Design, synthesis, and evaluation of cyclic amide/imide-bearing hydroxamic acid derivatives as class-selective histone deacetylase (HDAC) inhibitors

Article abstract of DOI:10.1016/j.bmc.2006.07.008

A series of hydroxamic acid derivatives bearing a cyclic amide/imide group as a linker and/or cap structure, prepared during our structural development studies based on thalidomide, showed class-selective potent histone deacetylase (HDAC)-inhibitory activity. Structure-activity relationship studies indicated that the steric character of the substituent introduced at the cyclic amide/imide nitrogen atom, the presence of the amide/imide carbonyl group, the hydroxamic acid structure, the shape of the linking group, and the distance between the zinc-binding hydroxamic acid group and the cap structure are all important for HDAC-inhibitory activity and class selectivity. A representative compound (30w) showed potent p21 promoter activity, comparable with that of trichostatin A (TSA), and its cytostatic activity against cells of the human prostate cell line LNCaP was more potent than that of the well-known HDAC inhibitor, suberoylanilide hydroxamic acid (SAHA).

Full text of DOI:10.1016/j.bmc.2006.07.008

Bioorganic & Medicinal Chemistry 14 (2006) 7625–7651
Design, synthesis, and evaluation of cyclic amide/imide-bearing
hydroxamic acid derivatives as class-selective histone deacetylase
(HDAC) inhibitors
Chihiro Shinji,a Satoko Maeda,b Keisuke Imai,c Minoru Yoshida,b,d
Yuichi Hashimotoa and Hiroyuki Miyachia,*
aInstitute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
bRIKEN, Saitama 351-0198, Japan
cLead Discovery Research Labs., Astellas Pharma Inc., 21 Miyukigaoka, Tsukuba-shi, Ibaraki 305-8585, Japan
dCREST Research Project, Japan Science and Technology Agency, Saitama 332-0012, Japan
Received 18 May 2006; revised 30 June 2006; accepted 1 July 2006
Available online 31 July 2006
Abstract—A series of hydroxamic acid derivatives bearing a cyclic amide/imide group as a linker and/or cap structure, prepared
during our structural development studies based on thalidomide, showed class-selective potent histone deacetylase (HDAC)-inhib-
itory activity. Structure–activity relationship studies indicated that the steric character of the substituent introduced at the cyclic
amide/imide nitrogen atom, the presence of the amide/imide carbonyl group, the hydroxamic acid structure, the shape of the linking
group, and the distance between the zinc-binding hydroxamic acid group and the cap structure are all important for HDAC-inhib-
itory activity and class selectivity. A representative compound (30w) showed potent p21 promoter activity, comparable with that of
trichostatin A (TSA), and its cytostatic activity against cells of the human prostate cell line LNCaP was more potent than that of the
well-known HDAC inhibitor, suberoylanilide hydroxamic acid (SAHA).
ꢀ 2006 Elsevier Ltd. All rights reserved.
1. Introduction
p21WAF1/CIP1
, which is a cyclin-dependent kinase
(CDK) inhibitor that inhibits the kinase activities of a
class of CDKs, leading to cell cycle arrest and dephos-
phorylation of Rb.5 Much evidence suggests that
p21WAF1/CIP1 plays an important role in determining
the fate of cells during growth and differentiation.
Therefore, compounds that inhibit HDAC activity
may depress expression of the p21WAF1/CIP1 gene, result-
ing in antiproliferative and antitumor effects.6 Natural
and synthetic HDAC inhibitors have been studied exten-
sively (see Fig. 1).
Reversible acetylation of the side-chain amino groups of
specific histone lysine residues plays an important func-
tion in modifying chromatin structure and regulating
gene expression. The key enzymes that modify histone
proteins and regulate gene expression are histone acetyl-
transferases (HATs) and histone deacetylases
(HDACs).1,2 Both of these acetylating/deacetylating
enzymes are included in large protein complexes con-
taining other proteins that are known to function in
transcriptional activation and/or repression.3,4 Such
complexes are recruited to specific regions in the DNA
and induce expression and/or silencing of the genes.
Many recent studies have shown that inhibition of
HDAC elicits anticancer effects in several lines of tumor
cells by inhibiting cell growth and inducing apoptosis. A
well-known target that is upregulated by HDAC is
Numerous biological studies indicated that HDACs are
heterogeneous, consisting of 18 isozymes, which can be
categorized into four classes (class I, class IIa, class
IIb, and class III). Class I and class II HDACs are
zinc-containing amidehydrolases, and class III HDAC
consists of NAD-dependent amidehydrolase. The bio-
logical function and distribution of each class of
HDACs have been extensively studied from a molecu-
lar-pharmacological viewpoint, but much remains to
be learnt. Although some class-selective HDAC inhibi-
tors are known, most of the reported HDAC inhibitors
Keywords: HDAC; HDAC inhibitor; Cyclic amide; Class selectivity;
Hydroxamic acid.
*
Corresponding author. E-mail: miyachi@iam.u-tokyo.ac.jp
0968-0896/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2006.07.008

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C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
O
reduction, followed by PDC oxidation, gave the N-ben-
zyl-6-formyl-2,3-dihydroisoindole-1-one derivative (8).
Compound (8) was treated with Horner–Emmons re-
agent in the presence of base, and subsequent alkaline
hydrolysis afforded the cinnamic acid derivative (10).
Compound (10) was condensed with O-(tetrahydro-
2H-pyran-2yl)hydroxylamine via the mixed anhydride
to give the protected N-hydroxybenzamide derivative
(11). Deprotection of 11 under acidic conditions affor-
ded the N-hydroxybenzamide derivative (12) (Chart 1).
Regioisomeric isoindolone-hydroxamic acid derivatives
were prepared from 4-nitro-2-methylbenzoic acid (13).
Compound (13) was etherified, brominated and treated
with benzylamine to afford the cyclic amide derivative
(16). Compound (16) was reduced with 10% Pd on car-
bon, followed by Meerwein arylation reaction, strong
base treatment and alkaline hydrolysis to afford the cin-
namic acid derivative (20). Compound (20) was deriva-
tized to hydroxamic acid (22) by means of procedures
similar to those employed for the preparation of com-
pound (12) (Chart 2).
H
N
OH
HN
O
O
O
N
H
HN
N
O
H
N
O
O
O
1
O
S
O
O
N
HO
2
4
O
O
O
H
N
OH
N
H
OH
N
H
N
O
3
Figure 1. Structures of representative natural HDAC inhibitors (1
(trapoxin A), 3 (TSA)), and synthetic HDAC inhibitors (2 (tubacin), 4
(SAHA)).
are non-class-selective, pan-inhibitors, and little is
known about the structure–activity relationships associ-
ated with class selectivity.
Our earlier study indicated the importance of the meta
position carbonyl group of phthalimide hydroxamic
acid derivatives. However, the synthetic route depicted
in Chart 2 is not convenient for the preparation of var-
ious types of amide nitrogen substituents, since the sub-
stituents are introduced at the first step of the sequence.
Therefore, we tried to develop another route that would
be suitable for incorporating diverse amide substituents.
This route is outlined in Chart 3. Methyl (or tert-butyl)
2-methyl-5-amino benzoate (23) was subjected to
Meerwein arylation reaction, followed by strong base
treatment to afford the methyl (or tert-butyl) cinnamate
derivatives (25). Compound (25) was brominated, and
treated with various kinds of benzylamines, followed
by alkaline hydrolysis to afford various cinnamic acid
derivatives (28). Compounds (28) were condensed with
tert-butylhydroxylamine or O-(tetrahydro-2H-pyran-
2yl)hydroxylamine by means of the mixed anhydride
method to give protected N-hydroxybenzamide deriva-
tives (29). Deprotection of 29 under acidic conditions
We have been engaged in structural development studies
of the multi-drug template thalidomide for the creation
of structurally novel drug leads7–12 and have already
reported the design and synthesis of potent HDAC
inhibitors with phthalimide structure.13 In this paper,
we report the design, synthesis, and structure–activity
relationship of novel HDAC inhibitors containing a cyc-
lic amide/imide structure. The in vitro p21 promoter
activity and cytostatic effects on the human prostate
cancer cell LNCaP of representative compounds are also
described.
2. Chemistry
Synthetic routes to the present series of cyclic amide
derivatives are outlined in Charts 1–7. Selective reduc-
tion of one (para) carbonyl group of the phthalimide
hydroxamic acid derivative (5) afforded 6, then BH3
O
O
O
O
O
OH
OH
OH
N
N
N
a)
b)
c)
O
5
8
6
9
7
O
O
O
O
O
O
H
OMe
OH
N
N
N
d)
e)
f)
10
O
O
O
O
N OTHP
N OH
N
N
H
H
g)
11
12
Chart 1. Reagents and conditions: (a) Sn, cHCl, AcOH, reflux; (b) BH3, THF, 0 ꢁC; (c) MnO2, CH2Cl2, rt; (d) (EtO)2POCH2CO2Me, t-BuOK, THF,
rt; (e) 1 mol/L NaOH, MeOH, 50 ꢁC; (f) 1—ethyl chloroformate, triethylamine, THF, 0 ꢁC; 2—O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, rt,
MeOH; (g) CSA, MeOH, rt.

