Note: Descriptions are shown in the official language in which they were submitted.
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TITLE OF THE INVENTION
INHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
BACKGROUND OF THE INVENTION
The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ra~)
are part of a ~ignalling pathway that links cell surface growth factor
receptors to nuclear signals initiating cellular proliferation. Biological
and biochemical studies of Ras action indicate that Ras functions like
a G-regulatory protein. In the inactive state, Ra,s is bound to G~P.
Upon growth factor receptor activation Ras is induced to exchange
GDP for GTP and undergoes a conformational change. The GTP-
bound form of Ra~s propagates the growth stimulatory signal until the
signal i~ terminated 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. Bi~chem. ~2:X51-~91 (1993)). Mutated
ras genes (Ha-~ as, Ki4a-7 as, Ki4b-ras and N-ra~) are found in many
human cancers, including colorectal carcinoma, exocrine pancreatic
carcinoma, and myeloid leukemias. The protein products of the~se
genes are defective in their GTPase activity and con~stitutively transmit
a growth stimulatory ~signal.
Ras must be localized to the pla~;ma membrane for
both normal and oncogenic functions. At least 3 post-translational
modification.s are involved with Ras membrane localization, and all
3 modification~ occur at the C-terminus of Ras. The Ras C-terminus
contains a sequence motif termed a "CAAX" or "Cys-Aaal-Aaa2-Xaa"
box (Cys is cy~teine? Aaa is an aliphatic amino acid, the Xaa i~s any
amino acid) (Willumsen et al., Natu~-e 310:5~3-5~6 (19~4)). Depend-
ing on the ~pecific ~sequence, this motif serves as a ~iignal se~uence for
the enzyme~i f~rnesyl-protein transferase or geranylgeranyl-protein
transferase~ which catalyze the alkylation of the cysteine residue of the
CAAX motif with a Cl~ or C2() i~oprenoid, re~ipectively. (S. Clarke.,
Ann. Rel~. Bioehenl. ~1:355-3~6 (1992); W.R. Schafer and J. Rine,
Ann. Rel!. Genelie~ 30:209-237 (1992)). The Ra~ protein i.~ one of
~ieveral protein~ that are known to undergo po~t-translational
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farnesylation. Other farnesylated proteins include the Ra.s-related GTP-
binding proteins such as Rho, fungal mating factors, the nuclear lamins,
and the gamma subunit of transducin. Jarnes, et al., J. Biol. Chem. 269,
14182 (1994) have identified a peroxisome associated protein Pxf which
is also farnesylated. James, et al., have also sugge.sted that there are
farnesylated proteins of unknown structure and function in addition
to those listed above.
Inhibition of farnesyl-protein transferase ha.s been
.shown to block the growth of Ras-transforrned cell.s in soft agar and
to modify other aspects of their transformed phenotype. It has also
been demonstrated that certain inhibitors of farnesyl-protein tran.sferase
.selectively block the processing of the Ras oncoprotein intracellularly
(N.E. Kohl et al., S~ience, 2~0:1934-1937 (1993) and G.L. James
et al., S~ienc~e, 2~0:1937-1942 (1993). Recently, it ha.s been shown
that an inhibitor of farnesyl-protein transfera.se blocks the growth of
ras-dependent tumors in nude mice (N.E. Kohl et al., p~AO(~. Natl. Ac~ad.
Sci U.S.A ., 91 :9141 -9145 (1994) and induces regression of mammary
and salivary carcinomas in ras transgenic mice (N.E. Kohl et al., Nature
Medicine, 1:792-797 (1995).
Indirect inhibition of farnesyl-protein transfera.se i~Z ViV(~
has been demonstrated with lovastatin (Merck & Co., Rahway, NJ)
and compactin (Hancock et al., ihid; Casey et al., ihid; Schafer et C~
S~ience 245:379 (19~9)). These drug.s inhibit HMG-CoA reductase, the
rate limiting enzyme for the production of polyisoprenoid.s including
farnesyl pyrophosphate. Farnesyl-protein tranlsferase utilize.s farne~syl
pyrophosphate to covalently modify the Cys thiol group of the Ra.s
CAAX box with a farnesyl group (Rei.ss et al., Cell, f)2:Xl -~ (1990);
Schaber etal., J. Bi~l. Chen1., 2~5:14701-14704 (1990); Schafer etal.,
Scie~lce, 249:1133-1139 (1990); Manne et al., Pr0c. Natl. Acad. Sci
USA. 87:7541-7545 (1990)). Inhibition of farnesyl pyrophosphate
biosynthesis by inhibiting HMG-CoA reductase block.s R~.s membrane
localization in cultured cell.s. However, direct inhibition of farne.syl-
protein transfera.se would be more specific and attended by fewer .side
effects than would occur with the re4uired dose of a general inhibitor
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of isoprene biosynthesis.
Inhibitors of farnesyl-protein transferase (FPTase) have
been described in four general classes (S. Graham, Expe7t Opinion
Ther. Patents, (1995) 5:1269-1285). The first are analogs of farne.syl
5 diphosphate (FPP), while a second clas.s of inhibitors is related to the
protein substrates (e.g., Ras) for the enzyme. Bisubstrate inhibitor.s and
inhibitors of farnesyl-protein transferase that are non-competitive with
the substrates have also been described. The peptide derived inhibitors
that have been described are generally cysteine containing molecules
10 that are related to the CAAX motif that is the signal for protein prenyl-
ation. (Schaber et ~11., ihid; Reiss et. al., il~id; Reiss ef f~l., PNAS,
f~8:732-736 (1991)). Such inhibitors may inhibit protein prenylation
while serving as alternate substrates for the farnesyl-protein trans-
ferase enzyme, or may be purely competitive inhibitors (U.S. Patent
15 5,141,~51, Univer.sity of Texas; N.E. Kohl et al., S~ien~, 260:1934-
1937 (1993); Graham, et al., J. Med. Chen1., 37, 725 (1994)).
In general, deletion of the thiol from a CAAX derivative has
been shown to dramatically reduce the inhibitory potency of the
compound. However, the thiol group potentially place.s limitations
20 on the therapeutic application of FPTase inhibitors with respect to
pharrnacokinetics, pharmacodynamics and toxicity. Therefore, a
functional replacement for the thiol i~s desirable.
It has recently been disclo.sed that certain tricyclic
compounds which optionally incorporate a piperidine moiety are
25 inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516).
Imidazole-containing inhibitors of farnesyl protein transferase have also
been disclo.sed (WO 95/09001 and EP 0 675 112 Al).
It has recently been reported that farnesyl-protein
transferase inhibitor.s are inhibitor.s of proliferation of vasculal- smooth
30 muscle cells and are therefore useful in the prevention and therapy of
arteriosclerosis and diabetic disturbance of blood vessels (JP H7-
112930).
It is, therefore, an object of this invention to develop
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low molecular weight compounds that will inhibit farnesyl-protein
transferase and thus, the post-translational farnesylation of proteins.
It is a further object of this invention to develop chemotherapeutic
compositions containing the compounds of this invention and methods
5 for producing the compounds of this invention.
SUMMARY OF THE INVENTION
The present invention comprises biheteroaryl-containing
compound~s which inhibit the farnesyl-protein transferase. Further
10 contained in this invention are chemotherapeutic compo~ition,s
containing these farnesyl transferase inhibitors and methods for their
production,
The compounds of this invention are illustrated by the
formula A:
R6a-d
R3
(Fl8)r /(~9~ b~a~e'
V - A1 (CR12)nA2(CR 12)n ~W~l - (CR22)p - X -(CR22~ R5
A
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DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention are useful in the inhibition
of farne~yl-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:
R6a-d
R3
( l 8)r ~R9~\ bà~e'
V - A (CR 2)nA (CR 2)ntW~ (CR22)p - X -(CR22~ R5
wherem:
a is N or C;
from 0-4 of b, c, d and e are independently N, NH, O and S, and the
rem~ining b, c, d and e atoms are independently CH, provided that if a
is C, then at least one of b, c, d or e is independently N, NH, O or S;
15 a' is N or C;
from 0-4 of b', c'. d' and e' are independently N. NH, O and S, and the
remaining b', c', d' and e' atoms are independently CH, provided that if
a' i,s C, then at least one of b', c', d' or e' i~ independently N, NH, O or
20 S;
~ - Rl ~nd R2 are independently selected from:
a) hydrogen,
b) aryl. heterocycle, C3-CIo cycloalkvl. C2-C6 alkenyl,
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C2-C6 alkynyl, R~0O-, Rl 1S(O)m-, Rl lC(o)O-,
R 1 0C(O)NR 10, (R 1 0)2NC(O)-, R 1 02N-C(NR 10), CN,
NO2, R 1 ~C(O)-, N3, -N(R 1~)2, or R I 1 OC(O)NR 10
c) unsubstituted or substituted Cl-C6 alkyl wherein the
S substituent on the ~substituted Cl-c6 alkyl is selected from
unsubstituted or substituted aryl, heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R100-, RllS(O)m-, RlOC(O)NR10-, (RlO)2Nc(o)
R 1 02N-C(NR 10), CN, R I ~C(O)-, N3, -N(R 1~)2,
and R 1 1 OC(O)-NR 10;
R3, R4 and R5 are independently selected from:
a) hydrogen,
b) unsub~stituted or sub~stituted aryl, unsubstituted or
~sub,stituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 ~Ikynyl, halogen, Cl-C6 perfluoroalkyl,
R120, Rl lS(o)m, Rloc(o)NRlo-~ Rl lc(o)o-,
(R 1 0)2NC(o)-, R 1 02N-C(NR 10), CN, NO2, R 1 ~C(O)-,
N3, -N(R 1~)2, or R I 1 OC(O)NR 10,
c) unsubstituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
substituted Cl-c6 alkyl is selected from unsubstituted or
substituted aryl, unsubstituted or substituted heterocyclic,
C~-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 1 S(O)m-~ R 1 0C(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R I ~C(O)-, N3, -N(R 1~)2, and
R l 10C(O)-NR10-;
R6a, R6b R6C and R6d are independently selected from:
a) hydrogen,
b) un.substituted or substituted aryl, unsubstituted or
~sub~stituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-c6 alkynyl, halogen, Cl-c6 perfluoroalkyl,
R 1 20, R I 1 S(O)m ~ R 1 0C(O)NR 10, R 1 I C(O)O-,
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(R 1 0)2NC(O)-, R 1 02N-C(NR 10), CN, NO2, R 1 ~C(O)-,
N3, -N(R10)2, or Rl IOC(O)NRl0-,
c) un,substituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
S ~ub~itituted Cl-C6 alkyl is ~elected from unsubstituted orsubstituted aryl, unsub~tituted or ~ubstituted heterocyclic,
C3-C1o cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R120-, Rl lS(O)m-, RlOC(O)NR10-, (R10)2NC(o)-,
R 1 02N-C(NR 10), CN, R 1 ~C(O)-, N3, -N(R 1~)2. and
1 0 R 1 1 OC(O) NR 10 ;
R7 i,s selected from: H; Cl 4 alkyl, C3-6 cycloalkyl, heterocycle, aryl,
aroyl, heteroaroyl, arylsulfonyl, heteroarylsulfonyl, un~iubstituted or
substituted with:
a) C1 4 alkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
e) ~ R
f) --SO2R
g) N(R10)2 or
h) C 1-4 perfluoroalkyl;
R~ is independently selected from:
a) hydrogen,
b) aryl, sub.stituted aryl, heterocycle, C3-Clo cycloalkyl,
C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl,
Br, R10O-, Rl lS(O)m-, R10C(o)NR10-, (R10)2NC(o)-~
R102N-C(NR10)-, CN, NO2, R10C(o)-, N3, -N(R10)2, or
R 1 1 OC(O)NR 10, and
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WO g7/36585 PCTtUS97/062!j9
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
cyanophenyl, heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br,
R 1 0O-, R 1 1 S(O)m ~ R 1 0C(O)NH-, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R I ~C(O)-, N3, -N(R 1~)2, or
R 1 0OC(O)NH-;
provided that when R~ is heterocycle, attachment of R8 to V is
through a substitutable ring carbon;
10 R9 is independently selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl,
F, Cl, Br, Rl lo-, Rl lS(O)m-, R10C(O)NRl0-~
(R 1 0)2NC(O)-, R 1 02N-C(NR 10), CN, NO2, R 1 ~C(O)-,
N3, -N(R 1~)2, or R 1 1 OC(O)NR 10, and
c) Cl-C6 alkyl unsub.stituted or substituted by perfluoroalkyl,
F, Cl, Br, R10O-, Rl lS(O)m-, R10C(O)NR10-,
(R 1 0)2NC(O)-, R 1 02N-C(NR 10) , CN, R 1 ~C(O)-, N3,
-N(R 1~)2, or R I 1 OC(O)NR 10-;
R10 i.s independently selected from hydrogen, C]-C6 alkyl, 2,2,2-
trifluoroethyl, benzyl and aryl;
Rl 1 is independently selected from Cl-C6 alkyl and aryl;
R12 is independently selected from hydrogen, Cl-c6 alkyl, Cl-C6
aralkyl, Cl-C6 sub~stituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 substituted heteroaralkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2 2-trifluoroethyl;
A 1 and A2 are independently selected from: a bond, -CH=CH-, -C-C-,
-C(O)-, -C(O)NR 10, -NR I ~C(O)-, O, -N(R 10) ,
-S(O)2N(R 10), -N(R I ~)S(O)2-, or S(O)m;
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- V i~ selected from:
a) hydrogen,
b) heterocycle,
S c) aryl,
d) Cl-C20 alkyl wherein from 0 to 4 carbon atom,s are
replaced with a a heteroatom selected from O, S, and N,
and
e) C2-C20 alkenyl,
10 provided that V is not hydrogen if Al i~i S(O)m and V i~s not hydrogen
if Al i~ a bond, n is 0 and A2 ;~s S(O)nl; and
provided that when V i~ heterocycle, attachment of V to R~ and to Al i~
through a ~substitutable ring carbon;
1~ W is a heterocycle;
X i.s a bond, -CH=CH-, O, -C(=O)-, -C(o)NR7-, -NR7C(o)-, -C(O)O-,
-OC(O)-, -C(o)NR7C(o)-, -NR7-, -S(O)2N(R 10),
-N(RIo)s(o)2- or -S(=O)m-, provided that if a i.s N, then
X i,s not O, -C(o)NR7-, -C(O)O-, -C(o)NR7C(o)-,
-S(0)2N(R10)- or-NR7-;
mis 0, 1 or2;
n i,~ independently 0, 1, 2, 3 or 4;
2~ p i!i independently 0, 1, 2, 3 or 4;
q i~ 0, 1, 2 or 3;
r i!i 0 to ~. provided that r i~ 0 when V i~i hydro~en; and
t i~ 0 or 1;
30 or the pharmaceutically acceptable ~alt~ thereof.
