Potent and selective non-cysteine-containing inhibitors of protein farnesyltransferase

Article abstract of DOI:10.1021/jm980298s

Potent and selective non-thiol-containing inhibitors of protein farnesyltransferase are described. FTI-276 (1) was transformed into pyridyl ether analogue 19. The potency of pyridyl ether 19 was improved by modification of the biphenyl core to that of an

Full text of DOI:10.1021/jm980298s

4288
J . Med. Chem. 1998, 41, 4288-4300
P oten t a n d Selective Non -Cystein e-Con ta in in g In h ibitor s of P r otein
F a r n esyltr a n sfer a se
David J . Augeri,* Stephen J . O’Connor,* Dave J anowick, Bruce Szczepankiewicz, Gerry Sullivan, J ohn Larsen,
Douglas Kalvin, J erry Cohen, Edward Devine, Haichao Zhang, Sajeev Cherian, Badr Saeed, Shi-Chung Ng, and
Saul Rosenberg
Departments of Cancer Research, D-47B, and Combinatorial Chemistry, D-4CP, Abbott Laboratories,
Abbott Park, Illinois 60064-3500
Received May 13, 1998
Potent and selective non-thiol-containing inhibitors of protein farnesyltransferase are described.
FTI-276 (1) was transformed into pyridyl ether analogue 19. The potency of pyridyl ether 19
was improved by modification of the biphenyl core to that of an o-tolyl substituted biphenyl
core to give 29. In addition to 0.4 nM in vitro potency, 29 displayed 350 nM potency in whole
cells as the parent carboxylic acid. The o-tolyl biphenyl core dramatically and unexpectedly
enhanced the potency of other compounds as exemplified by 46, 47, 48, and 49.
In tr od u ction
Oncogenic Ras proteins are found in 20-30% of all
human tumors including those of the lung (30%), colon
(50%), and pancreas (90%).1,2 These aberrant proteins
are constitutively activated and promote cell division
even in the absence of signals from diverse growth
factors.3 To function, Ras protein must undergo a series
of posttranslational modifications, of which farnesyla-
tion of a cysteine residue near the C-terminus by the
enzyme farnesyltransferase (FTase) is critical for
activity.4-8 Inhibition of this enzyme will render Ras
inactive and block the uncontrolled mitogenic signaling
pathway.
F igu r e 1.
Sch em e 1a
Several structurally distinct classes of farnesyltrans-
ferase inhibitors have recently been described.9-23 The
first reported inhibitors were designed to mimic the Ras
C-terminal tetrapeptide (CVFM) which is the minimum
substrate-derived inhibitor. FTI-276 (1, Figure 1) is one
of the most potent members of this class.24,25 This
compound incorporates a lipophilic 2-phenyl-4-ami-
nobenzoic acid as an isosteric replacement for the
internal Val-Phe dipeptide. The in vitro IC50 of 1 is 0.5
nM against farnesyl transferase, and the methyl ester
2 (FTI-277) potently blocks farnesylation of Ras in whole
cells (Ras processing, EC50 ) 0.1-0.2 µM).26 FTI-276
has also demonstrated significant in vivo activity against
human tumor xenografts in nude mice.24
Despite the impressive activity of 1 and 2, a number
of problems remained. The terminal cysteine residue
was highly susceptible to oxidative dimerization, neces-
sitating the use of dithiothreitol (DTT) for in vivo
evaluation. In addition, 1 (EC50 ∼1 µM) lacks the whole
cell potency of 2 (EC50 ) 0.1-0.2 µM), presumably due
to the lower membrane permeability of the acid. There-
fore, we sought to develop a compound that (1) lacks
the reactive sulfhydryl group and (2) is as active in
whole cells as the parent carboxylic acid.
a
(a) Pd(PPh3)4, Na2CO3, PhB(OH)2, toluene, reflux, 91%; (b)
KMnO4, 2:1 H2O/pyridine, reflux, 87%; (c) H2SO4, isobutylene,
dioxane, 70%; (d) 10% Pd/C, NH4CO2, MeOH, 99%; (e) i-Pr2NEt,
FMOC chloroformate, CH2Cl2, 92%; (f) TFA, CH2Cl2, 96%; (g)
L-Met-Wang resin, diisopropylcarbodiimide, HOOBt, NMP; (h) 33%
piperidine/DMF.
easy access to a diverse series of compounds. The core
FMOC-protected amino acid 5 was prepared from com-
mercially available 2-bromo-4-nitrotoluene 3. Palladium-
catalyzed aryl coupling using Suzuki conditions27 fol-
lowed by KMnO4 oxidation, formation of the tert-butyl
ester, and reduction of the nitro group gave 4 in 56%
yield. Protection of the amine with FMOC chlorofor-
mate and acid hydrolysis of the tert-butyl ester gave the
FMOC biphenyl acid 5 in 68% yield. Attachment of this
protected amine to a Wang resin28 containing the
requisite methionine residue and deprotection of the
FMOC group gave the desired substrate 6 suitable for
our library (Scheme 1).
Ch em istr y
We decided that a focused combinatorial library
derived from biphenylamine 5 would allow for quick and
The initial lead 7 resulted from this library and was
modified with more classical medicinal chemistry. The
10.1021/jm980298s CCC: $15.00 © 1998 American Chemical Society
Published on Web 09/19/1998

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4289
Sch em e 2a
a
(a) Ph-B(OH)2, aqueous Na2CO3, toluene, reflux, 97%; (b) aqueous KOH, MeOH/THF, rt, 70%; (c) BH3‚THF, rt, 98%; (d) KOH, aqueous
MeOH, reflux, 99%; (e) L-methionine, methyl ester hydrochloride, EDAC, HOBT, Et3N, DMF, rt, 79%; (f) C2O2Cl2/DMSO, Et3N, CH2Cl2,
-78 °C, 84%; (g) 3-aminopyridine, NaCNBH3, MeOH/AcOH, 73%; (h) LiOH, aqueous THF, 90%; (i) PBr3/LiBr, DMF, 96%; (j) K-O-(3-Pyr),
DME, cat. 18-C-6, 34%; (k) H2O2, TFAA, CH2Cl2, 94%; (l) NaOH, aqueous MeOH, reflux, 80-100%; (m) L-methionine methyl ester
hydrochloride, EDAC, HOBT, Et3N, DMF, rt, 70-80%; (n) LiOH, aqueous THF.
Sch em e 3a
F igu r e 2.
first change involved the removal of the nicotinamide
carbonyl to give 8. This was accomplished by using
3-pyridinecarboxaldehyde in a reductive amination
procedure (Figure 2).
The picolinate 9 and isonicotinate 10 were prepared
by standard coupling procedures. We also transposed
a
the carbon and nitrogen atoms of compound 8. We
investigated sulfur and oxygen linkers as well. The
preparation of these transposed compounds came from
the common precursor 13. From this alcohol we could
prepare the benzyl halide or the aldehyde (using Swern’s
conditions29), allowing for the assembly of a variety of
target compounds (Scheme 2).
(a) 3-Chloromethylpyridine‚HCl, KOH/aqueous DMF, 24%; (b)
Tf2O, pyridine, -10 °C to rt, 81%; (c) Ph-B(OH)2, aqueous Na2CO3,
toluene, reflux, 83%; (d) NaOH, aqueous MeOH, reflux, 90%; (e)
L-methionine methyl ester hydrochloride, EDAC, HOBT, Et3N,
DMF, rt, 78%; (f) LiOH, aqueous THF.
access to ortho, meta, and para substituted systems was
possible from intermediate 27. Use of iododimethyl-
terephthalate 11 for this purpose was precluded by
complete lack of selectivity in differentiation of the
carbonyl groups by reduction or hydrolysis. Therefore,
we decided to use commercially available dimethylami-
noterephthalate 25. We were able to realize a selective
reduction of the distal ester using DIBAL-H. This
selectivity is likely due to the vinylogous carbamate
character of the proximal ester. The resulting amino
alcohol was iodinated under the usual Sandmeyer
From aldehyde 14, we prepared a variety of amines
exemplified by compound 15. The intermediate benzyl
halide derived from alcohol 13 was used to prepare ether
and thioether analogues exemplified by compounds 19
and 20, respectively (Scheme 2). Retro-inversion of
oxygen analogue 19 to give oxygen analogue 24 was
carried out as shown in Scheme 3.
In addition to optimizing a cysteine replacement, we
investigated substitution of the biphenyl core. Efficient

4290 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Augeri et al.
Sch em e 4a
Ta ble 1. Results of Pyridine Modification of Lead Structure 7
compd
R
IC50 (nM)
EC50 (µM)
a
(a) DIBAL-H, THF/hexanes, -78 °C, 68%; (b) NaNO2, KI,
acetone, -15 °C, 83%; (c) SOCl2, LiCl, DMF, rt, 100%; (d)
3-hydroxypyridine potassium alkoxide, 18-crown-6, toluene, 48%;
(e) aryl boronic acids, Pd(PPh3)2Cl2, Cs2CO3, DMF, 80 °C, 61-96
%; (f) saturated aqueous LiOH, MeOH, reflux, 100%; (g) HOOBT/
EDC/DMF, L-methionine methyl ester HCl, 91%; (h) LiOH-THF-
H2O, 86%.
conditions to afford 26 in 56% yield over two steps.
Formation of the benzyl chloride followed by displace-
ment with the potassium alkoxide of 3-hydroxypyridine
gave the advanced intermediate 27 in 48% yield over
two steps. Aryl iodide 27 was utilized to access modi-
fications of the biphenyl core. o-, m-, and p-Tolyl boronic
acids and o-, m-, and p-methoxyphenyl boronic acids
were coupled to aryl iodide 27 using modified Suzuki
coupling conditions27 (Scheme 4). Biphenyl intermedi-
ates 28a -f were obtained in yields of 61-96%. Aryl
esters 28a -f were hydrolyzed using LiOH in refluxing
aqueous methanol to the corresponding acids which
were coupled to L-methionine methyl ester in 65-80%
yield over two steps. The methionine esters were
hydrolyzed with LiOH in THF-H2O to give the corre-
sponding acids 29-34. The o-Cl 35, o-CF3 36 , and o-Et
37 substituted biphenyls were prepared analogously.
To study amino acids other than methionine as
C-terminal substituents, we prepared amides derived
from most of the natural amino acids. These were
prepared by standard techniques (eq 1). Methionine
a
b
Methionine methyl ester. Methionine acid. Unless statistical
limits are given, the compounds were assayed once. The reliability
of the in vitro assay is (50%. The reliability of the cell-based assay
is (50-100%. Compounds that differ in potency by more than
3-fold should be considered statistically different.
Resu lts a n d Discu ssion
Directed combinatorial synthesis generated compound
7 as a 64 nM lead. However, this compound (as its
methyl ester) was only modestly active in our cell-based
assay (EC50 ) 50 µM). More traditional medicinal
chemistry was employed to modify this lead structure
to improve both in vitro and cellular activity (Table 1).
A 3-substituted pyridine, as originally discovered
through combinatorial chemistry, proved optimal (com-
pare 7, 9, and 10). Reduction of the amide carbonyl of
7 enhanced the potency 4-fold (8). Retro-inversion of
the nitrogen provided an additional 2-fold increase in
activity (15). Compounds incorporating ether (19) and
thioether (20) linking groups were similarly active
although the corresponding sulfone (21) showed a loss
in potency. The reversed oxygen analogue 19 was the
most potent, possessing an IC50 of 4 nM. However,
oxygen analogue 19 was still 10-fold less active than
FTI-276. In addition, it displayed similar characteris-
tics as FTI-276, requiring an ester for efficient whole
cell activity (EC50 methyl ester ) 3 µM).
(1)
Clearly, regiochemistry and, to a certain extent, the
electronic nature of the pyridine affect potency. Al-
though the exact function or binding mode of the
pyridine is unknown, one possibility is that it coordi-
nates with the zinc atom known to be present in the
sulfone and homoserine lactone analogues of the ami-
nopyridine were also prepared.