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7627
NO2
NO2
NO2
NO2
Br
MeO
N
HO
h)
MeO
i)
j)
O
O
O
O
13
14
15
16
O
O
NH2
OMe
OMe
N
N
N
m)
k)
l)
Br
O
O
O
17
18
19
O
O
O
OH
N OTHP
N OH
H
N
N
N
H
n)
o)
O
O
O
20
21
22
Chart 2. Reagents and conditions: (h) MeI, K2CO3, DMF, rt; (i) NBS, benzoyl peroxide, CCl4, reflux; (j) benzylamine, TEA, MeOH, reflux.; (k) 10%
Pd–C, H2, AcOEt, rt.; (k) 1—NaNO2, 47% HBr, acetone, MeOH, <5 ꢁC; 2—methyl acrylate, Cu2O, 35–40 ꢁC; (l) DBU, toluene, reflux; (m) 0.8 mol/L
HCl, AcOH, reflux; (n) 1—ethyl chloroformate, TEA, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, THF, 0 ꢁC-rt; (o) CSA, MeOH, rt.
O
O
O
O
O
NH2
3
3
p)
q)
O
r)
MeO
MeO
O
MeO
O
R
R
Br
24a R3 = Me, 24b R3 = t-butyl
23
25a R3 = Me, 25b R3 = t-butyl
O
O
O
O
O
3
MeO
Br
O
OR3
OH
s)
t)
R
R4
R4
N
N
26a R3 = Me, 24b R3 = t-butyl
27a-aa
R4
28a-aa
O
O
O
O
R5
O
OH
u)
v)
N
N
R4
N
N
H
H
29a-aa, R5 = THP or t-butyl
30a-aa
Chart 3. Reagents and conditions: (p) 1—NaNO2, 47% HBr, acetone, MeOH, <5 ꢁC; 2—methyl acrylate, Cu2O, 35–40 ꢁC; (q) DBU, toluene, reflux;
(r) NBS, benzoyl peroxide, CCl4, reflux; (s) benzylamine, TEA, MeOH, reflux; (t) 0.8 mol/L HCl, AcOH, reflux; (u) 1—ethyl chloroformate,
triethylamine, THF, 0 ꢁC; 2—NH2OTHP (or NH2OC(CH3)3), MeOH, rt; (v) CSA, MeOH, rt.
O
O
O
O
R2
R2
R2
OH
OTf
OMe
N
N
N
w)
x)
y)
R1
R1
R1
O
O
O
31a-l
32a-l
33a-l
R1
O
O
O
O
N
O
O
R2
R2
R2
O
OH
OH
N
N
N
N
H
H
z)
aa)
R1
R1
O
O
O
34a-l
35a-l
36a-l
Chart 4. Reagents and conditions: (w) trifluoromethanesulfonic anhydride, triethylamine, toluene, rt; (x) methyl acrylate, (P(Ph)3)2PdCl2,
triethylamine, DMF, reflux; (y) 0.5 mol/L HCl, AcOH, reflux; (z) 1—ethyl chloroformate, triethylamine, THF 0 ꢁC; 2—tert-butylhydroxylamine,
MeOH, rt; (aa) trifluoroacetic acid, CH2Cl2, rt.
afforded the desired N-hydroxybenzamide derivatives
(30) (Chart 3).
Compounds (32) were treated with methyl acrylate in
the presence of (P(Ph)3)2PdCl2 and triethylamine in
N,N-dimethylformamide, and then alkaline hydrolysis
afforded the cinnamic acid derivatives (34). Compounds
(34) were condensed with tert-butylhydroxylamine by
means of the mixed anhydride method to give protected
As previously described,13 5-hydroxyphthalimide deriv-
atives (31) were treated with trifluoromethanesulfonic
anhydride to give the Heck reaction substrates (32).

