tempelalule for 16hrs, diluted with EtOAc
and the organic layer washed with sat. aq NaHCO3, brine, dried
- (Na2S043~ and evaporated in vacuo. The residue was purified by .!4,
preparative HPLC (C-18; 95:5 to 5:95 water:CH3CN cont~ining 0.1%
. 30 trifluoroacetic acid; gradient elution). Lyophilisation of the collected
fractions afforded the title compound as a white solid.
lH NMR(CD30D, 400 MHz) o 9.00-8.90(1H, m), 8.05-7.94(2H,m),
7.94-7.40(10H,m), 5.60-5.40(2H,m), 5.00-2.80(9H,m), 1.90-
1.15(4H,m) and 1.05-0.40(5H, m)ppm.
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Anal. calc'd for C32H33N5O2 1.40 TFA; 0.55H20: C, 60.65; H, 5.19;
N, 10.16. Found: C, 60.66; H, 5.17; N, 10.06.
EXAMPLE 39
s
5(S)-n-Butyl- 1 -(2,3-dimethylphenyl)-4-(4-imidazolylmethyl)piperazin-
2-one ditrifluoroacetic acid salt
Step A: N-Methoxy-N-methyl 2(S)-(tert-butoxycarbonylamino)-
hexanamide
2(S)-(tert-Butoxycarbonylamino)hexanoic acid (24.6 g,
0.106 mol), N,O-dimethylhydroxylamine hydrochloride (15.5 g, 0.15
mol), EDC hydrochloride ( 22.3 g, 0.117 mol) and HOBT (14.3 g,
0.106 mol) were stirred in dry, de~a~se-l DMF (300 mL) at 20~C under
15 nitrogen. N-Methylmorpholine was added to obtain pH 7. The reaction
was stirred overni~ht, the DMF distilled under high vacuum, and the
residue partitioned between ethyl acetate and 2% potassium hydrogen
sulfate. The organic phase was washed with saturated sodium
bicarbonate, water, and saturated brine, and dried with magnesium
20 sulfate. The solvent was removed in vacuo to give the title compound.
Step B: 2(S)-(tert-Butoxycarbonylamino)hexanal
A mechanically stirred suspension of lithium alnminllm
hydride (5.00 g, 0.131 mol) in ether (250 mL) was cooled to -45~C
25 under nitrogen. A solution of the product from Step A (28.3 g, 0.103
mol) in ether (125 mL) was added, maintaining the tempelalu~e below
-35~C. When the addition was complete, the reaction was warmed to
5~C, then recooled to -45~C. A solution of potassium hydrogen sulfate
(27.3 g, 0.200 mol) in water was slowly added, m~int~ining the
30 temperature below -5~C. After quenching, the reaction was stirred at
room temperature for lh. The mixture was filtered through Celite, the
ether evaporated, and the remainder partitioned between ethyl acetate
and 2% potassium hydrogen sulfate. After wa~hin~; with saturated
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brine, dLrying over m~gnesium sulfate and solvent removal, the title
compound was obtained.
-
Step C: N-(2,3-Dimethylphenyl)-2(S)-(tert-butoxycarbonylamino)-
hex~n~mine
2,3-Dimethyl~niline (8.32 mL, 68.3 mmol) was dissolved
in dichloroethane under nitrogen. Acetic acid was added to obtain pH 5,
and sodium triacetoxyborohydride (17.2 g, 80.8 mmol) and crushed
molecular sieves (4 g) were added. A solution of the product from Step
B (13.3 g, 62.1 mmol) in dichloroethane (80 mL) was added slowly
dropwise at 20~C. The reaction was stirred overni~ht, then quenched
with saturated sodium bicarbonate solution. The aqueous layer was
removed, the organic phase washed with sa~ulated brine and dried over
magnesium sulfate. Cryst~lli7~tion from hexane gave the title
compound.
Step D: 4-tert-Butoxycarbonyl-5(S)-n-butyl-1-(2,3-
dimethylphenyl)piperazin-2-one
A solution of the product from Step C (8.50 g, 26.5 mmol)
in ethyl acetate (250 mL) was vigorously stirred at 0~C with saturated
sodium bicarbonate (150 mL). Chloroacetyl chloride (2.33 mL, 29.1
mmol) was added, and the reaction stirred at ) 0~C for lh. The layers
were separated, and the ethyl acetate phase was washed with saturated
brine, and dried over magnesium sulfate. The crude product was
dissolved in DMF (300 mL) and cooled to 0~C under nitrogen. Sodium
hydride (1.79 g, 60% dispersion in oil, 44.9 mmol) was added
portionwise to m~int~in moderate hydrogen evolution. After 30 min,
an additional amount of sodium hydride was added (0.8 g). The
reaction was stirred another 30 min, then quenched with saturated
" 30 ammonium chloride. The DMF was distilled zn vacuo, and the residue
t partitioned between ethyl acetate and water. The organic phase was
washed with water, saturated brine, and dried over magnesium sulfate.
The crude product was chromatographed on silica gel with 20-30%
ethyl acetate in hexane to obtain the title compound.
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Step E: 5(S)-n-Butyl-1-(2,3-dimethylphenyl)-4-[4-(1-
triphenylmethylimidazolyl)methyllpiperazin-2-one
A solution of the product from Step D (0.570 g, 1.58
5 mmol) in ethyl acetate (50 mL) was cooled to -15~C under nitrogen.
HCl gas was bubbled through for 15 min, and the reaction solution
warmed to 0~C for 2h. The solvent was removed in vacuo, and the
resulting solid was dissolved in dichloroethane (20 mL). Sodium
triacetoxyborohydride (0.502 g, 2.37 mmol) and 1-triphenylmethyl-4-
10 imida_olyl carboxaldehyde (0.534 g, 1.58 mmol) was added. Thereaction was stirred overnight at 20~C then poured into saturated
sodium bicarbonate solution. The organic phase was washed with
saturated brine and dried over magnesium sulfate. Silica gel
chromatography using 4% methanol in dichloromethane as eluant
15 yielded the title compound.
Step F: 5(S)-n-Butyl-1-(2,3-dimethylphenyl)-4-(4-
imidazolylmethyl)piperazin-2-one dill;rluoroacetic acid salt
To a solution of the compound from Step E (0.233 g, 0.40
20 mmol) in dichloromethane (6 mL) was added triethylsilane (0.254 mL)
and trifluoroacetic acid (2 mL) were added, and the reaction stirred at
20~C for 2h. The volatiles were removed in vacuo, and the residue
partitioned between hexane and water-methanol. The aqueous phase
was injected onto a preparative HPLC column and purified with a mixed
25 gradient of 15%-60% acetonitrile/0.1% TFA; 85%-40% 0.1% aqueous
TFA over 50 min. The title compound was isolated after lyophili7~tion.
FAB ms (m+l) 341. Anal. Calc. for C20H2gN4O 2.0 TFA: C, 50.80;
H, 5.15; N, 9.87. Found: C, 51.31; H, 5.41; N, 10.11.
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EXAMPLE 40
5(S)-n-Butyl-4-[1-(4-cyanobenzyl)imi~1~7Ol-5-ylmethyl]-1-(2,3-
dimethylphenyl)piperazin-2-one ditrifluoroacetic acid salt
4-Cyanobenzylbromide (0.043 g, 0.22 mmol) was added at
20~C to a solution of 5(S)-n-butyl-1-(2,3-dimethylphenyl)-4-[4-(1-
triphenylmethylimidazolyl)methyl]piperazin-2-one (0.120 g, 0.21
mmol) :~rom Example 39, Step E, in acetonitrile (10 mL). After 48 h,
the solvent was removed in vacuo, and the crude product dissolved in
dichloromethane (6 mL). Triethylsilane (0.13 mr ) and trifluoroacetic
acid (2 mL) were ~ e~1, and the reaction stirred at 20~C for 2h. The
volatiles were removed in vacuo, and the residue partitioned between
hexane and water-methanol. The aqueous phase was injected onto a
reverse phase preparative HPLC column and pllrifiP~l with a mixed
gradient of 30%-60% acetol~iLIile/0.1% TFA; 70%-40% 0.1% aqueous
TFA over 50 min. The title compound was isolated after lyophili7~tion
from water-acetonitrile solution. FAB ms (m+l) 456. Anal. Calc. for
C28H33N50 ~ 0.7 H2O ~ 2.0 TFA: C, 55.28; H, 5.13; N, 10.07. Found:
C, 55.27; H, 5.20; N, 10.41.
EXAMPLE 41
4-[1-(4-Cyanobenzyl)imicl~7Ol-5-ylmethyl]-1-(2,3-dimethylphenyl)-
5(S)-(2-methoxyethyl)piperazin-2-one diLIilluoroacetic acid salt
Step A: N-Methoxy-N-methyl 4-benzyloxy-2(S)-(tert-
butoxycarbonylamino)bllt~n~mide
4-Benzyloxy-2(S)-(tert-butoxycarbonylamino)butanoic acid
(1.00 g, 3.23 mmol) was converted to the title compound following the
procedure described in Example 39, Step A, using EDC ~ HCl (0.680 g,
3.55 mmol). HOBT (0.436 g, 3.23 mmol) and N,O-
dimethylhydroxylamine hydrochloride (0.473 g, 4.85 mmol) in DMF
(50 mL) at pH 7. After workup, the title compound was obtained as a
clear gum.
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Step B: 4-(1-Benzyloxyethyl)-2(S)-(tert-butoxycarbonylamino)
butanal
The title compound was obtained by lithium ~ mimlm
5 hydride reduction of the product of Step A using the procedure
described in Example 39, Step B.
Step C: N-(2,3-Dimethylphenyl)-4-(2-benzyloxyethyl)-2-(S)-(tert-
butoxycarbonylamino)bllt~n~rnine
The title compound was prepared from the product of Step
C according to the procedure described in Example 39, Step B, using
2,3-dimethyl~niline (0.505 mL, 4.14 mmol), sodium
triacetoxyborohydride (1.20 g, 5.65 mmol) and crushed molecular
sieves (1 g) at pH 5 in dichloroethane (20 mL). The title compound was
15 obtained after purification on silica gel, eluting with 15% ethyl acetate
in hexane.
Step D: 5(S)-(2-Benzyloxyethyl)-4-tert-butoxycarbonyl-1-(2,3-
dimethylphenyl)piperazin-2-one
The title compound was prepared from the product of Step
C according to the procedure described in Example 39, Step D, using
chloroacetyl chloride (0.21 mL, 2.57 mmol) in 60 mL 1:1 ethyl
acetate:saturated sodium bicarbonate, followed by reaction of the crude
product with sodium hydride (0.373 g, 60% dispersion in oil, 9.32
25 mmol) in D~F (30 mL). After workup, the crude product was
chromatographed on silica gel with 30% ethyl acetate in hexane to
obtain the title compound.
Step E: 4-tert-Butoxycarbonyl-1-(2,3-dimethylphenyl)-5(S)-(2-
hydroxyethyl)piperazin-2-one
The product from Step D was dissolved in methanol (40
mL) and 10% Pd/C was added (0.160 g). The reaction was shaken
under 60 psi hydrogen overnight. The catalyst was removed by
filtration, and the solvent evaporated to give the title compound.
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- Step F: 4-tert-Butoxycarbonyl-1-(2,3-dimethylphenyl)-5(S)-(2-
methoxyethyl)piperazin-2-one
The product from Step E (0.241 g, 0.688 mmol) was
dissolved in DMF (10 mL) cont~ining methyl iodide (0.21 mT, 3.44
mmol) and the stirred solution cooled to 0~C under nitrogen. Sodium
hydride (0.070 g, 60% dispersion in oil, 1.72 mmol) was added and the
reaction stirred for lh. The reaction was quenched with water, and the
DMF removed under vacuum. The residue was partitioned between
ethyl acetate and water, and the organic phase washed with saturated
brine and dried over m~gnesium sulfate. The crude product was
chromatographed on silica gel with 40% ethyl :~cet~te in hexane to give
the title compound.
Step G: 1-(2,3-Dimethylphenyl)-5(S)-(2-methoxyethyl)-4-[4-(1-
triphenylmethylimidazolyl)methyllpiperazin-2-one
The product from Step F (0.113 g, 0.312 mmol) was
converted to the title compound according to the procedure described
in Example 39, Step E, except using 30% trifluoroacetic acid in
dichloromethane (10 mL) for 1 h for the initial deprotection. The
volatiles were removed in vacuo, and the residue dissolved in
dichloroethane. Triethyl~mine was added to obtain pH 5. Sodium
triacetoxyborohydride (0.100 g, 0.468 mmol) and l-triphenylmethyl-4-
imidazolylcarboxaldehyde (0.1164 g, 0.343 mmol) was ~ le~l. The
reaction was stirred overnight at 20~C then poured into saturated
sodium bicarbonate solution. The organic phase was washed with
saturated brine and dried over magnesium sulfate. Silica gel
chromatography using 5% methanol in chloroform as eluant yielded the
title compound.
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Step H: 4-[1 -(4-Cyanobenzyl)imidazol-S-ylmethyl]- 1 -(2,3-
dimethylphenyl)-5(S)-(2-methoxyethyl)piperazin-2-one
ditrifluoroacetic acid salt
The product from Step G (0.182 g, 0.312 mmol) was
S converted to the title compound according to the procedure described in
Fx~mple 40, using 4-cyanobenzylbromide (0.061 g, 0.312 mmol) in
acelo~ e (10 mT.), followed by reaction of the crude imi(l~7olium salt
with triethylsilane (0.13 mL) and trifluoroacetic acid (2 mL) in
dichloromethane (6 mT ). Purification was accompli.~hed by reverse
phase preparative HPLC with a mixed gradient of 0%-70%
acetonitrile/0.1% TFA, 100%-30% 0.1% aqueous TFA over 60 rnin.
The title compound was isolated after lyophili7~tion from water. FAB
ms (m+l) 458. Anal. Calc. for C27H3lNso2 ~ 0.35 H20 ~ 2.0 TFA: C,
53.81, H, 4.91; N, 10.21. Found: C, 53.83, H, 4.95; N, 10.29.