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4291
Ta ble 2. SAR of Biphenyl Core Substitution
F igu r e 3.
Ta ble 3. Comparison of Unsubstituted and o-Tolyl Biphenyl
Derived FTase Inhibitors
compd
R
IC50 (nM)
EC50 (µM)
19
29
30
31
32
33
34
35
36
37
H
o-CH3
4.3 ( 0.5a
0.4 ( 0.2b
5.4
360
3.3
12
230
0.61
5a
0.35a
nd
m-CH3
p-CH3
o-OCH3
m-OCH3
p-OCH3
o-Cl
nd
10
nd
nd
0.4
0.3
<0.1
o-CF3
o-CH2CH3
0.50
0.35
compd
R1
R2
IC50 (nM)
fold increase
* n ) 3, ** n ) 5 Unless statistical limits are given, the
compounds were assayed once. The reliability of the in-vitro assay
is ( 50%. The reliability of the cell-based assay is ( 50-100%.
Compounds that differ in potency by more than 3-fold should be
considered statistically different.
active site. Another explanation is that the pyridine
may serve as a hydrogen bond acceptor.
To improve the activity of oxygen analogue 19, a
structure-activity study of substituents on the biphenyl
core was conducted. The underlying premise was that
the additional van der Waal interaction of a lipophilic
group appended to the unsubstituted phenyl ring might
enhance potency, though the optimal position and
nature of the substituent was unknown to us. As shown
in Table 2, o-methyl derivative 29 was 10-fold more
potent than parent structure 19. Compounds 35, 36,
and 37, bearing o-chloro, trifluoromethyl, and ethyl
substituents, respectively, were similarly 7- to 11-fold
more potent than 19. There was a sharp decline in
activity as the methyl substituent was moved first to
the meta and then to the para position. While the same
trend was observed for the o-, m-, and p-methoxy
substituted compounds, o-methoxy inhibitor 32 was
equipotent with unsubstituted compound 19.
and suggests 29 to be a mixture of rotational diaster-
eomers. The proton NMR spectrum of 37 also reveals
two methyl triplet signals. Variable temperature NMR
experiments of 29 showed that the two o-tolyl methyl
signals coalesced as the temperature was increased to
70 °C. Variable temperature NMR experiments of 37
displayed nearly identical results. This implies a
rotational barrier of 17.4 kcal about the biphenyl
carbon-carbon bond, which is, in essence, slow rotation
on the NMR time scale at room temperature. This may
be indicative of a conformational effect whereby the
ortho substituent sterically imparts an orthogonal re-
lationship between the two phenyl rings of the biphenyl
core. Apparently, the binding pocket is complementary
to the shape of an orthogonal biphenyl system. As
further evidence that 29 is a rotational diastereomer,
glycine analogue 38 (Figure 3) was prepared.
The EC50 of 29 for the inhibition of Ras processing in
the whole cell assay was 0.35 µM. Thus, we were able
to obtain a potent parent carboxylic acid that is as active
in whole cells as FTI-277.
The results of the biphenyl core modifications raised
some questions regarding the nature of the enhanced
potency. The Hansch π constants, in increasing order
of lipophilicity, for methoxy, methyl, chloro, trifluorom-
ethyl, and ethyl span 1 order of magnitude. The potency
of these molecules span 1 order of magnitude as well.
The o-methoxy is the least active of the ortho substitu-
tions and provides no potency increase over the parent
compound. There is considerable difference between the
electronic and lipophilic character of a chloro, trifluo-
romethyl, and an ethyl substituent. Yet, these mol-
ecules are equipotent. While there is some agreement
between the degree of lipophilicity and observed in vitro
potency (i.e., a beneficial van der Waal interaction may
help to enhance potency), there is clearly some other
factor responsible for the observed activity. The proton
NMR spectrum of 29 reveals two o-tolyl methyl signals
The NMR spectrum of 38 reveals one signal for the
o-methyl group. In addition, the NMR spectrum of the
o-methoxy substituted system 32 reveals one signal for
the methyl ether. Apparently, the intervening oxygen
allows for free rotation. Thus ortho substitution with

4292 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Augeri et al.
Ta ble 5. Physical Data and Synthetic Methods for FTase
Ta ble 4. Effects of Substituting Methionine with Different
Amino Acids
Inhibiting Compounds
compd
amino acid
R
IC50 (nM) ED50 (µM)
compda
formulab
7
9
C24H23N3SO4HCl‚2.15H2O
C
C
C
C
C
24H23N3SO4‚0.95H2O
24H23N3SO4‚1.00H2O
24H25N3SO3‚1.25TFA
24H24N2SO4‚1.14H2O
24H24N2S2O3
10
15
19
20
21
24
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
C24H24N2SO4‚0.45H2O
C
C
C
24H24N2SO4‚1.14H2O
25H26N2SO4‚0.60EtOAc
25H26N2SO4‚0.10CH2Cl2
C25H26N2SO4‚0.15CH2Cl2
C25H26N2SO5‚0.25H2O
C
C
25H26N2SO5‚0.55H2O
25H26N2SO5
C24H23N2SClO4
C25H23N2SO4F3‚0.65H2O
C
C
C
C
C
C
C
C
C
C
C
26H28N2SO4‚0.22H2O
22H20N2O4‚1.50H2O
25H27N3SO3‚0.65HCl
24H25N3SO5‚1.30H2O
24H25N3SO3‚0.90H2O
24H24N2O5‚0.50H2O
25H26N2O4‚0.50H2O
23H22N2SO4‚0.20H2O
24H23N3O5‚2.40HCl
25H25N3SO4HCl‚0.50H2O
25H27N3SO3
sterically demanding groups (methyl, ethyl, trifluorom-
ethyl, chloro) unexpectedly enhanced potency 7 to 12-
fold over 19, giving compounds that are equipotent with
FTI-276.
The o-tolyl methyl group has enhanced the in vitro
potency of every system studied. The minimum in-
crease was 10-fold (19 and 29), which occurred with the
most potent initial lead (Table 3). However, different
series of compounds exhibit much greater than a 10-
fold increase in potency when the o-tolyl is incorporated.
As a result of enhanced FTI activity, the EC50 data were
also dramatically improved (19 EC50 ) 5.0 µM, 29 EC50
) 0.35 µM).
Replacement of the C-terminal amino acid was less
successful. Only a few amino acids displayed good
activity (Table 4). Inversion of the methionine stereo-
chemistry (41) resulted in a ∼50-fold less potent com-
pound. The N-methyl-L-methionine31 (39) derivative
was equally less potent. Only methionine sulfone (40)
and glutamine (45) were viable substitutions. However,
the activity of these analogues was still lower than that
of the parent methionine. Some other noteworthy
compounds were O-methylhomoserine31 (42), norleucine
(43), and S-methylcysteine (44). Though very similar
in structure to the methionine analogues, these com-
pounds were significantly less active.
C25H27N3SO3‚0.30HCl
C
25H26N2S2O3
a
b
See Tables 1-4 for structures. Compounds were purified
using flash chromatography with methanol/methylene chloride or
methanol/ethyl acetate and were lyophilized from water/acetoni-
trile. Salt formation was necessary in some cases in order to obtain
well-behaved solids.
containing carboxylic acid 29 that is active in whole
cells. The o-tolylpyridyl ether 29 has 0.4 nM in vitro
potency against farnesyltransferase and has a measured
EC50 of 350 nM for inhibition of Ras processing in whole
cells. The o-tolyl biphenyl core has a dramatic effect
on the potency of every non-cysteine-containing farne-
syltransferase inhibitor that we have synthesized. We
conducted an exact comparison between many pairs of
compounds that contained either o-hydrogen or o-methyl
as the only structural difference. This effect was
illustrated by compounds 46, 47, 48, and 49 in Table 3.
It seems clear that the o-tolyl methyl group sterically
imparts an orthogonal relationship to the biphenyl core,
and this conformational change makes it more compli-
mentary to the shape of the protein binding site. This
discovery has laid the groundwork for the development
of even more potent inhibitors of protein farnesyltrans-
ferase. Reports of these studies will be published in due
course.
The active farnesyltransferase inhibitors described
herein (Table 5) possess little or no inhibitory activity
against GGTase I. All of these compounds possess an
IC50 > 10 µM against GGTase I, giving rise to ∼10 000-
fold selectivity. This is a significant accomplishment,
given that FTI-276 is only 50-fold selective. The ortho
substituted biphenyl compounds were also inactive as
inhibitors of GGTase I (IC50 > 10 µM), showing that the
increase in potency observed for FTase is not observed
in GGTase I.
Exp er im en ta l Section
Gen er a l. Proton magnetic resonance spectra were obtained
on a Nicolet QE-300 (300 MHz), a General Electric GN-300
(300 MHz), or a Varian Unity 500 (500 MHz) instrument.
Chemical shifts are reported as δ values (ppm) downfield
relative to Me4Si as an internal standard. Mass spectra were
obtained with a Hewlett-Packard HP5965 spectrometer; CI/
NH3 indicates chemical ionization mode in the presence of
ammonia. Combustion analyses were performed by Robertson
Microlit Laboratories, Inc., Madison, NJ . Melting points were
determined on a Buchi melting point apparatus with a silicone
oil bath and are uncorrected. Chromatographies were carried
out in flash mode using silica gel 60 (230-400 mesh) from E.
Merck. Compounds synthesized by combinatorial chemistry
Con clu sion
In summary, we were successful in transforming FTI-
276 1 into the potent and selective non-cysteine-