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C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
O
O
O
O
O
O
O
NH2
OH
O
O
bb)
cc)
dd)
MeO
MeO
O
MeO
O
MeO
Br
O
23
37
38
39
O
O
O
O
N
R1
O
O
O
R1
R1
OTHP
N
O
O
OH
N
N
H
ee)
ff)
gg)
40a-e
41a-e
42a-e
O
O
R1
OH
N
H
N
hh)
43a-e
Chart 5. Reagents and conditions: (bb) 1—NaNO2, 47% HBr, acetone, MeOH, <5 ꢁC; 2—Cu2O, H2O, reflux; (cc) 1—tert-bromoacetoacetate,
K2CO3, acetone, reflux; (dd) NBS, benzoyl peroxide, CCl4, reflux; (ee) substituted benzylamine, TEA, MeOH, reflux; (ff) TFA, rt; (gg) 1—ethyl
chloroformate, triethylamine, THF, 0 ꢁC; 2—O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, THF, 0 ꢁC-rt. (hh) CSA, MeOH, rt.
O
O
O
O
O
NO2
NO2
NH2
ii)
jj)
kk)
nn)
HO
N
N
H
N
H
O
H
Br
44
45
46
47
O
O
O
O
O
O
OTHP
ll)
N
H
O
mm)
N
OH
N
H
N
H
H
48
51
49
50
O
O
OH
oo)
N
N
H
H
Chart 6. Reagents and conditions: (ii) benzylamine, DMC, TEA, THF, 0 ꢁC; (jj) H2, 10% Pd–C, AcOET, rt; (kk) 1—NaNO2, 47% HBr, acetone,
MeOH, <5 ꢁC; 2—tert-butyl acrylate, Cu2O, 35–40 ꢁC; (ll) DBU, toluene, reflux; (mm) TFA, rt; (nn) 1—ethyl chloroformate, triethylamine, THF
0 ꢁC; 2—O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, THF, rt; (oo) CSA, MeOH, rt.
O
O
O
O
O
O
pp)
qq)
HO
H
N
N
H
N
OMe
tt)
54
53
52
O
O
O
O
OTHP
rr)
ss)
OH
N
N
H
55
56
O
O
OH
N
N
H
57
Chart 7. Reagents and conditions: (pp) benzylamine, DMC, TEA, THF, 0 ꢁC; (qq) (EtO)2POCH2CO2Me, t-BuOK, THF, rt; (rr) dil NaOH, MeOH,
60 ꢁC; (ss) 1—ethyl chloroformate, triethylamine, THF, 0 ꢁC; 2—O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, THF, rt; (tt) CSA, MeOH, rt.
N-hydroxybenzamide derivatives (35). Deprotection of
the tert-butyl group of 35 under acidic conditions affor-
ded the N-hydroxybenzamide derivatives (36) (Chart 4).
Hydroxymethyl-linker derivatives (43) were prepared
starting from methyl 2-methyl 5-aminobenzoate (23).
Compound (23) was subjected to Sandmeyer reaction
and subsequent treatment with tert-butyl bromoacetate
to afford compound (38). This was brominated and
treated with substituted benzylamines, then alkaline
hydrolysis afforded the phenoxyacetic acid derivatives
(41). Compounds (41) were derivatized to the desired
hydroxamic acids (43) by means of the procedures
similar to those employed for the preparation of
compounds (12) (Chart 5).
To examine the importance of the cyclic amide (isoindo-
lone) structure, two structural types of acyclic amide
derivatives, one with the C–N bond disconnected and

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7629
the other with the C–C bond disconnected, were pre-
pared as depicted in Charts 6 and 7. 2-Methyl-5-nitro-
benzoic acid (44) was amidated with benzylamine, and
subsequent reduction afforded N-benzyl 5-amino-2-
methylbenzamide (46). Compound (46) was subjected
to Meerwein arylation reaction, strong base treatment,
and subsequent alkaline hydrolysis to afford the cinnam-
ic acid derivative (49). Compound (49) was derivatized
to the desired C-N bond disconnected-type hydroxamic
acid (51) by means of the procedures described above
(Chart 6). The 3-formylbenzoic acid (52) was amidated
with N-methylbenzylamine, followed by Wittig–
Horner–Emmons reaction and subsequent alkaline
hydrolysis to afford the cinnamic acid (55). Compound
(55) was derivatized to the desired C–C bond discon-
nected-type hydroxamic acid (57) by means of the proce-
dures described above (Chart 7).
tube-like region of the HDAC active pocket are hydro-
phobic. At the beginning of our current study, we
planned to synthesize the desired compound by selective
reduction of the lead compound (II) with tin in an acidic
medium, but unfortunately, the reduction did not pro-
ceed regioselectively, and a 1:1 mixture of regioisomeric
decarbonyl products was obtained. Fortunately, howev-
er, the regioisomeric mixture exhibited more potent
HDAC-inhibitory activity than the imide lead (II), as as-
sessed with the HDAC inhibitor assay kit (data not
shown). Therefore, we prepared both regioisomers spe-
cifically via the synthetic routes depicted in Charts 1
and 2. The assay results indicated that the meta position
carbonyl group is important, that is, compound (12) was
about 3 times more potent than the regioisomer (22).
Furthermore, cyclic amide/imide structure was impor-
tant for potent HDAC-inhibitory activity, because the
acyclic amide derivatives 51 and 57 exhibited decreased
HDAC inhibitory activity (Fig. 3). Molecular modeling
studies of compounds 12 and 22 complexed with HDAC
818 indicated that there might be a weak hydrogen-
bonding interaction between the meta position carbonyl
group of 12 and the a-nitrogen of 151Gly, while there is
no apparent interaction between the para position car-
bonyl group of compound 22 and the HDAC8 back-
bone (Fig. 4). This molecular modeling study also
indicated that the benzene ring of 12 and 22, fused to
the cyclic amide skeleton, exhibits a p–p stacking inter-
action with the side-chain benzyl group of 208Phe of
HDAC8. The amino acids 151Gly and 208Phe are both
well conserved in HDACs. Therefore, compound 12 was
expected to exhibit potent HDAC-inhibitory activity for
all classes of HDAC.
3. Pharmacological results and discussion
X-ray crystallographic analysis of histone deacetylase-
like protein (HDLP)14 and HDAC 8,15 complexed with
trichostatin
A
(TSA, 3)16 and/or suberoylanilide
hydroxamic acid (SAHA, 4),17 has revealed that the
HDAC catalytic domain consists of a narrow tube-like
pocket spanning a length equivalent to a straight chain
of four to six carbons, with the zinc ion buried near
the bottom of the active site. Therefore, the structural
requirements for potent HDAC-inhibitory activity in-
volve three key regions, that is, (1) a zinc-binding motif,
which interacts with the active site zinc, (2) a linking do-
main, which occupies the channel, and (3) a surface rec-
ognition domain, which interacts with residues on the
rim of the active site. Previously we have reported novel
HDAC inhibitors based on a new structural scaffold.13
We used a phthalimide structure as a novel linker/cap
domain that might be suitable to occupy the narrow
channel of the HDAC active site. We expected that
the introduction of an aromatic ring might lead to favor-
able interaction with the side chains of aromatic amino
acids located at the active site. Therefore, we prepared
compounds of general formula (I) and found that the
lead compound (II) is a potent non-class-selective
HDAC inhibitor, as assessed with an HDAC inhibitor
assay kit. The potency of compound (II) was compara-
ble with that of SAHA, which is under phase III clinical
trial (Fig. 2).
We selected the new HDAC lead 12 for further study,
and performed chemical modifications, focusing on the
cyclic amide nitrogen substituents, in order to examine
the relationship of structure to HDAC class-selective
inhibition. The results are summarized in Tables 1 and
2. We selected HDAC 1, HDAC4, and HDAC6 as rep-
resentatives of class I, class IIa, and class IIb, respective-
ly, because sufficient amounts of these isozymes were
O
O
O
OH
OH
N
H
N
H
N
N
O
12 IC50 = 38 nM
22 IC50 = 105 nM
We next focused our attention on the phthalimide car-
bonyl groups of the lead compound (II). We considered
that reduction of one of the carbonyl groups to methy-
lene might afford a more potent HDAC inhibitor,
because most of the amino acids located around the
O
O
OH
N
N
H
O
(II) IC50 = 150 nM
O
O
O
O
O
O
O
H
OH
OH
OH
X
N
N
N
N
H
N
N
H
OH
N
H
H
R N
O
O
O
51 IC50 = 461 nM
57 IC50 = 416 nM
General formula (I)
Lead compound (II)
Figure 3. Chemical structures of our HDAC inhibitors with cyclic and/
or acyclic amide.
Figure 2. Chemical structures of our HDAC inhibitors.