EXAMPLE 42
Preparation of (S)-1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-5-
imitl~7olylmethyl]-5-[2-(methanesulfonyl)ethyl]-2-piperazinone
dihydrochloride
Step A: Preparation of l-triphenylmethyl-4-(hydroxymethyl)-
imidazole
To a solution of 4-(hydroxymethyl)imidazole
hydrochloride (35.0 g, 260 mmol) in 250 mL of dry DMF at room
temperature was added triethylamine (90.6 mL, 650 mmol). A white
solid precipitated from the solution. Chlorotriphenylmethane (76.1 g,
273 mmol) in S00 mL of DMF was added dropwise. The reaction
mixture was stirred for 20 hours, poured over ice, filtered, and washed
with ice water. The resulting product was slurried with cold dioxane,
filtered, and dried in vacuo to provide the titled product as a white solid
which was sufficiently pure for use in the next step.
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c StepB: Preparation of 1-triphenylmethyl-4-(acetoxymethyl)-
., imidazole
Alcohol from Step A (260 mmol, prepared above) was
suspendLed in 500 mL of pyridine. Acetic anhydride (74 mL, 780
5 mmol) was added dl~wise, and the reaction was stirred for 48 hours
during which it became homogeneous. The solution was poured into 2
L of EtOAc, washed with water (3 x 1 L), 5% aq. HCl soln. (2 x 1 L),
sat. aq. NaHCO3, and brine, then dried (Na2S04), filtered, and
concentrated in vacuo to provide the crude product. The acetate was
10 isolated as a white powder which was sufficiently pure for use in the
next reaction.
Step C: Preparation of 1-(4-cyanobenzyl)-5-(acetoxymethyl)-
imitl~701e hydrobromide
A solution of the product from Step B (85.8 g, 225 mmol)
and a-bromo-p-tolunitrile (50.1 g, 232 mmol) in 500 mL of EtOAc was
stirred at 60 ~C for 20 hours, during which a pale yellow precipitate
formed. The reaction was cooled to room temperature and filtered to
provide the solid imi~l~7.olium bromide salt. The filtrate was
20 concelltlated in vacuo to a volume 200 mL, reheated at 60 ~C for two
hours, cooled to room tempe~ ;, and filtered again. The filtrate was
concentrated in vacuo to a volume 100 mL, reheated at 60 ~C for
another two hours, cooled to room temperature, and concentrated in
vacuo to provide a pale yellow solid. All of the solid material was
25 combined, dissolved in 500 mL of methanol, and warmed to 60 ~C.
After two hours, the solution was reconcentrated in vacuo to provide a
white solid which was triturated with hexane to remove soluble
materials. Removal of residual solvents in vacuo provided the titled
product hydrobromide as a white solid which was used in the next step
30 without further purification.
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Step D: Preparation of 1-(4-cyanobenzyl)-5-(hydroxymethyl)-
imidazole
To a solution of the acetate from Step C (50.4 g, 150
mmol) in 1.5 L of 3:1 THF/water at 0 ~C was added lithium hydroxide
monohydrate (18.9 g, 450 mmol). After one hour, the reaction was
concentrated in vacuo, diluted with EtOAc (3 L), and washed with
water, sat. aq. NaHCO3 and brine. The solution was then dried
(Na2SO4), filtered, and concentrated in vacuo to provide the crude
product as a pale yellow fluffy solid which was suf~lciently pure for use
in the next step without further purification.
Step E: Preparation of 1-(4-cyanobenzyl)-5-
imidazolecarboxaldehyde
To a solution of the alcohol from Step D (21.5 g, 101
mmol) in 500 mL of DMSO at room temper~lule was added
triethylamine (56 mL, 402 mmol), then SO3-pyridine complex (40.5 g,
254 mmol). After 45 minutes, the reaction was poured into 2.5 L of
EtOAc, washed with water (4 x 1 L) and brine, dried (Na2SO4),
filtered, and concentrated in vacuo to provide the aldehyde as a white
powder which was sufficiently pure for use in a subsequent step (Step
L) without further purification.
Step F: Preparation of (S)-2-(tert-butoxycarbonylamino)-N-
methoxy-N-methyl-4-(methylthio)bllt~n~mide
L-N-Boc-methionine (30.0 g, 0.120 mol), N,O-
dimethylhydroxylamine hydrochloride (14.1 g, 0.144 mol), EDC
hydrochloride (27.7 g, 0.144 mol) and HOBT (19.5 g, 0.144 mol) were
stirred in dry DMF (300 mL) at 20~C under nitrogen. More N,O-
dimethylhydroxylamine hydrochloride (2.3 g, 23 mmol) was added to
obtain pH 7-8. The reaction was stirred overnight, the DMF distilled to
half the original volume under high vacuum, and the residue partitioned
between ethyl acetate and sat. NaHCO3 soln. The organic phase was
washed with saturated sodium bicarbonate, water, 10% citric acid, and
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brine, and dried with sodium sulfate. The solvent was removed in
vacuo to give the title compound.
Step G: Preparation of (S)-2-(tert-butoxycarbonylamino)-4-
(methylthio)butanal
A suspension of lithium alllminllm hydride (5.02 g, 0.132
mol) in ether (500 mL) was stirred at room tempe~atul~ for one hour.
The solution was cooled to -50 ~C under nitrogen, and a solution of the
product from Step F (39.8 g, ca. 0.120 mol) in ether (200 mL) was
added over 30 rnin, m~int~ining the temperature below -40 ~C. When
the addition was complete, the reaction was warmed to 5~C, then
recooled to -45~C. Analysis by tlc revealed incomplete reaction. The
solution was rewarmed to 5 ~C, stirred for 30 ~ lules~ then cooled to
-50 ~C. A solution of potassium hydrogen sulfate (72 g, 0.529 mol) in
200 mL water was slowly ~ltle~l, m~i"l~i"i"g the tempel~lule below -20
~C. The mixture was warmed to 5 ~C, ~lltered through Celite, and
concentrated in vacuo to provide the title aldehyde.
Step H: Preparation of (S)-2-(tert-butoxycarbonylamino)-N-(3-
chlorophenyl)-4-(methylthio)bllt~n~mine
To a solution of 3-chloroaniline (10.3 mL, 97.4 mmol), the
product from Step G (23.9 g, 97.4 mmol), and acetic acid (27.8 mT.,
487 mmol) in dichloroethane (250 mL) under nitrogen was added
sodium triacetoxyborohydride (41.3 g, 195 mmol). The reaction was
stirred overni~;ht then quenched with saturated sodium bicarbonate
solution. The solution was diluted with CHC13, and the organic phase
was washed with water, 10% citric acid and brine. The solution was
dried over sodium sulfate and concentrated in vacuo to provide the
crude product (34.8 g) which was chromatographed on silica gel with
20% ethyl acetate in hexane to obtain the title compound.
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Step I: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-S-r2-(methylthio)ethvllpiperazin-2-one
A solution of the product from Step H (22.0 g, 63.8 mmol)
in ethyl acetate (150 InT ) was vigorously stirred at 0~C with saturated
sodium bicarbonate (150 ml). Chloroacetyl chloride (5.6 ml, 70.2
mmol) was added dl~wise, and the reaction stirred at 0~C for 2h. The
layers were separated, and the ethyl ~cet~t.o phase was washed with 10%
citric acid and saturated brine, and dried over sodium sulfate. After
concentration in vacuo, the resulting crude product (27.6 g) was
dissolved in DMF (300 mL) and cooled to 0~C under argon. Cesium
carbonate (63.9 g, 196 mmol) was added, and the reaction was stirred
for two days, allowing it to warm to room tempelalule. Another
portion of cesium carbonate (10 g, 30 mmol) was added, and the
reaction was stirred for 16 hours. The DMF was distilled in vacuo, and
the residue partitioned between ethyl acetate and water. The organic
phase was washed with saturated brine, and dried over sodium sulfate.
The crude product was chromatographed on silica gel with 20-25%
ethyl acetate in hexane to obtain the title compound.
Step J: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-S-r2-(methanesulfonyl)ethyllpiperazin-2-one
A solution of the product from Step I (14.2 g, 37 mmol) in
methanol (300 mL) was cooled to 0 ~C, and a solution of magnesium
monoperoxyphth~l~te (54.9 g, 111 mmol) in 210 mL MeOH was added
over 20 minlltes. The ice bath was removed, and the solution was
allowed to warm to room temperature. After 45 minlltes, the reaction
was concentrated in vacuo to half the ori~:in~l volume, then quenched by
the addition of 2N Na2s2o3 soln. The solution was poured into EtOAc
and sat NaHCO3 solution, and the organic layer was washed with brine,
dried (Na2so4)~ filtered, and concentrated in vacuo to provide the .t
crude sulfone. This material was chromatographed on silica gel with
60-100% ethyl acetate in hexane to obtain the titled compound.
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Step K: Preparation of (S)-1-(3-chlorophenyl)-5-[2-
(methanesulfonvl)ethyllpi~el azin-2-one
Through a solution of Boc-protected piperazinone from
Step J (1.39 g, 3.33 mmol) in 30 mL of EtOAc at O ~C was bubbled
5 anhydrous HCl gas. The saturated solution was stirred for 35 minlltes7
then conce~ ated in vacuo to provide the hydrochloride salt as a white
powder. This material was suspended in EtOAc and treated with dilute
aqueous NaHC03 solution. The aqueous phase was extracted with
EtOAc, and the combined organic mixtllre was washed with brine, dried
10 (Na2S~4), filtered, and concentrated in vacuo. The resulting amine was
reconcentrated from toluene to provide the titled material suitable for
use in the next step.
Step L: Preparation of (S)-1-(3-chlorophenyl)-4-[1-(4-
cyanobenzyl)-5-imidazolylmethyl]-5-[2-(methanesulfonyl)-
ethyll-2-piperazinone dihydrochloride
To a solution of the amine from Step K (898 mg, 2.83
mmol) and imicl~701e carboxaldehyde from Step E (897 mg, 4.25
mmol) in 15 mT~ of 1,2-dichloroethane was added sodium
20 triacetoxyborohydride (1.21 g, 5.7 mmol). The reaction was stirred for
23 hours, then quenched at O ~C with sat. NaHC03 solution. The
solution was poured into CHC13, and the aqueous layer was back-
extracted with CHCl3. The combined organics were washed with brine,
dried (Na2S04), filtered, and concentrated in vacuo. The resulting
25 product was purified by silica gel chromatography (95:5:0.5-90:10:0
EtOAc:MeOH:NH4Cl), and the resultant product was taken up in
EtOAc/methanol and treated with 2.1 equivalents of 1 M HCl/ether
solution. After concentrated in vacuo, the product dihydrochloride was
isolated as a white powder.
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EXAMPLE 43
Preparation of (S )- 1 -(3 -chlorophenyl)-4- [1 -(4-cyanobenzyl)-5-
imidazolylmethyl]-5-[2-(ethanesulfonyl)ethyl]-2-piperazinone
5 dihydrochloride
Step A: Preparation of (S)-N-(tert-butoxycarbonyl)homoserine
lactone
To a solution of (S)-homoserine lactone hydrochloride
10 (11.0 g, 79.9 mmol) and di-tert-butylpyrocarbonate (19.2 g, 88.0
mmol) in 160 mL of dichloromethane at 0 ~C was added
diisopropylethylamine (13.9 mL, 79.9 mmol) over 3 min. The solution
was allowed to warm to room temp~ lulc;. After 3 hours, another
portion of di-tert-butylpyrocarbonate (1.75 g, 8.0 rnmol) and
15 diisopropylethylamine (0.70 mL, 4.0 mmol) were added, and the
mixture was stirred for an additional 2.5 hours. The solution was
washed with 10% citric acid, sat. NaHCO3, and brine, dried (Na2S04),
filtered, and concentrated in vacuo. The resulting material was purified
by silica gel chromatography (50% EtOAc/hexane) to provide pure
20 titled compound.
Step B: Preparation of (S)-N-(ter~-butoxycarbonyl)homoserine
lactol
To a solution of the lactone from Step A (7.0 g, 35 mmol)
25 in 175 mL of THF at -78 ~C was added diisobutylaluminum hydride
(72.0 mL, lM in THF, 72 mmol) dropwise, while m~int~inin~ the
reaction temperature below -72 ~C. After 3 hours, another portion of
diisobutylaluminum hydride (10.0 mL, 10 mmol) was ~ le~l, followed
by another after 1 hour (20.0 mL, 20 mmol). After an additional hour,
30 the reaction was quenched with EtOAc at -78 ~C, followed by sat. Na-
K-tartrate soln., then warmed to room temperature. The solution was
poured into EtOAc, washed with brine, dried (Na2SO4), filtered, and
concentrated in vacuo. The resulting material was purified by silica gel
chromatography (50% EtOAc/hexane) to give the titled lactol.
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Step C: Preparation of (S)-3-(tert-butoxycarbonylamino)-N-(3-
chlorophenyl)-4-hydroxy- 1 -bllt~n~mine
To a solution of lactol from Step B (4.49 g, 22.2 m~ol)
S and 3-chloro~niline (2.58 mL. 24.4 mmol) in 50 mL of
dichloromethane at room temperaLule was added acetic acid (1.27 ml.
22.2 m~nol). After 10 min~ sodium triacetoxyborohydride (6.59 g, 31.1
mmol) was ~ e-l7 and the solution was stirred for 1.5 hours. The
reaction was quenched with sat. aq. NaHCO3, ~lillltecl with CH2C12, and
10 the layers were separated. The organic material was dried (Na2SO4),
filtered, and concentrated in vacuo to provide a solid which was
purified by silica gel chromatography (EtOAc/hexane) to give the titled
amine.
~5 Step D: Preparation of (S)-N-[2-(tert-butoxycarbonylamino)-4-
hydroxybutyll -2-chloro-N-(3-chlorophenyl)acetamide
The ~niline derivative from Step C (5.29 g, 16.9 m~ol)
was dissolved in 60 mL of EtOAc and 60 ml of sat. NaHCO3 soln.,
then cooled to 0 ~C. With vigorous stirring~ chloroacetyl chloride (1.48
20 mT . 18.5 mmol) was added d~ wise. After 2 hours, the reaction was
diluted with water and EtOAc, and the organic layer was washed with
brine, dried (Na2SO4), filtered, and concentrated in vacuo to provide
the titled chloroacet~micle, which was used without further purification.