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4293
2H), 7.45-7.25 (m, 11H), 4.58 (d, 2H), 4.27 (t, 1H). CIMS:
techniques that displayed biological activity were resynthe-
sized by the project team in order to obtain pure compounds
for biological assay.
+
MNH4 453 C28H21NO4.
Gen er a l P r oced u r e: Dep r otection of F MOC-L-Met-
Wa n g Resin . FMOC-L-Met-Wang resin (8.80 g; 0.47 mmol/g
substitution MidWest Biotech Indianapolis, IN; 0.0041 mol)
was placed in a manual solid-phase synthesis vessel (100 mL
volume). The resin was suspended in N-methyl pyrrolidone
(NMP, 50 mL) and was rotated for 5 min on a 120° rotary
shaker. The solvent was then drained, and the resin was
treated with additional NMP (50 mL; 2×; 5 min). The swollen
resin was then resuspended in a 75 mL solution of 33%
piperidine/dimethylformamide (DMF) and was rotated for 1
h. The resin was drained and then washed as follows: acetone
(75 mL, 1×, 5 min); DMF (75 mL, 3×, 5 min); 2-propanol (75
mL, 5×, 5 min); DMF (75 mL, 3 × 5 min); diethyl ether (75
mL, 3 × 5 min); NMP (75 mL, 3×, 5 min).
4-Am in o-2-p h en ylben zoic Acid , ter t-Bu tyl Ester (4).
Commercially available 2-bromo-4-nitrotoluene 3 (14.0 g, 64.9
mmol) was dissolved in 300 mL of toluene and degassed with
nitrogen for 10 min. Pd(PPh3)4 (3.0 g, 2.6 mmol, 4.0 mol %)
was added. After an additional 10 min, Na2CO3 (2.0 M
aqueous solution, 150 mL) was added followed by a solution
of phenyl boronic acid (8.7 g, 71.4 mmol) in 60 mL of ethanol,
and the reaction was heated to reflux for 12 h. TLC analysis
indicated the reaction was complete (20% CH2Cl2/hexanes), the
reaction was poured into a separatory funnel, and the aqueous
layer was extracted with 2 × 100 mL portions of ethyl acetate.
The organic layers were combined, dried over Na2SO4, filtered,
evaporated, and purified by flash chromatography (50% ethyl
acetate/hexanes) to give 12.67 g (91%) of 4-nitro-2-phenyltolu-
ene. Rf ) 0.3 (20% CH2Cl2/hexanes). 1H NMR (300 MHz,
CDCl3): δ 8.25-8.15 (m, 2H), 7.52-7.38 (m, 4H), 7.35-7.28
(m, 2H), 2.37 (s, 3H). CIMS: MH+ 214 C13H11NO2.
Cou p lin g of 4-(9-F lu or en ylm eth yloxyca r bon yla m in o)-
2-p h en yl Ben zoic Acid (5) to l-Met-Wa n g Resin : Gen er a l
P r oced u r e. The deprotected L-Met-Wang resin was resus-
pended in NMP (30 mL). To the suspension were then added
4-FMOC-amino-2-phenyl-benzoic acid 5 (2.25 g; 0.0052 mol;
1.25 equiv), diisopropylcarbodiimide (0.65 g; 0.0052 mol; 1.25
equiv), and 3-hydroxy-1,2,3-benzotriazin-4(3H)-one (HOOBt)
(0.84 g; 0.0052 mol; 1.25 equiv). The suspension was rotated
for 20 h. The solution was drained, and the resin was washed
as follows: acetone (75 mL, 1×; 2 min); DMF (75 mL, 3×, 5
min); 2-propanol (75 mL, 5×, 5 min); DMF (75 mL, 3×, 5 min);
methanol (75 mL, 2×, 5 min); diethyl ether (75 mL, 2×, 5 min).
A portion (5 mg) of the resin was removed and subjected to
quantitative “FMOC-chromophore” analysis32a-b in order to
determine the amount of coupled FMOC-4-amino-2-phenyl-
benzoic acid present. Typically the substitution of the biaryl
amino acid ranged from 0.39 to 0.42 mmol/g of L-Met-Wang
resin.
Dep r otection of F MOC-4-a m in o-2-p h en yl-ben za m id e-
L-Met-Wa n g Resin (6). The coupled resin from the previous
step was swollen in DMF (75 mL), rotated for 5 min, and
drained. The drained resin was then washed with DMF (75
mL; 2×; 5 min). The resin was then suspended in 75 mL of a
33% piperidine/DMF solution and was rotated for 1 h. The
solution was drained, and the resin was washed as follows:
acetone (75 mL, 1×, 5 min); DMF (75 mL, 3×, 5 min);
2-propanol (75 mL, 5×, 5 min); DMF (75 mL, 3×, 5 min);
diethyl ether (75 mL, 2×, 5 min). The drained resin was then
dried in vacuo overnight at ambient temperature. The resin
was stored desiccated at ambient temperature until used.
Cou p lin g of Nicotin ic Acid to 4-Am in o-2-p h en yl-ben -
za m id e-L-Met-Wa n g Resin a n d Su bsequ en t Clea va ge.
4-Amino-2-phenyl-benzamide-L-Met-Wang resin (80 mg) (sub-
stitution 0.40 mmol/g, 0.000032 mol) was placed in a 5 mL
manual solid-phase synthesis reaction vessel. The resin was
suspended in NMP (1.0 mL), rotated for 3 min, and drained.
The resin was then treated with NMP (1.0 mL, 2×, 3 min). To
the now swollen resin were then added 0.200 mL of a 1.92 M
solution of diisopropylethylamine (DIEA)/NMP (15 equiv), 1.00
mL of a 0.180 mM NMP solution of nicotinic acid (5 equiv),
and finally 0.200 mL of a a 0.90 M bromo-tris-pyrrolidino
phosphonium hexafluorophosphate33 (PyBroP, 5 equiv)/NMP
solution. The reaction slurry was mixed for 6 h, and the
solution was drained. The resin was then washed as follows:
NMP (1.0 mL, 3×, 3 min); 2-propanol (1.0 mL, 5×, 3 min);
NMP (1.0 mL, 3×, 5 min); methanol (1.0 mL, 2×, 3 min); and
finally diethyl ether (1.0 mL, 2×, 3 min). The resin was then
dried in vacuo for several hours. The dried resin was then
suspended in a 1.50 mL solution of 95:5 trifluoroacetic acid/
water and was rotated for 1.5 h at ambient temperature. The
filtrate was removed from the spent polystyrene-resin and the
resulting cleavage solution was evaporated in vacuo. The
residue (typically 5-10 mg) was analyzed by reversed-phase
(C18 ) analytical HPLC and by electrospray MS. The desired
[4-(3-pyridylcarbonylamino)-2-phenylbenzoyl]methionine acid
7 was found to have an HPLC purity of 75%.
To a solution of 4-nitro-2-phenyltoluene (12.4 g, 58.2 mmol)
in 145 mL of 2:1 H2O/pyridine was added KMnO4 (46.0 g, 291
mmol, 5 eq.) portionwise over 60 min, and the reaction was
heated to reflux for 48 h and then cooled and filtered through
Celite. The filtrate was acidified to pH ) 2 with concentrated
aqueous HCl and extracted with 3 × 100 mL portions of ethyl
acetate. The organic layers were combined, dried over Na2-
SO4, filtered, and evaporated to 12.25 g (87%) of 4-nitro-2-
phenylbenzoic acid. Rf ) 0.2 (10% MeOH/CHCl3 w/ 0.25%
HOAc). 1H NMR (300 MHz, CD3OD): δ 8.26 (dd, 1H), 8.20
(d, 1H), 7.95 (d, 1H), 7.48-7.38 (m, 5H). CIMS: MH+ 244
C
13H9NO4.
To a solution of 4-nitro-2-phenylbenzoic acid (6.0 g, 34.7
mmol) in 100 mL of dioxane were added 2.4 mL of concentrated
sulfuric acid and 100 mL of isobutylene, and the reaction was
stirred in a closed pressure apparatus at 1 atm and at room
temperature for 6 days. The reaction was carefully vented,
and the dioxane was evaporated. The resulting oil was taken
up in 100 mL of ethyl acetate, washed with brine, dried over
Na2SO4, filtered, evaporated, and purified by flash chroma-
tography (25% ethyl acetate/hexanes) to give 5.16 g (70%) of
4-nitro-2-phenyl-tert-butylbenzoate. Rf ) 0.7 (50% ethyl acetate/
hexanes). 1H NMR (300 MHz, CDCl3): δ 8.25-8.20 (m, 2H),
7.9 (dd, 1H), 7.45-7.40 (m, 3H), 7.36-7.32 (m, 2H), 1.26 (s,
9H). CIMS: MH+ 300 C17H17NO4.
To a mixture of 4-nitro-2-phenyl-tert-butylbenzoate (4.84 g,
16.2 mmol) and 10% Pd/C (750 mg) in 60 mL of methanol was
added ammonium formate (5.11 g, 81.0 mmol, 5 equiv), and
the reaction was heated to reflux for 1 h. The reaction was
cooled, filtered through Celite, and evaporated to give 4.31 g
(99%) of 4-amino-2-phenylbenzoic acid, tert-butyl ester 4. Rf
) 0.5 (50% ethyl acetate/hexanes). 1H NMR (300 MHz, CD3-
OD): δ 7.55 (d, 1H), 7.35-7.25 (m, 3H), 7.18 (d, 1H), 6.62 (dd,
1H), 6.48 (d, 1H), 1.16 (s, 9H). CIMS: MH+ 270 C17H19NO2.
4-(9-Flu or en ylm eth yloxycar bon ylam in o)-2-ph en ylben -
zoic Acid (5). To a solution of 4 (4.31 g, 16.0 mmol) in 50
mL of CH2Cl2 were added diisopropylethylamine (4.52 g, 35.2
mmol) and FMOC chloroformate (9.1 g, 35.2 mmol), and the
reaction stirred for 18 h. The reaction was extracted with 50
mL of 1 N HCl, dried over Na2SO4, filtered, evaporated, and
purified by flash chromatography (25% ethyl acetate/hexanes)
to give 7.19 g (92%) of 4-(9-fluorenylmethyloxycarbonylamino)-
2-phenyl-tert-butylbenzoate. 1H NMR (300 MHz, CDCl3): δ
7.82-7.75 (m, 3H), 7.6 (d, 2H), 7.45-7.25 (m, 12H), 6.8 (bs,
1H), 4.55 (d, 2H), 4.26 (t, 1H), 1.22 (s, 9H). CIMS: MH+ 492
C
32H29NO4.
To a solution of 4-(9-fluorenylmethyloxycarbonylamino)-2-
phenyl-tert-butylbenzoate (7.19 g, 14.6 mmol) in 100 mL of
CH2Cl2 was added 50 mL of TFA. The reaction was complete
in 1 h and was evaporated and purified by flash chromatog-
raphy over silica gel (gradient of 50% ethyl acetate/hexanes
followed by 5% methanol/ethyl acetate) to give 6.1 g (96%) of
5 as a white solid. Rf ) 0.2 (50% ethyl acetate/hexanes). 1H
NMR (300 MHz, CDCl3): δ 7.95 (d, 1H), 7.78 (d, 2H), 7.60 (d,
Gen er a l P r oced u r e for th e Solu tion Syn th esis of 7, 9
a n d 10. [4-(3-P yr id ylca r bon yla m in o)-2-p h en ylben zoyl]-

4294 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Augeri et al.
m eth ion in e Acid (7). Nicotinic acid (345 mg, 2.8 mmol) was
suspended in 10 mL of CH2Cl2, and oxalyl chloride (2.8 mL of
a 2.0 M solution in methylene chloride) was added by syringe
followed by 1 drop of DMF. The reaction stirred at 25 °C for
2 h and was evaporated and azeotroped with toluene. The
acid chloride was then dissolved in CH2Cl2, and a solution of
the 4-amino-2-phenylbenzoylmethionine methyl ester (669 mg,
1.87 mmol) in 5 mL of CH2Cl2 was added followed by 4 mL of
saturated aqueous NaHCO3. The reaction was stirred at 25
°C for 3 h. The layers were separated, and the organic layer
was dried over Na2SO4, filtered, and evaporated to give 830
mg (96%) of [4-(3-pyridylcarbonylamino)-2-phenylbenzoyl]me-
thionine methyl ester as an oil. Analysis was performed on
the oil. Rf ) 0.3 (5% MeOH/EtOAc). 1H NMR (300 MHz,
CDCl3): δ 9.2 (d, 1H), 8.76 (d, 1H), 8.42 (bs, 1H), 8.2 (dt, 1H),
7.72-7.65 (m, 2H), 7.6 (dd, 1H), 7.45-7.35 (m, 5H), 6.02 (bd,
1H), 4.65 (dq, 1H), 3.68 (s, 3H), 2.20 (t, 2H), 2.01 (s, 3H), 1.98-
1.88 (m, 1H), 1.80-1.78 (m, 1H). CIMS: MH+ 464.
[4-(3-Pyridylcarbonylamino)-2-phenylbenzoyl]methionine
methyl ester (830 mg, 1.79 mmol) was dissolved in 8 mL of
THF and cooled to 0 °C, and LiOH monohydrate (226 mg, 5.38
mmol) was added followed by 2 mL of H2O. The reaction was
complete in 2 h and was evaporated and acidified to pH ) 3
with 1 N HCl, and the resulting precipitate was taken up in
EtOAc, washed with water, and dried over Na2SO4, and
evaporated. The resulting residue was crystallized from hot
ethanol to give 281 mg (32%) of the HCl salt of 7 as a white
crystalline solid. Rf ) 0.2 (10% MeOH/CHCl3 with 0.25%
HOAc). 1H NMR (300 MHz, CD3OD): δ 9.4 (d, 1H), 9.1 (d,
1H), 9.0 (d, 1H), 8.2 (dd, 1H), 7.85-7.80 (m, 2H), 7.58 (dd, 1H),
7.45-7.32 (m, 5H), 4.52-4.45 (m, 1H), 2.20-2.02 (m, 2H), 2.00
(s, 3H1.90-1.80 (m, 2H), 1.15 (t, 1H). CIMS: MH+ 450. Anal.
(C24H23N3SO4HCl‚2.15H2O): C, H, N.
[4-(3-P yr id ylm eth yla m in o)-2-p h en ylben zoyl]m eth ion -
in e Acid (8). 4-Amino-2-phenylbenzoylmethionine methyl
ester (1.15 g, 3.21 mmol) and 3-pyridine carboxaldehyde (361
mg, 3.37 mmol) were combined in 15 mL of MeOH, sodium
cyanoborohydride (302 mg, 4.81 mmol) was added followed by
crushed molecular sieves, the reaction was adjusted to pH )
6 with acetic acid and stirred at 25 °C. After 3 h the reaction
was concentrated and transferred directly to a column of silica
gel and purified by flash chromatography (5% MeOH/ EtOAc)
to give 1.38 g (95%) of the ester as an oil that solidified after
standing. Rf ) 0.15 (5% MeOH/EtOAc). 1H NMR (300 MHz,
CDCl3): δ 8.6 (d, 1H), 8.52 (dd, 1H), 7.72-7.65 (m, 2H), 7.45-
7.30 (m, 6H), 6.62 (dd, 1H), 6.48 (d, 1H), 5.72 (bd, 1H), 4.64
(dq, 1H), 4.42 (bs, 2H), 3.64 (s, 3H), 2.25-2.05 (m, 3H), 2.00
(s, 3H), 1.95-1.80 (m, 1H), 1.72-1.60 (m, 1H). CIMS: MH+
450.
The methyl ester (1.38 g, 3.07 mmol) was dissolved in THF,
LiOH monohydrate (387 mg, 9.22 mmol) was added in 2 mL
of H2O at 0 °C, and the reaction stirred at 0 °C for 2 h. The
reaction was complete in 2 h and was evaporated and acidified
to pH ) 3 with 1 N HCl. The resulting precipitate was taken
up in EtOAc, washed with water, dried over Na2SO4, and
evaporated. The resulting residue was crystallized from hot
ethanol to give 1.12 g (78%) of the HCl salt of 8 as a white
crystalline solid. Rf ) 0.2 (10% MeOH/CHCl3 with 0.25%
HOAc). 1H NMR (300 MHz, CD3OD): δ 8.82 (d, 1H), 8.75 (d,
1H), 8.70 (d, 1H), 8.08 (dd, 1H), 7.42-7.30 (m, 6H), 6.72-6.62
(m, 3H), 4.72 (bs, 2H), 4.42 (dd, 1H), 2.22-2.02 (m, 2H), 2.00
(s, 3H), 2.00-1.85 (m, 1H), 1.82-1.70 (m, 1H). CIMS: MH+
436. FAB HRMS: MH+ calcd for C24H26N3SO3, 436.1695;
found, 436.1688.
7.90 (d, 2H), 7.85 (m, 2H), 7.38-7.53 (m, 6H), 4.39 (ddd, 1H),
2.25 (m, 2H), 2.01 (s, 3H), 1.87 (m, 2H). CIMS (DCI, NH3):
450 (MH+), 467 (M + NH4)+. CIMS: MH+ 450 C24H23N3SO4.
Anal. (C24H23N3O4S‚1.00H2O): C, H, N.
Dim eth yl-2-p h en ylter ep h th a la te (12). To a stirred solu-
tion of 9.60 g (30.00 mmol) of dimethyl iodoterephthalate 11
in 75 mL of toluene at room temperature was added 1.74 g
(1.50 mmol) of Pd(Ph3)4. After the yellow solution was stirred
for 10 min, 4.02 g (33.00 mmol) of phenylboronic acid was
added in 33 mL of ethanol followed by 70 mL of 2 M aqueous
Na2CO3. The mixture was vigorously stirred and heated to
reflux for 3 h and then allowed to reach room temperature.
The mixture was diluted with 200 mL of ethyl ether, and the
layers separated. The organic phase was dried over Na2SO4,
filtered, and concentrated. The dark residue was purified by
column chromatography on silica gel (150 g, 2% ethyl acetate/
hexanes) to give 7.86 g (97%) as a yellow oil. 1H NMR (300
MHz, CDCl3): δ 8.08 (s, 1H), 8.05 (d, 1H), 7.85 (d, 1H), 7.61
(m, 3H), 7.53 (m, 2H), 3.96 (s, 3H), 3.65 (s, 3H). MS (DCI,
NH3): 288 (M+NH4)+ (100), 271 (MH+, 17).
Meth yl-4-Hyd r oxym eth yl-2-p h en yl Ben zoa te (13). To
a solution of 18.4 g (68.1 mmol) of dimethyl-2-phenyltereph-
thalate 12 in 70 mL of 1:1 THF/ methanol was added a solution
of 4.56 g (71.5 mmol) of KOH (Fisher 88%) in 30 mL of water
dropwise such that the internal temperature remained below
35 °C. The clear solution was stirred overnight, and the
organics were removed by evaporation on a rotary evaporator.
The aqueous solution was extracted with 2 portions of ethyl
acetate and then cooled in an ice bath. To this cold, stirred
solution was carefully added 10 mL of concentrated aqueous
HCl, and stirring continued for 30 min. The solid formed was
collected by vacuum filtration and recrystallized from 100 mL
of hot ethanol/water (3:1) to give 10.4 g (60%) of the desired
mono acid. The mother liquors were concentrated to dryness,
and the residue was chromatographed on 300 g of SiO2 (97:
2:1-96:3:1 CHCl3/MeOH/AcOH) to give an additional 1.7 g
(total yield 70%), mp 173-175 °C. 1H NMR (300 MHz, DMSO-
d6): δ 13.39 (bs, 1H), 8.02 (dd, 1H), 7.93 (d, 1H), 7.85 (d, 1H),
7.27-7.50 (m, 3H), 7.34 (m, 2H), 3.62 (s, 3H). MS (DCI,
NH3): 274 (M + NH4+, 100).
To a solution of 10.5 g (41.0 mmol) of the monoacid in 40
mL of dry THF (Aldrich, Gold Label) at 0 °C was added 82
mL of a 1.0 M solution of BH3-THF in THF dropwise such
that the internal temperature remained below 6 °C. The
reaction was stirred at 0 °C for 30 min after the addition was
complete, and the cooling bath was removed. After it was
stirred for 2 h at room temperature, the reaction mixture was
cooled to 0 °C in an ice bath and very carefully quenched by
the dropwise addition of 100 mL of 3 N aqueous HCl (ca u tion ,
vigorous evolution of H2). The mixture was stirred for 1 h at
room temperature, and the THF was removed on a rotary
evaporator. The residue was extracted with 3 × 100 mL of
ethyl acetate, and the combined organic phases were back
extracted with 2 × 1 N aqueous NaOH, 1× water, and 1× with
brine, dried over MgSO4, filtered, and concentrated to give 9.75
g (98%) of 13 that was used directly. An analytical sample
was prepared by flash chromatography on silica gel (50% ethyl
acetate/hexanes). 1H NMR (300 MHz, CDCl3): δ 7.85 (d, 1H),
7.33-7.44 (m, 4H), 7.31 (m, 2H), 4.79 (d, 2H), 3.63 (s, 3H),
1.78 (t, 1H). MS (DCI, NH3): 260 (M + NH4+, 100), 243 (MH+,
50).
Gen er a l Ben zoa te Hyd r olysis: 2-P h en yl-4-h yd r oxy-
m eth ylben zoic Acid . To a solution of 13 (9.75 g, 40.2 mmol)
in 50 mL of methanol at room temperature was added a
solution of 5.13 g (80.4 mmol) of KOH (Fisher, 88%) in 30 mL
of water. The mixture was brought to reflux and maintained
for 2.5 h. The mixture was cooled to room temperature and
the methanol removed on a rotary evaporator. The resulting
solution was extracted with 4 × 50 mL of ethyl acetate, and
the aqueous phase was then cooled to 0 °C and acidified by
the careful addition of 10 mL of concentrated aqueous HCl.
The mixture was extracted with 2 portions of ethyl acetate,
dried over MgSO4, filtered, and concentrated to give 9.16 g
(99%) of the desired compound as a waxy white solid. 1H NMR
[4-(2-P yr idylcar bon ylam in o)-2-ph en ylben zoyl]m eth ion -
in e Acid (9). Prepared according to procedure for 7. 1H NMR
(300 MHz, DMSO-d6): δ 10.9 (s, 1H), 8.76 (t, 1H), 8.50 (d, 1H),
8.18 (m, 1H), 8.11 (dt, 1H), 8.03-7.96 (m, 2H), 7.72 (m, 1H),
7.46-7.12 (m 6H), 4.30 (m, 1H), 2.35-2.15 (m, 2H), 2.00 (s,
3H), 1.91-1.77 (m, 2H). CIMS: MH+ 450 C24H23N3SO4. Anal.
(C24H23N3O4S‚0.95H2O): C, H, N.
[4-(4-P yr idylcar bon ylam in o)-2-ph en ylben zoyl]m eth ion -
in e Acid (10). Prepared according to procedure for 7. 1H
NMR (DMSO-d6): δ 10.70 (s, 1H), 8.82 (d, 2H), 8.49 (d, 1H),