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C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
Figure 4. Docking model structures of 12 and 22 into the HDAC8 binding pocket (see text).
Table 1. HDAC-inhibitory activity of the prepared compounds
O
O
OH
N
R
N
H
Compound
R
Inhibitory activity (EC50 (nM))
HDAC 4b
HDAC 1a
HDAC 6c
12
Benzyl
250 30
520 20
240 10
520 20
300 30
610 20
180 30
420 30
340 10
420 30
410 30
450 40
360 60
1480 590
280 30
250 20
640 20
350 30
250 40
300 80
350 40
290 30
500 10
420 50
590 70
750 110
490 20
990 20
30a
30b
30c
30d
30e
30f
30g
30h
30i
2-Phenylethyl
3-Phenylpropyl
Cyclohexylmethyl
1-Naphthylmethyl
(2-Cl)Benzyl
260
610 20
8
200
6
230
500
7
9
640 70
240 40
890 90
400 20
(3-Cl)Benzyl
(4-Cl)Benzyl
190 50
570 40
(2-CF3)Benzyl
(3-CF3)Benzyl
(4-CF3)Benzyl
(2-MeO)Benzyl
(3-MeO)Benzyl
(4-MeO)Benzyl
(2-Me)Benzyl
(3-Me)Benzyl
(4-Me)Benzyl
(2-Ph)Benzyl
(3-Ph)Benzyl
(4-Ph)Benzyl
(2-tBu)Benzyl
(4-tBu)Benzyl
390
4
170
7
230 20
230 30
590 20
160 70
99 11
30j
320 30
440 30
150 10
160 10
430 30
250 30
200 20
1310 220
330 50
220 30
480 60
230 20
30k
30l
30m
30n
30o
30p
30q
30r
30s
30t
30u
480 50
300 30
240 20
630 50
180 20
98
500 80
180
6
4
a–cAssays for inhibitory activities toward partially purified HDAC1, 4, and 6 were perfomed according to the reported methods.19,21
Table 2. HDAC-inhibitory activity of the prepared compounds
O
O
OH
N
N
H
R
Compound
R
Inhibitory activity (EC50 (nM))
HDAC 4b
HDAC 1a
HDAC 6c
30v
Phenyl
Benzyl
630 40
190 30
270 40
420 60
250 30
560 20
150 20
230 10
240 20
220 20
230 30
350 90
250 30
370 50
370 20
460 90
230 30
30w
30x
30y
30z
2-Phenylethyl
3-Phenylpropyl
(S)-Benzyloxymethyl
(R)-Benzyloxymethyl
30aa
250
5
a–cSee footnotes a–c in Table 1.

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7631
available for our purpose. We first examined the effect of
the distance between the distal benzene ring and amide
nitrogen atom (12, 30a, 30b, and 30d) and the impor-
tance of the aromatic ring (30c). Neither the methylene
chain length nor the presence of the aromatic ring was
critical, because all the compounds exhibited similar
inhibitory activity to each of the HDACs. Therefore,
we focused our attention on the effects of substituents
introduced on the distal benzene ring (Table 1). Broadly
speaking, a substituent at the meta position is preferable
for HDAC-inhibitory activity, whether it is electron-
withdrawing (30e–j) or electron-donating (30k–p). This
might mean that steric factors are more important than
electronic factors for the activity. In the cases of
HDAC1 and HDAC4, there seemed to be a tendency
for the introduction of an electron-donating group to
enhance the inhibitory activity. For HDAC6, the intro-
duction of a substituent at the ortho position tended to
increase the inhibitory activity, while the introduction
of a substituent at the para position tended to decrease
the activity. These results again suggested that steric fac-
tor(s) are important. We anticipated that the introduc-
tion of a bulky substituent at the ortho position might
afford HDAC6 selectivity, while the introduction of a
bulky substituents at the para position might afford
HDAC1/4 selectivity. In order to test this hypothesis,
we prepared phenyl-substituted and tert-butyl-substitut-
ed derivatives, and found that the 4-phenyl derivative
and the 4-tert-butyl derivative (30s, 30u) exhibited
apparent HDAC1/4 selectivity. However, the contribu-
tion of the bulkiness of the ortho-substituent was unclear
(30q, 30t).
used for further biological studies. Although our earlier
study indicated that the isoindolone skeleton is better
than the phthalimide skeleton for potent HDAC inhibi-
tors, there was little information on the HDAC class
selectivity of the phthalimide derivatives, and it is also
of interest to compare the class selectivity in these two
structural series, so we re-investigated the HDAC-inhib-
itory activity of the phthalimide derivatives (Table 3).
As a whole, these phthalimide derivatives tended to
show some preference for HDAC4. Compound 30h,
with an optically active, (S) form branched substituent,
exhibited the most HDAC4-selective inhibitory activity.
When we compared compounds in the isoindolone series
bearing the same substituents, the isoindolone and
phthalimide derivatives exhibited equipotent HDAC4-
inhibitory activity, while the phthalimide derivatives
exhibited somewhat lower inhibitory activities toward
HDAC1 and HDAC6, implying that the phthalimide
skeleton might impart a degree of HDAC4 selectivity.
Possibly the distal carbonyl group of phthalimide deriv-
atives, which could be positioned outside of the surface
recognition domain, has an unfavorable steric or elec-
tronic interaction with HDAC1 and HDAC6. Although
we have proposed that the cap structure is the primary
determinant for HDAC class selectivity, the above
results suggest that the linker structure, which connects
the cap structure and the zinc binding motif, is also
important for class selectivity. Therefore, we prepared
hydroxymethyl derivatives as another type of linker
(phenoxyacetamides), which was considered to be a bio-
isoster of a double bond (cinnamamides) (Table 4). It is
very interesting to note that although our lead
compound, the cinnamamide derivative 12, exhibited
non-class-selective pan-HDAC-inhibitory activity, the
phenoxyacetamide derivative 43a exhibited HDAC6-
selective inhibitory activity, that is, 43a exhibited equi-
potent HDAC6-inhibitory activity with 12, while its
HDAC1- and HDAC4-inhibitory activity was about
10-fold less. We speculated that the linker structure of
12 might interact more favorably with the narrow
tube-like pocket of both HDAC1 and HDAC4, as com-
pared with 43a. Although the HDAC class-inhibitory
In order to examine further the SAR for the para substi-
tuent, we prepared cyclic amide derivatives with
branched substituents at the amide nitrogen (Table 2),
but these compounds all exhibited non-class-selective
pan-HDAC inhibitory activity. This might indicate that
highly specific steric features are needed for HDAC class
selectivity. Among these compounds, however, the
N-(1,2-diphenylethyl) derivative (30w) was found to
exhibit potent pan-HDAC inhibitory activity, and was
Table 3. HDAC-inhibitory activity of the prepared compounds
O
O
R N
O
OH
N
H
Compound
R
Inhibitory activity (EC50 (nM))
HDAC 1a
HDAC 4b
HDAC 6c
36a
36b
36c
36d
36e
36f
36g
36h
36i
Phenyl
Benzyl
2310 280
530 20
900 60
360 20
440 30
N.T.d
1010 190
710 30
610 40
2-Phenylethyl
Diphenylmethyl
1130 220
N.T.d
N.T.d
2,2-Diphenylethyl
(S)-a-Methylbenzyl
(R)-a-Methylbenzyl
(S)-a-(Benzyloxymethyl)benzyl
(R)-a-(Benzyloxymethyl)benzyl
2640 160
1760 290
4040 580
1160 210
1510 90
910
2
1370 250
1080 100
2070 330
1230 250
1460 150
640 90
1300 100
270 30
530 80
a–cSee footnotes a–c in Table 1.
d N.T. means not tested.