~5 Step E: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-(2-hydroxyethyl)piperazin-2-one
To a solution of the chloroacetamide from Step D (6.32 g,
16.1 mmol) in 80 mL of DMF at 0 ~C was added cesium carbonate
(15.8 g, 48.3 mmol). The solution was stirred until tlc analysis
30 indicated con~uln~ion of the starting material (ca. 5 hours). The
solution was poured into EtOAc, washed with water and brine, dried
(Na2SO4), filtered, and concentrated in vacuo to provide the crude
product. This material was purified by silica gel chromatography
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(99:1:0-95:5:0.15 CHC13:MeOH:NH40H) to yield the product
cont~ining a minor amount of DMF i~ ulily.
Step F: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-[2-(methanesulfonyloxy)ethyl]piperazin-2-
one
To a solution of the alcohol from Step E (3.58 g, 10.1
mmol) in 50 mL of dichlororhethane at 0 ~C was added
diisopropylethylamine (3.5 mL, 20.2 mmol), followed by
methanesulfonyl chloride (0.936 mL, 12.1 mmol). The solution was
stirred for 45 minutes, then quenched with 10% citric acid. The
solution was washed with brine, dried (Na2S04), filtered, and
concentrated in vacuo to provide the crude product which was used in
the next step without further purification.
Step G: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-S-r2-(ethylthio)ethyllpiperazin-2-one
To a solution of the mesylate from Step F (3.6 g, 8.3
mmol) in 100 mL of DMF at 0 ~C was added sodium ethanethiolate (1.4
g, 16.6 mmol). After 2 hours, the reaction was poured into EtOAc,
washed with sat. NaHCO3 and brine, dried (Na2SO4), filtered, and
concentrated in vacuo to provide the crude product which was used in
the next step without further purification.
StepH: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-r2-(ethanesulfonyl)ethyl~piperazin-2-one
To solution of the product from Step G (3.12 g, 7.82
mmol) in methanol (50 mL) was added a solution of magnesium
monoperoxyphth~l~te (11.6 g, 23.5 mmol) in 50 mT. MeOH at room t
temperature. After 45 min~ltes, the reaction was quenched by the
addition of 2N Na2S203 soln. The solution was poured into EtOAc and
sat NaHCO3 solution, and the organic layer was washed with brine,
dried (Na2SO4), filtered, and concentrated in vacuo to provide the
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~ crude sulfone. This m~teri~l was chromatographed on silica gel wi~
2% methanol in chloroform to obtain the titled compound.
Step I: Preparation of (S)-1-(3-chlorophenyl)-5-[2-
(ethanesulfonyl)ethyllpiperazin-2-one
Through a solution of the Boc-protected piperazinone from
Step H (1.75 g, 4.06 mmol) in 20 mL of EtOAc at 0 ~C was bubbled
anhydrous HCl gas. The saturated solution was stirred for 30 minllte.s,
then concentrated in vacuo to provide the hydrochloride of title
compound as a white powder. This m~teri~l was suspended in EtOAc
and treated with dilute aqueous NaHCO3 solution. The aqueous phase
was extracted with EtOAc, and the combined orgallic mixtllre was
washed with brine, dried (Na2SO4), filtered, and concentrated in vacuo.
The resulting titled amine was reconcentrated from toluene to provide
the title compound suitable for use in the next step.
Step J: Preparation of (S)-1-(3-chlorophenyl)-4-[1-(4-
cyanobenzyl)-5-imi~1~7olylmethyl]-5-[2-
(ethanesulfonyl)ethyll-2-piperazinone dihydrochloride
To a solution of the amine from Step I (480 mg, 1.45
mmol), imicl~7ole carboxaldehyde from Step E of Fx~mple 42 (460 mg,
2.2 mmol), and acetic acid (0.415 rnL, 7.25 mmol) in 10 mL of 1,2-
dichloroethane was added sodium triacetoxyborohydride (615 mg, 2.9
mmol). The reaction was stirred for 18 hours, then quenched at 0 ~C
with sat. NaHCO3 solution. The solution was poured into CH2C12, and
the organic layer was washed with brine, dried (Na2SO4), filtered, and
concentrated in vacuo. The resulting product was purified by silica gel
chromatography (2-5% MeOH:CHC13), to give the desired product and
less polar boron complex. The latter compound was taken up in
dichloromethane (1 mL) and benzene (5 mL), treated with n-
propylamine (1 mL) for 18 hours, and concentrated in vacuo. The
residue was purified by silica gel chromatography (2-5%
MeOH:CHC13), combined with the former batch, taken up in
EtOAc/methanol, and treated with 2.1 equivalents of 1 M HCl/ether
=
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solution. After concentrated in vacuo, the product dihydrochloride was
isolated as a white powder.
EXAMPLE 44
s
Preparation of (S)- 1 -(3-chlorophenyl)-4- [1 -(4-cyanobenzyl)-5-
imidazolylmethyl]-5-[2-(ethanesulfonyl)methyl]-2-piperazinone
dihydrochloride
~0 Step A: Preparation of (S)-2-(tert-butoxycarbonylamino)-N-(3-
chlorophenyl)-3- r(triphenylmethyl)thiol- 1 -prop~n~mine
To a solution of 3-chloro~niline (0.709 mT . 6.70 mrnol) in
30 mT of dichloromethane at room tempel~ure was added 1.2 g of
crushed 4A molecular sieves. Sodium triacetoxyborohydride (3.55 g,
15 16.7 mmol) was ~-lcle~l, followed by dlu~wise addition of N-
methylmorpholine to achieve a pH of 6.5. L-S-Trityl-N-Boc-cysteinal
(3.15 g, 7.04 mmol) (prepared according to S.L. Graham et al. J. Med.
Chem., (1994) Vol. 37, 725-732) was added, and the solution was
stirred for 48 hours. The reaction was quenched with sat. aq. NaHCO3,
20 diluted with EtOAc, and the layers were separated. The organic
material was washed with brine, dried (Na2SO4), filtered, and
concellllated in vacuo to provide an oil which was purified by silica gel
chromatography (15% EtOAc/hexane) to give the title amine.
~5 Step B: Preparation of (S)-N-t2-(tert-butoxycarbonylamino)-3-
((triphenylmethyl)thio)propyl] -2-chloro-N-(3-
chlorophenyl)acetamide
The aniline derivative from Step A (2.77 g, 4.95 mmol)
was dissolved in 73 mL of EtOAc and 73 mL of sat. NaHCO3 soln.,
30 then cooled to 0 ~C. With vigorous stirring, chloroacetyl chloride
(0.533 mL. 6.69 mmol) was added dropwise. After 3 hours, the
reaction was diluted with water and EtOAc, and the organic layer was
washed with brine, dried (Na2so4)~ filtered, and concentrated in vacuo
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.
to provide crude titled chloroacetamide which was used without further
purification.
Step C: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-[S-(triphenylmethyl)thiomethyl]piperazin-
2-one
To a solution of chloroacetamide from Step B (3.29 g
crude, theoretically 4.95 mmol) in 53 mL of DMF at 0 ~C was added
cesium carbonate (4.84 g, 14.85 mmol). The solution was stirred for
48 hours, allowing it to warm to room temperature. The solution was
poured into EtOAc, washed with water and brine, dried (Na2SO4),
filtered, and concentrated in vacuo to provide the crude product as an
oil. This m~t~ was purified by silica gel chromatography (20%
EtOAc/hexane) to yield the product as a white solid.
Step D: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-(thiomethyl)piperazin-2-one
A solution of piperazinone from Step C (625 mg, 1.04
mmol) in degassed EtOAc (38 mL) and EtOH (12 mL) was warmed to
30 ~C. A solution of AgNO3 (177 mg, 1.04 mmol) and pyridine (0.084
mL, 1.04 mmol) in 8 mL of EtOH was added, and the solution was
heated to reflux. After 45 minutes, the reaction was concentrated in
vacuo, then redissolved in 26 mL of degassed EtOAc. Through this
solution was bubbled H2S gas for 2.5 minutes, then activated charcoal
was added after 4 minutes. The material was filtered through celite and
rinsed with degassed EtO~c, concentrated in vacuo, then reconcentrated
from degassed CH2C12 to provide the crude product which was used
without further purification.
Step E: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-S-r(ethylthio)methyllpiperazin-2-one
A solution of the thiol from Step D (ca. 1.04 mmol) in 3
mL of THF was added via cannula to a suspension of NaH (51.4 mg,
60% disp. in mineral oil, 1.28 mmol) in 2 mL THF at 0 ~C. After 10
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minutes, iodoethane was added (0.079 mL, 0.988 mmol), and the
solution was stirred for 1.5 hours. The reaction was poured into
EtOAc, washed with sat. NaHCO3 and brine, dried (Na2SO4), filtered,
and concentrated in vacuo to provide the crude product. This material
5 was purified by silica gel chromatography (l~o MeOH/CH2C12) to yield
the titled product.
Step F: Preparation of (S)-4-(tert-butoxycarbonyl)- 1-(3-
chlorophenyl)-S-r(ethanesulfonyl)methyllpiperazin-2-one
To a solution of the sulfide from Step E (217 mg, 0.563
mmol) in 3 mL of MeOH at 0 ~C was added a solution of m~nesium
monoperoxyphth~l~te (835 mg, 1.69 mmol) in 2 rnL MeOH. The
reaction was stirred overnight, allowing it to warm to room
temperature. The solution was cooled to 0 ~C, quenched by the addition
15 of 4 mL 2N Na2S2O3 soln., then concentrated in vacuo. The residue
was partitioned between EtOAc and sat NaHCO3 solution, and the
organic layer was washed with brine, dried (Na2SO4), filtered, and
concentrated in vacuo to provide the crude sulfone as a white waxy
solid.
Step G: Preparation of (S)- 1-(3-chlorophenyl)-4-[1-(4-
cyanobenzyl)-5-imidazolylmethyl]-5-[2-
(ethanesulfonyl)methyll-2-piperazinone dihvdrochloride
To a solution of the Boc-protected piperazinone from Step
F (224 mg, 0.538 mmol) in 5 mL of dichloromethane at 0 ~C was added
2.5 mL of trifluoroacetic acid (TFA). After 45 minutes, the reaction
was concentrated in vacuo, then azeotroped with benzene to remove the
excess TFA. The residue was taken up in 4 mL of 1,2-dichloroethane
and cooled to 0 ~C. To this solution was added 4A powdered molecular .
sieves (340 mg), followed by sodium triacetoxyborohydride (285 mg,
1.34 mmol) and several drops of triethylamine to achieve pH = 6. The
imidazole carboxaldehyde from Step E of Example 42 (125 mg, 0.592
mmol) was added, and the reaction was stirred at 0 ~C. After 2 days,
the reaction was poured into EtOAc, washed with dilute aq. NaHCO3,
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and brine, dried (Na2so4)~ filtered, and concentrated in vacuo. The
- crude product was taken up in methanol and injected onto a preparative
HPLC column and purified with a mixed gradient of 15%-50%
acetonitrile/0.1% TFA; 85%-50% 0.1% aqueous TFA over 60 min~tes.
After concentration in vacuo, the resultant product was parti~ioned
betweell dichloromethane and aq. NaHCO3 soln., and the aqueous phase
was extracted with CH2cl2~ The organic solution was washed with
brine, dried (Na2SO4), filtered, and concentrated to dryness to provide
the product free base, which was taken up in CH2C12 and treated with
2.1 equivalents of 1 M HCl/ether solution. After concentrated in vacuo,
the product dihydrochloride was isolated as a white powder.
EXAMPLE 45
Preparation of (S)-1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-5-
imid~7.olylmethyl]-5-[N-ethyl-2-acetamido]-2-piperazinone
dihydrochloride
Step A: Preparation of (S)-4-(tert-butoxycarbonyl)-1-(3-
chlorophenyl)-5-r2-(oxo)ethyllpipe.~zin-2-one
To a solution of oxalyl chloride (0.608 mL, 6.97 mmol) in
dichloromethane (40 mL) at -78 ~C was added DMSO (0.990 mL, 13.9
mmol) over 2-3 minlltes. The solution was stirred for 10 minutes, then
a solution of the alcohol from Step E of Example 43 (2.06 g, 5.81
mmol) in 10 mL of dichloromethane was added over 5 minlltes, keeping
the reaction temperature below -70 ~C. The reaction was stirred for 10
minutes, then triethylamine (2.43 mL) was ~lderl, and the reaction was
stirred at -78 ~C for 5 minutes, then allowed to warm to room
temperature. After 45 minutes, the solution was poured into
dichloromethane, and washed with sat. NH4Cl soln., 10% citric acid
soln., water, and brine. The solution was dried (Na2SO4), filtered, and
concentrated in vacuo to provide the titled aldehyde.
,
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Step B: Preparation of (S)-4-(tert-butoxycarbonyl)-5-
(carboxymethyl)- 1 -(3-chlorophenyl)piperazin-2-one
To a solution of the aldehyde from Step A (1.52 g, 4.31
mmol) in 2-methyl-2-propanol (50 mL) and 2-methyl-2-butene (10 mL)
5 at room temperature was added a solution of sodium chlorite (585 mg,
5.17 mmol) and sodium dihydrogenphosphate (595 mg, 4.31 mmol) in
10 mL of water. The reaction turned yellow, then slowly faded to light
pink over 45 minutes. The solution was poured into EtOAc, and
washed with sat. sodium bisulfite soln. The aqueous layer was acidified
10 to pH 3 with 2.75 M KHSO4 soln., and extracted several times with
EtOAc. The combined organic extracts were dried (Na2SO4), filtered,
and concentrated in vacuo to provide the titled carboxylic acid.
Step C: Preparation of (S)-4-(tert-butoxycarbonyl)-5-[N-ethyl-2-
acetamidol-1-(3-chlorophenyl)piperazin-2-one
The product from Step B (200 mg, 0.56 mmol), ethyl~rninç
hydrochloride (114 mg, 1.4 mmol), EDC hydrochloride (140 mg, 0.73
mmol) and HOBT hydrate (113 m g, 0.84 mmol) were stirred in dry
DMF (3 mL) at 0~C under nitrogen. After one hour, the solution was
20 warmed to room temperature, and stirred overnight. The DMF was
removed in vacuo, and the residue was partitioned between ethyl acetate
and water. The organic phase was washed with 10% citric acid,
saturated sodium bicarbonate, water, and brine, and dried with sodium
sulfate. The solvent was removed in vacuo to give the title compound.