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4295
(300 MHz, CDCl3): δ 7.97 (d, 1H), 7.31-7.46 (m, 6H), 4.81 (s,
2H). MS (DCI, NH3): 246 (M + NH4+, 100).
judged complete by TLC analysis. The solution was concen-
trated to remove the organic fractions and diluted with water,
and the pH of the solution adjusted to ∼4 with aqueous HCl.
If a solid precipitate formed, the product was collected by
filtration. If an oil formed, the aqueous mixture was extracted
with ethyl acetate, and the extracts were dried over sodium
sulfate, filtered, and concentrated. If a solid was not isolable
by filtration, the material was extracted into ethyl acetate and
the organic phase dried over Na2SO4, filtered, and concen-
trated. These procedures generally gave material of suitable
purity. If not, the compound was purified by column chroma-
tography on silica gel or reversed-phase preparative HPLC.
N-4-[(3-P yr idylam in om eth yl)-2-ph en yl]ben zoylm eth ion -
in e (15). The amino ester was hydrolyzed according to the
general procedure for hydrolysis of amino esters above and
purified by HPLC using acetonitrile/0.1% aqueous trifluoro-
acetic acid as the mobile phase. The product was obtained by
lyophilization. 1H NMR (300 MHz, DMSO-d6): δ 8.48 (d, 1H),
8.17 (m, 1H), 7.97 (m, 1H), 7.56 (m, 2H), 7.29-7.44 (m, 7H),
4.51 (bd, 2H), 4.48 (ddd, 1H), 2.24 (m, 2H), 1.99 (s, 3H), 1.85
(m, 2H). CIMS (NH3): C24H25N3O3S 436, (MH+), 418, 319, 287,
194, 165. Anal. (C24H25N3O3S‚1.26TFA): C, H, N.
4-Br om om eth yl-2-p h en ylben zoic, Meth yl Ester . A so-
lution of 1.81 g (7.47 mmol) of alcohol 13 and 0.66 g (7.47
mmol) lithium bromide in DMF (10 mL) was chilled in an ice-
water bath, and then 0.71 mL (7.47 mmol) phosphorus
tribromide was added. After 2 h the reaction was partitioned
between water (200 mL) and Et2O (200 mL). The aqueous
layer was extracted with Et2O (2 × 50 mL), and then the
combined Et2O layers were washed with water (2 × 50 mL),
brine, and dried over Na2SO4. Filtration and concentration
resulted in 2.20 g (96%) of an oil that solidified on standing.
1H NMR (300 MHz, CDCl3): δ 7.82 (d, 1H), 7.35-7.47 (m, 4H),
7.27-7.34 (m, 2H), 4.52 (s, 2H), 3.64 (s, 3H). CIMS (NH3):
322/324 (M + NH4+).
2-P h en yl-4-(3-p yr id yloxym eth yl)ben zoic Acid , Meth yl
Ester (16). A solution of 3-hydroxypyridine 9.51 g (0.10 mol)
in 50 mL of methanol was treated with 50.0 mL of 2.0 M
aqueous KOH, and the solution was concentrated to dryness.
The resulting solid was dried in a vacuum oven at over 100
°C to provide 13.25 g (99%) of the potassium salt. A suspen-
sion of 1.73 g (13.00 mmol) of this salt and 1.19 g (4.50 mmol)
of 18-crown 6 in 20 mL of DME was cooled to -10 °C, and a
solution of 3.05 g (10.00 mmol) of 4-bromomethyl-2-phenyl-
benzoic methyl ester in 10 mL of DME was added dropwise.
Stirring continued for 24 h during which time the bath melted.
The reaction mixture was poured into water and extracted 3
times with ethyl acetate, and the combined organic layers were
washed with brine, dried over Na2SO4, filtered, and concen-
trated. The residue was purified by column chromatography
on silica gel (100 g, 60% ethyl acetate/hexanes) to provide 1.10
g (34%) of 16. 1H NMR (300 MHz, CDCl3): δ 8.41 (m, 1H),
8.25 (dd, 1H), 7.87 (d, 1H), 7.42-7.50 (m, 2H), 7.35-7.42 (m,
3H), 7.27-7.41 (m, 3H), 7.19-7.26 (m, 2H), 5.18 (s, 2H), 3.64
(s, 3H). CIMS (NH3): 320 (MH+), 337 (M + NH4+).
Gen er a l Am in o Acid Cou p lin g P r oced u r e: N-[4-Hy-
d r oxym eth yl-2-p h en ylben zoyl]-L-Meth ion in e, Meth yl Es-
ter . A solution of 3.42 g (15.00 mmol) 2-phenyl-4-hydroxym-
ethylbenzoic acid in 30 mL of DMF was treated sequentially
with 2.20 g (16.30 mmol) of HOBt, 3.12 g (16.3 mmol) of EDAC,
and 3.90 g (19.50 mmol) of L-methionine methyl ester hydro-
chloride. The suspension was stirred for 15 min, 4.00 mL
(27.00 mmol) of triethylamine was added, and the mixture
stirred for 24 h. The reaction was quenched by pouring into
300 mL of ethyl acetate and extracting with 3 portions of 1 N
aqueous HCl, 2 portions of water, 1 portion of saturated
aqueous NaHCO3, and 1 portion of brine. The solution was
then dried over Na2SO4, filtered, and concentrated. The
residue was purified by column chromatography on silica gel
(120 g, 70:30 ethyl acetate/hexanes) to provide 4.40 g (79%) of
N-[4-hydroxymethyl-2-phenylbenzoyl]-L-methionine methyl es-
ter as an oil that solidified upon standing. 1H NMR (300 MHz,
CDCl3): δ 7.68 (d, 1H), 7.34-7.48 (m, 5H), 5.90 (bd, 1H), 4.78
(s, 2H), 4.66 (ddd, 1H), 3.67 (s, 3H), 2.06 (m, 2H), 2.02 (s, 3H),
1.85-1.98 (m, 1H), 1.65-1.79 (m, 1H). ESI MS: 374 (MH)+,
372 (M-H).
N-[4-F or m yl-2-p h en ylben zoyl]-L-Meth ion in e, Meth yl
Ester (14). A solution of 1.50 mL (17.70, mmol) of oxalyl
chloride in 50 mL of CH2Cl2 was cooled to -75 °C, and a
solution of 1.70 mL (23.60 mmol) of DMSO in 10 mL of CH2-
Cl2 was added such that the internal temperature did not
exceed -60 °C. After the mixture was stirred for 20 min, a
solution of 4.40 g (11.80 mmol) of N-[4-hydroxymethyl-2-
phenylbenzoyl]-L-methionine methyl ester in 25 mL of CH2-
Cl2 was added via cannula such that the internal temperature
did not exceed -65 °C. After it was stirred for 2 h, the mixture
was quenched by the dropwise addition of 6.60 mL (47.20 mL)
of Et3N, and after 10 min of stirring, the cooling bath was
removed. The solution was allowed to warm to ∼10 °C and
then was poured into a separatory funnel. The mixture was
extracted with 2 portions of water and 2 portions of 3 N
aqueous HCl. The combined aqueous phases were back
extracted with 25 mL of CH2Cl2, and the combined organic
phases were washed with saturated aqueous NaHCO3, dried
over Na2SO4, filtered, and concentrated. The residue was
purified by column chromatography on silica gel (120 g, 40%-
60% ethyl acetate/hexanes) to give 3.68 g (84%) of the 14 as a
light yellow oil that solidified on standing. 1H NMR (300 MHz,
CDCl3): δ 10.11 (s, 1H), 7.93 (dd, 1H), 7.90 (d, 1H), 7.84 (d,
1H), 7.46 (m, 5H), 5.99 (bd, 1H), 4.69 (ddd, 1H), 3.69 (s, 3H),
2.07 (m, 2H), 2.01 (s, 3H), 1.84-2.00 (m, 1H), 1.70-1.83 (m,
1H). MS (DCI, NH3): 372 (MH+, 80), 389 (M + NH4+, 100).
N-4-[(3-P yr idylam in om eth yl)-2-ph en yl]ben zoylm eth ion -
in e, Meth yl Ester . A solution of 2.50 g (6.73 mmol) of the
aldehyde 14 in 15 mL of methanol at room temperature was
treated with 0.94 g (10.00 mmol) of 3-aminopyridine, and then
5 mL of acetic acid was added. After the mixture was stirred
for 1 h, 0.85 g (13.50 mmol) of NaCNBH3 was added. After 2
h of stirring an additional 0.42 g (6.73 mmol) of NaCNBH3
was added, and stirring continued for an additional hour. The
mixture was poured into 100 mL of 1 N aqueous NaOH and
extracted with 3 portions of ethyl acetate. The combined
organic phases were washed twice with saturated NaHCO3, 3
times with water, once with brine, and then dried over Na2-
SO4, filtered, and concentrated. The residue was purified by
column chromatography on silica gel (150 g, ethyl acetate) to
give 2.20 g (73%) of N-4-[(3-pyridylaminomethyl)-2-phenyl]-
benzoylmethionine methyl ester as a thick oil. 1H NMR (300
MHz, CDCl3): δ 8.08 (bs, 1H), 8.00 (bd, 1H), 7.70 (d, 1H), 7.41
(m, 6H), 7.36 (d, 1H), 7.07 (dd, 1H), 6.86 (ddd, 1H), 5.88 (bd,
1H), 4.68 (ddd, 1H), 4.44 (d, 2H), 4.21 (bt, 1H), 3.65 (s, 3H),
2.07 (m, 2H), 2.01 (s, 3H), 1.84-1.98 (m, 1H), 1.65-1.79 (m,
1H). CIMS (NH3): 450 (MH+, 100).
2-P h en yl-4-(3-p yr id yloxym eth yl)ben zoic Acid . A solu-
tion of 1.40 g (4.38 mmol) of 16 in 10 mL of methanol at room
temperature was treated with 1.2 mL of a 4 N aqueous NaOH
solution (5.26 mmol), and the mixture was heated to reflux
for 20 h. The solution was cooled to ambient temperature and
concentrated to remove the methanol. The solution was
diluted with water and the pH adjusted to 5 with aqueous HCl.
The solid was collected by filtration and dried in the air to
give 1.31 g (98%) of 2-phenyl-4-(3-pyridyloxymethyl)benzoic
acid. 1H NMR (300 MHz, DMSO-d6): δ 12.79 (bs, 1H), 8.41
(d, 1H), 8.21 (dd, 1H), 7.76 (d, 1H), 7.29-7.36 (m, 2H), 7.47 (s,
1H), 7.19-7.24 (m, 7H), 5.21 (s, 2H). CIMS (NH3): 306 (MH+),
323 (M + NH4+).
N-4-[(3-P yr id yloxym eth yl)-2-p h en yl]ben zoylm eth ion -
in e, Meth yl Ester . Using the general amino acid coupling
procedure 2-phenyl-4-(3-pyridyloxymethyl)benzoic acid (225
mg, 0.74 mmol) provided 247 mg (74%) of the desired com-
pound. 1H NMR (300 MHz, CDCl3): δ 8.41 (m, 1H), 8.24 (dd,
1H), 7.75 (d, 1H), 7.48 (dd, 1H), 7.36-7.46 (m, 6H), 7.19-7.27
Gen er a l P r oced u r e for Hyd r olysis of Am in o Ester s.
The methionine ester was dissolved in 3:1 THF/methanol and
cooled in an ice bath. The solution was treated with 200 mol
% of aqueous lithium hydroxide, and the mixture stirred until