A preferred embodiment of the compound~; of this
invention is illu~strated by the following formula A:
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- 10 -
R6a-d
(I )r ~RW~
V - A (CR 2)nA (CR 2)n - (CR22)p - X -(CR22~ R5
wherein:
a is N or C;
from 0-4 of b, c, d and e are independently N, NH, O and S, and the
rem~ining b, c, d and e atoms are independently CH, provided that if a
is C, then at least one of b, c, d or e is independently N, NH, O or S;
a' is N or C;
from 0-4 of b', c', d' and e' are independently N, NH, O and S, and the
remaining b', c'. d' and e' atoms are independently CH, provided that if
a' is C, then at least one of b', c', d' or e' i.s independently N, NH, O or
S;
Rl is independently selected from: hydrogen, C3-Clo cycloalkyl,
RlOO-, -N(R10)2, F or Cl-C6 alkyl;
R2 is independently selected from:
a) hydro~en,
b) aryl. heterocycle, C3 -C I o cycloalkyl. R l OO-, -N(R 1 ~)2
F Ol C~-C6 alkenyl,
c) un.~iubstituted or substituted Cl-C6 alky] wherein the
sub~tituent on the ~ubstituted Cl-C6 alkyl is ~;elected from
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unsubstituted or sub.stituted aryl, heterocycle, C3-Clo
Gycloalkyl, C2-C6 alkenyl, RlOO- and -N(R10)2;
R3~ R4 and R5 are independently selected from:
5 a) hydrogen,
b) unsubstituted or substituted aryl, un~substituted or
substituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen~ Cl-c6 perfluoroalkyl,
R 1 20, R 1 1 S (~)m-, R 1 OC(O)NR 10 , (R 1 0)2NC(O)-,
R1o2N-c(NRlo)-~ CN, NO2, RIOC(o)-~ N3~ -N(R10)2,
or R I 1 OC(O)NR 10
c) unsub,stituted Cl-C6 alkyl;
d) .substituted Cl-C6 alkyl wherein the sub.stituent on the
sub~stituted Cl-C6 alkyl is ~selected from unsubstituted or
substituted aryl, unsubstituted or substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 I S(O)m-~ R 1 OC(O)NR 10 , (R 1 0)2NC(o)-,
R102N-C(NR10)-, CN, RlOC(O)-, N3, -N(R10)2, and
R 1 1 OC(O)-NR 10;
R6a, R6b, R6c ~nd R6d are independently .selected *om:
a) hydrogen,
b) unsub.stituted or substituted aryl, unsubstituted or
substituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 20, R 1 1 S(O)m-~ R 1 OC(O)NR 10 (R 1 0)2NC(o)-
R 1 02N-C(NR 10), CN, NO2, R 1 ~C(O)-, N3, -N(R 1~)2,
or R 1 1 OC(O)NR 10
c) un.sub,stituted Cl-C6 alkyl;
30 d) .substituted Cl-C6 alkyl wherein the substituent on the
substituted Cl-C6 alkyl i~s selected from unsub.stituted or
substituted aryl, unsubstituted or substituted heterocyclic, C3-Clo
cycloalkyl, C2-c6 alkenyl, C2-C6 alkynyl, R 1 20, R 1 1 S(O)m ~
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- 12 -
R 1 0C(O)NR 10, (R 1 0)2NC(O)-, R 1 02N-C(NR 10) , CN~
R 1 ~C(O)-, N3, -N(R 1~)2, and R 1 1 OC(O)-NR 10;
R7 is selected from: H; Cl 4 alkyl, C3-6 cycloalkyl, heterocycle, aryl,
5 aroyl, heteroaroyl, arylsulfonyl, heteroarylsulfonyl, unsublstituted or
~ubstituted with:
a) Cl 4 alkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
o
f) --SO2R
g) N(R 1~)2 or
h) C 1_4 perfluoroalkyl;
R2 is independently selected from:
a) hydrogen,
b) aryl, stituted aryl, heterocycle, Cl-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl,
R 1 0O, R 1 0C(O)NR 10, CN, NO2, (R 1 0)2N-C(NR 10
R 1 ~C(O)-, -N(R 1~)2, or R 1 1 OC(O)NR 10-, and
c) Cl-C6 alkyl .~ubstituted by Cl-C6 perfluoroalkyl, R10O-,
R 1 0C(O)NR 1 0-, (R 1 0)2N-C(NR 1 0)-, R 1 ~C(O)-,
-N(R 1~)2, or R I I OC(O)NR 10-;
provided that when R~ heterocycle, attachment of R~ to V i~
through a ~substitutable ring carbon;
R9 is ~elected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F~
Cl, Rl lo-, Rl lS(O)m-, R10C(O)NR10-, (Rl0)2Nc(o)
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CN, NO2, (R 1 0)2N-C(NR 10), R 1 ~C(O)-, -N(R 1 ~)2 or
Rl lOC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6
perfluoroalkyl, F, Cl, R 1 0O-, R 1 1 S(O)m-~ R 1 0C(O)NR 10,
S (R 1 0)2NC(o)-, CN, (R 1 0)2N-C(NR 10), R 1 ~C(O)-,
-N(R 1~)2, or R 1 1 OC(O)NR 10;
R10 is independently selected from hydrogen, Cl-C6 alkyl~ 2,2,2-
trifluoroethyl. benzyl and aryl;
Rl I is independently l~ielected from Cl-C6 alkyl and aryl;
R12 j~ independently selected from hydrogen, Cl-C6 alkyl, Cl-C6
aralkyl, Cl-C6 .substituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 sub~tituted heteroaralkyl, aryl, ~ubstituted aryl,
heteroaryl, sub~tituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluoroethyl;
A ~ and A2 are independently selected from: a bond, -CH=CH-, -C_C-,
-C(O)-, -C(O)NR 1 0-, O, -N(R 1 0)-? or S(O)m,
V is ~ielected from:
a) hydrogen,
b) heterocycle l~elected from pyrrolidinyl, imidazolyl,
imidazolinyl, pyridinyl, thiazolyl, oxazolyl, indolyl,
quinolinyl, i.soquinolinyl, triazolyl and thienyl,
c) aryl,
d) Cl-C20 alkyl wherein from 0 to 4 carbon atom,s are
replaced with a a heteroatom .~elected from O, S, and N,
and
e) C2-C20 alkenyl, and
provided that V is not hydrogen if Al i~ S(O)m and V i~ not hydrogen
if Al i.~i a bond, n i~ 0 and A2 i,~; S(O)m;
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WO 97136585 PCT~US97/062S9
- 14 -
provided that when V i.s heterocycle, attachment of V to R~ and to Al i~s
through a substitutable ring carbon;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, imidazolinyl,
5 pyridinyl, thiazolyl, oxazolyl, indolyl, ~luinolinyl, triazolyl or
isoquinolinyl;
X i.s a bond, 0, -C~=0)-, -CH=CH-, -C(o)NR7-, -NR7C(o)-, -NR7-~
-S(0)2N(R10)-, -N(R10)s(o)2- or-S(-O)m-; provided
I0 that if a is N, then X i.s not 0~ -C(o)NR7-~-s(o)2N(Rlo)
or -NR7;
mis 0, 1 or2;
n is independently 0, 1, 2, 3 or 4;
15 p is independently 0, 1, 2, 3 or 4;
clis 0,1,20r3;
r i.s 0 to 5, provided that r is 0 when V is hydrogen; and
t is 0 or 1 ;
20 or the pharmaceutically acceptable salts thereof.
A preferred embodiment of the compounds of thi~
invention are illustrated by the formula B:
R6a-d
( lR8)r R9~ R à~
V ~ Al(CR12)nA2(CR12)~N,~ \~ à~R4
R (C R22)F X
wherein:
a i.s N or C;
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W O 97/36S85 PCTAJS97/062S9
from 0-4 of b, c, d and e are independently N, NH, O and S, and the
~ rem~ining b. c, d and e atoms are independently CH, provided that if a
is C, then at least one of b, c, d or e i.s independently N, NH, O or S;
S a' is N or C:
from 0-4 of b', c', d' and e' are independently N, NH, O and S, and the
remaining b~ c', d' and e' atoms are independently CH, provided that if
~' is C, then at lea.st one of b'. c'~ d' or e' is independently N. NH, O or
10 S;
Rl i.s independently selected from: hydrogen, C3-Clo cycloalkyl,
RlOO-, -N(R10)2, F or Cl-C6 alkyl;
15 R2 is independently .~elected from:
a) hydrogen,
b) aryl~ heterocycle, C3-C I o cycloalkyl, R I OO-, -N(R 1 0)2,
F or C2-C6 alkenyl,
c) unsubstituted or substituted Cl-C6 alkyl wherein the
~ubstituent on the substituted Cl-C6 alkyl is ~selected from
un~ubstituted or sub.stituted aryl, heterocycle, C3-Clo
cycloalkyl, C2-C6 alkenyl, RIOO- and -N(R10)2;
R3 and R4 ~re independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
~ub.stituted heterocycle, C3-clo cycloalkyl, C2-C6
~Ikenyl, C2-C6 alkynyl~ halogen, Cl-C6 perfluoroalkyl,
R120-, RllS(O)m-, RIOC(O)NR10-, (R10)2NC(o)-,
R10~N-C(NRlO)-, CN, NO2, RIOC(o)-~ N3, -N(R10)2,
or R l l OC(O)NR l o-
c) un,sub~stituted Cl-C6 alkyl,
d) ~ubstituted Cl-c6 alkyl wherein the ~ub,stituent on the
~ub.stituted Cl-c6 alkyl i,~ .selected from un,sub.stituted or
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~substituted aryl, unsubstituted or ,substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R I I S(O)m-, R 1 0C(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R 1 ~C(O)-, N3, -N(R 1~)2, and
R1 1OC(O) NR10;
R6a, R6b, R6C and R6d are independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsub~tituted or
sub~stituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C:2-C6 alkynyl halogen, Cl-C6 perfluoroalkyl,
R 1 20, R I 1 S(O)m-~ R 1 0C(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, NO2, R I ~C(O)-, N3, -N(R 1~)2,
or R 1 1 OC(O)NR 10
c) un.substituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
sub~stituted Cl-C6 alkyl is selected from unsub~tituted or
substituted aryl, unsub,stituted or substituted heterocyclic,
C3-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R I I S(O)m-, R 1 0C(O)NR 1 0-, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R I ~C(O)-, N3, -N(R 1 ~)2, and
R11OC(O) NR10;
R~ is independently selected from:
a) hydrogen,
b) aryl, substituted aryl, heterocycle, Cl-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl. Cl-C6 perfluoroalkyl, F, Cl,
R 1 ~O-, R 1 0C(O)NR 10 CN. NO2, (R 1 0)2N-C(NR 10),
R I ~C(O)-, -N(R 1~)2, or R I 1 OC(O)NR 10, and
c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R10O-,
R 1 0C(O)NR 10, (R 1 0)2N-C(NR 10), R 1 ~C(O)-,
-N(R10)2, or Rl IOC(O)NRl0-;
provided that when R~ i.s heterocycle, attachment of RX to V i.s
through a sub~stitutable ring carbon;
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R9a and R9b are independently hydrogen, C I -C6 alkyl, trifluoromethyl
and halogen;
R10 i~i independently ,selected from hydrogen, Cl-C6 alkyl, 2,2,2-
trifluoroethyl, benzyl and aryl;
R l I is independently selected from Cl -C6 alkyl and aryl;
~0 R12 is independently ~selected from hydrogen, Cl-C6 alkyl, Cl-C6
aralkyl, Cl-c6 ~sub~tituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 ,sub~tituted heteroaralkyl, aryl, ~ub~tituted aryl,
heteroaryl, ~sub.stituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluoroethyl;
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 ,~elected from pyrrolidinyl, imidazolyl,
imidazolinyl, pyridinyl, thiazolyl, oxazolyl, indolyl,
quinolinyl, isoquinolinyl, triazolyl and thienyl,
c) aryl,
d) Cl-C20 alkyl wherein from 0 to 4 carbon atom~ are
replaced with a a heteroatom ~elected from O, S, and N,
and
e) C2-C20 alkenyl~ and
provided that V i.~ not hydrogen if A~ S(O)m and V i~ not hydl-ogen
30 if A1 i~ a bond, n i~ 0 and A2 i~ S(O)m;
provided that when V i~ heterocycle, att~chment of V to Rg and to A
through a ,~ub.stitutable ring carbon;
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X is a bond, -CH=CH-, -C(O)NR 10, -NR 1 ~C(03-, -NR 10, 0 or
-C(=O)-;
provided that if a is N, then X is not -C(O)NR10-, -NR10-, or 0;
5 mi.s 0, 1 or2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or 4; and
r is 0 to 5, provided that r i.s 0 when V is hydrogen;
10 or the pharmaceutically acceptable ~alt,s thereof.