Step D: Preparation of (S)- 1 -(3-chlorophenyl)-4-[ 1-(4-
cyanobenzyl)-5-imidazolylmethyl]-5-[N-ethyl-2-acetamido]-
2-piperazinone dihydrochloride
The titled product was prepared from the product of Step C
(184 mg, 0.47 mmol) in analogy to Example 42, Steps K and L. The J
product was isolated as a white solid.
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EXAMPLE 46
-
Preparation of (+)-5-(2-butynyl)-1-(3-chlorophenyl)-4-[1-(4-
cyanobenzyl)-5-imidazolylmethyll-2-piperazinone dihydrochloride
s
Step A: Preparation of l-(methanesulfonyl)-2-butyne
To a solution of 2-butynol (10.0 mL, 134 rnmol) in 200
mL of dichloromethane at 0 ~C was added methanesulfonyl chloride
(23.4 g, 134 mmol), followed by dropwise addition of
10 diisopropylethylamine (30 mL, 174 mmol). After 1.5 hours, the
solution was poured into 0.5 N KHSO4 soln, and the organic layer was
washed with brine. The solution was dried over Na2SO4, filtered, and
concentrated in vacuo to provide the titled product.
15 Step B: Preparation of (+)-ethyl 2-[(phenylmethyl)imino]-4-
hexynoate
To a solution of glycine ethyl ester hydrochloride (10.11 g,
72.4 mmol) in 200 mL of dichloromethane was added ben7~ ehyde
(7.36 mL, 72.4 mmol), triethylamine (20.0 mL, 143 mmol), and
20 magnesium sulfate (6g). The solution was stirred at room temperature
for 16 hours, filtered through a glass frit, and concentrated in vacuo.
The residue was partitioned between ether and water, and the organic
layer was washed with brine. The solution was dried over Na2SO4,
filtered, and concentrated in vacuo to provide a pale yellow oil. A
25 portion of this oil (9.90 g, 51.8 mmol) was dissolved in 200 mL of THF
and cooled to -78 ~C under nitrogen atmosphere. A solution of
potassium tert-butoxide in THF (51.8 mL of 1 M, 51.8 mmol) was
added dropwise to produce a bright red solution. After 20 minutes, a
solution of the mesylate from Step A (8.05 g, 54,4 mmol) in 20 mL of
30 THF was added dropwise via c~nmll~, and the solution was allowed to
warm to room temperature. After 2 hours, the reaction was poured
into EtOAc and washed with sat. NaHCO3 soln. and brine, dried
(Na2SO4), filtered, and concentrated in vacuo to provide the titled
product.
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Step C: Preparation of (+)-ethyl 2-~(tert-butoxycarbonyl)amino]-4-
hexvnoate
A solution of the product from Step B (ca. 51.8 mmol) was5 stirred at room temperature in 5% aqueous HCl solution (100 mL).
After 12 hours, the solution was concentrated in vacuo to give an
orange oil. This product was taken up in 50 mL of THF and sat.
NaHCO3 soln. was added (50 mL), followed by di-tert-
butylpyrocarbonate (11.3 g, 51.8 mmol) at room temperature. After 6
10 hours, the reaction was poured into EtOAc and washed with sat.
NaHCO3 soln. and brine, dried (Na2SO4), filtered, and concentrated in
vacuo to provide the titled product.
Step D: Preparation of (+)-2-[(tert-butoxycarbonyl)amino]-4-
hexynoic acid
To a solution of the product from Step C (ca. 51.8 mmol)in THF (100 mL) and water (20 mL) was added at 0 ~C a solution of
lithium hydroxide monohydrate (6.5 g, 155 mmol). The solution was
stirred for 1 hour at 0 ~C, then warmed to room temperature. After 48
20 hours, the solution was concentrated in vacuo. The aqueous mixture
was extracted with EtOAc, acidified at 0 ~C with 10% aq. HCl soln.,
then extracted with three portions of dichloromethane. The combined
dichloromethane extracts were dried (Na2SO4), filtered, and
concentrated in vacuo to provide the titled product as an orange oil.
Step E: Preparation of (+)-2-(tert-butoxycarbonylamino)-N-
methoxy-N-methyl-4-hexynamide
The product from Step D (10.58 g, 46.6 mmol), N,O-
dimethylhydroxylamine hydrochloride (9.09 g, 93.2 mrnol), HOBT
30 hydrate (9.44 g, 69.9 mmol) and triethylamine (13.0 mL, 93.2 mmol)
were stirred in dry DMF (150 mL) at 0~C under nitrogen. EDC
hydrochloride (11.5 g, 60.6 mmol) was added, and the reaction was
stirred for 3 hours. The solution was partitioned between 2:1 ethyl
acetate:hexane and water, washed with water, 10% aq. HCl, sat.
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..
NaHCO3 soln and brine, then dried with sodium sulfate. The solvent
- was removed in vacuo to give the title compound as an orange oil
Step F: Preparation of (+)-2-(tert-butoxycarbonylamino)-4-hexynal
A suspension of lithium aluminum hydride (1.56 g, 41.1
mmol) in ether (150 mL) was stirred at room temperature for 30
minutes. The solution was cooled to -55 ~C under nitrogen, and a
solution of the product from Step E (11.10 g, 41.1 mmol) in ether (150
mL) was added over 15 min, m~int~inin~; the temp~ ure below -50 ~C.
When the addition was complete, the reaction was warmed to 5 ~C, then
recooled to -40 ~C. A solution of potassium hydrogen sulfate (21.8 g)
in 25 mL water was slowly added, m~int~inin~ the tempelalule below
-35 ~C. The mixture was warmed to room temperature and stirred for
one hour, filtered through Celite, and concentrated in vacuo to provide
the title aldehyde.
Step G: Preparation of (+)-2-(tert-butoxycarbonylamino)-N-(3-
chlorophenyl)-4-butyn~mine
To a 0 ~C solution of 3-chloro~niline (4.33 mL, 40.9
mmol), the product from Step F (ca. 41 mmol), and crushed 4 A
molecular sieves (10 g) in dichloroethane (100 mL) under nitrogen was
added sodium triacetoxyborohydride (12.9 g, 61.5 mmol). The reaction
was stirred for one hour, then warmed to room tempel~alule. After 3
hours, the solution was poured into EtOAc and washed with water, sat.
NaHCO3 soln. and brine. The solution was dried over sodium sulfate
and concentrated in vacuo to provide the crude product.
Step H: Preparation of (+)-N-[2-(tert-butoxycarbonylamino)-4-
hexynyll-2-chloro-N-(3-chlorophenyl)acetamide
. 30 A solution of the product from Step G (1.68 g, 5.22 mmol)
and triethylamine (1.20 mL, 8.61 mmol) in 15 mL of CH2C12 was
cooled to 0 ~C. Chloroacetyl chloride (0.457 mT, 5.74 mmol) was
added dropwise, and the reaction was m~int~ined at 0 ~C with stirring.
After 30 minutes, another portion of chloroacetyl chloride (0.20 mL)
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and triethylamine (0.5 mL) was added dropwise. After 30 min, the
reaction was poured into EtOAc and washed with 10% aq. HCl, sat. aq.
NaHCO3 soln., and brine. The solution was dried (Na2SO4), filtered,
and concentrated in vacuo to provide a brown oil This material was
purified by silica gel chromatography (20-35% EtOAc/hexane) to yield
the titled product.
Step I: Preparation of (+)-4-(tert-butoxycarbonyl)-5-(2-butynyl)-
1 -(3 -chlorophenyl)-2-piperazinone
To a solution of the chloroacetamide from Step H (1.68 g,
4.23 mmol) in 15 mL of dry DMF at 0 ~C was added CS2CO3 (3.08 g,
9.48 mmol). The solution was stirred for 30 minlltes, then allowed to
warm to room temperature. After 14 hours, the reaction was poured
into 50% EtOAc/hexane, washed with water and brine, dried (Na2SO4),
filtered, and concentrated in vacuo to provide the crude product. This
m~teri~l was purified by silica gel chromatography (20-40%
EtOAc/hexane) to yield the titled product.
Step J: Preparation of (+)-5-(2-butynyl)- 1 -(3-chlorophenyl)-4-[1 -
(4-cyanobenzyl)-5-imidazolylmethyl]-2-piperazinone
dihydrochloride
To a solution of the Boc-protected piperazinone from Step
I (1.03 g, 2.85 mmol) in 7 mL of dichloromethane at 0 ~C was added 4
mL of trifluoroacetic acid (TFA). After 4 hours, the reaction was
warmed to room temperature, and stirred an additional 6 hours. The
solution was concentrated in vacuo, then azeotroped with benzene to
remove the excess TFA. A portion of the residue (255 mg, 0.678
mmol) was taken up in 6 mL of 1,2-dichloroethane and cooled to 0 ~C.
To this solution was added 4A powdered molecular sieves (600 mg),
followed by sodium triacetoxyborohydride (214 mg, 1.02 mmol). The
imidazole carboxaldehyde from Step E of Example 42 (186 mg, 0.881
mmol) was added, and the reaction was stirred at 0 ~C. After 24 hours,
the reaction was poured into EtOAc, washed with dilute aq. NaHCO3,
and brine, dried (Na2so4)~ filtered, and concentrated in vacuo. This
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material was purified by silica gel chromatography (2-5%
MeOH:CH2Cl2) to yield a white foam which was taken up in CH2C12
and treated with 2.1 equivalents of 1 M HCl/ether solution. After
concentrated in vacuo, the product dihydrochloride was isolated as a
white powder.
EXAMPLE 47
Preparation of 1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)imidazolyl-
methyll-2-piperazinone dihydrochloride
Step A: Preparation of N-(3-chlorophenyl)ethylene~ mine
hydrochloride
To a solution of 3-chloroaniline (30.0 mL, 284 mmol) in
500 mL of dichloromethane at 0 ~C was added dlo~wise a solution of 4
N HCl in 1,4-dioxane (80 mL, 320 mmol HCl). The solution was
warmed to room tempeldlul~, then concentrated to dryness in vacuo to
provide a white powder. A mixture of this powder with 2-
oxazolidinone (24.6 g, 282 mmol) was heated under nitrogen
atmosphere at 160 ~C for 10 hours, during which the solids melted, and
gas evolution was observed. The reaction was allowed to cool, forming
the crude ~ mine hydrochloride salt as a pale brown solid.
Step B: Preparation of N-(tert-butoxycarbonyl)-N'-(3-
chlorophenvl)ethylenediamine
The amine hydrochloride from Step A (ca. 282 mmol,
crude material prepared above) was taken up in 500 mL of THF and
500 mL of sat. aq. NaHCO3 soln., cooled to 0 ~C, and di-tert-
butylpyrocarbonate (61.6 g, 282 mmol) was added. After 30 h, the
reaction was poured into EtOAc, washed with water and brine, dried
(Na2SO4), filtered, and concentrated in vacuo to provide the titled
carbamate as a brown oil which was used in the next step without
further purification.
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Step C: Preparation of N-[2-(tert-butoxycarbamoyl)ethyl]-N-(3-
chlorophenyl)-2-chloroacetamide
A solution of the product from Step B (77 g, ca. 282
mmol) and triethylamine (67 mL, 480 mmol) in 500 mL of CH2C12 was
5 cooled to 0 ~C. Chloroacetyl chloride (25.5 mT, 320 mmol) was added
dropwise, and the reaction was m~int~ined at 0 ~C with stirring. After
3 h, another portion of chloroacetyl chloride (3.0 mL) was added
dl~wise. After 30 min, the reaction was poured into EtOAc (2 L) and
washed with water, sat. aq. NH4Cl soln, sat. aq. NaHCO3 soln., and
10 brine. The solution was dried (Na2SO4), filtered, and concentrated in
vacuo to provide the chloroacetamide as a brown oil which was used in
the next step without further purification.
Step D: Preparation of 4-(tert-butoxycarbonyl)-1-(3-chlorophenyl)-
2-piperazinone
To a solution of the chloroacetamide from Step C (ca. 282
mmol) in 700 mL of dry DMF was added K2CO3 (88 g, 0.64 mol).
The solution was heated in an oil bath at 70-75 ~C for 20 hours, cooled
to room temperature, and concentrated in vacuo to remove ca. 500 mL
of DMF. The rem~ininp material was poured into 33~o EtOAc/hexane,
washed with water and brine, dried (Na2SO4), ~lltered, and
concentrated in vacuo to provide the product as a brown oil. This
m~teri~l was puri~led by silica gel chromatography (25-50%
EtOAc/hexane) to yield pure product, along with a sample of product
(ca. 65% pure by HPLC) cont~ining a less polar impurity.
Step E: Preparation of 1-(3-chlorophenyl)-2-piperazinone
Through a solution of Boc-protected piperazinone from f
Step D (17.19 g, 55.4 mmol) in 500 mL of EtOAc at -78 ~C was
bubbled anhydrous HCl gas. The saturated solution was warmed to 0
~C, and stirred for 12 hours. Nitrogen gas was bubbled through the
reaction to remove excess HCl, and the mixture was warmed to room
temperature. The solution was concentrated in vacuo to provide the
hydrochloride as a white powder. This material was taken up in 300
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- mL of CH2C12 and treated with dilute aqueous NaHCO3 solution. The
aqueous phase was extracted with CH2C12 (8 x 300 m~ ) until tlc
analysis indicated complete extraction. The combined organic mixture
was dried (Na2SO4), filtered, and concentrated in vacuo to provide the
S titled free amine as a pale brown oil.