4296 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Augeri et al.
(m, 2H), 5.91 (d, 1H), 5.18 (s, 3H), 4.57 (ddd, 1H), 3.66 (s, 3H),
2.08 (m, 2H), 2.02 (s, 3H), 1.87-1.98 (m, 1H), 1.66-1.80 (m,
1H). APCI MS: 451 (MH+).
1H), 7.67 (m, 2H), 7.28-7.44 (m, 4H), 7.10-7.18 (m, 3H), 4.95
(s, 2H). DCIMS (NH3): 354 (MH+, 55), 371 (M + NH4+, 100).
N -4-[(3-P yr id ylsu lfon ylm e t h yl)-2-p h e n yl]b e n zoyl-
m eth ion in e, Meth yl Ester . 2-Phenyl-4-(3-pyridylsulfonyl-
methyl)benzoic acid was coupled to L-methionine methyl ester
HCl using the general amino acid coupling procedure, mp 152-
154 °C. 1H NMR (300 MHz, CDCl3): δ 8.95 (d, 1H), 8.87 (dd,
1H), 7.94 (ddd, 1H), 7.66 (d, 1H), 7.43 (m, 4H), 7.30 (m, 2H),
7.21 (dd, 1H), 7.10 (d, 1H), 5.91 (bd, 1H), 4.66 (ddd, 1H), 4.42
(s, 2H), 3.68 (s, 3H), 2.08 (t, 2H), 2.02 (s, 3H), 1.93 (m, 1H),
1.75 (m, 1H). MS (DCI, NH3) 516 (M + NH4+), 499 (MH+).
Anal. Calcd for C25H26N2O5S2: C, 60.22; H, 5.25; N, 5.62.
Found: C, 60.28; H, 4.94; N, 5.56.
N -4-[(3-P yr id ylsu lfon ylm e t h yl)-2-p h e n yl]b e n zoyl-
m eth ion in e (21). Prepared using the general amino ester
hydrolysis procedure. 1H NMR (300 MHz, DMSO-d6): δ 12.68
(bs, 1H), 8.92 (m, 2H), 8.59 (bd, 1H), 8.18 (ddd, 1H), 7.68 (m,
1H), 7.32 (m, 7H), 7.18 (d, 1H), 4.94 (s, 1H), 4.29 (ddd, 1H),
2.22 (m, 2H), 1.99 (s, 3H), 1.85 (m, 2H). MS (DCI, NH3): 502
(M + NH4+), 485 (MH)+. Anal. (C24H24N2O5S2‚0.45H2O): C,
H, N.
4-(3-P yr id ylm eth yloxy)-2-h yd r oxyben zoic Acid , Meth -
yl Ester . A solution of methyl 2,4-dihydroxybenzoate 22
(16.82 g, 0.10 mol) and 3-chloromethylpyridine hydrochloride
(19.68 g, 0.12 mol) in 150 mL of DMF was treated with KOH
(14.66 g, 0.23 mol) in 50 mL of water, and the mixture was
heated to 55-60 °C for 18 h. The mixture was treated with
an additional 16.40 g (0.10 mol) of 3-chloromethylpyridine
hydrochloride and KOH (9.67 g, 0.15 mol) in 20 mL of water,
and heating continued for 6 h. The mixture was cooled and
poured into 1.5 L of water, and the solid was collected by
filtration. Recrystallization from 1:1 ethanol/water (∼125 mL)
provided 6.15 g (24%) of 4-(3-pyridylmethyloxy)-2-hydroxym-
ethyl benzoate. 1H NMR (300 MHz, CDCl3): δ 10.98 (s, 1H),
8.69 (bs, 1H), 8.61 (bd, 1H), 7.77 (d, 2H), 7.34 (dd, 1H), 6.53
(s, 1H), 6.50 (dd, 1H), 5.10 (s, 2H), 3.93 (s, 3H). MS (DCI,
NH3): 260 (MH+).
4-(3-P yr id ylm et h yloxy)-2-t r iflu or om et h a n esu lfon yl-
oxyben zoic Acid , Meth yl Ester . 4-(3-Pyridylmethyloxy)-2-
hydroxymethyl benzoate (560 mg, 2.16 mmol) was dissolved
in 3 mL of dry pyridine and cooled in an ice/acetone bath to
-10 °C. Triflic anhydride (0.73 mL, 4.32 mmol) was added
dropwise such that the temperature remained below 5 °C. The
bath was removed after completion of the addition, and the
mixture stirred at ambient temperature for 96 h. The mixture
was quenched by pouring into 20 mL of water, and the pH
was made basic by the addition of 2 N aqueous NaOH. The
solution was extracted with 3 portions of ethyl acetate, and
the combined organic extracts were washed with water and
brine, dried over Na2SO4, filtered, and concentrated. The
residue was purified by column chromatography on silica gel
(30 g, 1:1 ethyl acetate/hexanes) to give 685 mg (81%) of 4-(3-
pyridylmethyloxy)-2-trifluoromethanesulfonyloxybenzoic acid,
methyl ester. 1H NMR (300 MHz, CDCl3): δ 8.70 (d, 1H), 8.64
(dd, 1H), 8.09 (d, 1H), 7.77 (dt, 1H), 7.36 (dd, 1H), 7.04 (dd,
1H), 6.88 (d, 1H), 5.15 (s, 2H), 3.94 (s, 3H). MS (DCI, NH3):
392 (MH+), 409 (M + NH4+).
N-4-[(3-P yr id yloxym eth yl)-2-p h en yl]ben zoylm eth ion -
in e (19). Using the general amino ester hydrolysis procedure,
the acid was prepared. 1H NMR (300 MHz, DMSO-d6):
δ
12.66 (bs, 1H), 8.58 (d, 1H), 8.38 (d, 1H), 8.17 (dd, 1H), 7.30-
7.56 (m, 10H), 5.29 (s, 2H), 4.29 (ddd, H), 2.23 (m, 2H), 1.98
(s, 3H), 1.84, (m, 2H). CIMS (NH3): 437 (MH+), 454 (M +
NH4+). Anal. (C24H24N2O4S‚1.14H2O): C, H, N.
3-Mer ca p top yr id in e, Sod iu m Sa lt. A solution of 3-(N,
N-dimethylaminocarbonyl)mecaprtopyridine (1.23 g, 6.70 mmol)
in 10 mL of methanol was treated with 3.5 mL of 2 N aqueous
NaOH (7.0 mmol), and the mixture was heated to reflux for 2
h. After cooling to room temperature, the mixture was
concentrated to dryness to give the sodium salt of 3-mercap-
topyridine which was used directly.30
2-P h en yl-4-(3-p yr id ylth iom eth yl)ben zoic Acid , Meth yl
Ester (17). A solution of 4-bromomethyl-2-phenylmethyl
benzoate (916 mg, 3.00 mmol) in 10 mL of DME was cooled in
an ice/acetone bath and treated with sodium 3-mercaptopyri-
dine30 (450 mg, 3.25 mg), and the mixture was stirred
overnight. The brown mixture was poured into water and
extracted with 3 portions of ethyl acetate. The combined
organic fractions were washed with brine, dried over Na2SO4,
filtered, and concentrated. The residue was purified by column
chromatography on silica gel, 40% ethyl acetate/hexanes gave
611 mg (60%) of 17. 1H NMR (300 MHz, CDCl3): δ 8.55 (d,
1H), 8.45 (dd, 1H), 7.77 (d, 1H), 7.56 (dq, 1H), 7.33-7.42 (m,
3H), 7.16-7.31 (m, 5H), 4.13 (s, 2H), 3.63 (s, 3H). DCIMS
(NH3): 336 (MH+, 100), 353 (M + NH4+, 40).
N-4-[(3-P yr id ylth iom eth yl)-2-p h en yl]ben zoylm eth ion -
in e, Meth yl Ester . Using the general benzoate hydrolysis
protocol, the title compound was prepared. The resulting aryl
acid was coupled to L-methionine methyl ester HCl using the
general amino acid coupling procedure. 1H NMR (300 MHz,
CDCl3): δ 8.56 (m, 1H), 8.45 (dd, 1H), 7.66 (d, 1H), 7.38 (ddd,
1H), 7.30-7.47 (m, 6H), 7.21 (m, 2H), 5.87 (bd, 1H), 4.65 (ddd,
1H), 4.14 (s, 2H), 3.67 (s, 3H), 2.06 (m, 2H), 2.01 (s, 3H), 1.92
(m, 1H), 1.74 (m, 1H). DCIMS (NH3): 467 (MH+). Anal.
Calcd for C25H26N2O3S2: C, 64.35; H, 5.62; N, 6.00. Found:
C, 64.21; H, 5.61; N, 6.00.
N-4-[(3-P yr id ylth iom eth yl)-2-p h en yl]ben zoylm eth ion -
in e (20). N-4-[(3-Pyridylthiomethyl)-2-phenyl]benzoylme-
thionine, methyl ester, was converted to 20 by the general
amino ester hydrolysis procedure. 1H NMR (300 MHz, DMSO-
d6): δ 8.54 (m, 1H), 8.39 (dd, 1H), 7.83 (m, 2H), 7.29-7.47 (m,
8H), 4.39 (s, 2H), 4.24 (m, 1H), 2.25 (m, 2H), 1.98 (s, 3H), 1.85
(m, 2H). DCIMS (NH3): 453 (MH+). Anal. (C24H24N2O3S2):
C, H, N.
2-P h en yl-4-(3-p yr id ylsu lfon ylm et h yl)b en zoic Acid ,
Meth yl Ester (18). Trifluoroacetic anhydride (2.5 mL, 17.9
mmol) was dissolved in 10 mL of CH2Cl2, and the solution
cooled in an ice bath. Hydrogen peroxide (0.56 mL of a 30%
aqueous solution, 5.4 mmol) was slowly added (ca u tion : a
vigorous exothermic reaction occurs), and the mixture stirred
until the internal temperature returned to <5 °C. A solution
of pyridylthioether 17 (600 mg, 1.79 mmol) in 5 mL of CH2Cl2
was added dropwise, and stirring continued for 1 h at 0 °C
and 30 min at room temperature. The mixture was diluted
with 50 mL of ethyl ether and extracted with 50 mL of 2 N
aqueous NaOH. The aqueous phase was extracted with an
additional portion of ether, and the combined organic phases
were extracted with 20% aqueous Na2SO3, water, brine. The
solution was dried over Na2SO4, filtered, and concentrated to
give 620 mg (94%) of 18 as an off-white solid. 1H NMR (300
MHz, CDCl3): δ 8.91 (dd, 1H), 8.85 (dd, 1H), 7.91 (dq, 1H),
7.77 (d, 1H), 7.34-7.46 (m, 4H), 7.22 (dd, 1H), 7.14-7.19 (m,
2H), 7.08 (d, 1H), 4.43 (s, 2H), 3.63 (s, 3H). DCIMS (NH3):
368 (MH+, 90), 385 (M + NH4+, 100).
4-(3-P yr id ylm eth yloxy)-2-p h en ylben zoic Acid , Meth yl
Ester (23). Using the same palladium-catalyzed coupling
procedure used to prepare 12, 4-(3-pyridylmethyloxy)-2-trif-
luoromethanesulfonyloxybenzoic acid, methyl ester, was con-
verted to 23 in 83% yield after chromatography on silica gel.
1H NMR (300 MHz, CDCl3): δ 8.71 (d, 1H), 8.62 (dd, 1H), 7.90
(d, 1H), 7.78 (dt, 1H), 7.24-7.43 (m, 6H), 6.98 (dd, 1H), 6.94
(d, 1H), 5.14 (s, 2H), 3.62 (s, 3H). MS (DCI, NH3): 320 (MH+),
337 (M + NH4+).
4-(3-P yr id ylm et h yloxy)-2-p h en ylb en zoic Acid . Pre-
pared using the general benzoate hydrolysis. 1H NMR (300
MHz, DMSO-d6): δ 12.40 (bs, 1H), 8.69 (d, 1H), 8.56 (dd, 1H),
7.89 (dt, 1H), 7.78 (d, 1H), 7.28-7.48 (m, 6H), 7.09 (dd, 1H),
6.96 (d, 1H), 5.27 (s, 2H). MS (DCI, NH3): 306 (MH+), 323
(M + NH4+).
2-P h en yl-4-(3-pyr idylsu lfon ylm eth yl)ben zoic Acid. Us-
ing the general aryl acid hydrolysis, 18 was converted to
2-phenyl-4-(3-pyridylsulfonylmethyl)benzoic acid. 1H NMR
(300 MHz, DMSO-d6): δ 8.91 (dd, 1H), 8.85 (dd, 1H), 8.13 (dq,
N-[4-(3-P yr id ylm eth yloxy)-2-p h en yl]ben zoylm eth ion -
in e (24). Employed the general amino acid coupling procedure