Another preferred embodiment of the compound.s of thi.s
invention are illu,strated by the forrnula C:
6a-d
(R8)r R3 1
V- Al(cRl2)nA2(cR12)n~/~ à~e'
C R9b (CR22)F)(/
.
wherem:
a i,s N or C;
from 0-4 of b, c, d and e are independently N, NH, 0 and S, and the
rem~ining b, c, d and e atoms are independently CH, provided that if a
20 is C, then at least one of b, c. d or e i~s independently N, NH, 0 or S;
a' is N or C;
from 0-4 of b', c'. d' ~nd e' are independently N, NH, 0 and S, and the
25 remaining b'. c', d' and e' atoms are independently CH, provided that if
ar i,s C, then at lea,st one of b', c', d' or e' i.s independently N, NH, 0 or
S;
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R1 is independently selected from: hydrogen, C3-clo cycloalkyl,
R10O-, -N(R10)2, F or C1-C6 alkyl;
5 R2 i~s independently ~elected from:
a) hydrogen,
b) aryl, heterocycle, C3-Clo cycloalkyl, RIOO-~ -N(R10)2,
F or C2-C6 alkenyl,
c) unsubstituted or ~ubstituted Cl-C6 alkyl wherein the
~ub~tituent on the substituted Cl-C6 alkyl i.~ selected from
unsubstituted or substituted aryh heterocycle, C3-clo
cycloalkyl, C2-C6 alkenyl, R100- ;md -N(R10)2;
R3 and R4 are independently ~ielected from:
a) hydrogen,
b) unsubstituted or substituted aryl, un~ub~tituted or
substituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen~ Cl-C6 perfluoroalkyl,
R 1 20 R 1 1 S(O)m ~ R 1 OC(O)NR 10, CN(R I 0)2NC(O)-,
R 1 02N-C(NR 1 0)-, CN, NO2, R l OC(O)-, N3 ? -N(R I o)27
or R 1 1 OC(O)NR 10
c) unsubstituted Cl-C6 alkyl,
d) ~ubstituted Cl-C6 alkyl wherein the ~substituent on the
~ubstituted Cl-C6 alkyl i~ elected from un~sub~tituted or
.substituted aryl, unsubstituted or substituted heterocyclic,
C~-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R I 1 S(O)m ~ R 1 OC(O)NR 10, (R 1 0)2NC(O)-,
R102N-C(NR10)-, CN, RlOC(O)- N~, -N(R10)2~ and
RllOc(o)NRlo;
R6a R6b R6C and R6d are independently ,~elected from:
a) hydrogen,
b) unstituted or ~iub~tituted aryl. un~iub.stituted or
.~ub.~tituted heterocycle. C3-Clo cycloalkyl, C2-C6
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alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 20 , R 1 I S(O)m-~ R 1 0C(O)NR 1 0-, CN(R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, NO2, R 1 ~C(O)-, N3, -N(R 1 ~)2,
or R 1 1 OC(O)NR 10
c) un.substituted Cl-C6 alkyl,
d) .substituted Cl-C6 alkyl wherein the ~substituent on the
substituted Cl-C6 alkyl is .~ielected from unsubstituted or
.substituted aryl, un~ubstituted or substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20 R 1 1 S(O)m-, R 1 0C(O)NR 1 O-? (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R l Oc(O)-? N3, -N(R 1 ~)2, and
RllOc(o) NRlO;
R~ independently selected from:
a) hydrogen,
b) aryl, .substituted aryl, heterocycle, Cl-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl,
R 1 0O-, R 1 0C(O)NR 10, CN, NO2, (R 1 0)2N-C(NR 10
R 1 ~C(O)-, -N(R 1~)2, or R 1 1 OC(O)NR 10 ~ and
c) Cl-C6 alkyl sub,~tituted by Cl-C6 perfluoroalkyl,
R 1 0O R 1 0C(O)NR 10 , (R 1 0)2N-C(NR 10) , R 1 ~C(O)-,
-N(R 1~)2, or R 1 1 OC(O)NR 10;
provided that when Rg is heterocycle, attachrnent of R~ to V is
through a sub,stitutable ring carbon;
R9a and R9b are independently hydrogen, Cl-C6 alkyl, trifluoromethyl
and halogen;
R10 i~ independently selected from hydrogen, Cl-C6 alkyl, 2,2,2-
trifluoroethyl, benzyi and aryl;
Rl I is independently selected from Cl-C6 alkyl and aryl;
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R12 is independently selected from hydrogen, Cl-C6 alkyl, Cl-c6
aralkyl, Cl-C6 sub,stituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 substituted heteroaralkyl, aryl, substituted aryl,
heteroaryl, sub~stituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluoroethyl;
Al and A2 are independently ,selected from: a bond, -CH=CH-, -C_C-,
-C(O)-, -C(O)NR 1 0-, O, -N(R I 0)-, or S(O)m;
10 V i~ selected from:
a) hydrogen,
b) heterocycle selected from pyrrolidinyl, imidazolyl,
imidazolinyl, pyridinyl, thiazolyl, oxazolyl, indolyl,
Lluinolinyl, i,soquinolinyl, triazolyl and thienyl,
c) aryl,
d) Cl-C20 alkyl wherein from 0 to 4 carbon atom.~ are
replaced with a a heteroatom ~ielected from O, S, and N,
and
e) C2-C20 alkenyl, and
20 provided that V i~ not hydrogen if Al is S(O)m and V is not hydrogen
if Al i~ a bond, n i~ 0 and A2 is S(O)m;
provided that when V i~ heterocycle, attachment of V to RX and to A
through a .substitutable ring carbon;
2~ X i.~ a bond, -CH=CH-, -C(O)NR 10, -NR 1 ~C(O)-, -NR 10-, O or
-C(=O)-;
provided that if a i~ N, then X i.s not -C(O)NR10-, -NR10- or O;
mis 0, 1 or2;
ni~; 0, 1, 2, 3 or4;
p i.~i 0~ 1, 2, 3 or 4, provided that p i~ not 0 if X i~ a bond,
-NR 10 or O; and
r i~ 0 to ~, provided that r i~ 0 when V i~i hydrogen;
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or the phalmaceutically acceptable .salts thereof.
In a more preferred embodiment of this invention, the
inhibitors of farnesyl-protein transferase are illustrated by the formula
D:
6a-d
(CR12)l~--N ~ ~J
9b (C R2 ) X
R8 D
wherein:
a is N or C;
from 0-4 of b, c, d and e are independently N, NH, O and S, and the
rem~ining b, c, d and e atoms are independently CH, provided that if a
i,s C, then at least one of b, c, d or e is independently N, NH, O or S;
a' is N orC;
from 0-4 of b', c', d' and e' are independently N, NH, O and S, and the
remaining b', c', d' and e' atoms are independently CH, provided that if
a' is C, then at least one of b', c', d' or e' is independently N, NH, O or
S;
Rl is independently selected from: hydrogen, C3-Clo cycloalkyl
Cl-C6 alkyl;
R2 i,s independently .selected from:
a) hydrogen.
b) aryl. heterocycle, C3-CIo cycloalkyl, RlOO-, -N(R10)2,
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F or C2-C6 alkenyl,
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, C3-C1o cycloalkyl, C2-C6 alkenyl, R100-, or
-N(R 1 ~)2;
R3 i~s ~selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, un~substituted or
substituted heterocycle, C3-clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 20, R 1 I S(O)m-, R I OC(O)NR 1 0-, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, N02, R I ~C(O)-, N3, -N(R 1~)2,
or R I 1 OC(O)NR 10
c) unsubstituted Cl-C6 alkyl,
d) ~ub~stituted Cl-C6 alkyl wherein the ~ubstituent on the
substituted Cl-C6 alkyl i~; selected from unsubstituted or
substituted aryl, un~substituted or sub,stituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R120-, Rl lS(O)m-, RlOC(O)NR10-, (R10)2NC(o)-,
R102N-C(NR10)-, CN, RlOC(O)-, N3, -N(R10)2, and
R 1 1 OC(O)-NR 10;
R4 is selected from H, halogen, C1-C6 alkyl and CF3;
25 R6a, R6b, R6C and R6d are independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
substituted heterocycle, C3-clo cycloalkyl, C2-C6
alkenyl, C2-c6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
Rl~O-, Rl lS(O)m-, RlOC(O)NR10-, (R10)2Nc(o)-~
R10~N-C(NRlO)-, CN, N02, RIOC(O)-~ N3. -N(R10)2,
or R 1 1 OC(O)NR 10,
c) un.sub~stituted Cl-C6 alkyl,
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- 24 -
d) sublstituted Cl-C6 alkyl wherein the substituent on the
substituted C~-C6 alkyl is selected from unsubstituted or
substituted aryl, unsubstituted or substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 1 S(O)m ~ R 1 OC(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 1 0)-, CN, R l OC(O)-, N3, -N(R 1 0)2, and
Rl IOC(O)-NRlO;
RX i~s independently ,selected from:
a) hydrogen,
b) aryl, substituted aryl, heterocycle, Cl-C6 alkyl, C2-C6
allcenyl, C2-C6 alkynyl, C~-C6 perfluoroalkyl, F, Cl,
R l 0O, R 1 OC(O)NR 10, CN, N02, (R 1 0)2N-C(NR 1 0),
R 1 ~C(O)-, -N(R 1~)2, or R 1 1 OC(O)NR 10 , and
c~ Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R100-,
R 1 OC(O)NR 1~-, (R 1 0)2N-C(NR 10), R 1 ~C(O)-,
-N(R10)2, or Rl lOC(O)NR10-;
provided that when R8 is heterocycle, attachment of R~ to V is
through a substitutable ring carbon;
R9a and R9b are independently hydrogen~ halogen, CF3 or methyl;
R10 is independently selected from hydrogen, Cl-C6 alkyl, 2,2,2-
trifluoroethyl, benzyl and aryl;
Rl 1 is independently selected from Cl-C6 alkyl and aryl;
Rl'~ i.s independently selected from hydrogen, Cl-c6 alkyl, Cl-c6
aralkyl, Cl-C6 substituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 ,sublstituted heteroaralkyl, aryl, substituted aryl,
heteroaryl, .sub,stituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluo1oethyl;
Al i.s Iselected from: a bond, -C(O)-, 0~ -N(R10)-, or S(O)m;
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X is a bond, -CH=CH-, -C(O)NR10-, -NRlOC(O)-, -NR10-, O or
-C(=O)-, provided that if a i~s N, then X is not -C(O)NR10-, -NR10-
or O;
n is 0 or 1; provided that n is not 0 if A l is a bond, O,
-N(R10)-, or S(O)m;
m is 0, 1 or 2; and
pis 0, 1, 2, 3 or4;
or the pharmaceutically acceptable salt,s thereof.