Step F: Preparation of 1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-5-
imidazolylmethyll-2-piperazinone dihydrochloride
To a solution of the amine from Step E (55.4 mmol,
10 prepared above) in 200 mL of 1,2-dichloroethane at 0 ~C was added 4A
powdered molecular sieves (10 g), followed by sodium
triacetoxyborohydride (17.7 g, 83.3 mmol). The imidazole
carboxaldehyde from Step E of Example 42 (11.9 g, 56.4 mmol) was
added, and the reaction was stirred at 0 ~C. After 26 hours, the reaction
15 was poured into EtOAc, washed with dilute aq. NaHCO3, and the
aqueous layer was back-extracted with EtOAc. The combined orgar~ics
were washed with brine, dried (Na2SO4), filtered, and concentrated in
vacuo. The resulting product was taken up in 500 mL of 5:1
benzene:CH2C12, and propyl~mine (20 mL) was added. The mixture
20 was stirred for 12 hours, then concentrated in vacuo to afford a pale
yellow foam. This material was purified by silica gel chromatography
(2-7% MeOH/CH2C12), and the resultant white foam was taken up in
CH2C12 and treated with 2.1 equivalents of 1 M HCVether solution.
After concentrated in vacuo, the product dihydrochloride was isolated
25 as a white powder.
EXAMPLE 48
5(S)-Butyl-4-[ 1 -(4-cyanobenzyl)-2-methyl-5-imidazolylmethyl]- 1-(2,3-
30 dimethylphenyl)-piperazin-2-one dihydrochloride
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Step A: 4-Bromo-2-methylimidazole-5-carboxaldehyde
4-Bromo-5-hydroxymethyl-2-methylimidazole was
prepared according to the procedure described by S. P. Watson,
Synthetic Communications, 22, 2971-2977 (1992). A solution of 4-
S bromo-5-hydroxymethyl-2-methylimidazole (4.18 g, 21.9 mmol) was
refluxed with m~ng~nese dioxide (16.1 g) in 1:1 methylene
chloride:dioxane (200 mL) for 16 h. The cooled reaction was filtered
through celite and concentrated to yield the title compound as a pale
yellow solid. lH NMR (CDCl3, 300 MHz) ~ 9.57 (lH, s), 2.52 (3H, s).
Step B: 4-Bromo-1-(4-cyanobenzyl)-2-methylimidazole-5-
carboxaldehyde
4-Cyanobenzylbromide (1.05 g, 5.39 mmol) was added to a
solution of 4-bromo-2-methylimidazole-5-carboxaldehyde (1.02 g, 5.39
mmol) in dimethylacet~ e (15 mL). The solution was cooled to
-10~C and powdered potassium carbonate (0.745 g, 5.39 mmol) ~ltle-l
The reaction was stirred at -10~C for 2 h, and a further 4 h at 20~C.
The reaction was diluted with water and extracted with ethyl acetate.
The organic phase was washed with water, saturated brine, and dried
over m~gnesium sulfate. Solvent evaporation yielded a white solid. IH
NMR (CDCl3, 400 MHz) o 9.68 (lH, s), 7.64 (2H, d, J=7 Hz), 7.15
(2H, d, J=7Hz) 5.59 (2H, s), 2.40 (3H, s).
Step C: 1-(4-Cyanobenzyl)-2-methylimidazole-5-carboxaldehyde
A solution of 4-bromo-1-(4-cyanobenzyl)-2-
methylimidazole-5-carboxaldehyde (1.33 g, 4.37 mmol) and imidazole
(0.600 g, 8.74 mmol) in 1:1 ethyl acetate-alcohol (150 mL) was stirred
with 10% palladium on carbon (0.020 g) under 1 atm hydrogen. After
2 h, the reaction was filtered through celite and concentated to give the
title compound as a white solid. IH NMR (DMSO-d6 400 MHz) ~ 9.62
(1 H, s), 7.90 (lH, s), 7.81 (2H, d, J=8 Hz), 7.20 (2H, d, J=8 Hz), 5.64
(2H, s), 2.33 (3H, s).
-
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Step D: 5(S)-Butyl-4-[1-(4-cyanobenzyl)-2-methyl-5-
imidazolylmethyl]- 1 -(2,3-dimethylphenyl)piperazin-2-one
dihydrochloride
Sodium triacetoxyborohydride (0.265 g, 1.25 mmol) was
5 added to a solution of 1-(4-cyanobenzyl)-2-methylimidazole-5-
carboxaldehyde (0.190 g, 0.843 mmol), 5(S)-butyl-1-(2,3-
dimethylphenyl)piperazin-2-one hydrochloride (0.250 g, 0.843 mmol),
N-methylmorpholine (0.093 mL, 0.843 mmol) in dichloroethane (10
m~ ), according to the procedure described in Example 39, Step E. The
10 title compound was purified by preparative HPLC using a gradient of
70 to 45% Solvent A. The free base was isolated and converted to
dihydrochloride salt. The title compound was obtained as a white solid.
FAB ms: 470 (M+l). Anal. Calc for C2gH3sN5O ~ 2 HCl - 1.45 H2O, C,
61.25; H, 7.07; N,12.32. Found: C, 61.56; H, 6.99; N, 11.32.
EXAMPLE 49
4- [1 -(2-(4-Cyanophenyl)-2-propyl)-5-imidazolylmethyl] - 1 -(3-
chlorophenyl)-5(S)-(2-methylsulfonylethyl)piperazin-2-one
20 dihydrochloride
Step A: 4-(tert-Butyldimethylsilyloxymethyl)-l-
triphenylmethylimidazole
tert-Butyldimethylsilylchloride (2.83 g, 18.76 mmol) was
25 added to a suspension of 4-hydroxymethyl-1-triphenylmethylimidazole
(5.80 g, 17.05 mmol) in DMF (200 mL) containing imidazole (3.48 g,
51.1 mmol). After 15 min, a clear colorless solution was obtained
which was stirred at room temperature. When reaction was complete,
the DMF was removed in vacuo and the residue patitioned between ethyl
30 acetate and water. The organic phase was washed with water, saturated
brine, and dried over magnesium sulfate. The title compound was
obtained as a clear gum.
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Step B: S-tert-Butyldimethylsilyloxymethyl-1-(4-
cyanobenzyl)imidazole
A solution of 4-(tert-butyldimethylsilyloxymethyl)-1-
triphenylmethylimidazole (4.66 g, 10.26 mmol) and 1-bromomethyl-4-
cyanobenzene (2.01 g, 10.26 mmol) in acetonitrile (50 mL) was
rei~luxed 4 h. The reaction was cooled, acetonitrile removed in vacuo,
and the residue dissolved in methanol (30 mL). This solution was
refluxed for 2h, cooled and methanol evaporated. The residue was
partitioned between ethyl acetate and saturated sodium bicarbonate. The
crude product was chromatographed on silica with 3% methanol in
chloroform. The title compound was obtained as a white solid.
Step C: S-tert-Butoxydimethylsilyloxymethyl-1-[2-(4-
cyanophenyl)-2-propyl)limidazole
A solution of 5-tert-butyldimethylsilyloxymethyl-1-(4-
cyanobenzyl)imidazole (1.005 g, 3.07 mmol) in THF (25 mL) under
nitrogen was cooled to -78~C. A solution of lithium
hexamethylfli~ 7.ide (4.61 mL, 1 M in THF) was added and the reaction
stirred at -78~C for 1 h, then warmed to -60~C over 30 min. The
reaction was cooled to -78~C, methyl iodide added (0.287 mL, 4.61
mmol), and stirring continued at -78~C for an additional 2 h, before
w~rmin~ to 0~C over 2 h. After 30 min, the reaction was cooled to
-78~C, and lithium hexamethyl~ 7ide (4.61 mL, 1 M in THF) ~ 1e~1
After 1 h, methyl iodide was added (0.287 mL, 4.61 mmol) and the
reaction allowed to warm to room temperature overnight. The reaction
was quenched with water, extracted with ethyl acetate, and the organic
phase washed with saturated brine. The crude product was
chromatographed on silica gel with 6:4 ethyl acetate: methylene
chloride. The title compound was obtained as a golden oil. lH NMR
(CDCl3, 400 MHz) o 7.78 (lH, s), 7.61 (2H, d, J=8Hz), 7.15 (2H, d,
J=8Hz), 7.02, (lH, s), 4.00 (2H, s), 1.99 (6H, s), 0.79 (9H, s), -0.74
(6H, s).
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Step D: 1-[2-(4-Cyanophenyl)-2-propyl)]-5-hydroxymethyl-
imidazole
Tetra-N-butylammonium fluoride (2.99 mL, lM in THF)
was added to a solution of 5-tert-butoxydimethylsilyloxymethyl-1-[2-(4-
S cyanophenyl)-2-propyl)]imidazole (0.750 g, 2.72 mmol) in THF (10
mL). After 2 h at room temperature, the reaction was poured into ethyl
acetate and extracted with saturated sodium bicarbonate solution. The
organic phase was extracted with saturated brine, and dried over
m~n~,sium sulfate. The crude product was chromatographed on silica
10 gel with 3% methanol in ethyl acetate. The title compound was obtained
as a semi-solid. lH NMR (CDCl3, 400 MHz) ~ 7.77 (lH, s), 7.62 (2H,
d, J=8Hz), 7.19 (2H, d, J=8Hz), 7.02, (lH, s), 4.01 (2H, s), 2.57 (lH,
br s), 2.01 (6H, s).
Step E: 1-[2-(4-Cyanophenyl)-2-propyl)]imidazole-5-
carboxaldehyde
A solution of 1-[2-(4-cyanophenyl)-2-propyl)]-5-
hydroxymethylimidazole (0.450 g, 1.87 mmol) was refluxed in dioxane
20 (20 mL) with m~ng~nese dioxide (1.62 g, 18.7 mmol) for Sh. The
reaction was cooled, filtered through celite and concellLIated. The
crude product was purified by chromatography on silica gel; the title
compound was isolated as a semi-solid. lH NMR (CDCl3, 400 MHz) o
9.37 (lH, s), 8.11 (lH, s), 7.92 (lH, s), 7.58 (2H, d, J=8Hz), 7.15 (2H,
d, J=8Hz), 2.01 (6H, s).
Step F: 4-[1-(2-(4-Cyanophenyl)-2-propyl)-5-imi(1~7.olylmethyl]-l-
(3-chlorophenyl)-5(S)-(2-methylsulfonylethyl)piperazin-2-
one dihydrochloride
,s
The title compound was prepared according to the
procedure described in Example 39, Step E, except using 1-(3-
chlorophenyl)-5(S)-(2-methylsulfonylethyl)piperazin-2-one
hydrochloride (0.297 g, 0.84 mmol), 1-[2-(4-cyanophenyl)-2-
propyl)]imidazole-5-carboxaldehyde (0.200 g, 0.84 mmol), N-
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methylmorpholine (0.092 mL, 0.84 mmol) and sodium
triacetoxyborohydride (0.267 g, 1.26 mmol). The crude product was
purified by preparative HPLC with an 80 to 58% Solvent A gradient.
The pure fractions were combined and converted to the hydrochloride
5 salt, yielding the title compound. FAB ms: 540 (M+l). Anal. Calc for
C27H30ClNsO3S ~ 2 HCl ~ 3 H2O, C, 48.62; H, 5.74; N,10.50. Found: C,
48.62; H, 5.73; N, 9.89.
EXAMPLE 50
5(S)-n-Butyl-4-~1 -(4-cyanobenzyl)-5-imidazolylmethyl]-1 -(2-
methylphenyl)piperazin-2-one dihydrochloride
~tep A: N-(2-Methylphenyl)-2(S)-(tert-butoxycarbonylamino)-
hex~n~mine
The title compound was prepared according to the
procedure described in Example 39, Step C, except using o-tol~ line
(0.32 mL, 3.00 mmol), 2(S)-(tert-butoxycarbonylamino)hexanal (0.538,
2.50 mmol), sodium triacetoxyborohydride (0.795 g, 3.75 mmol) in
dichloroethane (10 mT.) The crude product was purified by column
chromatography to yield the title compound.
Step B: 4-tert-Butoxycarbonyl-5(S)-n-butyl-1-(2-
methylphenyl)piperazin-2-one
The title compound was prepared essentially according to
the procedure described in Example 39, Step D, except using N-(2-
methylphenyl)-2(S)-(tert-butoxycarbonylamino)hex~n~mine (0.506 g,
1.65 mmol), chloroacetyl chloride (0.158 mL, 1.98 mmol) in ethyl
acetate-saturated sodium bicarbonate at 0~C. The crude product thus
obtained was dissolved in DMF (15 mL), cooled to 0~C under nitrogen,
and treated with cesium carbonate (1.61 g, 4.95 mmol). The reaction
was stirred at 0~C for 2h, and at room temperature for 2h. The
reaction was quenched with saturated ammonium chloride, and
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extracted with ethyl acetate. The extracts were dried and evaporated to
give the title compound.
Step C: 5(S)-n-Butyl-4-[1-(4-cyanobenzyl)-5-
imidazolylmethyl]- 1 -(2-methylphenyl)piperazin-2-one
dihydrochloride
The product from Step B (0.534 g, 1.50 mmol) was
deprotected with trifluoroacetic acid (4 mL) in methylene chloride (10
mL). The title compound was prepared according to the procedure
described in Example 39, Step E, except using 5(S)-n-butyl-1-(2-
methylphenyl)piperazin-2-one ditrifluoroacetic acid salt, 1-(4-
cyanobenzylirnidazole)-5-carboxaldehyde (0.317 g, 1.50 mmol), and
sodium triacetoxyborohydride (0.477 g, 2.25 mmol) in dichloroethane
(15 mL). The crude product was injected onto a preparative HPLC
column and purified with a mixed gradient of acetonitrile/0.1% TFA
and 0.1% aqueous TFA. The pure fractions were combined and
converted to the HCl salt. The title compound was obtained as a white
solid. FAB ms (m+l) 442. Anal. Calc. for C27H31N5O ~ 2.5 HCl -
2.05 H;2O: C, 56.95; H, 6.66; N, 12.30. Found: C, 56.93; H, 5.75; N,
1 1.55.