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4297
1
and the general methionine hydrolysis protocol to give 24. H
NMR (300 MHz, DMSO-d6): δ 8.69 (bs, 1H), 8.55 (bd, 1H),
8.39 (d, 1H), 7.88 (dt, 1H), 7.40 (m, 6H), 7.07 (dd, 1H), 7.03 (d,
1H), 5.17 (s, 2H), 4.28 (ddd, 1H), 2.25 (m, 2H), 2.00 (s, 3H),
1.84 (m, 2H). MS (CI, NH3): 454 (M + NH4+), 437 (MH+),
419, 320, 288. Anal. (C24H24N2O4S‚0.23H2O): C, H, N.
Meth yl-4-Hyd r oxym eth yl-2-iod o ben zoa te (26). To a
solution of dimethylaminoterphthalate 25 (3.07 g, 14.7 mmol)
in 30 mL of a 2:1 mixture of THF/Et2O at -78 °C was added
neat DIBAL (6.27 g, 44.1 mmol, 3.0 equiv), and the reaction
was warmed to 0 °C for 4 h and quenched with 5 mL of MeOH
followed by 5 mL of saturated aqueous Na-tartrate. The
reaction mixture stirred overnight and was taken up in EtOAc,
and the layers were separated. The EtOAc layer was washed
with 20 mL of saturated aqueous NaHCO3 and brine and then
dried over Na2SO4, filtered, evaporated to an oil, and purified
by flash chromatography over silica gel (50% EtOAc/hex) to
give 1.03 g (39%) of 4-hydroxymethyl-2-amino-methyl benzoate
as a colorless oil. 1H NMR (300 MHz, CDCl3): δ 7.82 (d, J )
8.09 Hz, 1H), 6.68 (bs, 1H), 6.60 (dd, J ) 8.46 Hz, 1.84, 1H),
4.62 (s, 2H), 3.86 (s, 3H). CIMS: C9H11NO3 MH+ 182.
To a stirred solution of 4-hydroxymethyl-2-amino-methyl
benzoate (152 mg, 0.84 mmol) in 20 mL of acetone and 20 mL
of 3 N H2SO4 at -15 °C was added NaNO2 (1.34 g, 19.4 mmol)
in 10 mL of H2O dropwise by addition funnel. After the
addition was complete, urea (210 mg, 3.52 mmol) was added
followed by KI (5.11 g, 30.8 mmol) in 5 mL of H2O, the ice
bath was removed, and the reaction warmed to ambient
temperature. The reaction was complete at 2 h and was
quenched with 10 mL of saturated aqueous NaHSO3, the
acetone was then evaporated, and the aqueous layer was
extracted with EtOAc 50 mL × 3. The EtOAc layers were
combined and dried over Na2SO4, filtered, evaporated to an
oil, and purified by flash chromatography (25% EtOAc/hex)
to give 4.31 g (84%) of the 4-hydroxymethyl-2-iodo-methyl
benzoate 26 as a light yellow oil. 1H NMR (300 MHz, CDCl3):
δ 8.02 (d, J ) 2 Hz, 1H), 7.82 (d, J ) 8 Hz, 1H) 7.38 (dd, J )
8, 2 Hz, 1H), 4.72 (s, 2H), 3.86 (s, 3H). CIMS: C9H9O3I MH+
292.
loxymethyl)-2-(2-trifluoromethylphenyl)methyl benzoate as an
oil.
1H NMR (300 MHz, CDCl3): δ 8.40 (bs, 1H), 8.26 (bs, 1H),
8.09 (d, J ) 8.14 Hz, 1H) 7.72 (dd, J ) 8.48, 0.69 Hz, 1H),
7.60-7.44 (m, 4H), 7.36-7.23 (m, 3H) 5.21 (s, 2H), 3.62 (s,
3H). CIMS: C21H16O3NF3 MH+ 388.
4-(3-P yr id yloxym et h yl)-2-(2-t r iflu or om et h ylp h en yl)-
b en zoic Acid . 4-(3-Pyridyloxymethyl)-2-(2-trifluorometh-
ylphenyl)methyl benzoate (241 mg, 0.62 mmol) was dissolved
in 5 mL of MeOH, 1 mL of saturated aqueous LiOH was added,
and the reaction was heated to reflux for 1 h. The reaction
was then evaporated, and 1 mL of formic acid was added to
acidify the crude product to pH ) 3. The reaction was
evaporated again to remove formic acid, and 5 mL of EtOAc
and 1 mL of H2O were added to completely solubilize the
reaction mixture. The aqueous layer was extracted with
EtOAc (5 mL × 3), and the EtOAc layers were combined and
dried over Na2SO4, filtered, and evaporated to give 231 mg
(100%) of 4-(3-pyridyloxymethyl)-2-(2-trifluoromethylphenyl)-
benzoic acid as an oil. 1H NMR (300 MHz, CDCl3): δ 8.41
(bs, 1H), 8.26 (bs, 1H), 8.15 (d, J ) 8.13 Hz, 1H), 8.11-8.06
(m, 1H), 7.70 (d, J ) 7.46 Hz, 1H), 7.55-7.40 (m, 3H), 7.40-
7.20 (m, 3H), 5.21 (s, 2H). CIMS: C20H14O3NF3 MH+ 374.
4-(3-P yr id yloxym et h yl)-2-(2-t r iflu or om et h ylp h en yl)-
ben zoylm eth ion in e, Meth yl Ester . 4-(3-Pyridyloxymethyl)-
2-(2-trifluoromethylphenyl)benzoic acid (231 mg, 0.62 mmol)
was dissolved in 4 mL of DMF, and HOOBT (152 mg, 0.93
mmol) was added followed by methionine methyl ester HCl
(185 mg, 0.93 mmol), EDC (179 mg, 0.93 mmol), and TEA (0.18
mL, 1.24 mmol). The reaction stirred for 12 h at 25 °C and
was taken up in EtOAc and washed 3 × 10 mL with H2O and
3 × 10 mL with saturated aqueous NaCl. The EtOAc layer
was dried over Na2SO4, filtered, evaporated to an oil, and
purified by radial chromatography (25% EtOAc/hexanes to 50%
EtOAc/hexanes to 5% MeOH/EtOAc) to give 291 mg (91%) of
4-(3-pyridyloxymethyl)-2-(2-trifluoromethylphenyl)benzoyl-
methionine methyl ester as an oil. 1H NMR (300 MHz,
CDCl3): δ 8.38 (bs, 1H), 8.25 (bs, 1H), 8.02 (m, 3 H)7.84-7.73
(m, 2H), 7.57-7.53 (m, 3H), 7.36-7.34 (m, 1H), 7.24-7.22 (m,
1H), 6.16 (m, 1H), 5.18 (s, 2H), 4.63-4.58 (m, 1H), 3.78 (s, 3H),
2.56-2.53 (m, 1H), 2.30-2.16 (m, 1H), 1.98-1.88 (m, 4H
contains methionine SMe), 1.80-1.70 (m, 1H). CIMS:
Meth yl-4-(3-P yr id yloxym eth yl)-2-iod o ben zoa te (27).
To a solution of 26 (6.01 g, 20.6 mmol) in 30 mL DMF were
added SOCl2 and LiCl. The reaction was stirred at 25 °C and
was complete in 5 min. The reaction was taken up in EtOAc
(100 mL), washed with H2O (20 mL × 3) and saturated
aqueous NaCl (10 mL × 4), dried over Na2SO4, filtered, and
evaporated to give 4-chloromethyl-2-iodo-methyl benzoate as
an oil that was carried on to the displacement reaction. The
benzyl chloride (6.39 g, 20.6 mmol) was dissolved in toluene,
18-crown-6 (8.17 g, 30.9 mmol) was added followed by the
potassium alkoxide of 3-hydroxypyridine, and the reaction was
heated to reflux. The reaction was complete in 2 h, and the
mixture was cooled, transferred to a separatory funnel, washed
with H2O (50 mL × 3), dried over Na2SO4, filtered, evaporated
to an oil, and purified by flash chromatography over silica gel
(gradient of 50% EtOAc/ hexanes to 75% EtOAc/ hexanes) to
give 3.01 g (40%) of the pyridyl ether 27. Rf ) 0.3 (50% EtOAc/
hex). 1H NMR (300 MHz, CDCl3): δ 8.39 (bs, 1H), 8.28 (t, J
) 2.94 Hz, 1H), 8.08 (d, J ) 1.11 Hz, 1H), 7.84 (d, J ) 7.72
Hz, 1H), 7.47 (dd, J ) 8.08, 1.83 Hz, 1H), 7.25 (d, J ) 2.58
Hz, 1H), 5.18 (s, 2H), 3.94 (s, 3H)1.86 (bs, 1H). CIMS:
C
24H25O4N2SF3 MH+ 519.
4-(3-P yr id yloxym et h yl)-2-(2-t r iflu or om et h ylp h en yl)-
ben zoylm eth ion in e (36). 4-(3-Pyridyloxymethyl)-2-(2-trif-
luoromethylphenyl)benzoyl] methionine methyl ester (291 mg,
0.56 mmol) was dissolved in 4 mL of THF, 1 mL of saturated
aqueous LiOH and 1 mL of water were added, and the reaction
stirred at room temperature. After 1 h, the reaction was
thoroughly evaporated, and formic acid was added until pH
) 3 was obtained at which time the reaction was evaporated
to dryness and 10 mL of EtOAc was added followed by a
minimum quantity of H2O (∼1 mL) to completely solubilize
the free acid and the water soluble salts, respectively. The
layers were separated, and the aqueous layer was extracted
with EtOAc (5 mL × 3). The EtOAc layers were combined,
dried over Na2SO4, filtered, evaporated, and then lyophilized
to give 242 mg (86%) of the final acid 36 as an amorphous
white solid. Rf ) 0.1 (10% MeOH/CHCl3 with 1.0% HOAc).
1H NMR (300 MHz, CD3OD): δ 8.30 (bs, 1H), 8.14 (m, 1H),
7.76-7.33 (m, 9H), 5.28 (s, 2H), 4.87-4.40 (m, 1H), 2.40-2.06
(m, 2H), 2.04-1.94 (m, 4H contains methionine SMe), 1.92-
1.80 (m, 1H). CIMS: MH+ 505. Anal. (C25H23O4N2SF3‚
0.65H2O): C, H, N. HRMS FAB Calcd m/z MH+ for C25H23O4N2-
SF3: 505.1409. Found: 505.1408.
C
14H12O3NI MH+ 370.
Gen er a l P r oced u r e for P r ep a r a tion of Com p ou n d s
w ith Mod ified Bip h en yl Cor e (Com p ou n d s 29-37). 4-(3-
P yr id yloxym et h yl)-2-(2-t r iflu or om et h ylp h en yl)b en zo-
ic Acid , Meth yl Ester (28h ). To a solution of the aryl iodide
27 (365 mg, 0.96 mmol) in 4 mL of DMF at 25 °C was added
the catalyst PdCl2(PPh3)2 (67 mg, 0.096 mmol, 10 mol %)
followed by 2-trifluoromethyl boronic acid (366 mg, 1.93 mmol),
and Cs2CO3 (629 mg, 1.93 mmol) and the reaction was heated
to 80 °C for 12 h. The reaction was then cooled and taken up
in 50 mL of EtOAc and washed with H2O (5 × 10 mL), dried
over Na2SO4, filtered, evaporated to an oil, and purified by
radial chromatography using a gradient of 25% EtOAc/hexanes
to 75% EtOAc/hexanes to give 261 mg (70%) of 4-(3-pyridy-
4-(3-P yr id yloxym et h yl)-2-(2-m et h ylp h en yl)b en zoyl-
m eth ion in e (29). Compound 29 was prepared according to
the general procedure for the preparation of compounds with
modified biphenyl core as compd 36. Rf ) 0.1 (10% MeOH/
CHCl3 with 1.0% HOAc). 1H NMR (300 MHz, CD3OD): δ 8.30
(d, 1H), 8.15 (dd, 1H), 7.68 (bd, 1H), 7.58-7.48 (m, 2H), 7.40-
7.30 (m, 2H), 7.26-7.16 (m, 4H), 5.25 (s, 2H), 4.50-4.40 (m,
1H), 2.20-2.02 (m, 5H, contains both signals for o-tolyl methyl
group), 2.00 (s, 3H. methionine methyl), 2.00-1.90 (m, 1H),