In another more preferred embodiment of this invention,
the inhibitors of farne~yl-protein transferase are illustrated by the
formula E:
R6a-d
R
2)n ~;/ \ à~R4
9b (CR22)p X
~¦~ E
R8
wherein:
a i.s N or C;
20 from 0-4 of b~ c, d and e are independently N, NH, O and S, and the
rem~ining b, c, d and e atoms are independently CH, provided that if a
i,s C. then at lea~st one of b, c, d or e i.s independently N, NH, O or S;
a' i!i N or C;
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from 0-4 of b', c', d' and e' are independently N, NH, O and ~, and the
remaining b', c', d' and e' atoms are independent]y CH, provided that if
a' is C, then at least one of b', c', d' or e' i,s independently N, NH, O or
S;
Rl is independently .selected from: hydrogen, R10O-, -N(R10)2, F,
C3-C l o cycloalkyl or C l -C6 alkyl;
R2 is independent]y selected from:
a) hydrogen,
b) aryl, heterocycle, C3-C I o cycloalkyl. R 1 ~O-~ -N(R 1 ~)2
F or C2-C6 alkenyl,
c) Cl-C6 alkyl unsubstituted Ol substituted by aryl,
heterocycle, C3-CIo cycloalkyl, C2-C6 alkenyl, R10O-, or
1 :S -N(R 1 ~)2;
R3 is selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
.~ubstituted heterocycle, C3-CIo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen, Cl-C~ perfluoroalkyl,
R 1 20, R 1 1 S(O)m ~ R 1 0C(O)NR 1 0-, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10)-, CN, NO2, R I ~C(O)-, N3, -N(R 1~)2,
or R 1 1 OC(O)NR 10-,
c) unsubstituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the ~;ubstituent on the
.substituted Cl-C6 alkyl i,s ,selected from unsub.stituted or
sub,stituted aryl, un,sub~stituted or ~substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R I I S(~)m-, R 1 0C(O)NR 1{~ , (R 1 0)2NC(O)-,
R102N-c(NRlo)-~ CN, R10C(O)-~ N3, -N(RIo)2~ and
R I 1 OC(O)-NR 10;
R4 is ,~ielected from H~ halogen, Cl-C6 alkyl and CF3;
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R6a R6b, R6C and R6d are independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
substituted heterocycle, C3-clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 20, R 1 1 S(O)m-, R 1 OC(O)NR 10, (R 1 0)2NC(O)-,
R102N-C(NR10)-. CN, N02, RlOC(O)-, N3, -N(R10)2,
or R 1 1 OC(O)NR 10
c) unsubstituted Cl-C6 alkyl,
d) ~substituted Cl-C6 alkyl wherein the sub.stituent on the
substituted Cl-C6 alkyl is selected from unstituted or
substituted aryl, unsubstituted or substituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R120-, Rl lS(O)m-, RlOC(O)NR10-, (RlO)2Nc(o)-~
R102N-C(NR10)-, CN, RlOC(O)-, N3, -N(R10)2, and
R 1 1 OC(O) NR 10;
R~ is independently selected from:
a) hydrogen,
b) aryl, .substituted aryl, heterocycle, Cl-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl,
R100-, RlOC(O)NR1O-, CN, N02, (R10)2N-c(NRlo)
R 1 ~C(O)-, -N(R 1~)2, or R 1 1 OC(O)NR 10, and
c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R100-,
RlOC(O)NR10-, (R10)2N-C(NR10)-, RlOC(O)-,
-N(R10)2, or Rl lOC(O)NR10-;
provided that when R~ is heterocycle, attachment of R~s to V is
through a substitutable ring carbon~
R9~ and R9b ~re independently hydrogen, halo~en. CF;3 or methyl;
R10 is independently selected from hydrogen. Cl-c6 alkyl? 2,2,2-
trifluoroethyl, benzyl and aryl;
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- 2~ -
Rl 1 i.s independently selected from Cl-C6 alkyl and aryl;
R12 is independently selected from hydrogen, Cl-c6 alkyl, Cl-C6
S aralkyl, Cl-C6 substituted aralkyl, Cl-c6 heteroaralky],
C1-C6 substituted heteroaralkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, Cl-c6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluoroethyl;
10 X is a bond. -CH=CH-, -(~(O)NR 10, -NR 1 ~C(O)-, -NR 10, o or
-C(=O)-;
provided that if a is N, then X is not -C(O)NR10-, -NR~ ~- or O;
nis Oor 1;
m is 0. 1 or 2; and
p is 0, 1, 2, 3 or 4, provided that p is not O if X is a bond,
-NR10- or 0;
or the pharmaceutically acceptable salts thereof.
In a further embodiment of this invention, the inhibitors of
farne~syl-protein tran.sfera~se are illustrated by the formula F:
R6a-d
R9a~ R3 l~it/
CR12--N\~ R9b
(CR22)p X
NC F
- wherein:
a is N or C;
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- 29 -
from 0-4 of b, c, d and e are independently N, NH, O and S, and the
remaining b, c, d and e atoms are independently CH, provided that if a
is C, then at least one of b, c, d or e i,s independently N, NH, O or S;
5 a' is N or C;
from 0-4 of b', c', d' and e' are independently N, NH, O and S, and the
remaining b', c', d' and e' atom~s are independently CH, provided that if
a' is C, then at least one of b', c', d' or e' is independently N, NH, O or
10 S;
Rl is independently ~elected from: hydrogen, C3-Clo cycloalkyl or
Cl-C6 alkyl;
1~ R2 is independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-C I o cycloalkyl, R 1 0O-, -N(R I o)2 or
F,
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocycle, C3-C~o cycloalkyl, R10O-, or-N(R10)2;
R3 is selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
substituted heterocycle, C3-Clo cycloalkyl, C2-C6
alkenyl, C2-C6 alkynyl, halo~en, Cl-C6 perfluoroalkyl,
R120-,RllS(O)m-,RIOC(O)NR10-,(R10)2NC(o)-,
R10~N-C(NR10)-, CN, NO2, R10C(o)-, N3, -N(R10)2
or R I I OC(O)NR 10-,
c) unsubstituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
substituted Cl-C6 alkyl is .selected from unstituted or
sub,stituted aryl, unsubstituted or sub,stituted heterocyclic,
C~-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
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- 30 -
R120, Rl lS(o)m, Rloc(o)NRlo-7 (R10)2NC(o)-,
R102N-C(NR10)-, CN, RlOC(0)-, N3, -N(R10)2, and
RllOC(O) NR10;
S R4 is .selected from H, halogen, CH3 and CF3;
R6a, R6b, R6C and R6d are independently .selected from:
a) hydrogen,
b) un.substituted or.substituted aryl? un,substituted or
Isubstituted heterocycle, C3-Clo cycloalkyl~ C2-C6
alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 20, R 1 I S(O)m-, R 1 OC(O)NR 10, (R 1 0)2NC(0)-,
R 1 02N-C(NR 10) , CN, N02, R 1 ~C(0)-, N3, -N(R 1~)2,
or R 1 1 OC(O)NR 10
c) un,substituted Cl-C6 alkyl,
d) ,substituted Cl-c6 alkyl wherein the Isubstituent on the
,sub,stituted Cl-C6 alkyl i,s selected from un.substituted or
sub.stituted aryl, unsubstituted or sub,stituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 I S(O)m-~ R 1 OC(O)NR 1 0-, (R 1 0)2NC(0)-,
RlO2N-c(NRlo)-7 CN, RIOC(o)-~ N3, -N(RIo)2~ and
R 1 1 0C(O)-NR 10;
R9a and R9b are independently hydrogen, halogen, CF3 or methyl;
R10 is independently ~elected from hydrogen, Cl-C6 alkyl, 2,2,2-
trifluoroethyl, benzyl and aryl;
R 1 1 is independently ~selected from Cl -C6 alkyl and aryl:
R12 i~ independently ~ielected from hydrogen, Cl-C6 alL~yl, Cl-C6
aralkyl, Cl-c6 .sub,stituted aralkyl, Cl-C6 heteroaralkyl.
Cl-C6 substituted heteroaralkyl, aryl, .substituted aryl,
heteroaryl, sub.stituted heteroaryl, Cl-c6 perfluoroalkyl?
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- 31 -
2-aminoethyl and 2,2,2-trifluoroethyl;
X is a bond, -CH=CH-, -C(O)NR 10, -NR 1 ~C(O)-, -NR 10 , O or
-C(=O)-;
5 provided that if a i~ N, then X is not -C(O)NR10-, -NR10- or O;
m i~ 0. 1 or 2; and
pi~ 0. 1, '', 3 or4;
10 or the pharmaceutically acceptable salts thereof.
In a further embodiment of thi.s invention, the inhibitor.~ of
farne~yl-protein tran,~ifera~e are illustrated by the formula G:
R6a-d
R à~'
9b~ R4e
NC~--A (CR 2)n R
wherein:
a i,s C;
from 0-4 of b~ c, d and e are independently N, NH, O and S, and the
rem~ining b, c~ d and e atom~s are independently CH, provided that if a
20 i,~ C, then at lea~t one of b, c, d or e i,s independently N, NH, O or S;
a' is N or C;
from 0-4 of b'~ c', d' and e' are independently N, NH, O and S, and the
25 remaining b'~ c'~ d' and e' atoms are independently CH, provided that if
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W O 97/36585 PCT~US97/06259
a' is C, then at lea~t one of b', c', d' or e' i.s independently N, NH, O or
S;
K l is independently selected from: hydrogen, R1 00-, -N(R10)2, F,
C3-CIo cycloalkyl or Cl-C6 alkyl;
R2 i,s independently ~elected from:
a) hydrogen,
b) aryl, heterocycle or C3-CIo cycloalkyl,
c) C]-C6 alkyl un,~ubstituted or substituted by aryl,
heterocycle, C3-Clo cycloalkyl, C2-C6 alkenyl, R100-, or
-N(R I ~)2;
R3 i~ ~elected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or
substituted heterocycle, C3-clo cycloalkyl, C2-C6
alkenyl, C2-c6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R120-, RllS(O)m, RlOC(o)NRlo- (R10)2NC(o)
R 1 02N-C(NR 10), CN, N02, R 1 ~C(O)-, N3, -N(R 1~)2,
or Rl 10c(o)NRlo
c) un~ubstituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
Isubstituted C1-C6 alkyl is ~selected from unsubstituted or
substituted aryl, unsub~tituted or Isubstituted heterocyclic,
C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 I S(O 3m-~ R 1 OC(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 10), CN, R 1 ~C(O)-, N3, -N(R 1~)2, and
R 1 1 OC(O)-NR 10;
R4 i~ ~ielected from H, halogen, CH3 and CF~;
R6a R6b R6c and R6d are independently !;elected from:
a) hydrogen,
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b) unsubstituted or substituted aryl, unsubstituted or
~ub~stituted heterocycle, C3-clo cycloalkyl, C2-c6
alkenyl, C2-C6 alkynyl, halogen, Cl-C6 perfluoroalkyl,
R 1 2O, R 1 1 S(O)m-~ R 1 OC(O)NR 10, (R 1 0)2NC(O)-,
R 1 02N-C(NR 1 0)-, CN, NO2, R l OC(O)-, N3, -N(R 1 0)2,
or R 1 1 OC(O)NR 10
c) un,substituted C1-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the substituent on the
substituted Cl-C6 alkyl i.s selected from unsubstituted or
~sub~stituted aryl, unsubstituted or ,sub,stituted heterocyclic,
C~-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
R 1 20, R 1 I S(O)m-~ R 1 OC(O)NR 10 , (R 1 0)2NC(O)-,
R 1 02N-C(NR 1 0)-, CN, R l OC(O)-, N3, -N(R 1 0)2, and
R l l OC(O) NR 10;
R9~ and R9b are independently hydrogen, halogen, CF3 or methyl;
R10 is independently ~selected from hydrogen, Cl-C6 alkyl, 2,2,2-
trifluoroethyl, benzyl and aryl;
Rl 1 is independently selected from Cl-C6 alkyl and aryl;
R12 is independently selected from hydrogen, Cl-C6 alkyl, Cl-C6
2;~ aralkyl, Cl-C6 substituted aralkyl, Cl-C6 heteroaralkyl,
Cl-C6 substituted heteroaralkyl, aryl~ substituted aryl,
heteroaryl, ,sub,stituted heteroaryl, Cl-C6 perfluoroalkyl,
2-aminoethyl and 2,2,2-trifluoroethyl;
30 AI is selected from: a bond, -C(O)-, O. -N(R10)-, or S(O)m;
m i~ 0, 1 or 2; and
n is 0 or I ;
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- 34 -
or the pharmaceutically acceptable salts thereof.
Specific examples of the compounds of the invention are:
I -(5-(Thien-2'-yl)thien-2-ylmethyl)-5 -(4-cyanobenzyl)imidazole
NC~,~
ii
~ N~
I -(4-Cyanobenzyl)-~-(5-( 1 -methylpyrrol-2-yl)-thiazol-2-
ylmethyl~imidazole
NC ~,fi~,
H3C~
~N~"~ 5>~
10 or the pharmaceutically acceptable Isaltls thereof.
The compounds of the pre~sent invention may have
asymmetric centers and occur a.s racemates, racemic mixtures, and a.
individual diastereomers, with all possible isomers, including optical
isomers, being included in the present invention. Also, combinations
15 of substituents/or variable.s are permissible only if such combinations
result in stable compounds.
A.s u~ed herein, "alkyl" and the alkyl portion of aralkyl and
similar term.<;~ i.s intended to include both branched and straight-chain
saturated aliphatic hydrocarbon groups having the .specified number of
20 carbon atom.s; "alkoxy" represent.s an ;llkyl group of indicated number
of carbon atoms attached through an oxygen bridge.
A.s u.sed herein, "cycloalkyl" is intended to include non-
aromatic cyclic hydrocarbon group~ having the specified number of
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carbon atoms. Examples of cycloalkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl and the like.
"Alkenyl" group~ include those groups having the specified
number of carbon atoms and having one or several double bonds.
5 Examples of alkenyl groups include vinyl, allyl, isopropenyl, pentenyl,
hexenyl, heptenyl, cyclopropenyl, cyclobutenyl, cyclopentenyl,
cyclohexenyl, l-propenyl, 2-butenyl, 2-methyl-2-butenyl, i.soprenyl,
farne~yl, geranyl, geranylgeranyl and the like.
"Alkynyl" groups include tho,se group.s having the specified
10 number of carbon atoms and having one triple bond.s. Examples of
alkynyl groups include acetylene. 2-butynyl, 2-pentynyl. 3-pentynyl and
the like.
"Halogen" or "halo" a.s u.sed herein means fluoro, chloro,
bromo and iodo.
A.s used herein, "aryl," and the aryl portion of aralkyl,
i~ 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.
Examples of such aryl elements include phenyl, naphthyl,
tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or
20 acenaphthyl.