7632
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
Table 4. HDAC-inhibitory activity of the prepared compounds
O
O
O
OH
N
H
N
R
Compound
R
Inhibitory activity (EC50 (nM)
HDAC 4b
HDAC 1a
N.T.
HDAC 6c
36f
36g
36h
36i
36j
Phenyl
(S)-Methyl
N.T.
N.T.
1760 290
4040 580
1160 210
1510 90
640 90
1300 10
270 30
530 80
1080 100
2070 330
1230 250
1460 150
(R)-Methyl
(S)-Benzyloxymethyl
(R)-Benzyloxymethyl
a–cSee footnotes a–c in Table 1.
profiles of 12 and 43a are quite different, it is interesting
that the influence of a substituent introduced on the ben-
zene ring of N-benzyl group appears to be the same.
That is, the introduction of a bulky phenyl group or
tert-butyl group at the ortho position (43b, 43d)
increased HDAC6-inhibitory activity. However, the
introduction of a phenyl group or tert-butyl group at
the para position (43c, 43e) specifically increased
HDAC1- and HDAC4-inhibitory activity, causing these
compounds to become pan-inhibitors of HDAC.
discover novel low-molecular-weight, class-selective
HDAC inhibitors.
As mentioned above, HDAC inhibitors have been
reported to upregulate the expression of p21WAF1/CIP1
and to downregulate cyclin D1 in many types of tumor
cells, in parallel with cell cycle arrest in the G1 phase.
Activation of the p21WAF1/CIP1 gene is associated with
the inhibition of proliferation and induction of differen-
tiation and/or apoptosis of tumor cells, in vitro and
in vivo.5,6 Therefore, we examined the ability of
representative compounds of the present series to upreg-
ulate the expression of p21WAF1/CIP1, by means of
reporter gene assay (Table 5).19 Among the basic struc-
tures (lead (II), 12, 22, and 43a), compound 12 was
found to exhibit the most potent activity. Although
the rank order of the activity was 12 > 22 > lead
Our SAR study clearly indicated that the structural fea-
tures of not only the cap part, but also the linker part of
HDAC inhibitors are critical for HDAC class-selective
inhibitory activity. Further chemical modification stud-
ies combined with X-ray crystallographic and/or molec-
ular modeling studies should make it possible to
Table 5. p21 promoter-activating activity and LNCaP cell growth-inhibitory activity of representative compounds
H
N
R2
X
A
B
OH
N
O
R1
Compound
R1
R2
A
B
X
p21 promoter assay LNCaP growth
inhibition
EC1000 (nM)a
IC50 (nM)b
Lead (II)
12
22
H
H
CO
CO
CH2
CO
CO
CO
CO
CO
CO
CO
CO
CO
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
O–CH2
3290 640
230 80
1460 310
870 260
180 110
140 50
130 60
130 10
96 21
40% (@10 lM)
H
H
CH2
CO
100
690
886
101
108
131
202
75
H
H
30q
30s
2-Phenyl
4-Phenyl
3-MeO
4-MeO
3-CF3
H
H
CH2
CH2
CH2
CH2
CH2
CH2
CO
H
30l
H
30m
30i
H
H
Benzyl
30w
36k
H
(S)-Benzyloxymethyl
H
1730 920
8160 880
N.T.c
570
7000
161
17
43a
SAHA
TSA
H
CH2
12
5
a Assay for P21 promoter activation was performed according to the reported methods.19,21
b Exponentially growing cells in RPMI1640 medium supplemented with 10% fetal bovine serum were adjusted to 2 · 104 cells/mL and 100 lL
aliquots were plated in 96-well plates and incubated for 24 h at 37 ꢁC under an atmosphere of 5% CO2 in air. After incubation, various
concentrations of test compounds were added and incubation was continued for a further 4 days. Viable cells were counted with a cell counting kit
(Dojindo). n = 3.
c N.T. means not tested.

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7633
0.2
0
0.2
0
y = 0.981x - 0.1711 n = 29 r = 0.613
0.2
0
y= 0.5349x - 0.1718 n = 29 r = 0.831
y = 0.8247x - 0.1182 n = 29 r = 0.593
-1.2
-1
-0.8
-0.6
-0.4
-0.2
0
0.2
-1.2
-1
-0.8
-0.6
-0.4
-0.2
0
0.2
-1.2
-1
-0.8
-0.6
-0.4
-0.2
0
0.2
-0.2
-0.4
-0.6
-0.8
-1
-0.2
-0.4
-0.6
-0.8
-1
-0.2
-0.4
-0.6
-0.8
-1
-1.2
-1.2
-1.2
HDAC1 log IC
50
HDAC4 log IC
HDAC6 log IC
50
50
Figure 5. Correlation between LNCaP cell growth-inhibitory activity and HDAC-inhibitory activity (left, HDAC1; middle, HDAC4; right,
HDAC6). Horizontal bars indicate the IC50 values (log(lM)) for LNCaP cell growth inhibition. Vertical bars indicate the IC50 values (log(lM)) for
inhibition of representatives of each HDAC class.
(II) > 43a, all the compounds exhibited micromolar to
sub-micromolar values of EC1000 for p21 expression
upregulation activity. The reason for this is not known
at present, but differences in the ability of the com-
pounds to penetrate the cell membrane might be in-
volved, at least to some extent. Among the compounds
tested, we found that 30w exhibited the most potent
activity, being comparable in potency to the natural
product TSA. These results indicated that compounds
of the present series exhibit significant HDAC-inhibito-
ry activity not only in vitro, but also at the cellular level.
Therefore, we further examined their inhibitory activity
on the growth of the human prostate cancer cell line
LNCaP. These results are also summarized in Table 5.
The rank order of effect was similar to that noted above,
and compound 12 was the most potent among the lead
(II), 12, 22, and 43a. Compound 30w was the most po-
tent in the present series of compounds. Its LNCaP
growth-inhibitory activity was more potent than that
of SAHA, which is currently under phase III clinical
evaluation, and comparable with that of TSA. As can
be seen in Table 5, compound 30s, which exhibited
HDAC1/4-selective inhibitory activity, showed potent
LNCaP cell growth-inhibitory activity, while com-
pounds 36k (HDAC4-selective), 30q, and 43a (both
HDAC6-selective) showed weak LNCaP cell growth-
inhibitory activity. Since inhibitory activity toward
LNCaP cell growth might be associated with inhibition
of a certain class of HDAC, possibly class I, we investi-
gated the correlation between the LNCaP cell growth-
inhibitory activity and the HDAC class selectivity of
our compounds by means of multivariate analysis.
(Fig. 5). That is, the LNCaP cell growth inhibition
was most highly correlated with HDAC1 inhibition.
Furthermore, we found a strong correlation between
LNCaP cell growth inhibition and p21 promoter activity
(correlation coefficient r = 0.752). We therefore
speculated that the mechanism of LNCaP cell growth
inhibition by our compounds may involve HDAC1
class-selective inhibition, and subsequent transcriptional
augmentation of p21 expression, at least in part. To seek
support for this idea, we examined the dose dependency
and the time courses of both the LNCaP cell growth
inhibition and the degree of expression increase of
p21, using 30w (Figs. 6 and 7). The p21 gene expression
Contl.
0.1 μM
1 μM
5 μM
160000
120000
80000
40000
0
0
24
48
72
Time (h)
30w (1μM)
8 24 48
DMSO
24
(hr)
0
8
48
p21
β-actin
The values of the correlation coefficients between the
LNCaP cell growth-inhibitory activity and the inhibito-
ry activities toward HDAC1, HDAC4, and HDAC6
were r = 0.831, r = 0.593, and r = 0.613, respectively
Figure 7. Time courses of LNCaP cell growth inhibition and
augmentation of p21 message expression by 30w.
a
24 hr
0.5
b
0
1
100
30 w (μM)
75
50
25
0
p21
β-actin
Contl
0.05
0.1
0.5
1
Conc. of 30w
Figure 6. Dose dependency of LNCaP cell growth inhibition (a) and augmentation of p21 message expression (b) by 30w.