4298 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Augeri et al.
1.80-1.68 (m, 1H). CIMS: MH+ 451. Anal. (C25H26N2O4S‚
0.60EtOAc): C, H, N.
7.53 (dd, J ) 8.14, 1.7 Hz, 1H), 7.46 (ddd, J ) 4.41, 3.05, 1.35
Hz, 1H), 7.34 (dd, J ) 8.48, 5.08 Hz, 1H), 7.28 (d, J ) 1.0 Hz,
1H), 7.21-7.06 (m, 4H), 5.27 (s, 2H), 3.68 (d, 5.8 Hz, 2H)2.04
(s, 3H). CIMS: MH+ 377. Anal. (C22H20O4N2‚1.50H2O): C,
H, N.
N -Me t h y l-N -4-(3-p y r id y la m in o m e t h y l)-2-p h e n y l-
ben zoylm eth ion in e (39). This amino acid modification was
accomplished using the 3-aminopyridyl analogue of 23 followed
by the general benzoate hydrolysis procedure, the general
amino acid coupling procedure, and the general amino ester
hydrolysis procedure. 1H NMR (300 MHz, CD3OD): δ 7.94
(d, 1H), 7.78 (d, 1H), 7.40 (m, 7H), 7.20 (m, 1H), 7.11 (m, 1H),
5.20, 4.52 (both m, total 1H), 4.47 (s, 2H), 2.85-1.66 (m, 10H).
MS (DCI/NH3): 450 (MH+). Anal. (C25H27N3O3S‚0.65HCl): C,
H, N.
N -4-(3-P y r i d y la m i n o m e t h y l)-2-p h e n y l)b e n z o y l-
m eth ion in e Su lfon e (40). This amino acid modification was
accomplished using the 3-aminopyridyl analogue of 23 followed
by the general benzoate hydrolysis procedure, the general
amino acid coupling procedure, and the general amino ester
hydrolysis procedure. 1H NMR (300 MHz, D2O): δ 7.95 (m,
1H), 7.92 (m, 1H), 7.40-7.64 (m, 10H), 4.58 (s, 2H), 4.22 (ddd,
1H), 3.01 (s, 3H), 2.71 (m, 1H), 2.48 (m, 1H), 2.17 (m, 1H),
1.93 (m, 1H). FAB MS: 468 (MH+); FAB(-) 466 (MH-). Anal.
(C24H25N3O5S‚1.30H2O): C, H, N.
N -4-(3-P y r id y la m in o m e t h y l)-2-p h e n y l)b e n zo y l-D -
m eth ion in e (41). This amino acid modification was ac-
complished using the 3-aminopyridyl analogue of 23 followed
by the general benzoate hydrolysis procedure, the general
amino acid coupling procedure, and the general amino ester
hydrolysis procedure. 1H NMR (300 MHz, DMSO-d6): δ 1.75-
1.91 (m, 2H), 1.98 (s, 3H), 2.16-2.27 (m, 2H), 4.27 (m, 1H),
4.39 (d, J ) 6.4 Hz, 2H), 6.62 (t, J ) 6.4 Hz, 1H), 6.90 (ddd, J
) 1.4, 2.7, 8.5 Hz, 1H), 7.03 (dd, J ) 4.6, 8.3 Hz, 1H), 7.30-
7.41 (m, 8H), 7.40 (d, J ) 4.1 Hz, 1H), 7.97 (d, J ) 2.7 Hz,
1H), 8.50 (d, J ) 7.8 Hz, 1H), 12.65 (br s, 1H). MS (DCI): m/e
436 (MH+). Anal. (C24H25N3O3S‚0.90H2O): C, H, N.
N-4-(3-P yr id yloxym eth yl)-2-p h en ylben zoyl-O-m eth yl-
h om oser in e (42). This amino acid modification was ac-
complished beginning with 23 followed by the general benzoate
hydrolysis procedure, the general amino acid coupling proce-
dure, and the general amino ester hydrolysis procedure. 1H
NMR (300 MHz, DMSO-d6): δ 8.57 (d, 1H), 8.43 (d, 1H), 8.22
(d, 1H), 7.60 (m, 1H), 7.53 (m, 2H), 7.40 (m, 7H), 5.33 (s, 2H),
4.26 (m, 1H), 3.08 (s, 3H), 3.07 (m, 2H), 1.90 (m, 1H), 1.75 (m,
1H). MS (APCI): 421 (MH+). Anal. (C24H24N2O5‚0.50H2O):
C, H, N.
N -4-(3-P yr id yloxym e t h yl)-2-p h e n ylb e n zoyln or le u -
cin e (43). This amino acid modification was accomplished
beginning with 23 followed by the general benzoate hydrolysis
procedure, the general amino acid coupling procedure, and the
general amino ester hydrolysis procedure. 1H NMR (300 MHz,
DMSO-d6): δ 8.60 (d, 1H), 8.53 (d, 1H), 8.37 (d, 1H), 7.90 (dd,
1H), 7.70 (dd, 1H), 7.52 (d, 1H), 7.51 (s, 1H), 7.42 (m, 3H),
7.38 (m, 3H), 5.38 (s, 2H), 4.16 (m, 1H), 1.60 (m, 2H), 1.20 (m,
2H), 1.10 (m, 2H), 0.82 (t, 3H). MS (DCI/NH3): 419 (MH+).
Anal. (C25H26N2O4‚0.50H2O): C, H, N.
N-4-(3-P yr id yloxym eth yl)-2-p h en ylben zoyl-S-m eth yl-
cystein e (44). This amino acid modification was accom-
plished beginning with 23 followed by the general benzoate
hydrolysis procedure, the general amino acid coupling proce-
dure, and the general amino ester hydrolysis procedure. 1H
NMR (300 MHz, DMSO-d6): δ 8.77 (d, 1H), 8.59 (d, 1H), 8.35
(d, 1H), 7.85 (dd, 1H), 7.63 (dd, 1H), 7.50 (m, 5H), 7.35 (m,
3H), 5.38 (s, 2H), 4.42 (m, 1H), 2.89 (dd, 1H), 2.72 (dd, 1H),
2.04 (s, 3H). MS (DCI/NH3): 423 (MH+). Anal. (C23H22N2O4S‚
0.20H2O): C, H, N.
4-(3-P yr id yloxym et h yl)-2-(3-m et h ylp h en yl)b en zoyl-
m eth ion in e (30). Compound 30 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. Rf ) 0.1 (10%
MeOH/CHCl3 with 1.0% HOAc). 1H NMR (300 MHz, CD3-
OD): δ 8.30 (d, 1H), 8.15 (d, 1H), 7.68-7.48 (m, 6H), 7.40-
7.16 (m, 4H), 5.25 (s, 2H), 4.50-4.40 (m, 1H), 2.40 (s, 3H),
2.18-1.75 (m, 7H). CIMS: MH+ 451. Anal. (C25H26N2O4S‚
0.10CH2Cl2): C, H, N.
4-(3-P yr id yloxym et h yl)-2-(4-m et h ylp h en yl)b en zoyl-
m eth ion in e (31). Compound 31 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. 1H NMR (300
MHz, CD3OD): δ 8.30 (d, 1H), 8.15 (d, 1H), 7.58-7.44 (m, 4H),
7.40-7.28 (m, 3H), 7.24-7.10 (m, 3H), 5.25 (s, 2H), 4.42 (dd,
1H), 2.10-1.90 (m, 6H), 1.84-1.70 (m, 1H). CIMS: MH+ 451.
Anal. (C25H26N2O4S‚0.15CH2Cl2): C, H, N.
4-(3-P yr id yloxym eth yl)-2-(2-m eth oxyp h en yl)ben zoyl-
m eth ion in e (32). Prepared according to the general proce-
dure for the preparation of compounds with
a modified
biphenyl core as in compound 36. 1H NMR (300 MHz, CD3-
OD): δ 8.30 (d, 1H), 8.15 (d, 1H), 7.68 (bd, 1H), 7.54-7.50 (m,
2H), 7.38-7.32 (m, 3H), 7.22 (dd, 1H), 7.04-6.98 (m, 2H), 5.25
(s, 2H), 4.42 (dd, 1H), 3.74 (s, 3H), 2.16-2.08 (m, 2H), 2.00 (s,
3H), 1.98-1.86 (m, 1H), 1.78-1.64 (m, 1H). CIMS: MH+ 467.
Anal. (C25H26N2O5S‚0.25H2O): C, H, N.
4-(3-P yr id yloxym eth yl)-2-(3-m eth oxyp h en yl)ben zoyl-
m eth ion in e (33). Compound 33 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. 1H NMR (300
MHz, CD3OD): δ 8.34 (s, 1H), 8.15 (d, 1H), 7.60-7.54 (m, 4H),
7.38-7.24 (m, 3H), 7.02-6.90 (m, 3H), 5.25 (s, 2H), 4.44 (dd,
1H), 3.82 (s, 3H), 2.18-1.90 (m, 6H), 1.92-1.82 (m, 1H).
CIMS: MH+ 467. Anal. (C25H26N2O5S‚0.55H2O): C, H, N.
4-(3-P yr id yloxym eth yl)-2-(4-m eth oxyp h en yl)ben zoyl-
m eth ion in e (34). Compound 34 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. 1H NMR (300
MHz, CD3OD): δ 8.34 (s, 1H), 8.15 (bs, 1H), 7.72-7.42 (m,
6H), 7.40-7.35 (m, 2H), 6.96-6.90 (m, 2H)5.25 (s, 2H), 4.44
(dd, 1H), 3.84 (s, 3H), 2.20-1.90 (m, 6H), 1.88-1.76 (m, 1H).
CIMS: MH+ 467. Anal. (C25H26N2O5S): C, H, N.
4-(3-P yr id yloxym e t h yl)-2-(2-ch lor op h e n yl)b e n zoyl-
m eth ion in e (35). Compound 35 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. Rf ) 0.1 (10%
MeOH/CHCl3 with 1.0% HOAc). 1H NMR (500 MHz, CD3-
OD): δ 8.31 (bs, 1H), 8.14 (d, J ) 4.4 Hz, 1H), 7.70-7.34 (m,
9H), 5.29 (s, 2H), 4.48-4.45 (m, 1H), 2.30-2.22 (m 1 H), 2.20-
2.15 (m, 1H), 2.05-1.95 (m, 4H,contains SMe), 1.86-1.76 (m,
1H). CIMS: MH+ 471. Anal. (C24H23O4N2SCl): C, H, N.
HRMS FAB Calcd m/z MH+ for C24H23O4N2SCl: 471.1145.
Found: 471.1165.
4-(3-P yr id yloxym e t h yl)-2-(2-e t h ylp h e n yl)b e n zoyl-
m eth ion in e (37). Compound 37 was prepared according to
the general procedure for the preparation of compounds with
a modified biphenyl core as in compound 36. Rf ) 0.1 (10%
MeOH/CHCl3 with 1.0% HOAc). 1H NMR (300 MHz, CD3-
OD): δ 8.30 (bs, 1H), 8.14 (d, J ) 4.4 Hz, 1H), 7.71-7.17 (m,
9H), 5.29 (s, 2H), 4.87-4.43 (m, 1H), 2.54-2.37 (m 2 H), 2.24-
1.84 (m, 7H, contains SMe), 1.90-1.82 (m, 1H), 1.04 and 0.97
(rotameric triplets, J ) 7.3 Hz, 3H). CIMS: MH+ 465. Anal.
(C26H28O4N2S‚022H2O): C, H, N. HRMS FAB Calcd m/z MH+
for C26H28O4N2S: 465.1848. Found: 465.1865.
4-(3-P yr id yloxym et h yl)-2-(2-m et h ylp h en yl)b en zoyl-
glycin e (38). Compound 38 was prepared according to the
general procedure for the preparation of compounds with a
modified biphenyl core as in compound 36 except that glycine
methyl ester HCl was substituted for L-methionine methyl
ester HCl in the coupling. Rf ) 0.15 (10% MeOH/CHCl3 with
1.0% HOAc). 1H NMR (500 MHz, d6DMSO): δ 8.37-8.33 (m,
2H), 8.17 (dd, J ) 4.7, 1.4 Hz, 1H), 7.59 (d, J ) 8.14 Hz, 1H),
N -4 -( 3 -P y r i d y l o x y m e t h y l ) -2 -p h e n y l b e n z o y l -
glu ta m in e (45). This amino acid modification was ac-
complished beginning with 23 followed by the general benzoate
hydrolysis procedure, the general amino acid coupling proce-
dure, and the general amino ester hydrolysis procedure. 1H
NMR (300 MHz, DMSO-d6): δ 1.79 (m, 1H), 1.95 (m, 1H), 2.09
(m, 2H), 4.18 (m, 1H), 5.42 (s, 2H), 6.80 (bs, 1H), 7.25 (m, 2H),