The term heterocycle or heterocyclic~ a~ ulsed herein,
repre,sents a stable 5- to 7-membered monocyclic or stable ~- to
1 l-membered bicyclic heterocyclic ring which is either saturated
or unsaturated, and which consists of carbon atoms and from one to
25 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 ring~s i~ fused to a benzene ring. The heterocyclic ring
may be attached at any heteroatom or carbon atom which re~ult~; in the
creation of a stable structure. Example~ of ~uch heterocyclic element~i
30 include, but are not limited to, azepinyl, benzimidazolyl, benzil~oxazolyl,
benzofurazanyl, benzopyranyl, benzothiopyranyh benzofuryl,
benzothiazolyl. benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothiopyranyl ~ulfone, furyl, imidazolidinyl~ imidazolinyl,
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- 36 -
imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl,
isothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl,
naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-
oxopyrrolidinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl,
S pyridazinyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, cluinazolinyl,
quinolinyl, 4uinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl,
tetrahydro~uinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide,
thiazolyl~ thiazolinyl, thienofuryl, thienothienyl, and thienyl.
As ulsed herein, "heteroaryl" i,s intended to mean any .stable
lO 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. Example,s of .such heterocyclic elements
include, but are not limited to, benzimidazolyl, benzisoxazolyl,
15 benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl,
isochromanyl, isoindolinyl, isoquinolinyl, isothiazolyl, naphthyridinyl,
20 oxadia~olyl, pyridyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl,
pyrrolyl quinazolinyl, quinolinyl, 4uinoxalinyl,
tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiazolyl, thienofuryl,
thienothienyl~ triazolyl and thienyl.
As used herein in the definition of R3, R4, R5 and R6a-d,
25 the term "the sub,stituted group" i,s intended to mean a substituted Cl g
alkyl, substituted C2 ~ alkenyl, substituted C2 X alkynyl, substituted aryl
or .substituted heterocycle from which the ,sub~stituent(s) R3, R4, RS and
R6a-e are selected.
As used herein in the definition of R7, the substituted C
30 alkyl, substituted C3-6 cycloalkyl, substituted aroyl, substituted aryl,
substituted heteroaroyl, substituted arylsulfonyl, substituted heteroaryl-
sulfonyl and substituted heterocycle include moieties containing from l
to 3 ,substituellt.s in addition to the point of attachment to the rest of the
compound.
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As used herein, when no specific substituent,s are set forth,
the terms "substituted aryl", "substituted heterocycle" and "substituted
cycloalkyl" are intended to include the cyclic group which i~ substituted
on a substitutable ring carbon atom with 1 or 2 substitutents ,selected
5 from the group which include.s 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-C6 alkyl)C(O)NH-, H2N-C(NH)-, (Cl-C6
alkyl)C(O)-, (Cl-C6 alkyl)OC(O)-, N3,(Cl-C6 alkyl)OC(O)NH-,
phenyl, pyridyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thienyl,
10 furyl, isothiazolyl and Cl-C20 alkyl.
The moieties designated by the following structures
bà~)/d b'~,
repre,sent aromatic 5-membered heterocyclic ring~s and includes the
following ring system.s:
~N ~ N N~N N--N~N
N~ N~\ N~ N--~
N ' N ' N 11 N
N~/ N~/ N-N~ N--N
[C~ N ~ ¢~ N~
~ N_ N> N~ S,> 'N ~
N~ ~ N - N/> N~O>
Preferably the aromatic :~-membered heterocyclic ring is selected from:
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- 3?s -
~N
N~
~ ~> N~>
It i.s under.stood that .such ring sy.stem,s may be independently selected
and that
bl~),d and b ~,
5 may be the same or different in the instant compound~.
Lines drawn into the rin~ systems from sub~stituents (,such
as ~rom R3, R4 etc.) means that the indicated bond may be attached to
any of the sub,stitutable ring carbon or nitrogen atoms.
Preferably, Rl and R2 are independently selected from:
10 hydrogen, R 1 1 C(0)0-, -N(R 1 ~)2, R I OC(O)NR 10, R l ~O or
unsubstituted or substituted Cl-C6 alkyl wherein the sub~stituent on the
substituted Cl-C6 alkyl is selected from un~ubstituted or,substituted
phenyl, -N(R 1 ~)2, R l ~O and R 1 OC(O)NR 10 .
Preferably, R3 is selected from:
a) hydrogen,
b) C~-C I o cycloalkyl, halogen, C l -C6 perfluoroalkyl, R 1 20,
CN, N02, R l OC(0)- or -N(R 1 0)2,
c) un~ub.stituted Cl-C6 alkyl,
d) substituted Cl-C6 alkyl wherein the ~ub.~titutent on the
2Q ~iub,stituted Cl-C6 ~Ikyl i~s selected from unsub.stituted or
Isub~stituted aryl, un.substituted or .substituted heterocyclic,
C~-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,
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R120-, Rl lS(O)m-, RlOC(O)NR10-, (R10)2NC(o)-,
- R102N-C(NR10)-, CN, RlOC(0)-, N3, -N(R10)2, and
R 1 1 OC(0)-NR 10
Preferably, R4 is selected from: hydrogen, halogen,
5 trifluoromethyl, trifluoromethoxy and Cl-C6 alkyl.
Preferably, R5 is hydrogen.
Preferably R6a, R6b, R6C and R6d are independently
selected from:
a) hydrogen,
b) C3-CIo cycloalkyl, halogen, C~-c6 perfluoroalkyl, R120-,
R l l S(O)m-, CN, N02, R I ~C(0)- or -N(R l O)2,
c) unsub~tituted Cl-C6 alkyl;
d) .~iubstituted Cl-c6 alkyl wherein the sub~titutent on the
substituted Cl-C6 alkyl is selected from unsub.stituted or
substituted aryl, C3-Clo cycloalkyl, R120, Rl lS(O)m-,
R 1 ~C(0)- or -N(R 1~)2.
Preferably, R~ is independently selected from:
a) hydrogen, and
b) aryl, substituted aryl, heterocycle, substituted heterocycle,
C 1 -C6 perfluoroalkyl or CN.
Preferably, R9 i~, hydrogen, halogen, CF3 or methyl.
Preferably, R10 is selected from H, Cl-C6 alkyl and
benzyl.
Preferably, Al and A2 are independently selected from:
25 a bond, -C(O)NR 10, -NR 1 ~C(0)-, 0, -N(R 10), -S(0)2N(R 10) and
-N(R 1 ~)S(0)2-.
Preferably, V i~ selected from hydrogen, heterocycle and
aryl. More preferably, V i~ phenyl.
Preferably, W is selected from imidazolinyl, imidazolyl,
30 oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl. More
preferably, W i~ .~,elected from imidazolyl and pyridyl.
Preferably, n and r are independently 0, 1, or 2.
Preferably s i~, 0.
Preferably t i~i 1.
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- 40 -
It is intended that the definition of any sub~tituent or
variable (e.g., R1, R2, R9, n, etc.) at a particular location in a molecule
be independent of it,s definitions elsewhere in that molecule. Thus,
-N(R10)2 represents -NHH, -NHCH3, -NHC2H5, etc. lt is understood
5 that substituents and ,substitution patterns on the compounds of the
instant invention can be selected by one of ordinary skill in the art to
provide compounds that are chemically stable and that can be synthe-
sized by technique,s known in the art, as well as those methods set forth
below. from readily available starting materials.
The pharmaceutically acceptable salt~ of the compounds of
this invention include the conventional non-toxic salts of the compounds
of thi.s invention as fonned, e.g., from noll-toxic inorganic or organic
acids. For example, such conventional non-toxic salts include those
derived from inorganic acids such as hydrochlolic, hydrobromic,
15 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,
phenylacetic, glutamic, benzoic, salicylic, ,sulfanilic, 2-acetoxy-benzoic,
fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic,
20 isethionic, trifluoroacetic and the like.
The pharmaceutically acceptable .~;alt,~i of the compounds
of this invention can be .synthesized from the compounds of this
invention which contain a basic moiety by conventional chemical
methods. Generally, the salts are prepared either by ion exchange
25 chroma-tography or by reacting the fiee base with stoichiometric
amounts or with an exces,s of the desired ~salt-forming inorganic or
organic acid in a suitable solvent or various combination.s of solvents.
Reactions used to generate the compounds of this invention
are prepared by employing reactions a.s shown in the Schemes 1-24,
30 in addition to other ,standard manipulations such as e,ster hydrolysis,
cleavage of protecting groups, etc., a~s may be known in the literature
or exemplified in the experimental procedures. Substituents R3, R6
and RX. as ~shown in the Schemes, repre,sent the sub.stituent.s R3, R4, R5,
R6a R6b. R6C, R6d ~nd R~; although only one such R6 or RX i.s present
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- 41 -
in the intermediates and products of the schemes, it i~ understood that
the reactions .shown are also applicable when such aryl or heteroaryl
moietie~i contain multiple substituents.
These reactions may be employed in a linear sec~uence
to provide the compounds of the invention or they may be used to
synthesize fragment,s which are ~subsequently joined by the alkylation
reaction,s de,scribed in the Scheme~i. Other reactions useful in the
preparation of heteroaryl moieties are described in "Comprehensive
Organic Chemi~try, Volume 4: Heterocyclic Compounds" ed. P.G.
Samme~i, Oxford (1979) and references therein. Aryl-aryl coupling
i~s generally de.scribed in "Comprehensive Organic Functional Group
Transformations," Katritsky et al. ed,s., pp 472-473, Pergamon Pre,ss
(1995).
Synop~i,s of Scheme.s 1-24:
The requisite intermediates are in some cases commercially
available, or can be prepared according to literature procedures, for the
most part. Scheme~ 1- 14 illustrate synthesis of the instant diheteroaryl
compound which incorporate a preferred benzylimidazolyl sidechain.
Thus, in Scheme 1, for example, the comInercially available dithienyl-
carboxylic acid I may be reduced. The triflate of the interrnediate
alcohol II may be formed in situ and coupled to a ~uitably sub,stituted
benzylimidazolyl III to provide, after deprotection, the in~tant
compound IV.
Those biheteroaryl intermediates that are not commercially
;~vailable may be synthesized by methods known in the art a~s well as
tho~e reaction,s illu~strated in Schemes 2-6. Thus, a~ illustrated in
Scheme 2 a thienyl boronic acid V may be reacted under Suzuki
coupling condition.~ (Pur~ Appl. Chem., 63:4l9 (1991)) with a liuitably
~ub~tituted halogenated heteroaryl moiety, ~such a.s 2-bromo, to provide
the biheteroaryl carboxylic acid VI, which can then be processed a~
de~cribed in Scheme 1. Scheme 3 illu~trate~ the analogou~s serie.s of
reaction.~; .starting with the halogenated heteroarylaldehyde.
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- 42 -
Scheme 4 illustrates the reaction wherein the "telTninal"
heteroaryl moiety is employed in the Suzuki coupling a,s the halogenated
reactant. Such a coupling reaction is also compatible when one of the
reactants incorporates a suitably protected hydroxyl functionality a~
5 illustrated in Scheme 5.
Negishi chemi~stry (07g. Synth., 66:67 (1988)) may also be
employed to form the biheteroaryl component of the instant compounds,
as shown in Scheme 6. Thus, a zinc bromide adduct, .such as 3-thieny]
zinc bromide, may be coupled to a suitably ~ub,stituted heteroaryl halide
10 in the pre,sence of nickel (Il) to provide the biheteroaryl VII. The
heteroaryl halide and the zinc bromide adduct may be selected ba.sed
on the availability of the ,starting reagents.
As illu~trated in Scheme 7, the ,~e4uence of coupling
reactions may be modified .such that the heteroaryl-heteroaryl bond
1~ is formed la~t. Thus, a suitably substituted imidazole may first be
alkylated with a heteroarylmethyl halide to provide intermediate VIII.
Intermediate VIII can then undergo Suzuki type coupling to a ~suitably
substituted heteroaryl boronic acid.
Scheme g illustrate.s the ~synthesis of a thiazole containing
20 instant compound from the acyclic precursor.~. Similar strategies may
be utilized to prepare other bisheteroatom moietie,~.
Scheme 9 illustrates synthe~i,s of an instant compound
wherein a non-hydrogen R9b i.~ incorporated in the instant compound.
Thus, a readily available 4-substituted imidazole IX may be selectively
25 iodinated to provide the 5-iodoimidazole X. That imidazole may then
be protected and coupled to a suitably substituted benzyl moiety to
provide intermediate XI. Intermediate XI can then undergo the
alkylation reactions that were de.scribed hereinabove.
Scheme 10 illustrate.s .synthe.si.s of in.stant compounds
30 that incorporate a preferred imidazolyl moiety connected to the
biheteroaryl via an alkyl amino, .sulfonamide or amide linker. Thu.~,
the 4-aminoalkylimidazole XII, wherein the primary amine is protected
als the i-lhthalimide, i.s ~selectively alkylated then deprotected to provide
the amine XIII. The amine XIII may then react under condition~ well
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WO 97136585 PCT/[JS97/06259
- 43 -
known in the art with various activated biheteroaryl moieties to provide
the instant compounds shown.
Compounds of the instant invention wherein the
A 1 (CR 1 2)nA2(CR 1 2)n linker is oxygen may be synthesized by
methods known in the art, for example as shown in Scheme 11.
The suitably substituted phenol XIV may be reacted with methyl
N-(cyano)methanimidate to provide the 4-phenoxyimidazole XV.