7634
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
of LNCaP cells was augmented after exposure of the
cells to compound 30w for 8 h, and reached the maxi-
mum after 24 h. Inhibition of LNCaP cell growth was
also observed after treatment with 30w for 24 h. This
evidence strongly suggests that the enhancement of
p21 expression is involved in the initial inhibition of
LNCaP cell growth by 30w.
anhydrous MgSO4, and evaporated. The residue was
purified by silica gel column chromatography (eluant
CHCl3) to afford 145 mg (54%) of the title compound
1
as an oil. H NMR (500 MHz, CDCl3) d 7.83 (s, 1H),
7.48 (d, 1H, J = 7.7 Hz), 7.27 (m, 6H), 4.74 (s, 4H),
4.18 (s, 2H); FAB MS m/z 254 (M+H)+.
5.1.3. 2-Benzyl-3-oxoisoindoline-5-carbaldehyde (8). A
mixture of 7 (191 mg, 0.75 mmol), PCC (230 mg,
1.07 mmol), and 10 mL of CH2Cl2 was stirred for 1 h
at room temperature under Ar. The reaction mixture
was filtered through Celite, and the filtrate was washed
with brine, dried over anhydrous MgSO4, and evaporat-
ed. The residue was purified by silica gel column chro-
matography (eluant n-hexane/AcOEt 1:1 v/v) to afford
142 mg (76%) of the title compound as an oil. 1H
NMR (500 MHz, CDCl3) d 10.05 (s, 1H), 8.31 (s, 1H),
8.02 (d, 1H, J = 7.9 Hz), 7.49 (d, 1H, J = 7.9 Hz), 7.27
(m, 5H), 4.77 (s, 2H), 4.31 (s, 2H); FAB MS m/z 252
(M+H)+.
4. Conclusion
We have designed and synthesized various structural
types of cyclic amide/imide HDAC inhibitors, which
show characteristic HDAC class-selective inhibition.
SAR study indicated that both the cap structure and
the linker structure of the present series of compounds
are critical for HDAC class-selective inhibition. Further
chemical modification studies based on our present SAR
should provide even more potent and more class-selec-
tive HDAC inhibitors. Representative compounds of
the present series are potent HDAC inhibitors, effective
at the cellular level, so these compounds should be use-
ful not only as selective tools to investigate the func-
tion(s) of each HDAC class, but also as lead (or
candidate) compounds for the treatment of HDAC-
related diseases, which include cancer, cranial nerve dis-
ease, immune disorders, and so on.
5.1.4. (E)-Methyl 3-(2-benzyl-1-oxoisoindolin-6-yl)acry-
late (9). A mixture of methyl diethylphosphonoacetate
(0.11 mL, 0.61 mmol), t-BuOK (68 mg, 0.61 mmol),
and 10 mL of anhydrous THF was stirred for 10 min
at room temperature, then 8 (140 mg, 0.56 mmol) in
3 mL of anhydrous THF was added, and stirring was
continued for a further 4 h at room temperature. The
reaction mixture was washed with brine, dried over
anhydrous MgSO4, and evaporated. The residue was
purified by silica gel column chromatography (eluant
n-hexane/AcOEt 1:1 v/v) to afford 54 mg (31%) of the
5. Experimental
5.1. General
1
title compound as an oil. H NMR (500 MHz, CDCl3)
Melting points were determined by using a Yanagimoto
hot-stage melting point apparatus and are uncorrected.
NMR spectra were recorded on a JEOL JNM-GX500
(500 MHz) spectrometer. Chemical shifts are expressed
in ppm relative to tetramethylsilane. Mass spectra were
recorded on a JEOL JMS-DX303 spectrometer.
d 8.05 (d, 1H, J = 1.2 Hz), 7.74 (d, 1H, J = 6.1 Hz),
7.64 (dd, 1H, J = 1.2, 7.9 Hz), 7.39 (d, 1H, J = 7.9 Hz),
7.30 (m, 5H), 6.52 (d, 1H, J = 6.1 Hz), 4.80 (s, 2H),
4.28 (s, 2H), 3.81 (s, 3H); FAB MS m/z 308 (M+H)+.
5.1.5. (E)-3-(2-Benzyl-1-oxoisoindolin-6-yl)acrylic acid
(10). A mixture of 9 (160 mg, 0.52 mmol), 8 mL of gla-
cial acetic acid, and 8 mL of 0.5 mol/L of HCl was stir-
red overnight at 85 ꢁC. The mixture was extracted with
AcOEt, and the extract was washed with water and
brine, dried over anhydrous MgSO4, and evaporated
to afford 149 mg (98%) of the title compound as a color-
5.1.1. 2-Benzyl-3-oxoisoindoline-5-carboxylic acid (6).
Compound 5 (461 mg) and Sn (523 mg, 4.41 mmol) were
suspended in a mixture of 5 mL of concentrated (concd)
HCl and 5 mL of glacial acetic acid and stirred for 9 h at
room temperature. The reaction mixture was filtered
through Celite and extracted with AcOEt. The extract
was washed with water and brine, dried over anhydrous
MgSO4, and evaporated. The residue was purified by sil-
ica gel column chromatography (eluant CHCl3) to
afford 282 mg (66%) of the title compound as a yellow
oil. 1H NMR (500 MHz, CDCl3) d 8.68 (d, 1H,
J = 1.5 Hz), 8.28 (dd, 1H, J = 1.5, 8.0 Hz,), 7.50 (d,
1H, J = 8.0 Hz), 7.33 (m, 5H), 4.84 (s, 2H), 4.35 (s,
2H); FAB MS m/z 268 (M+H)+.
1
less solid. H NMR (500 MHz, CD3OD) d 7.99 (d, 1H,
J = 1.5 Hz), 7.80 (dd, 1H, J = 1.5, 7.9 Hz), 7.75 (d, 1H,
J = 6.1 Hz), 7.53 (d, 1H, J = 7.9 Hz,), 7.31 (m, 5H),
6.58 (d, 1H, J = 6.1 Hz), 4.80 (s, 2H), 4.38 (s, 2H);
FAB MS m/z 294 (M+H)+.
5.1.6. (E)-3-(2-Benzyl-1-oxoisoindolin-6-yl)-N-(tetrahy-
dro-2H-pyran-2-yloxy)acrylamide (11). To a mixture of
10 (60 mg, 0.27 mmol), triethylamine (0.086 mL,
0.819 mmol) and 5 mL of anhydrous THF was added
ethyl chloroformate (0.029 mL, 0.41 mmol) at 0 ꢁC,
and the whole was stirred for 1 h. Then, O-(tetrahy-
dro-2H-pyran-2-yl)hydroxylamine (53 mg, 0.45 mmol)
was added and stirring was continued for 4 h at room
temperature. The reaction mixture was washed with satd
NaHCO3, dil HCl, and brine, dried over anhydrous
MgSO4, and evaporated. The residue was purified by
silica gel column chromatography (eluant n-hexane/
5.1.2. 2-Benzyl-6-hydroxymethyl-isoindoline-1-one (7).
To a mixture of 6 (282 mg, 1.06 mmol) and 5 mL of
anhydrous THF was added dropwise 2.35 mL of
1 mol/L BH3–THF complex, and the whole was stirred
for 1 day at room temperature. To the mixture were
added 3 mL of water and 870 mg of K2CO3, and the
mixture was washed with saturated (satd) NaHCO3
solution, dilute (dil) HCl, and brine, then dried over