Non-Cysteine-Containing Inhibitors of Protein FTase
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4299
7.35 (m, 3H), 7.45 (m, 2H), 7.55 (m, 3H), 7.86 (dd, J ) 5.2, 8.5
Hz, 1H), 8.10 (d, J ) 7.7 Hz, 1H), 8.46 (d, J ) 4.4 Hz, 1H),
8.69 (d, J ) 8.1 Hz, 1H), 8.71 (bs, 1H). MS (DCI): m/e 434
(MH+). Anal. (C24H23N3O5‚2.40HCl): C, H, N.
identical fashion to 15 except with the o-tolyl biphenyl core
present in 29. 1H NMR (300 MHz, DMSO-d6): δ 8.08 (d, 1H),
7.97 (d, 1H), 7.72 (d, 1H), 7.46 (d, 1H), 7.43 (dd, 1H), 7.17 (m,
3H), 7.10 (m, 2H), 7.03 (dd, 1H), 6.89 (m, 1H), 6.55 (t, 1H),
4.37 (d, 2H), 4.20 (m, 1H), 2.00-2.24 (m, 2H), 1.99 (bs, 3H),
1.93 (s, 3H), 1.63-1.88 (m, 2H). CIMS: 450 (MH+). Anal.
(C25H27N3O3S‚0.30HCl): C, H, N.
N-4-(3-P yr id ylth iom eth yl)-2-(2-m eth ylp h en yl)ben zoyl-
m eth ion in e (49). Compound 49 was synthesized in identical
fashion to 20 except with the o-tolyl biphenyl core present in
29. 1H NMR (300 MHz, DMSO-d6): δ 12.55 (bs, 1H), 8.50 (d,
1H), 8.37 (dd, 1H), 8.11 (d, 1H), 7.78 (dt, 1H), 7.43 (s, 2H),
7.30 (dd, 1H), 7.18 (m, 2H), 7.11 (m, 2H), 7.04 (bs, 1H), 4.36
(s, 2H), 4.20 (ddd, 1H), 2.00-2.22 (m, 2H), 1.96 (bs, 3H), 1.94
(s, 3H), 1.63-1.88 (m, 2H). CIMS: 467 (MH+). Anal.
(C25H26N2O3S2): C, H, N.
In Vitr o En zym e Assa ys. In vitro IC50 data were deter-
mined against FTase and GGTase 1 (purified from bovine
brain) using the SPA assay (scintillation proximity assay,
Amersham, Arlington Heights). The substrates used were 3H-
farnesyl pyrophosphate and a biotin-linked Kras(B) decapep-
tide (KKSKTKCVIM, for FTase) or the CVLL decapeptide for
GGTase 1. The radioactivity captured by the SPA beads were
counted by a Packard Topcount, and data were stored and
analyzed in an Oracle-based database.
Cellu la r Assa ys for In h ibition of Ha -Ra s P r ocessin g:
Subconfluent NIH3T3 Ras transformed cells were used for the
Ras processing assay. Briefly, cells were dosed with various
compounds, and lysates were prepared. They were boiled for
5 min in the Laemmli sample buffer, and proteins were
resolved on a 15% Tris-glycine gel (Bio-Rad, Richmond, CA).
Proteins were then transferred to nitrocellulose membranes.
The blots were probed with antibody Y13-238 to Ras purified
from hybridoma. Ras bands were visualized by the ECL
technique (ECL kit, Amersham, Arlington Heights, IL), and
signals were quantified by densitometry using the image
analysis program Image-Pro Plus (Media Cybernetics, Silver
Spring, MD).
4-(3-P y r id y lc a r b o n y la m in o )-2-(2-m e t h y lp h e n y l)-
m eth ion in e (46). To a stirred solution of 4-amino-2-(2-
methylphenylbenzoyl)methionine methyl ester (85 mg, 0.23
mmol) in CH2Cl2 (5 mL) were added nicotinic acid chloride
hydrochloride (81 mg, 0.46 mmol) and saturated NaHCO3 (2
mL). The reaction was stirred at room temperature for 2 h.
TLC (CH2Cl2/ MeOH 20:1) showed the formation of new
products. The reaction was diluted with CH2Cl2 (10 mL), the
layers separated, and the organic layer was washed with
saturated NaHCO3 (5 mL), dried (MgSO4), and concentrated
in vacuo. Flash chromatography (CH2Cl2/MeOH 50:1) and
crystallization from EtOAc gave 87 mg (80%) of a white
powder. 1H NMR (300 MHz, CDCl3): δ 9.10 (dd, 1 H, J ) 2.4,
1.0 Hz), 8.80 (dd, 1 H, J ) 4.7, 1.7 Hz), 8.21 (ddd, 1 H, J )
7.8, 2.4, 1.7 Hz), 8.09-8.00 (m, 2 H), 7.71-7.66 (m, 1 H), 7.64-
7.61 (m, 1 H), 7.46 (ddd, 1 H, J ) 7.8, 4.7, 1.0 Hz), 7.35-7.20
(m, 4 H), 5.92 (bd, J ) 7.5 Hz), 4.67-4.57 (m, 1 H), 3.66 (s, 3
H), 2.23-2.01 (4s and m, 8 H), 2.13-2.00 (m, 1 H), 1.65-1.52
(m, 1 H). MS: m/z 478 (MH+, 100). To a stirred solution of
the methionine methyl ester (140 mg, 0.29 mmol) in THF (6
mL) was added a solution of LiOH‚H2O (37 mg, 88 mmol) in
H2O (1 mL), and the resulting solution stirred for 2 h at
ambient temperature. TLC analysis (20:1 CH2Cl2/MeOH)
showed the formation of new products. The reaction was
concentrated in vacuo, 1 N HCl was added to the residue, and
the resulting precipitate was filtered and washed with H2O.
Lyophilization gave 87 mg (59%) of a white powder. 1H NMR
(300 MHz, DMSO-d6, 90 °C): δ 9.12 (d, J ) 2.4 Hz, 1 H), 8.74
(dd, J ) 4.9, 1.9 Hz, 1 H), 8.31 (dt, J ) 7.9, 1.8 Hz, 1 H), 7.84
(dd, J ) 7.9, 1.8 Hz, 1 H), 7.63 (s, 1 H), 7.61 (d, J ) 2.4 Hz, 1
H), 7.54 (dd, J ) 7.9, 4.9 Hz, 1 H), 7.45 (d, J ) 7.9 Hz, 1 H),
7.23-7.21 (m, 2 H), 7.19-7.15 (m, 2H), 4.30-4.26 (m, 1 H),
2.28-2.22 (m, 1 H), 2.20-2.14 (m, 1 H), 2.11 (s, 3 H), 1.98 (s,
3 H), 1.88-1.81 (m, 1 H), 1.75-1.68 (m, 1 H). MS: m/z 464
(MH+, 100), 446. Anal. (C25H25N3O4S‚HCl‚0.5H2O): C, H, N.
4-(3-P yr id ylm eth yla m in o)-2-(2-m eth ylp h en yl)ben zoyl-
m et h ion in e (47). 4-Amino-2-(2-methylphenylbenzoyl)me-
thionine methyl ester (180 mg, 0.48 mmol) and 3-pyridine
carboxaldehyde (55 mg, 0.51 mmol) were combined in 4 mL of
MeOH, and sodium cyanoborohydride (48 mg, 0.77 mmol) was
added followed by 100 mg of crushed molecular sieves. The
reaction was adjusted to pH ) 6 with acetic acid and stirred
at 25 °C. After 3 h the reaction was concentrated and
transferred directly to a column of silica gel and purified by
flash chromatography (5% MeOH/ EtOAc) to give 182 mg (82%)
of 4-(3-pyridylmethylamino)-2-(2-methylphenyl)benzoylme-
thionine methyl ester as an oil that solidified after standing.
Rf ) 0.19 (5% MeOH/EtOAc). 1H NMR (300 MHz, CD3OD):
δ 8.54 (d, J ) 2.4 Hz, 1H), 8.40 (dd, J ) 5.1, 1.3 Hz, 1H), 7.84
(bd, J ) 8.4 Hz, 1H), 7.65-7.55 (m, 1H), 7.40 (dd, J ) 7.8, 4.7
Hz, 1H), 7.30-7.10 (m, 4H), 6.66 (dd, J ) 8.8, 2.3 Hz, 1H),
6.37 (d, J ) 2.3 Hz, 1H), 4.45 (s, 2H), 3.64 (s, 3H), 2.10-1.98
(m, 8H), 1.90-1.78 (m, 1H), 1.65-1.55 (m, 1H). CIMS: MH+
464. HRMS FAB Calcd m/z MH+ for C26H29O3N3S: 464.2008.
Found: 464.2023.
The methionine methyl ester (102 mg, 0.22 mmol) was
converted to the methionine acid using the general amino ester
hydrolysis protocol and then lyophilized as the HCl salt from
MeCN/H2O to give 68 mg (68%) of 47 as a light tan solid. Rf
) 0.18 (10% MeOH/CHCl3 with 0.25% HOAc). 1H NMR (300
MHz, CD3OD): δ 8.81 (bs, 1H), 8.76 (bd, J ) 11.8 Hz, 1H),
8.64-8.61 (m, 1H), 8.07 (dd, J ) 8.5, 6.1 Hz, 1H), 7.65-7.58
(m, 1H), 7.28-7.18 (m, 4H), 6.70 (dd, J ) 8.5, 2.4 Hz, 1H),
6.40 (d, J ) 2.3 Hz, 1H), 4.68 (s, 2H), 4.44-4.38 (m, 1H), 2.14-
1.99 (m, 8H), 1.90-1.80 (m, 1H), 1.65-1.55 (m, 1H). CIMS:
MH+ 450. Anal. (C25H28O3N3SCl‚1.10H2O and 0.80 HCl): C,
H, N. HRMS FAB Calcd m/z MH+ for C25H27O3N3S: 450.1851.
Found: 450.1864.
Ack n ow led gm en t. The authors thank Professors A.
D. Hamilton and S. M. Sebti for fruitful discussions
during the course of this research. The authors also
thank Dr. Paul West, Dr. Tom Pagano, Dr. Xiaolin
Zhang, and Ms. Carmina Presto of the structural
chemistry department for their assistance in obtaining
NMR and mass spectra, Mr. William Arnold for large-
scale preparation of several key intermediates, and Dr.
Stefan Loren for preparing several boronic acids used
in the biphenyl core modification study.
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