After selective protection of one of the imidazolyl nitrogens, the
intermediate XVI can undergo alkylation reactions as described for
the benzylimidazole~s hereinabove.
Scheme 12 illustrates an analogous .series of reactions
wherein the (CR22)pX(CR22)p linker of the instant compounds i.s
oxygen. Thus, a suitably ,substituted haloheteroaryl alcohol, such as, is
reacted with methyl N-(cyano)methanimidate to provide intermediate
XVI. lntermediate XVI is then protected and, if de.~ired to forrn a
compound of a preferred embodiment, alkylated with a ,suitably
protected benzyl. The intermediate XVII can then be coupled to a
second heteroaryl moiety by Suzuki chemistry to provide the instant
compound.
Compounds of the instant invention wherein the
A 1 (CR 1 2)nA2(CR 1 2)n linker is a substituted methylene may be
synthesized by the method,s ~;hown in Scheme 13. Thus, the N-protected
imidazolyl iodide XVIII i~s reacted, under Grignard condition~ with a
suitably protected benzaldehyde to provide the alcohol XIX. Acylation,
followed by the alkylation procedure illustrated in the Scheme,s above
(in particular, Scheme 1 ) provides the instant compound XX. If other
R 1 ~sub~stituents are desired, the acetyl moiety can be manipulated as
illustrated in the Scheme.
Addition of various nucleophile,s to an imidazolyl aldehyde
may also be employed to form a sub,stituted alkyl linker between the
bihetero~ryl moiety and the prel'erred W (imidazolyl) as shown in
Scheme 14. Thus a bishalogenated five membered heteroaryl, such a~
2,4-dibromothiophene. may undergo metal halogen exchange followed
by reaction with a suitably substituted imidazolyl aldehyde and
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- 44 -
acteylation to form a regioisomeric mixture of the acetyl intermediates.
The halogenated regioisomeric mixture may be chromatographically
separated at this stage, if convenient. Suzuki coupling with a suitably
substituted 5-membered heteroaryl boronic acid affords the instant
5 acetoxy compound, which can be treated with lithium hydroxide to
remove the acetyl group. Then, similar substituent manipulation a,s
,shown in Scheme 13 may be performed on a fully functionalized
compound which incorporate.s an R2 hydroxyl moiety.
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- 45 -
SCHEME 1
~OH rOH
,~ LiAlH
11
Tr Tr~
N~ NiCI2(PPh3)2, <~N
/~~ ~J
R8 R8 111
OH
S--~ (CF3SO2)20,-78~C
NEtiPr2-78~G 20~C
C H2CI2
Il T~
<\_N'~ 5~ r
~J 55~C, C H30H
R8
-~
IV
R8
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- 46 -
SCHEME 2
(HO)2B~
HO ~ Br
~r Pd(PPh3)4
HO ~ '~) 4
O Vl
HO~ ¢~
SCHEME 3
(HO)2B
Pd(PPh3)4
,¢~ NaBH4
d
A ~ S
HO ~
- CA 022S0204 1998-09-24
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- 47 -
SCHEME 4
Br
MeO~ ~ ~(OH)2
Pd(PPh3)4
MeO ~ LiAlH~
o
HO~ '~
SCHEME ;~
P3SiO ~/~B(OH)2
Pd(PPh3)4
R3SiO ~ ~ Bu 4N F
HO~
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- 48 -
SCHEME 5 (continued)
R3SiO~_~ (HOkB /¢~
Pd(P Ph3~4
R3SiO ~ J~'r~ Bu4NF
HO ~¢~
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- 49 -
SCHEME 6
BrZn~
R3SiO~,~Br NiCI2(PPh3)2
R3SiOJt~ 4
HO~,¢~r~
R3SiO~ ~ Br~
NiCI2(PPh3)2
R3SiO ~ Bu4N F
HO~
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PCTtUS97tO6259
- 50 -
SCHEME 7
~J~Br
li. MeOH
~J reflux
R8
<~_N~ ~r~Br (H~)
~J Pd(PPh3)4
R8 Vlll
<~N
R8/
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SCHEME ~s
R8 ~ NH3/EtOH/RT
~ y'COOCH3
N
R8 ~ Lawesson' s Reagent
N 1,4-dioxane
~ ~CONH2 80~C
N~
8 1~1
R
~CSNH2
N
-
CA 02250204 lg9X-09-24
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- 52 -
SCHEME ~s (continued)
Me Me
Py.HBr.Br
O O
N~
N NH2
R8
<N3~\~JN, Me
/ R6
R8
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- 53 -
SCHEME 9
H H
R9b~? Nal, NaHCO3,!2 Rgb~ TrCI, NEt~
IX X
9b~ ~I C ,lPPh3l2 ~ ~N
b,,~/, R6
Tr b~a~ e'
~/ i. -78~C-20~C
R8/~J ii. MeOH, reflux
Xl
R9b b~
~ N
- R8
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- ~4 -
SCHEME 10
0~
~~Br
<\ ~ ~ 55~C,CH~CN
N N~i. EtOH,80~C, NH2NH2
Xll N
<~ ~I
N----NH2
R8 ~/
~,
Xlll
acylation, sulfonylation <i 3~
or alkylatiorl R8 ~J ~ R6
</ ~ O~ "~
~/ e~o~P~ R6
</ 11
- R6 ~ N~,,~R6
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WO 97136585 PCT/US971062S9
SCHEME 1 1
OH i. Na, MeOH
NC~/ ii. 120~C
XIV H3C o
N--\~N
H Tr
N N
~ TrCI, NEt3, ~N
NC ~ NC
XV XVI
Tr~N~ b~a~ 78~C-20~C
NC~ OTf ii. MeOH reflux
XVI
R6
~_N ~ a~e, d
~0
NC ~'
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- 56 -
SCHEME 12
H
R2 N
~\~OH i, Na, MeOH ~_
~S ii 1 20~C ,~ O
Br
N--~N XVI
R8
Tr,
N~
TrCI, NEt3, R2 <\_N OTf
78~C-20~C
B ~5 ii. MeOH reflux
R8 ~B(~H)2
R2 ~N , ¢~ A~\JR6
~\3 0 DMF, Pd(PPh~)4
)~S K3PO4, 80~C
Br XVI I
R8
2 ~ q J ~
~\~0
~-S
A~\jJ
- R6
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SCHEME 13
Tr~ ~N~
~N~ EtMgBr ~N
~N ~ ~('OH
XVIII R R8 XIX
Ac O py ~N ~ ~z
OAc (C F3SO2)20, -78~C
R8/ NEtiPr2,CH2cl2
b,r~,, R 6
~ bl~a LiOH
~ <
~/ OAc
R8 XX
R8
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- 5~ -
SCHEME 13 (continued)
b,~/, R 6
N ~ b~ a ~ e '
~ ~ Cl
R8 l R6
b~
<~N~, a(~ R~
~,J NH2
R8
N~ abl,~d/'R6
b~
OMe
R8
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59 _
SC~IEME 14
Br BuLi Br Br
~ Br ;
N
1. </ 11
(i~ OAc
2. AcCI
R8
~ R6
a'_ e~''
(HO)2B LiOH
Pd(PPh3)4
~J R6
R8
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Schemes 15-24 illustrate reaction,s wherein the moiety
(l 8)r /'T9~
V ~ A1(CRl2~nA2(CR12)n~;~W,~ ~ (CR22)p-X
incorporated in the compounds of the in.stant invention i.s repre,sented by
other than ~ Isubstituted imidazole-containing group.
Thu.s, the intermediates who.se synthesis are illu.strated in
Scheme~s hereinabove and other biheteroalyl intermediate.s obtained
commercially or readily synthesized, can be coupled with a variety of
aldehyde.s. The aldehydes can be prepared by .~itandard procedures? .such
as that described by O. P. Goel, U. Krolls, M. Stier and S. Kesten in
10 Or~anic Synthelses, 19X8, ~7, 69-75, from the appropriate amino acid
(Scheme 15). Metal halogen exchange chemistry (Scheme 14) may be
employed when manipulating the aldehydes. Alternatively, Grignard
chemistry may be utilized, as shown in Scheme 15. Thu.s, Suzul~i
coupling provides, for example, the pyrrole containing biheteroaryl
15 XXI. Reaction of the intermediate XXI with a Grignard reagent
provides the N-pyrrylmagnesium derivative XXIa, which i~s then
reacted with an aldehyde to provide the C-alkylated instant compound
XXII. The product XXII can be deoxygenated by methods known in
the art, such as a catalytic hydrogention, then deprotected with
20 trifluoroacetic acid in methylene chloride to give the final compound
XXlla. The final product XXII may be isolated in the salt forrn, for
example. as a trifluoroacetate, hydrochloride or acetate salt, among
others. The product diamine XXII can further be selectively protected
to obtain XXIII, which can subse4uently be reductively alkylated with
25 second aldehyde to obtain XXIV. Removal of the protecting group, and
conversion to cyclized product.s such a,s the dihydroimidazole XXV can
- be accomplished by literature procedures.
Scheme 16 illustrates the u.se of in .situ forrnation of a
lithium anion of a ,suitably ,sub~stituted N-alkyl pyrrole to provide the
30 C-alkylated compound of the instant invention.
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If the biheteroaryl ,subunit is reductively alkylated with an
aldehyde which al~so has a protected hydroxyl group, ~uch a,~ XXVI in
Reaction Scheme 17, the protecting groups can be subsequently removed
to unmask the hydroxyl group (Reaction Schemes 17, l~). The alcohol
5 can be oxidized under standard condition,s to e.g. an aldehyde, which
can then be reacted with a variety of organometallic reagents such as
Grignard reagents, to obtain secondary alcohol,s such als XXX. In
addition, the fully deprotected amino alcohol XXXI can be reductively
alkylated (under condition,s de~scribed previously) with a variety of
10 aldehyde~ to obtain ~secondary amine~ uch a~ XXXII (Reaction
Scheme I X), or tertiary amine~;.
The Boc protected amino alcohol XXVIII can al,so be
utilized to ,synthesize 2-aziridinylmethylbiheteroaryl ~uch a~s XXXIII
(Reaction Scheme l9). Treating XXVIII with l,l'-sulfonyldiimidazole
1 ~S and ,sodium hydride in a ,solvent ~iuch a,s dimethylformamide led to the
formation of aziridine XXXIII . The aziridine i~s reacted with a
nucleophile, ,such a~s a thiol, in the pre~ience of ba~se to yield the ring-
opened product XXXIV .
In addition, the biheteroaryl ,subunit can be reacted with
20 aldehyde~s derived from amino acid~s ,such as O-alkylated tyro.sine~,
according to ~standard procedure~ to obtain compound~ ~uch a.~ XL, as
.shown in Reaction Scheme 20. When R' i~ an aryl group, XL can fir~;t
be hydrogenated to unmask the phenol, and the amine group deprotected
with acid to produce XLI. Alternatively, the amine protecting ~roup in
25 XL can be removed, and O-alkylated phenolic amines such as XLII
produced.
Scheme~ 21-24 illustrate .synthe,ses of ,suitably .substituted
aldehyde~i u~seful in the synthe.se~s of the in.stant compounds wherein the
variable W i,s pre~ent as a pyridyl moiety. Similar .synthetic strategie~
30 for preparing alkanols that incorporate other heterocyclic moietie~ for
variable W are al~o well known in the art.