EXAMPLE 51
4-[1-(4-Cyanobenzyl)-5-imidazolylmethyl]-5(S)-(2-fluoroethyl)-1-(3-
chlorophenyl)piperazin-2-one dihydrochloride
Step A: 4-tert-Butoxycarbonyl-1-(3-chlorophenyl)-5(S)-(2-
fluoroethyl)-piperazin-2-one
.~ 30 4-tert-Butoxycarbonyl-1-(3-chlorophenyl)-5(S)-(2-
methylsulfonyloxyethyl)- piperazin-2-one (0.433 g, 1.00 mmol) and
tert-butylammonium fluoride (3.0 mL, lM in THF) were stirred at
room temperature in acetonitrile (5 mL) for 72 h. The reaction was
quenched with saturated sodium bicarbonate and extracted with ethyl
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acetate. The organic e~tracts were dried, concentrated and purified by
column chromatography using 20% ethyl acetate in hexane. The title
compound was obtained as a thick oil.
Step B: 4-[1-(4-Cyanobenzyl)-5-imidazolylmethyl]-5(S)-(2-
fluoroethyl)- 1 -(3-chlorophenyl)piperazin-2-one
dihydrochloride
4-tert-Butoxycarbonyl- 1 -(3-chlorophenyl)-5(S)-(2-
10 fluoroethyl)-piperazin-2-one (0.191 g, 0.54 mmol) was deprotected
with trifluoroacetic acid (4 mL) in methylene chloride (10 mL). The
title compound was prepared according to the procedure described in
Example 39, Step E, except using 1-(3-chlorophenyl)-5(S)-(2-
fluoroethyl)piperazin-2-one ditrifluoroacetic acid salt, 1-(4-
15 cyanobenzylimidazole)-5-carboxaldehyde (0.114 g, 0.54 rnmol), and
sodium triacetoxyborohydride (0.172 g, 2.25 mmol) in dichloroethane
(5 mL). The crude product was injected onto a preparative HPLC
column and purified with a mixed gradient of acetonitrile/0.1% TFA
and O. l~o aqueous TFA. The pure fractions were combined and
20 converted to the HCl salt. The tide compound was obtained as a white
solid. FAB ms (m+l) 452. Anal. Calc. for C24H23ClFN50 ~ 2. HCl
1.7 H2O: C, 51.90; H, 5.15; N, 12.61. Found: C, 52.22; H, 5.10; N,
12.22.
EXAMPLE 52
4-[3-(4-Cyanobenzyl)pyridin-4-yl]- 1-(3-chlorophenyl)-5(S)-(2-
methylsulfonylethyl) -piperazin-2-one
~0 Step A: 3-(4-Cyanobenzyl)pyridin-4-carboxylic acid methyl ester
A solution of 4-cyanobenzyl bromide (625 mg, 3.27 mmol)
in dry THF (4mL) was added slowly over ~3 min. to a suspension of
activated Zn (dust; 250 mg) in dry THF (2 mL) at Oo under an argon
atmosphere.
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- The ice-bath was removed and the slurry was stirred at
room temperatul~ for a further 30 min. Then 3-bromopyridin-4-
carboxylic acid methyl ester (540 mg. 2.5 mmol) followed by
dichlorobis(triphenylphosphine)nickel (II) (50 mg). The resultant
re~ h-brown mixture was stirred for 3h at ~40-450c. The mixture
was cooled and distributed between EtOAc (100 ml) and 5% aqueous
citric acid (50 mL). The organic layer was washed with H20 (2X50
mL), dried with Na2SO4. After evaporation of the solvent the residue
was purified on silica gel, eluting with 35% EtOAc in hexane to give
420 mg as a clear gum. FAB ms (M+l) 253.
Step B: 3-(4-Cyanobenzyl)-4-(hydroxymethyl)pyridine
The title compound was obtained by sodium borohydride
(300 m~) reduction of the ester from Step A (415 mg) in methanol (5
mL) at room tempel~lure. After stirring for 4 h the solution was
evaporated and the product was purified on silica gel, eluting with 2%
methanol in chloroform to give the title compound. FAB ms (M+l)
225.
Step C: 3-(4-Cyanobenzyl)-4-pyridinal
The title compound was obtained by activated manganese
dioxide (l.Og) oxidation of the alcohol from Step B (240 mg, 1.07
mmol) in dioxane (10 mL) at reflux for 30 min. Filtration and
evaporation of the solvent provided title compound, mp 80-830C.
Step D: 4-[3-(4-Cyanobenzyl)pyridine-4-yl]-1-(3-chlorophenyl)-
5(S)-(2-methylsulfonylethyl)-piperazin-2-one.
This compound was prepared essentially by the same
methods as described in Example 39, Step E, except that the imidazolyl
t carboxaldehyde was replaced by an equal amount of product from Step
C (vide supra) and the piperazinone was replaced by an equal amount of
1-(3-chlorophenyl)-5(S)-(2-methylsulfonylethyl)-piperazin-2-one, there
was obtained the title compound. FAB ms 523. Anal. Calc. for
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C27H27ClN403S-0.15 CHCl3:C, 60.28; H, 5.06; N, 10.36 Found: C,
60.37; H, 5.03; N, 10.64.
EXAMPLE 53
4-[5-(4-Cyanobenzyl)- 1 -imidazolylethyl] - 1 -(3-chlorophenyl)piperazin-
2-one.
Step A: l-Trityl-4-(4-cyanobenzyl)-imidazole.
To a suspension of activated zinc dust (3.57g, 54.98mmol)
in THF (50ml)was added dibromoethane (0.315ml, 3.60mmol) and the
reaction stirred under argon at 20~C. The suspension was cooled to 0~C
and o~-bromo-p-tolunitrile (9.33g, 47.6mmol) in THF (lOOml) was
added d~c~wise over a period of 10 min. The reaction was then allowed
to stir at 20~C for 6hr and bis(triphenylphosphine)Nickel II chloride
(2.4g, 3.64mmol) and 5-iodotrityl imidazole (15.95g, 36.6mmol) was
added in one portion.The resulting mixt~lre was stirred 16hr at 20~C
and then quenched by addition of saturated NH4Cl solution (lOOml) and
the mixture stirred for 2 hours. Saturated NaHC03 solution was added
to give a pH of 8 and the solution was extracted with EtOAc (
2x250ml), dried MgS04 and the solvent evaporated in vacuo. The
residue was chromatographed (sio2~ 0-20% EtOAc/CH2C12 to afford
the title compound as a white solid.
1H NMR o CDCl3 (7.54 (2H,d, J=7.9Hz), 7.38(1H,s), 7.36-7.29
(llH,m), 7.15-7.09(6H,m), 6.58(1H,s), and 3.93(2H,s)ppm.
Step B: l-Methyl acetyl-5-(4-cyanobenzyl)-imidazole.
To a solution of 1-trityl-4-(4-cyanobenzyl)-imidazole (3.01
g, 6.91 mmol) in acetonitrile (SOml) was added methyl bromoacetate
(0.687ml, 7.26mmol) and the mixture heated at 55~C for 16 hours, and f
then the solvent was evaporated in vacuo. The solids were triturated
with EtOAc, collected by filtration and dissolved in methanol (60ml).
This suspension was heated at refux for 20min, cooled and evaporated
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to dryness. The residue was triturated with EtOAc and the title
compound was obtained as a white solid by filtration
1H NMR o CDCl3 (7.61 (2H,d, J=7.9Hz), 7.53(1H,s),
7.27(2H,d,J=7.9Hz), 6.89(1H,s), 4.47(2H,s), 3.98(2H,s) and
5 3.66(3H,s)ppm.
Step C: 1-Hydroxyethyl-5-(4-cyanobenzyl)-imi(l~zole.
A solution of 1-methyl acetyl-5-(4-cyanobenzyl)-imicl~7ole
(0.113 g, 0.472 mmol) in methanol (2ml) at 0~C was treated with
10 sodium borohydride (80.7mg, 2.1mmol). After 1 hour the reaction was
quenched by addition of saturated NH4Cl solution (2ml). Saturated
NaHCO3 was added and the mixture extracted with ethyl acetate
(3x25ml1), dried MgSO4 and evaporated in vacuo. The title compound
was isolated by chromatography (SiO2, 10%MeOH in CH2Cl2) as a
15 white solid. 1H NMR o CDCl3(7.61 (2H,d, J=7.9Hz), 7.55(1H,s),
7.27(2H,d,J=7.9Hz), 6.83(1H,s), 4.05(2H,s), 3.87(2H,t, J=5.lHz) and
3.74(2H,t, J=5.1Hz)ppm.
20 StepD: 2-(-5-(4-Cyanobenzyl)-imidazolvl)ethyl methanesulfonate
A solution of 1-hydroxyethyl-5-(4-cyanobenzyl)-imidazole
(0.532 g, 2.34 mmol) in methylene chloride (70ml) at 0~C was treated
with Hunigs base (0.489ml, 2.81mmol) and methane sulfonyl chloride
(0.219ml, 2.81mmol). After 2 hour the reaction was quenched by
25 addition of saturated NaHCO3 solution (SOml) and the mixture
extracted with methylene chloride (50ml), dried MgSO4 and the solvent
evaporated in vacuo. The title compound was used without ru~ u
purification. 1H NMR ~ CDCl3(7.62 (2H,d, J=7.9Hz), 7.54(1H,s),
7.29(2H,d,J=7.9Hz), 6.87(1H,s), 4.25(2H,t), 4.10-4.00(4H,m),
30 3.74(2H,t, J=5.lHz) and 2.90(3H,s)ppm.
-
Step E: 4-[5-(4-Cyanobenzyl)- 1 -imidazolylethyl]- 1-(3-
chlorophenyl)piperazin-2-one.
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A solution of 2-(-5-(4-cyanobenzyl)-imidazolyl)ethyl
methanesulfonate (24 mg, 0.079 mmol) in DMF (0.2ml) was added to 3-
(chlorophenyl)piperazin-2-one( 17.7mg, 0.084mmol), sodium iodide
(50mg, 0.336mmol) and Hunigs base (0,0146ml, 0.084mmol). The
5 mixt~lre was stirred at 55~C for 12 hours, and the solvent evaporated in
vacuo. The residue was purified by preparative tlc eluting with 10%
saturated ammonia/acetonitrile to afford the title compound.
1H NMR o CDC13(7.61 (2H,d, J=8.4Hz), 7.56(1H,s), 7.35-7.20 (7H,m),
7.16(1H,d, J=8Hz), 6.85(1H,s), 4.03(2H,s),, 3.83(2H,t, J=6.5Hz),
10 3.61(2H,t, J=5.4Hz), 3.27(2H,s), 2.68(2H,t, J=5.4Hz) and 2.57(2H,t,
J=6.5Hz)ppm.
EXAMPLE 54
15 In vitro inhibition of ras farnesyl transferase
Assays of farnesyl-protein transferase. Partially purified
bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and Ras-CAIL)
were prepared as described by Schaber et aL, J. Biol. Chem. 265: 14701-
14704 (1990), Pompliano, et al., Biochemistry 31:3800 (1992) and
20 Gibbs et al., PNAS U.S.A. 86:6630-6634 (1989), respectively. Bovine
FPTase was assayed in a volume of 100 ,ul cont~ining 100 mM N-(2-
hydroxy ethyl) piperazine-N'-(2-ethane sulfonic acid) (HEPES), pH 7.4,
5 mM MgCl2, 5 mM dithiothreitol (DTT), 100 mM [3H]-farnesyl
diphosphate ([3H]-FPP; 740 CBq/mmol, New Fn.~l~n~l Nuclear), 650
25 nM Ras-CVLS and 10 llg/ml FPTase at 31 ~C for 60 min. Reactions
were initiated with FPTase and stopped with 1 ml of 1.0 M HCL in
ethanol. Precipitates were collected onto filter-mats using a TomTec
Mach II cell harvestor, washed with 100% ethanol, dried and counted in ..
an LKB ~-plate counter. The assay was linear with respect to both
30 substrates, FPTase levels and time; less than 10% of the [3H]-FPP was
tili7e~1 during the reaction period. Purified compounds were dissolved
in 100% dimethyl sulfoxide (DMSO) and were diluted 20-fold into the
assay. Percentage inhibition is measured by the amount of
incorporation of radioactivity in the presence of the test compound
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when compared to the amount of incorporation in the absence of the test
- compoundL.
Human FPTase was prepared as described by Omer et al.,
Biochemistry 32:5167-5176 (1993). Human FPTase activity was
5 assayed as described above with the exception that 0.1 % (w/v)
polyethylene glycol 20,000, 10 ,llM ZnCl2 and 100 nM Ras-CVIM were
added to the reaction mixture. Reactions were performed for 30 min.,
stopped with 100 ,ul of 30% (v/v) trichloroacetic acid (TCA) in ethanol
and processed as described above for the bovine enzyme.
The compounds of the instant invention described in the
above Examples and in the Tables hereinafter were tested for inhibitory
activity against hllm~n FPTase by the assay descril~ed above and were
found to have IC50 of < 50 ,uM.
EXAMPLE 55
In vivo ras farnesylation assay
The cell line used in this assay is a v-ras line derived from
either ~atl or NIH3T3 cells, which expressed viral Ha-ras p21. The
assay is performed essentially as described in DeClue, J.E. et aL, Cancer
Research 51:712-717, (1991). Cells in 10 cm dishes at 50-75%
confluency are treated with the test compound (final concentration of
solvent, methanol or dimethyl sulfoxide, is 0.1%). After 4 hours at
37~C, the cells are labelled in 3 ml methionine-free DMEM supple-
meted with 10% regular DMEM, 2% fetal bovine serum and 400
mCi[35S]methionine (1000 Ci/mmol). After an additional 20 hours, the
cells are lysed in 1 ml lysis buffer (1% NP40/20 mM HEPES, pH 7.5/5
mM M~C12/lmM DTT/10 mg/ml aprotinen/2 mg/ml leupeptin/2 mg/ml
antipainl0.5 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 immunoprecipitated with the ras-specific
monoclonal antibody Y13-259 (Furth, M.E. et al., J. Virol. 43:294-304,
(1982)). Following a 2 hour antibody incubation at 4~C, 200 ml of a
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25% suspension of protein A-Sepharose coated with rabbit anti rat IgG
is added for 45 min. The immunoprecipitates are washed four times
with IP buffer (20 nM HEPES, pH 7.5/1 mM EDTA/1% Triton X-
100Ø5% deoxycholate/0.1%/SDS/0.1 M NaCl) boiled in SDS-PAGE
sample buffer and loaded on 13% acrylamide gels. When the dye front
reached the bottom, the gel is fixed, soaked in Fnlightening, 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.