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7635
AcOEt 1:3 v/v) to afford 52 mg (49%) of the title com-
pound as an oil. H NMR (500 MHz, CDCl3) d 7.98
1H), 8.04 (d, 1H, J = 8.2 Hz), 7.32 (m, 5H), 4.83 (s,
2H), 4.38 (s, 2H); FAB MS m/z 269 (M+H)+.
1
(s, 1H), 7.72 (d, 1H, J = 5.5 Hz), 7.71 (d, 1H,
J = 7.9 Hz), 7.53 (d, 1H, J = 7.9 Hz), 7.28 (m, 5H),
6.52 (d, 1H, J = 5.5 Hz), 5.05 (m, 1H), 4.77 (s, 2H),
4.24 (s, 2H), 3.99 (m, 1H), 3.61 (m, 1H), 1.81 (m, 3H),
1.55 (m, 3H); FAB MS m/z 393 (M+H)+.
5.1.11. 5-Amino-2-benzylisoindolin-1-one (17). A mixture
of 16 (180 mg, 0.67 mmol), 10% Pd–C (18 mg), and
10 mL AcOEt was hydrogenated at room temperature
for 6 h. The catalyst was removed by filtration through
Celite and the filtrate was evaporated to afford 131 mg
(82%) of the title compound. 1H NMR (500 MHz,
CDCl3) d 7.63 (d, 1H, J = 8.0 Hz), 7.26 (m, 5H), 6.67
(dd, 1H, J = 1.5, 8.0 Hz), 6.56 (d, 1H, J = 1.5 Hz), 4.72
(s, 2H), 4.11 (s, 2H), 3.73 (s, 2H); FAB MS m/z 239
(M+H)+.
5.1.7. (E)-3-(2-Benzyl-1-oxoisoindolin-6-yl)-N-hydroxy-
acrylamide (12). A mixture of 11 (26 mg, 0.066 mmol),
d-10-camphorsulfonic acid (18 mg, 0.073 mmol), and
5 mL of methanol was stirred for 1 h at room tempera-
ture under Ar. After evaporation of the solvent, the res-
idue was redissolved in AcOEt, then the solution was
washed with satd NaHCO3 and brine, dried over anhy-
drous MgSO4, and evaporated. The residue was recrys-
tallized from a mixed solvent of acetone, ethanol, and
AcOEt to afford 1.6 mg (7.9%) of the title compound.
Mp 172–174 ꢁC; 1H NMR (500 MHz, DMSO-d6) d
10.77 (s, 1H), 9.09 (s, 1H), 7.86 (s, 1H), 7.76 (d, 1H,
J = 7.3 Hz), 7.57 (d, 1H, J = 7.3 Hz), 7.53 (d, 1H,
J = 5.8 Hz), 7.35 (m, 2H), 7.28 (m, 3H), 6.56 (d, 1H,
J = 5.8 Hz), 4.72 (s, 2H), 4.38 (s, 2H); HR FAB MS:
(M+H)+ calcd for C18H17N2O3, 309.1239. Found:
309.1259.
5.1.12.
Methyl
3-(2-benzyl-1-oxoisoindolin-5-yl)-2-
bromopropanoate (18). A mixture of 17 (270 mg,
1.13 mmol), 3 mL of 47% HBr, 6 mL of methanol, and
15 mL of acetone was cooled to below 5 ꢁC, and sodium
nitrite solution (94 mg/3 mL) was added dropwise. After
30 min, methyl acrylate (1.01 ml, 11.3 mmol) was added
and the whole was heated rapidly to 40 ꢁC. Cu2O
(10 mg, 0.068 mmol) was added portionwise and the
whole was stirred for 7 h at 35–40 ꢁC. The reaction mix-
ture was evaporated and the residue was redissolved in
AcOEt, washed with dil NaOH, dil HCl and brine, dried
over anhydrous MgSO4, and evaporated. The residue
was purified by silica gel column chromatography
(eluant n-hexane/AcOEt 6:1 to 4:1 v/v) to afford 90 mg
(21%) of the title compound. 1H NMR (500 MHz,
CDCl3) d 7.79 (d, 1H, J = 7.9 Hz), 7.28 (m, 6H), 7.20
(s, 1H), 4.74 (s, 2H), 4.37 (dd, 1H, J = 7.3 Hz, 8.0 Hz),
4.20 (s, 2H), 3.68 (s, 3H), 3.49 (dd, 1H, J = 7.3 Hz,
4.3 Hz), 3.27 (dd, 1H, J = 7.3 Hz, 4.3 Hz); FAB MS
m/z 432, 434 (M+H)+.
5.1.8. Methyl 2-methyl-4-nitrobenzoate (14). A mixture
of 2-methyl-4-nitrobenzoic acid (1.00 g, 5.5 mmol),
iodomethane (863 mg, 6.08 mmol), K2CO3 (1.14 g,
8.28 mmol), and 10 mL of DMF was stirred for 2 h at
room temperature, then poured into water and extracted
with AcOEt. The extract was washed with water and
brine, dried over anhydrous MgSO4, and evaporated
to afford 1.05 g (98%) of the title compound as an oil.
1H NMR (500 MHz, CDCl3) d 8.11 (s, 1H), 8.07 (d,
1H, J = 8.6 Hz), 8.03 (d, 1H, J = 8.6 Hz), 3.95 (s, 3H),
2.69 (s, 3H); FAB MS m/z 196 (M+H)+.
5.1.13. (E)-Methyl 3-(2-benzyl-1-oxoisoindolin-5-yl)
acrylate (19). A mixture of 18 (90 mg, 0.23 mmol),
DBU (0.042 mL, 0.28 mmol), and 10 mL of toluene
was refluxed for 5 h. The reaction mixture was evaporat-
ed and the residue was redissolved in AcOEt. This solu-
tion was washed with dil HCl and brine, dried over
anhydrous MgSO4, and evaporated. The residue was
purified by silica gel column chromatography (eluant
n-hexane/AcOEt 1.5:1 v/v) to afford 68 mg (96%) of
the title compound. 1H NMR (500 MHz, CDCl3) d
7.82 (d, 1H, J = 7.5 Hz), 7.68 (d, 1H, J = 6.0 Hz), 7.58
(d, 1H, J = 7.5 Hz), 7.46 (s, 1H), 7.26 (m, 5H), 6.45 (d,
1H, J = 6.0 Hz), 4.76 (s, 2H), 4.24 (s, 2H), 3.77 (s,
3H); FAB MS m/z 308 (M+H)+.
5.1.9. Methyl 2-bromomethyl-4-nitrobenzoate (15). A
mixture of 14 (985 mg, 5.05 mmol), NBS (989 mg,
5.56 mmol), benzoyl peroxide (15 mg, 0.06 mmol), and
20 mL of carbon tetrachloride was heated at 85 ꢁC for
8 h. Further NBS (92 mg, 0.52 mmol) was added and
the whole was refluxed for 1 h. The mixture was washed
with satd NaHCO3 and brine, dried over anhydrous
MgSO4, and evaporated. The residue was purified by sil-
ica gel column chromatography (eluant n-hexane/AcOEt
3:1 v/v) to afford 1.35 g (98%) of the title compound as
an oil. 1H NMR (500 MHz, CDCl3) d 8.30 (d, 1H,
J = 2.1 Hz), 8.20 (dd, 1H, J = 2.1, 6.2 Hz), 8.10 (d, 1H,
J = 6.2 Hz), 4.96 (s, 2H), 4.00 (s, 3H); FAB MS m/z
273, 275 (M+H)+.
5.1.14. (E)-3-(2-Benzyl-1-oxoisoindolin-5-yl)acrylic acid
(20). This compound was prepared from 19 by means of
a procedure similar to that used for 10 (97%). 1H NMR
(500 MHz, CD3OD) dd 7.82 (d, 1H, J = 8.0 Hz), 7.75 (s,
1H), 7.74 (d, 1H, J = 8.0 Hz), 7.73 (d, 1H, J = 6.5 Hz),
7.31 (m, 5H), 6.58 (d, 1H, J = 6.5 Hz), 4.81 (s, 2H),
4.40 (s, 2H); FAB MS m/z 294 (M+H)+.
5.1.10. 2-Benzyl-5-nitroisoindolin-1-one (16). A mixture
of 15 (300 mg, 1.10 mmol), benzylamine (0.13 mL,
1.21 mmol), triethylamine (0.17 mL, 1.21 mmol), and
10 mL of methanol was refluxed for 24 h. The mixture
was diluted with AcOEt, washed with dil HCl and brine,
dried over anhydrous MgSO4, and evaporated. The res-
idue was purified by silica gel column chromatography
(eluant n-hexane/AcOEt 3:1 to 1:1 v/v) to afford
255 mg (86%) of the title compound. 1H NMR
(500 MHz, CDCl3) d 8.35 (d, 1H, J = 8.2 Hz), 8.25 (s,
5.1.15. (E)-3-(2-Benzyl-1-oxoisoindolin-5-yl)-N-(tetrahy-
dro-2H-pyran-2-yloxy)acrylamide (21). This compound
was prepared from 20 by means of a procedure similar
1
to that used for 11 (52%). H NMR (500 MHz, CDCl3)
d 9.18 (s, 1H), 7.81 (d, 1H, J = 7.9 Hz), 7.72 (d, 1H,