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- 62 -
SCHEME 15
~3 + ,b~ R
b~ EtMgBr
a e
H XXI
b~R
B M~ XXla
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- 63 -
SCHEME 15 (continued)
R6 Boc N H
ab~ e~d~ Boc NH CHO
~;~ Et20
BrMg
b~ 1. catalytic
a~ d~ hydrogenation
HO h~ 2. CF3CO2H
Boc NH~ ) 'N CH2CI2
NHBoc 6
XXII ab'~e,,d/'R Boc20
NH2~_~ CH2CI
XXlla b~
a,~Jd' ~,CHO
BocN H <~
\ < H NaBH(OAc)3
NH2 Et3N, CICH2CH2CI
XXIII
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SCHEME 15 (continued)
~;
BocNH~ C~3CO2H,
= NH NaHCO3
~;~ XXIV R6
b~
NH2 ~ ~ NC
~= NH AgCN
b~R
N
/ H
N~N~
XXV
~3
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SCHEME 16
~3 (HO) B/a e
alkyl Boc NH~
~R6 BuLi Boc NH CHO
a TMEDA Et20
alkyl b~
a~ d
Boc NH ) ~
~ I
alkyl
NHBoc
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- 66 ~
SCHEME 17
~2 Et~<)
BocNH CHO
XXVI
BnO~ b~ a'(~ 20% Pd(oH)2 H2
R2 C H3CO2H
NHBoc
HO ~ ~ R6
e R2 DMSO CH2C12
NHBoc (C2Hs~3N
XXVI I
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- 67 -
SCHEME 17 (CONTINUED~
OH~ ~ b'$~ R6
NHBoc
XXIX
HO '~ a;(~
R')~ e R2
NHBoc
XXX
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PCT~US97/06259
- 6~ -
SCHEME 18
HO b b~ R6
e 2 C H2C12
NHBoc
XXVIII
HO ~ b b~,R6
R2 NaBH(OAc)3
NH2 CICH2CH2
XXXI
~- a~
NH
R'CH2 XXXII
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SCHEME 19
6 N =~ / N
H0 a~ 2
\f R2NaH, DMF0~C
NHBoc
XXVIII
b~ b~R R~SH
<,~ a~d e' CH30H
H
XXXIII
b,~ R6
,~_ a~, d'
R"S ~ a~<d e
\~
NH2
XXXIV
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- 70 -
SCHEME 20
Hl~ 1 ) Boc20, K2CO3 )~
H2NCO2H2) CH2N2, EtOAc
XXXV XXXVI
HO,~
LiAlH4 ,,b,~ R"'CH2X
THF 1 Cs2CO3
0-20~C BocNH CH2OH DMF
XXXVII
R"'CH20~ R"'CH20
DMSON
BocNH CH2OH 20~C BocNH CHO
XXXVI I I IXL
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SCHEME 20 (continued)
R"'CH~3 g ~2
BocN H C HO
~~~~ R"' is not aryl
1. Et20
Et20
2. 20% Pd(OH)2, H2
CH30H, CH3CO2H
3. HCI, EtOAc \
~ ,b~ a'~(~
R"'CH20 R2
NHBoc
1) 20% Pd(~H)2~ H2
CHq,OH,CH3CO2H/
S~ a~
NH2
~H R2 XLI I
XLI
-
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SCHEME 21
,~ CH3 1) HNo2~Br2 ~CO2CH3
~I 2) KMnO4 , ~
H2N N 3) MeOH,H+ Br N
R6
~\MgCI R~ CO CH
NaBH4 (excess)
SO Py Et3N ~ 3 CHO
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SCHEME 22
1. EtO(CO)CI R6
Zn C~C~
N 3. S, xylene, heat N
R6 R6
NaBH4 ~ SO3Py, Et3N ~
(excess) ~; 3"CH20H DMSO ~CHO
R6 R
Br~"CO2CH3 [~\ 9 ¢~
NZnCI2, NiC12(Ph3P)2 l~Co2cH3
R6 R6
'
NaBH4 I SO3 Py, Et3N
~CH20H ~ ~CHO
(excess) N DMSO N
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- 74 -
SCHEME 23
CO2CH3
Br~ 1. LDA, CO2 Br~
N2. MeOH. H+ N
R6
~\ MgCI ~ co2CH3
ZnCI2, NiC12(Ph3P)2 N
R6
NaBH4 ~excess) ~32~H SO3 Py, Et3N
DMSO
N
R6
CHO
~3
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- 75 -
SCHEME 24
[~ 1. LDA, CO2 ~N,CBHr3
2. (CH3)3SiCHN2
R6 ~\ Br R6 ~
Zn, NiCI2(Ph3P)2 N~CO2CH3
R6 ~q
excess NaBH4 l~ SO3Py, Et3N
N~CH20H DMSO
R6 1~
N ~,CHO
The in~tant compounds are u.seful als pharmaceutical a
5 gent.s for m~mnl:~l.s, especially for humans. These compounds may be
administel-ed to patient.~ for u~e in the treatment of cancer. Example.~i
of the type of cancel which may be treated with the compounds of this
invention include, but are not limited to, colorectal carcinoma~ exocrine
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- 76 -
pancreatic carcinoma, myeloid leukemia,s and neurological tumors.
Such tumors may arise by mutations in the ras genes them.selves,
mutations in the proteins that can regulate Ras activity (i.e.,
neurofibromin (NF-I), neu, scr, abl, Ick, fyn) or by other mechanisms.
The compounds of the instant invention in~ibit farnesyl-
protein transferase and the farne,sylation of the oncogene protein Ras.
The instant compound,s may also inhibit tumor angiogenesis, thereby
affecting the growth of tumors (J. Rak et al. Can~er Research,
55:4575-45X0 (1995)). Such anti-angiogenesils properties of the
10 in.stant compounds may also be u~seful in the treatment of certain
forms of blindne~ss related to retinal va.scularization.
The compounds of thi~s invention ale also useful for
inhibiting other proliferative disea~ies, both benign and malignant,
wherein Ra~i proteins are aberrantly activated a,s a result of oncogenic
15 mutation in other genes (i.e., the Ras gene itself i~s not activ~ted by
mutation to an oncogenic form) with said inhibition being accomplished
by the administration of an effective amount of the compounds of the
invention to a m~mm~l in need of ,such treatment. For example, a
component of NF-I is a benign proliferative disorder.
The instant compound,s may also be u,seful in the treatment
of certain ~iral infections, in particular in the treatment of hepatitis
delta and related viru~e~ (J.S. Glenn et al. S~ n~c, 256:1331-1333
( I 992).
The compound,s of the instant invention are al,so u,seful in
25 the prevention of re,steno~is after percutaneous transluminal coronary
angioplalsty by inhibiting neointimal forrnation (C. Indolfi et al. Natu~
medic~ e~ 1:541-545(1995).
The instant compounds may also l~e useful in the tre~tment
and prevention of polycy,stic kidney disease (L L. Schaffner et al.
30 Ame)i~an Jourllal (~f Path~logy, 142:1051-1060 (1993) ~nd B. Cowley,
Jr. et al.~AS~B J~ ,rnal, 2:A3160 (19~
The instant compound~s m~y ~]so be useful for the treatment
of fungal infection.~.
The compound,s of thi,s invention may be adminis~ered
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to m~mmals, preferably humans, either alone or, preferably, in
combination with pharmaceutically acceptable carriers or diluent~,
optionally with known adjuvant~s, such as alum, in a pharmaceutical
composition, according to standard pharmaceutical practice. The
5 compounds can be administered orally or parenterally, including the
intravenous, intramuscular, intraperitoneal, subcutaneou.s, rectal and
topical routes of administration.
For oral use of a chemotherapeutic compound according
to this invention, the selected compound may be administered, for
10 example, in the form of tablet.~ or capsule.~, or as an aqueous solution
or suspension. ln the case of tablet~i for oral use~ carriers which are
commonly u.sed include lactose and corn starch, and lubricating agent~.
such as magnesium stearate, are commonly added. For oral
administration in capsule form, useful diluents include lactose and dried
15 corn ~starch. When aqueous suspen~sions are re(luired for oral use, the
active ingredient is combined with emul,sifying and suspending agents.
If desired, certain sweetening and/or flavoring agents may be added.
For intramuscular, intraperitoneal, subcutaneous and intravenou~ use,
sterile solutions of the active ingredient are usually prepared, and the
20 pH of the solution~s should be suitably adjusted and buffered. For
intravenous use, the total concentration of solutes ~hould be controlled
in order to render the preparation i~otonic.
The compounds of the instant invention may also be
co-~lministered with other well known therapeutic agents that are
25 selected for their particular usefulness again,st the condition that is
being treated. For example, the instant compounds may be useful in
combination with known anti-cancer and cytotoxic agent~i. Similarly~
the instant compounds may be u.seful in combination with agents that
are effective in the treatment and prevention of NF-l, restinosis,
30 polycystic kidney disease, infections of hepatitis delta and related
viruses and fungal infections.
If formulated as a fixed dose, such combination product~
employ the compound~ of thi.'i invention within the do~sage ran~e
de.~cribed below and the other pharm~ceutically active a~ent(.~) within
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its approved dosage range. Compound~s of the instant invention may
alternatively be used sequentially with known pharmaceutically
acceptable agent(s) when a combination formulation is inappropriate.
The present invention also encompa,sses a pharmaceutical
5 composition useful in the treatment of cancer, comprising the
arlministration of a therapeutically effective amount of the compounds
of thi~ invention, with or without pharmaceutically acceptable carriers
or diluents. Suitable compositions of this invention include aqueou.s
~solutions comprising compoundls of this invention and pharmacolo-
10 ~ically acceptable carrier~s, e.g.t saline, at a pH level, e.g.. 7.4. The~olutions may be introduced into a patient'~ blood-stream by local bolu,~
injection.
A~ used herein, the term "composition" is intended to
encompass a product comprising the specified ingredients in the specific
15 amounts, as well a.s any product which result~i, directly or indirectly,
from combination of the specific ingredients in the .specified amounts.
When a compound according to this invention is
administered into a human subject, the daily dosage will normally
be determined by the prescribing phy.~ician with the dosa~,e generally
20 varying according to the age, weight, and response of the individual
patient, as well as the severity of the patient's symptoms.
In one exemplary application, a suitable amount of
compound is administered to a m~mm~l undergoing treatment for
cancer. Administration occurs in an amount between about 0.1 mg/kg
25 of body weight to about 60 mg/kg of body weight per day, preferably
of between 0.~ mg/kg of body weight to about 40 mg~g of body weight
per day.
The compounds of the instant invention are al.~o useful
as a component in an a~s~iay to rapidly determine the presence and
30 quantity of farnesyl-protein tran.sferase (FPTa.se) in a composition.
Thus the composition to be te~;ted may be divided and the two
portions contacted with mixture.s which compri.se a known sub.strate
of FPTase (for example a tetrapeptide having a cy.steine at the amine
terminus) and farnesyl pyropho.sphate and, in one of the mixture.s,
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- 79 -
a compound of the instant invention. After the as.say mixtures are
incubated for an ,sufficient period of time, well known in the art, to
allow the FPTa.se to farnesylate the substrate, the chemical content
of the assay mixtures may be determined by well known immuno-
5 logical, radiochemical orchromatographic technique~s. Becausethe compound.s of the in.stant invention are selective inhibitors of
FPTa,se, ab.sence or quantitative reduction of the amount of substrate
in the assay mixture without the compound of the instant invention
relative to the presence of the unchanged .substrate in the assay
10 containing the instant compound is indicative of the presence of
FPTa.se 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 u.seful in identifying
tissue samples which contain farnesyl-protein transferase and quantitat-
15 ing the enzyme. Thus, potent inhibitor compound.s of the in.stantinvention may be used in an active ~site titration assay to determine the
quantity of enzyme in the sample. A serie,s of samples composed of
aliquots of a tissue extract containing an unknown amount of farnesyl-
protein transferase, an exces.s amount of a known strate of FPTase
20 (for example a tetrapeptide having a cysteine at the amine terminu.s) and
farnesyl pyropho,sphate are incubated for an appropriate period of time
in the presence of varying concentrations of a compound of the instant
invention. The concentration of a .~ufficiently potent inhibitor (i.e., one
that has a Ki substantially smaller than the concentration of enzyme in
25 the assay vessel) required to inhibit the enzymatic activity of the sample
by 50% is approximately e~lual to half of the concentration of the
enzyme in that particular .sample.
EXAMPLES
Example.~ provided are intended to assist in a further
under.standing of the invention. Particular material~ employed, specie.s
and condition~s are intended to be further illu.strative of the invention
and not limitative of the reasonable scope thereof.
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- ~0 -
EXAMPLE I
I -(5-(Thien-2'-yl)thien-2-ylmethyl)-5 -(4-cyanobenzyl)imidazole
5 trifluoroacetic acid salt
Step A: l-Trity]-4-(4-cyanobenzyl)-imidazole
To a suspension of activated zinc dust (3.57g,
54.9~ mmol) in THF (50 mL) was added dibromoethane (0.315 mL,
10 3.60 mmol) and the reaction stirred under argon for 45 minute,s, at
20~C. The su~pension wa.s cooled to 0~C and a-bromo-p-tolunitrile
(9.33g, 47.6 mmol) in TH~ (100 mL) wa~ added dropwi~e over a
period of 10 minutes. The reaction wa~ then allowed to xtir at 20~C
for 6 hour~ and bis(triphenylphosphine)Nickel II chloride (2.40g,
15 3.64 mmol) and 4-iodotritylimidazole (15.9Sg, 36.6 mmol) were
added in one portion. The resulting mixture wa~i stirred 16 hours
at 20~C and then quenched by addition of saturated NH4CI .solution
(100 mL) and the mixture stirred for 2 hour.s. Saturated aq.
NaHCO3 solution was added to give a pH of 8 and the solution was
20 extracted with EtOAc (2 x 250 mL), dried (MgSO4) and the solvent
evaporated in vacuo. The residue wa.s chromatographed (Silica ge],
0-20% EtOAc incH2cl2) to afford the title compound a~i a white
solid.
IH NMR (CDC13, 400Mz) ~ (7.54 (2H, d, J=7.9Hz), 7.3X(lH, s),
25 7.36-7.29 (1 IH. m), 7.I5-7.09(6H, m), 6.5~(1H, ~) and 3.93(2H, ,s)
ppm.
Step B: S-'Thien-2-yl)-thiophene-2-carboxaldehyde
To a ~olution of thien-2-ylboronic acid (0.939 g,
30 7.34 mmol) and Na2CO3 (2.40 g, 22.6 mmol) in water (75 mL)
i.~ added p-dioxane (75 mL). Thi.~ mixture i~ treated ,~equentially
with 5-carboxy-2-thiophenecarboxaldehyde (1.43g, 7.4~ mmol)
and palladium (Il) acetate (151 mg, 0.673 mmo]) and allowed to
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stir at ambient temperature for 16 hour~s. The ~solvent i~i evaporated
in vacuo. To the residue i.s added EtOAc (400 mL) and water (300
mL). The aqueous layer is acidified to pH I with 1.0 N a~l. HCl.
The aqueous layer is extracted with EtOAc (2 x 200 mL). The
organic extracts are combined, washing with brine (200 mL), 5~O aq.
Na2S~O3 (200 mL), sat. aq. NaCI (200 mL), drying (Na2SO4), and
the solvent evaporation in vacuo affords the title compound.