EXAMPLE 56
In vivo ~rowth inhibition assay
To determine the biological consequences of FPTase
inhibition, the effect of the compounds of the instant invention on the
anchorage-independent growth of Ratl cells transformed with either a
v-ras, v-raf, or v-mos oncogene is tested. Cells transformed by v-Raf
and v-Mos maybe included in the analysis to evaluate the specificity of
in.c~nt compounds for Ras-induced 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
appropriate concentration of the instant compound (dissolved in
methanol at 1000 times the final concentration used in the assay). The
cells are fed twice weekly with 0.5 ml of medium A cont~ining 0.1%
methanol or the concentration of the instant compound.
Photomicrographs are taken 16 days after the cultures are seeded and
comparisons are made.
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EXAMPLE 57
Tables 1-18 show other compounds of the instant invention
that were prepared by the procedures described in Examples 1-53.
5 These compounds are meant to be illustrative and are not meant to be
limiting. In Table 2, the stereochemistry of the arnino-cont~inin~
carbon atom is as indicated (R or S) or, if unknown, the two separated
stereoisomers are delineated by an "A" or "B" de~i~n~tion.
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Tablel
~ O
R-S ~ N N
NH2 ~J
R FAB ms(m+1) Formula
CH2 C33H39N3OS2-3 TFA0-9 H2O
526 C,56.16;H,5.40;N,7.22.
C,56.12;H,5.37;N,7.39.
C30H39N3O2S2-55 TFA0.55 H2O
CH3O ~ CH2 506 C,53.19;H,5.46;N,5.36.
C,53.17;H,5.46;N,5.60.
C29H36N3OS2-25 TFA0.35 H2O
F ~ CH2 494 C,53.19;H,5.18;N,5.55.
C,53.16;H,5.18;N,5.68.
C30H39N30S-2 TFA0-9 H2O
CH2CH2 490 C,55.64;H,5.88;N,5.73.
C,55.57;H,5.81;N,5.83.
~ C29H35F2N3OS2-4 TFA0.6 H2O
</ \~ CH2 512 C,50.99;H,4.89;N,5.28.
~ C,50.98;H,4.86;N,5.56.
F
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;
Table 2
r
y
u~
FAB mass
amine spectrum
stereo. X Y (M+l) Analysis (calcd.. found)
A n-Bu 2-O(CH2)13CH3 642.5 C42H63N3O2-2.55HCl
C, 68.46; H, 8.97; N, 5.70.
C, 68.40; H, 8.70; N, 5.61.
l S B n-Bu 2-0(CH2) 13CH3 642.6 C42H63N302-2.65HCl
C, 68.63; H, 8.99; N, 5.72.
C, 68.66; H, 8.90; N, 5.70.
A n-Bu 2-O(CH2)11CH3 614.4 C40H59N302-2.8HCl
C, 67.09; H, 8.70; N, 5.87.
C, 67.03; H, 8.66; N, 5.76.
B n-Bu 2-O(CH2)11CH3 614.4 C40H59N3o2-2~8sHcl-o~osH2o
C, 66.84; H, 8.69; N, 5.85.
C, 67.20; H, 8.70; N, 5.45.
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FAB mass
amine spectrum
stereo. X Y (M~l) Analysis (calcd.~ found)
S A n-Bu 2-O(CH2)9CH3 586.5 C3gH5sN3O2-3HCl-0.25hexane
C, 66.18; H, 8.65; N, 5.86.
C, 66.14; H, 8.44; N, 5.50.
B n-Bu 2-O(CH2)9CH3 586.5 C3gH55N302-3HCl-0.15hexane
C, 65.97; H, 8.55; N, 5.93.
C, 66.04; H, 8.41; N, 5.57.
A n-Bu 2-o(cH2)7cH3 558 C36H5lN3o2-2.lsHcl-o.6sH2
C, 66.73; H, 8.47; N, 6.49.
C, 66.74; H, 8.46; N, 6.52.
B n-Bu 2-o(cH2)7cH3 558 C36H5lN3o2-2.l5Hcl-o.65H2o
C, 66.82; H, 8.42; N, 6.49.
C, 66.85; H, 8.43; N, 6.52.
A n-Bu 2-o(cH2)3ph 564 C37H45N302-2HC1-1.05H20
C, 67.78; H, 7.55; N, 6.41.
C, 67.78; H, 7.52; N, 6.26.
B n-Bu 2-O(CH2)3Ph 564 C37H45N3o2-2Hcl-l.o5H2o
C, 67.78; H, 7.55; N, 6.41.
C, 67.78; H, 7.44; N, 6.33.
A n-Bu 3-OCH2Ph 536 C3sH41N3O2-3.55HCl
C, 63.19; H, 6.75; N, 6.32.
C, 63.12; H, 6.61; N, 6.47.
B n-Bu 3-OCH2Ph 536 C3sH41N302-3.55HCl-0.4H20
C, 62.52; H, 6.80; N, 6.25.
C, 62.14; H, 6.69; N, 6.65.
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r
FAB mass
amine spectrum
stereo. X Y (M+l) Analysis (calcd.~ found)
s
A n-Bu 3-O(CH2)13CH3 642 C42H63N302-2.4HCl
C, 69.15; H, 9.04; N, 5.76.
C, 69.08; H, 9.08; N, 5.73.
B n-Bu 3-o(cH2)l3cH3 642.5 C42H63N3o2-2.3sHcl-o.25H2o
C, 69.15; H, 9.04; N, 5.76.
C, 69.08; H, 9.08; N, 5.73.
R n-Bu 4-O(CH2)13CH3 642 C42H63N302-2.2HCl-lH20
C, 68.14; H, 9.15; N, 5.68.
C, 68.13; H, 9.03; N, 5.68.
S n-Bu 4-O(CH2)13CH3 642 C42H63N302-2.8HCl
C, 67.79; H, 8.91; N, 5.65.
C, 67.77; H, 9.80; N, 5.64.
S nL-Bu 4-OCH2Ph 536 C35H41N302-2.15HCl-lH20
C, 66.49; H, 7.20; N, 6.65.
C, 66.48; H, 7.15; N, 6.62.
R/S H 2-o(cH2)l3cH3 586 C38HssN3o2-l.sHcl-o.3H2o
C, 66.88; H, 8.58; N, 6.16.
C, 66.82; H, 8.64; N, 6.15.
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Table 3
Me~
NC ~ ~N N~
s
FAB mass
spectrum
Y (M+l) Analysis (calcd.~ found)
3-SO2Me 506 C27H31N5O3S-3.0 HCl-0.10 H2O
C, 52.61; H, 5.59; N, 11.36.
C, 52.63; H, 5.41; N, 10.72.
3-OCF3 512.23 C27H2gN5O2F3-2.30 HCl-0.50 H2O
C, 53.66; H, 5.22; N, 11.59.
C, 53.73; H, 5.23; N, 10.86.
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~ Table 4
X ~_~ Y
NC~N~ \~
N
FAB mass
S spec~um
X Y (M+l) Analysis (calcd.~ found)
CH2OCH2Ph CF3 574 C32H30Nso2F3-l.4oHcl-o.3oH2o
C,61.02; H,5.12; N,11.12.
C,61.01; H,5.10; N,10.78.
CH2SO2E~ CF3 560 C27H28Nso3F3s
~3.20HCl-O. lOH20
C,47.86; H,4.67; N,10.34.
C,47.89; H,4.49; N,9.92.
CH2SO2Ph CF3 608 C3lH2gNsO3F3S-2.10HCl
C,54.42; H,4.43; N,10.24.
C,54.43; H,4.42; N,9.98.
CH2SOzMe Cl 512 C25H26N5O3ClS
~2.0HCl-0.25CHC13-0.25H20
C,48.97; H,4.68; N,11.31.
C,48.95; H,4.64; N,11.51.
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Table 4 (continued)
NC~ ~\
FAB mass
spectrum
X Y (M+1) Analysis (calcd.. found)
CH2SO2Et Cl 526 C26H2gN503ClS-2HCl-0.25CHCl3
C, 50.14; H, 4.85; N, 11.14.
C, 50.20; H, 4.91; N, 10.93.
CONHMe CF3 511 C26H25N6O2F3-1.90HCl-1.70H20
C, 51.20; H, 5.01; N, 13.78.
C, 51.17; H, 4.99; N, 13.36.
CONHEt CF3 525 C27H27N6O2F3-1.60HCl-0.10H20
C, 55.54; H, 4.97; N, 14.39.
C, 55.57; H, 4.87; N, 15.28.
CH2SO2Ph Cl 574 C30H28N5o3cls-2Hcl-o.3oH2o
C, 55.23; H, 4.73; N, 10.73.
C, 55.25; H, 4.57; N, 10.61.
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Table 4 (continued)
-
NC~3~N~ \~
FAB mass
S spectrum
X Y (M+l) Analysis (calcd.. found)
CONHMe Cl 477 C25H25N602Cl-2HCl
~0. lOSCHC13-0.85H20
C, 52.24; H, 5.03; N, 14.56.
C, 52.21; H, 5.07; N, 14.98.
CONHEt Cl 491 C26H27N6O2Cl-2HCl-0.25CHC13
C, 53.10; H, 4.97; N, 14.15
C, 53.32; H, 5.21; N, 13.77.
CONHc-Pr Cl 503 C27H27N6O2Cl-2HCl-0.40CHC13
C, 52.77; H, 4.75; N, 13.47.
C, 53.01; H, 4.99; N, 13.32.
CONHc-Pr CF3 537 C28H27N6O2F3
~2.45HCl-0.55dioxane
C, 53.81; H, 5.06; N, 12.47.
C, 53.76; H, 5.09; N, 12.44.
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Table 4 (continued)
NC~ ~N~N~
FAB mass
spectrum
X Y (M+l) Analysis (calcd.~ found)
NHCOMe Cl 477 C25H25N6O2Cl
~2HCl-0.60CHC13-2-0H20
C, 46.76; H, 4.84; N, 12.78.
C, 46.79; H, 4.46; N, 12.37.
CONMe2 CF3 525 C27H27N6O2F3-2.0HCl-0.10H2O
C, 54.12; H, 4.91; N, 14.02.
C, 54.10; H, 4.96; N, 13.79.
SO2Et Cl 512 C25H26N5O3ClS-2.0HCl-0.10H20
C, 51.18; H, 4.84; N, 11.94.
C, 51.24; H, 5.19; N, 11.10.
CH2SMe Cl 480 C25H26N5OClS
~0. lSCHC13-0.05H20
C, 52.83; H, 4.98; N, 12.25.
C, 52.82; H, 5.36; N, 11.85.
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- Table 4 (continued)
b
NC~ ~N N~
FAB mass
S spectrum
X Y (M+l) Analysis (calcd.. found)
(+) C=CMe Cl 458 C26H24N5OCl-2.0HCl-l.OOH20
C, 56.89; H, 5.14; N, 12.76.
C, 56.99; H, 5.20; N, 12.42.
.
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Table S
NC~, 5X--N~N~
FAB mass
spectrum
X Y (M+l) Analysis (calcd.. found)
CH2CH2 H 386 C23H23N5O-1.40HCl-0.40H20
C, 62.29; H, 5.73; N, 15.79.
C, 26.26; H, 5.71; N, 15.43.
CH2CO H 400 C23H21N5O2-2.6HCl-1.70H2O
C, 52.71; H, S.l9; N, 13.36.
C, 52.82; H, 5.21; N, 13.04.
CH2 H 372 C22H21Nso-2.oHcl.2.6oH2o
C, 53.80; H, 5.79; N, 14.26.
C, 52.86; H, 5.98; N, 13.92.
CH2 3-C1 406 C22H20N5ocl-2.5oHcl-o.9oH2o
C, 51.48; H, 4.77; N, 13.65.
C, 51.55; H, 4.75; N, 13.34.
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Table 5 (continued)
NC~ X--N N~
FAB mass
spectrum
X Y (M+1) Analysis (calcd.~ found)
NHSO~ H C2lH2oN6o3s-l~oHcl-2.6oH2o
C, 48.53; H, 5.08; N, 16.17
C, 48.60; H, 5.19; N, 15.80.
CH2CH2CO 3-C1 448 C24H22N5O2Cl-l.OHCl-l.lOH2O
C, 57.17; H, 5.04; N, 13.89.
C, 57.22; H, 4.94; N, 13.47.
CH2 2,3-C12 440 C22HlgN5OC12-2.0HCl-0.60H20
C, 50.42; H, 4.27; N, 13.36.
C, 50.51; H, 4.56; N, 12.18.
CH2 2-Br 450 C22H20NsoBr-2.oHcl-o.3oH2o
C, 49.98; H, 4.31; N, 13.25.
C, 49.94; H, 4.47; N, 12.53.
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Table S (continued)
NC ~ ,~3X--N~N~
FAB mass
S spectrum
X Y (M+l) Analysis (calcd.~ found)
CH2 3-CF3 440 C23H20N50~3-l.70HC1-0.40H20
C, 54.40; H, 4.47; N, 13.79.
C, 54.45; H, 4.49; N, 13.82.
CH2 4-Cl C22H2oN5ocl-l.soHcl-o.8oH2o
C, 55.71; H, 4.91; N, 14.77.
C, 55.81; H, 4.94; N, 14.11.
CH2CO 3-Cl C23H20Nso2cl-l.4oHcl-l.loH2o
C, 54.73; H, 4.71; N, 13.87.
C, 54.80; H, 4.73; N, 13.46.
CH2 3-F C22H20N5oF-2Hcl
~0.35CHC13-0.95H20
C, 51.50; H, 4.69; N, 13.44.
C, 51.56; H, 4.73; N, 13.30.
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Table 5 (continued)
t
NC ~ X--N N~
N
FAB mass
S spectrum
X Y (M+l) Analysis (calcd.~ found)
CH2 3-Br C22H20NsoBr-l.4oHcl-l.3oH2o
C, 50.35; H, 4.61; N, 13.34.
C, 50.36; H, 4.63; N, 12.84.