Step C: S-(Thien-2-vl)-2-hydroxymethylthiophene
To a solution of ~-(thien-2'-yl)-thiophen-2-
ylcarboxaldehyde (1.11 g, 5.73 mmol) in THF (25 mL) at 0~C i.s
added 1.0 M lithium aluminum hydride in tetrahydrofuran (12.0
mL, 12.0 mmol) over 10 minutes. The reaction i~ allowed to ~;tir at
ambient temperature for 3 hours. The reaction i~ cooled to 0~C, and
1~ water (0.5 mL), 4 N aq. NaOH (0.5 mL), and water (1.5 mL) are
added sequentially. The reaction i.s filtered through a pad of Celite
and the filtrate is evaporated in vacuo. The residue ils
chromatographed to afford the title compound.
20 Step D: 1-(5-(Thien-2-yl)thien-2-ylmethyl)-5-(4-
cyanobenzyl)imidazole trifluoroacetic acid salt
To a solution of 5-(thien-2'-yl)-2-hydroxymethyl
thiophene (2~s3 mg, 1.44 mmol) and diisopropylethylamine (0.260
mL, 1.49 mmol) in dichloromethane (6.0 mL) at -7g~C is added
2~ trifluoromethanesulfonic anhydride (0.250 mL, 1.49 mmol) and
the mi~ture stirred at -7~i~C for I hour. To this mixture i!i added
a .solution of l-trityl-4-(4-cyanobenzyl)imidazole (613 m~ . 1.44
mmol) in dichloromethane (6.0 mL). The mixture is allowed to
warm to ambient temperature and stirred ~or 2 hours. The solvent i~
30 evaporated in vacuo. The residue i,~ di~s.~olved in methanol (15 mL),
heated at reflux for I hour~ and the solvent i~i evaporated in vacuo.
The residue i~i partitioned between C~12C12 and ~sat. aq. NaHCO3
~olution. The organic layer i.s dried, (Na2S04) and the solvent i,~;
evaporated in vacuo. The residue i~i purified by preparative HPLC,
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(gradient elution, 95 :5 to 5:95% water:acetonitrile containing 0.1%
trifluoroacetic acid) to afford the title compound a.s a trifluoroacetic
acid salt.
EXAMPLE 2
I -(4-Cyanobenzyl)-5-(5-( 1 -methylpyrrol-2-yl)-thiazol-2-
ylmethyl)imidazole
Step A: IH-Imidazole-4-acetic acid methyl ester hydrochloride
A solution of 1 H-imidazole-4-acetic acid hydrochloride
(4.00g, 24.6 mmol) in methanol (100 ml) was saturated with
gaseous hydrogen chloride. The resulting solution was allowed
to stand at room temperature for lX hours. The solvent was
evaporated in vacuo to afford the title compound as a white solid.
lH NMR (CDC13, 400MHz) ~ 5(lH, s), 7.45(1H, ~;), 3.~9(2H, s)
and 3.75(3H, s) ppm.
Step B: I-(Triphenylmethyl)-lH-imidazol-4-ylacetic acid methyl
ester
To ~ solution of the product from S~ep A (24.~5 g,
0.141 mol) in DMF (115 mL) wals added triethylamine (57.2 mL,
0.412 mol) and triphenylmethyl bromide (55.3 g, 0.171 mol) and
the suspension was stirred for 24 hours. After this time, the reaction
mixture wa,s diluted with EtOAc and water. The organic phase wa~
washed with sat. aq. NaHCO3 dried, (Na2SO4) and the solvent
evaporated in vacuo. The residue wa~ purified by chromatography
(Silica gel, 0-100% EtOAc in hexanes) to provide the title compound
a~ a white solid.
I H NMR (CDC13, 400MHz) ~ 7.35(1 H, ~i), 7 .3 1 (9H, m), 7 .22(6H,
m), 6.76(1H, ~), 3.6~(3H, s) and 3.60(2H, ~) ppm.
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Step C: [1 -(4-Cyanobenzyl)- 1 H-imidazol-5-yl]acetic acid methyl
ester
To a solution of the product from Step B (8.00g, 20.9
mmol) in acetonitrile (70 mL) was added 4-cyanobenzyl bromide
5 (4.10g, 20.92 mmol) and heated at 55~C for 3 hour.s. The reaction
was cooled to room temperature and the resulting imidazolium salt
was collected by filtration. The filtrate was heated àt 55~C for lX
hours. The reaction mixture was cooled to room temperature and
evaporated in vacuo. To the residue was added EtOAc (70 mL) and
10 the relsulting precipitate collected by filtration. The precipitated
imidazolium .salt~ were combined, suspended in methanol (100 mL)
and heated to reflux for 30 minute.s. After thi.s time, the solvent was
removed in vacuo. The resulting residue was suspended in EtOAc
(75 mL) and the solid i.solated by filtration and washed with EtOAc.
15 The solid was treated with sat. aq. NaHCO3 solution (300 mL) and
CH2C12 (300 mL) and .stirred at room temperature for 2 hours. The
organic layer was separated, dried, (MgSO4) and evaporated in
vacuo to afford the title compound as a white .solid
lHNMR (CDC13, 400MHz) ~ 7.65(1H, d, J=~Hz), 7.53(1H, s),
7.15(1H, d, J=~Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and
3.45(2H, s) ppm.
Step D: 4-[5-(Aminocarbonylmethyl)imidazol- I -
ylmethyllbenzonitrile
To a 100 mL gla.~s pres~sure vessel with a stirring bar
was addedl-(4-Cyanobenzyl)-lH-imidazol-5-yl]acetic acid methyl
ester (6.00g, 23.5 mmol) and absolute ethanol (50 mL). Thi,s
well stirred solution was cooled to -7~~C and 50 mL of anhydrous
ammonia was condensed in. The vessel was sealed and the mixture
warmed to ambient temperature. This solution wa.s .stirred 24 hours
at ambient temperature. The exces.~ ammonia wa.s allowed to
evaporate and the ethanol wa~s removed in vacuo. The solid residue
was triturated with EtOAc and collected on a frit. This material wa~
dried in vacuo to give the title compound as a white ~iolid.
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lH NMR (DMSO-d6, 400MHz) ~ 3.25(s, 2H), 5.32(s, 2H), 6.~(s,
lH), 6.96(s,1H), 7.24(d, j-8Hz,2H), 7.42(s,1H), 7.6~(s,1H), 7.~3(d,
j=8Hz, 2H).
5 Step E: 1-(4-Cyanobenzyl)-5-(aminothiocarbonyl)methyl-lH-
imidazole
To a 50 mL round bottomed flask with a stirring bar,
reflux condenser and an argon inlet was added 4-15-(aminocarbonyl-
methyl)imidazol-l-ylmethyl]benzonitrile (0.36g, 1.49 mmol),
10 Lawes,son's reagent ~0.73g, 1 .X mrnol) and 1 ,4-dioxane ( 10 mL).
This well stirred mixture wa~s heated at X0~C for 24 hours. The
cooled reaction mixture was concentrated in vacuo and the residue
wa.s chromatographed (silica gel, 10% 2-propanol in ammonia
saturated CHC13). The title compound was obtained as a yellow,
1~ crystalline solid.
lH NMR (DMSO-d6, 400MHz) ~ 3.66(s, 2H), ;S.41(s, 2H), 6.~5(s,
lH), 7.24(d. j-~Hz,2H), 7.70(s,1H), 7.~2(d, j=8Hz, 2H), 9.21(s, lH),
9.56(.s, lH).
~0 Step F: 1-(4-Cyanobenzyl)-5-~5-(1-methylpyrrol-2-yl)-thiazol-2-
ylmethyllimidazole
To a 25 ml round bottomed flask with a stirring bar
reflux condenser and an argon inlet is added 1-(4-cyanobenzyl)-5-
(aminothiocarbonyl)methyl-lH-imidazole (0.12g, 0.468 mmol), dry
25 THF (10 mL). and N-methyl-a-bromoacetylpyrrole (0.098g, 0.491
mmol). This mixture i.s heated at 50~C for 7 hours. The cooled
reaction mixture is diluted with EtOAc and washed suces.sively with
aq. NaHCO3 ~ water, and brine. The organic extract is dried,
-(MgSO4) and the solvent is evapor~ted in vacuo. The residue is
30 purified by chromatography to afford the title compound.
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EXAMPLE 3
In vitrf) inhibition of ras farne,syl transferase
Assay~ of fa) nesy~-prc)tein tran~fèrclse. Partially purified
S bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and Ras-CAIL)
were prepared a~ described by Schaber et al., J. Biol . Chem. 26~: 14701 -
14704 (1990), Pompliano,_ah, Biochemi,stry 31:3~00 (1992) and
Gibbs et ah, PNAS U.S.A. ~s6:6630-6634 (19~9), respectively. Bovine
FPTase wa,s assayed in a volume of 100 ~I containing 100 mM N-(2-
10 hydroxy ethyl) piperazine-N'-(2-ethane ~sulfonic acid) (HEPE~), pH
7.4, 5 mM MgC12, 5 mM dithiothreitol (DTT). 100 mM [3H]-farnesyl
diphosphate (~3H]-FPP; 740 CBq/mmol, New England Nuclear), 650
nM Ras-CVLS and 10 ~g/ml FPTa,se at 31 ~C for 60 min. Reaction.s
were initiated with FPTase and stopped with I ml of 1.0 M HCL in
15 ethanol. Precipitate,s 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 re,spect to both
substrates, FPTase levels and time; less than 10% of the [3H]-FPP was
utilized during the reaction period. Purified compounds were dissolved
20 in 100% dimethyl ,sulfoxide (DMSO) and were diluted 20-fold into the
assay. Percenta~e inhibition i,s measured by the amount of incorpora-
tion of radioactivity in the pre,sence of the test compound when
compared to the amount of incorporation in the absence of the test
compound.
Human FPTase was prepared a,s described by Omer
et ah, Biochemistry 32:5167-5176 (1993). Human FPTase activity
was assayed a,s described above with the exception that 0.1 % (w/v)
polyethylene glycol 20,000, 10 ~lM ZnCl~ and 100 nM Ra,s-CVIM were
added to the reaction mixture. Reaction.s were performed for 30 min.,
stopped with 100 ~1 of 30% (v/v) trichloroacetic acid (TCA) in ethanol
and processed as described above for the bovine enzyme.
The compounds of the instant invention are tested for
inhibitory activity against human FPTase by the as~ay de.scribed above.
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EXAMPLE 4
In vivo ras farne.sylation assay
The cell line used in this assay is a v-ras line derived
5 from either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21.
The assay i~ performed e.ssentially as described in DeClue, J.E. et ah,
Cancer Re~search 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 hourls at
10 37~C, the cells are labelled in 3 ml methionine-free DMEM supple-
meted with 10% regular DMEM, 2% fetal bovine Iserum and 400
mCi[35S]methionine (1000 Ci/mmol). After an addition~l 20 hours, the
cells are Iysed in 1 ml Iysis buffer (1% NP40/20 mM HEPES, pH 7.5/5
mM MgC12/lmM DTT/10 mg/ml aprotinen/2 mg/ml leupeptin/2 mg/ml
15 antipain/0.5 mM PMSF) and the lysate~s cleared by centrifugation at
100,000 x g for 45 min. Aliquot.s of Iysates containing equal numbers
of acid-precipitable counts are bought to 1 ml with IP buffer (lysi,s
buffer lacking DTT) and immunoprecipitated with the ras-specific
monoclonal antibody Y13-259 (Furth, M.E. et al., J. Virol. 43:294-304,
20 (19~2)). Following a 2 hour antibody incubation at 4~C, 200 ml of a
25% .suspension of protein A-Sepharo~;e coated with rabbit anti rat IgG
is added for 45 min. The immunoprecipitates are washed four time~
with IP buffer (20 nM HEPES, pH 7.5/1 mM EDTA/I % Triton X-
100Ø5% deoxycholate/0.1%/SDS/0.1 M NaCI) boiled in SDS-PAGE
25 sample buffer and loaded on 13% acrylamide gel~. When the dye front
reached the bottom, the gel i.s fixed~ soaked in Enlightening, dried and
autoradiographed. The intensitie.s of the bands corresponding to
farne.sylated and nonfarnesylated ras protein,s are compared to
determine the percent inhibition of farne.syl transfer to protein.
-
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EXAMPLE S
In viv(~ .rowth inhibition assay
To determine the biological consequences of FPTase
5 inhibition, the effect of the compounds of the instant invention on the
anchorage-independent growth of Ratl cell,s tran,sformed with either a
v-7~as, v-ra~; or v-mos oncogene is te.sted. Cells transformed by v-Raf
and v-Mos maybe included in the analysis to evaluate the specificity of
instant compounds for Ras-induced cell transformation.
Rat I cells transformed with either v-ras, v-raf, or v-mos
are seeded at a density of 1 x 104 cells per plate (~5 mm in diameter)
in a 0.3% top agarose layer in medium A (Dulbecco's modified Eagle'~
medium supplemented with 10% fetal bovine serum) over a bottom
agarose layer (0.6%). Both layer.s contain 0.1% methanol or an
15 appropriate concentration of the in~stant compound (dissolved in
methanol at 1000 time.s the final concentration u~sed in the assay).
The cell~s are fed twice weekly with 0.5 ml of medium A containing
0.1% methanol or the concentration of the in,stant compound.
Photomicrographs are taken 16 days after the cultures are seeded
20 and comparison~s are made.