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Table 6
MeO2S
X~ N~z N N~
S FAB mass
spectrum
X Y Z (M+l) Analysis (calcd.~ found)
4-CN CH2 S-CH2CH2CO 554 C27H2gN5O4ClS- 1.30HCl- 1.20H20
C, 52.05; H, 5.13; N, 11.24.
C, 52.09; H, 5.15; N, lO.9S.
3-CN CH2 S-CH2 512 C25H26N5O3ClS-
2.0HCl-O. lOCHC13-0.60H20
C, 49.61; H, 4.86; N, 11.52.
C, 49.63; H, S.01; N, 11.14.
2-CN CH2 S-CH2 512 C25H26N5O3ClS-2HCl-0.35H2O
C, 50.79; H, 4.89; N, 11.84. 4
C, 50.82; H, 5.29; N, 11.90.
4-CN CH2 4-CH2CH2CO 554 C27H2gN5O4ClS
~2.oHcl-o.socHcl3-o.6oH2o
C, 47.36; H, 4.58; N, 10.04.
C, 47.35; H, 4.60; N, 9.66.
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Table 6 (continued)
~.
MeO2S
~N ~
S FAB mass
spectrum
X Y Z (M+l) Analysis (calcd.. found)
4-CN ~CH2)2 4-CH2CH2CO C2gH30N5O4ClS
~1 .OHCl-0.40CHC13-0.95H20
C, 50.96; H, 5.01; N, 10.46.
C, 50.91; H, 5.02; N, 10.13.
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Table 7
Y~ Cl
Xl'/. ,~ ~
FAB mass
spectrum
X Y (M+1) Analysis (calcd.~ found)
4-Me CH2CH2SO2Me 526 C26H2gN5O3ClS
~2HCl-0.35CHC13-0.85H20
C, 48.24; H, 4.92; N, 10.67.
C, 48.25; H, 4.93; N, 10.36.
4-Me H 420 C23H22NsOCl-2.0Hcl.o.goH2o
C, 54.27; H, 5.11; N, 13.76.
C, 54.21; H, 5.37; N, 12.97.
2-Me H 420 C23H22N5OCl-3.20HCl-O.10H20
C, 51.31; H, 4.76; N, 13.01.
C, 51.30; H, 4.74; N, 12.87.
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TABLE 8
n-C4H~
NC ~ ~O
Fo~nula
FAB ms
Y (m+l) Analysis C~c
Found
3-CF3 496 C27H2gF3N5O 2HCl 1.55 H2C
C,54.36; H,5.59; N,11.74
C,54.40; H,5.29; N,11.26
3-CH3 442 C27H3lNsO 2 HCl
C,63.01;H,6.47;N,13.61
C,63.45; H,6.71; N,13.53
2,3-(CH2)4 482 C30H3sNsO 5.2 HCl 0.1 H2C
C,53.66; H,6.06; N,10.43
C,53.62; H,5.30; N,9.35
3-OCH3 458 C27H3lNs~2 2 HCl
t C, 61.13; H,6.27; N,13.20
C,62.43; H,6.53; N,13.24
;
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TABLE 8 (continued)
n-C4Hg
~ ~,y ~r
NC ~~ ~N N~
Formula
y FAB ms Analysis Found
3-Cl 463C26H28ClNsO 2HCl 0.75 H2C
C, 56.94; H, 5.79; N, 12.77
C, 56.92; H, 5.59; N, 12.30
2-CH3 3-Cl 477C27H30ClNsO2 2HCl 3.1 H2O
C, 53.62; H, 6.37; N, 11.58
C, 53.44; H, 5.37; N, 10.76
H 428C26H29NsO 3.9 HCl 2 H2O
C,51.63;H,6.51;N, 11.58
C, 51.59; H, 5.10; N, 10.35
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TABLE 8 (continued)
..
n-C4Hg
NC ~~ ~\ ~Y
Formula
Y (m+l) Analysis Found
3-F 446C26H2sFN50 2 HCl 2.55 H2C
C, 55.87; H, 6.22; N, 12.53
C, 55.83; H, 5.84; N, 11.71
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TABLE 9
R CF3
NC~ ~N N~
Formula
FAB ms
R (m+1) Analysis C~c
Found
CH2CH2OCH2CF3 566 C27H2sF6NsO2 2 HCl 0.67 H2O
C,49.85; H,4.39; N,10.77
C,49.86; H,4.26; N,10.33
OCH2CH2 524 C28H28F3NsO2 2 HCl
C,56.38; H,5.07; N,11.74
C,56.21;H,5.27;N,11.46
CH2CH2N3 509 C2sH23F3NgO2 2 HCl 2.35 H2O
C,48.14; H,4.80; N,17.96
C,48.79; H,4.38; N,16.99
CH2CH2NHCOCH3 525 C27H27F3N6O2 2 HCl 0.7 H2O
C,53.16; H,5.02; N,13.78
C,53.30; H,5.07; N,13.39
CH2CH2NHCOC2Hs 539 C2gH2gF3N6O2 2 HCl 3.66 H2O
C,52.43; H,5.39; N,13.10
C,52.44; H,5.20; N,12.48
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TABLE 9 (continued)
~.
F~ ~ CF3
NC ~\
Formula
FAB ms
R (m+l)Analysis Calc
Found
CH2CH2SO2CH(CH3)2 574C2gH3oF3NsO3S 2HCl 1.35 H2C
C, 50.13; H, 5.21; N, 10.44
C, 50.10; H, 4.88; N, 10.08
CH2CH2SCH(CH3)2 542 C2sH3oF3Nsos 2 HCl 0.75 H2O
C, 53.55; H, 5.38; N, ll.Sl
C, 53.57; H, 5.33; N, 11.04
CH2CH2S(O)CH(CH3)2 558 C2sH3oF3Nso2s 2 HCl 0-7 H2O
C, 52.29; H, 5.23; N, 10.89
C, 52.27; H, 5.08; N, 10.35
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TABLE 10
n~C4Hs CH3 CH3
~\ )=~ '
R--N N~
R FAB ms Formula
(m+l)Analysis Calc
Found
02N~N/~ 462C26H3lNsO3 2 CF2CO2H 2.8 H2C
C, 48.69; H, 5.26; N, 9.46
C,48.70;H,4.42;N,9.12
NC ~ CH3 470 0 6 Cs Hs 2 HCl 0.3 CH2C12
C, 63.76; H, 7.48; N, 11.30
CH2 C, 64.77; H, 7.07; N, 11.26
431 C27H34N4O 2 HCl
C,64.41;H,7.21;N, 11.13
C, 66.35; H, 7.28; N, 11.25
~CH2 C28H36N4O 2 HCl 1-3 H20
C, 62.17; H, 7.56; N, 10.36
NC~ C, 62.15; H, 7.09; N, 9.83
N CH2 470 C29H3sNsO 2HCl 3.05 H20
C, 58.30; H, 7.27; N, 11.72
N C, 58.30; H, 6.28; N, 10.90
~¢~\N~ 548, SS0 C2gH34BrNsO 2 HCl 0.1 H2O
NC ~'N~ Br C, 55.89; H, 5.85; N, 11.24
C, SS.91; H, 5.96; N, 10.73
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TABLE 11
H3C- S02
., ~ Y
~ ~<
R - N N~
Fonnu1a
R Y (m+1) ~nalysis FC~Cd
Cl 511 C26H27ClN4O3s
N 2 HCl 2.85 H2O
C,49.16;H,5.51;N,8.82
C,49.17; H,4.74; N,8.87
~ F 496 C2sH26FNso3s
NC N 2 HCl 1.65 H2O
C,50.20; H,5.27; N,11.71
C,50.19; H,4.90; N,11.44
CH3
~N--~CH2 Cl 527 C26H28ClNsO3S
NC ~J ~ ~ 2 HCl 0.55 H20
N C,51.29;H,5.15;N,11.50
C,51.32; H,5.39; N,11.31
r P
r~ ~
HN N- C --<\ />-- CN Cl 530 C26H32ClNsO3S
H2 ~ Y 3 HCl 0.6 HCl
C,48.02; H,5.61; N,10.77
C,48.30; H,5.91; N,9.78
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TABLE 12
H3C- S02
\ Cl
R--N N~
o
Fonnula
FAB ms
R (m+1) An~ysis C~c
CH3 Found
~ CH2 C26H30ClN504S
H2N~ N~ 545 2 HCl 0.75 H2O
~ C,49.53; H,5.36; N,11.11
C,49.56; H,5.58; N,10.65
552 C2gH2sClNsO3S
N C, 53.90; H, S.01; N,11.22
C,54.06; H,5.78; N,10.67
396 Cl7H20clN4O3s
N C,42.41 ; H,4.90; N,11.64
CH2 C, 42.98; H,5.49; N,11.06 L
HN N-C~CN 544 C26H30clNsO4s
--/ O 2 HCl 0.75 H20
C,49.51;H,5.36;N,11.11
C,49.74; H,5.66; N,10.11
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TABLE 12 (continued)
H3C- S02
R--N N~
Fonnula
FAB ms
R (m+l)~nalysis Calc
Found
N~
~CH2
< 524 C27H27ClN4O3S 0.55 CHCl3
P~
C,58.67; H,4.92; N,10.03
CN C, 58.71; H,4.94; N,9.95
~,,CH2
~,
1 523 C28H28ClN3O3S 0.1 CHCl
0.2 CH30H
C,62.89; H,5.39; N,7.78
CN C, 62.87; H,5.37; N,7.84
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TABLE 13
H3C- SO2
~ Cl
~ ~<
R--N N~
o Fonn~a
R (m+l) Analysis FC~cd
~ 563 C3~H3lClN403S
Ph N 2 HCl 1.8 H2O
C,53.90; H,5.52; N,8.38
C,53.90; H,5.38; N,7.82
~ 540 C27H30clNso3s
NC ~ 2HCl 1.35 H2O
C,50.88; H,5.49; N,10.99
C,51.37;H,5.51;N,10.20
~CH2 512 C25H26ClNsO3s
NC 2 HCl 0.3 H2O
C,50.86; H,4.88; N,11.86
CH2 C,51.06; H,5.33; N,10.87
HN NH 415 C28H27ClN4O3s
C,36.25;H,6.42;N,9.39
CH2 C,38.12;H,5.91;N,7.41
HN N - S ~ CN 580 C25H30clNsoss2
2 HCl 0.85 H2O
C,44.93; H,5.08; N,10.48
C,44.96; H,5.08; N,9.96
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TABLE 14
"
.. R2 Cl
R1_N N~
~ Formula
FAB ms
Rl R2 (m+l)Analysis Calc
Found
N~ ~CH H 355ClgHl9clN4O
2 HCl 0.5 H2O
C,52.25; H,5.08; N,12.83
CH2 C,52.31; H,5.14; N,12.23
N~ CH2CH2F 452C24H23ClFNso
~= N 2 HCl 1.70 H20
~ C,51.90; H,5.15; N,12.61
NC/-- C, 52.22; H,5.10; N,12.22
~ ~ O C24H22ClNsO2 1 HCl
HN~ H 4480.30 CHCl3 0-85 H20
C,54.50; H,4.71; N,13.08
C,54.51; H,4.69; N,12.87
,~
~, ~
NC
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TABLE 15
R--N N~ '
Formula
R Y FAB ms Analysis Calc
(m+l) Found
CH2
N~ 2,3-(CH2)4 426 C26H27Ns~
/=<-- \=N 2 HCl ~ 0.15 H20
~ C, 62.31; H, 5.89; N, 13.97
NC C,62.38;H,6.18;N, 13.27
CH2
--N~ 2-CH3, 3-C1 420 C23H~ClN5o
\=N 2 HCl 1.0 H20
~ C, 54.08; H, 5.13; N, 13.71
NC C, 54.57; H, 5.77; N, 12.92
0~
3-C1 420 C22HlsClNsO2
N> 2 HCl 0.5 H20
<\ /~ C, 56.78; H, 4.33; N, 15.05
~ C, 56.79; H, 4.69; N, 13.47
NC
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TABLE 16
n-C4H~
Formula
R FAB ms Analysis Calc
Found
,~ , CH2CH2
NC J~ 506 C32H3sNsO 3-2 CF3CO2H
C, 54.14; H, 5.75; N, 9.15
C, 54.15; H, 5.74; N, 9.18
~CH2
~,N~ N 484 C29H33NsO2 4 HCl 3 H20
C, 51.03; H, 6.35; N, 10.26
C, 51.02; H, 5.66; N, 9.67
NC
CH2
506 C32H35N5O 2 HCl 1-7 H2C
N
C, 63.09; H, 6.68; N, 10.26
C,51.02;H,5.66;N,9.67
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Table 17
/--\ /=\ r
R--N N~
Formula
FAB ms
R (m+l) Analysis Found
N N- CH2CH2
347 C21H22N4O 0.2 CHCl3
<~ 0.4 CH30H
C, 67.71; H, 6.26; N, 14.62
C, 67.71; H, 6.26; N, 14.53
~CH2
N~
,~, 358 C23H23N3O 0.55 CHCl3
C, 66.85; H, 5.61; N, 9.93
C, 66.87; H, 5.70; N, 10.03
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Table 18
.,
FAB mass
spectrum
(M+l) Analysis (calcd.~ found)
Me
Me Me
~_3
C29H33N502- 1.OHCl-
0.25cHcl3-o-7oH2o
C, 62.45; H, 6.39; N, 12.45.
C, 62.45; H, 6.46; N, 12.37.
H~ N~
406 C22H20Nsocl-2.6oHcl
C, 53.97; H, 4.78; N, 14.30
C, 53.97; H, 5.21; N, 13.10.
MeO25 ~ ~Cl
NC ~ ~N N
.. N
526 C26H2gN503ClS-2.0HCl- 1.20H20
C, 50.32; H, 5.26; N, 11.28
C, 50.31; H, 5.31; N, 10.72.
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Table 18 (continued)
FAB mass
spectrum
(M+l) Analysis (calcd..... found)
N~ \J ~
420 C23H22N50Cl-2-OHCl-2- 1~H2O
C, 52.06; H, 5.36; N, 13.20
C, 51.99; H, 5.36; N, 12.57.