7636
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
J = 5.5 Hz), 7.55 (d, 1H, J = 7.9 Hz), 7.41 (s, 1H), 7.26
(m, 5H), 6.46 (d, 1H, J = 5.5 Hz), 5.00 (m, 1H), 4.76
(s, 2H), 4.22 (s, 2H), 3.95 (m, 1H), 3.61 (m, 1H), 1.80
(m, 3H), 1.56 (m, 3H); FAB MS m/z 393 (M+H)+.
5.1.22. (E)-Methyl 5-(2-(tert-butoxycarbonyl)vinyl)-2-
bromomethylbenzoate (26b; R3 = tert-butyl). This com-
pound was prepared from 25b by means of a procedure
1
similar to that used for 15 (36%). H NMR (500 MHz,
CDCl3) d 8.10 (d, 1H, J = 1.8 Hz), 7.60 (dd, 1H,
J = 1.8, 8.3 Hz), 7.56 (d, 1H, J = 6.2 Hz), 7.47 (d, 1H,
J = 8.3 Hz), 6.42 (d, 1H, J = 6.2 Hz), 4.95 (s, 2H), 3.96
(s, 3H), 1.54 (s, 9H); FAB MS m/z 355, 357 (M+H)+.
5.1.16. (E)-3-(2-Benzyl-1-oxoisoindolin-5-yl)-N-hydroxy-
acrylamide (22). This compound was prepared from 21
by means of a procedure similar to that used for 12
(68%). Mp 186–188 ꢁC; H NMR (500 MHz, DMSO-
d6) d 11.33 (s, 1H), 9.08 (s, 1H), 7.73 (d, 1H,
J = 8.0 Hz), 7.72 (s, 1H), 7.67 (d, 1H, J = 8.0 Hz), 7.52
(d, 1H, J = 5.8 Hz), 7.35 (m, 2H), 7.28 (m, 3H), 6.54
(d, 1H, J = 5.8 Hz), 4.72 (s, 2H), 4.38 (s, 2H); HR
FAB MS: (M+H)+ calcd for C18H17N2O3, 309.1239.
Found: 309.1259.
1
5.1.23. (E)-Methyl 3-(1-oxo-2-phenethylisoindolin-6-
yl)acrylate (27a). This compound was prepared from
26a by means of a procedure similar to that used for
1
16 (11%). H NMR (500 MHz, CDCl3) d 8.03 (d, 1H,
J = 1.2 Hz), 7.76 (d, 1H, J = 5.8 Hz), 7.65 (dd, 1H,
J = 1.2, 7.6 Hz), 7.40 (d, 1H, J = 7.6 Hz), 7.27 (m, 5H),
6.53 (d, 1H, J = 5.8 Hz), 4.23 (s, 2H), 3.90 (t, 1H,
J = 7.3 Hz), 3.84 (s, 3H), 3.03 (t, 2H, J = 7.3 Hz); FAB
MS m/z 322 (M+H)+.
5.1.17. Methyl 5-(2-(methoxycarbonyl)-2-bromoethyl)-2-
methylbenzoate (24a; R3 = Me). This compound was
prepared from 23 by means of a procedure similar to
1
that used for 18 (40%). H NMR (500 MHz, CDCl3) d
5.1.24. (E)-Methyl 3-(1-oxo-2-(3-phenylpropyl)isoindolin-
6-yl)acrylate (27b). This compound was prepared from
26a by means of a procedure similar to that used for
7.76 (s, 1H), 7.25 (d, 1H, J = 8.0 Hz), 7.19 (d, 1H,
J = 8.0 Hz), 4.39 (dd, 1H, J = 7.3, 8.0 Hz), 3.89 (s,
3H), 3.74 (s, 3 H), 3.45 (dd, 1H, J = 8.0, 14.0 Hz), 3.23
(dd, 1H, J = 7.3, 14.0 Hz), 2.57 (s, 3H); FAB MS m/z
315, 317 (M+H)+.
1
16 (85%). H NMR (500 MHz, CDCl3) d 8.00 (d, 1H,
J = 1.5 Hz), 7.74 (d, 1H, J = 5.9 Hz), 7.64 (dd, 1H,
J = 1.5, 7.9 Hz), 7.43 (d, 1H, J = 7.9 Hz), 7.26 (m, 2H),
7.18 (m, 3 H), 6.52 (d, 1H, J = 5.9 Hz), 4.36 (s, 2H),
3.81 (s, 3H), 3.67 (t, 2H, J = 7.3 Hz), 2.69 (t, 2H,
J = 7.8 Hz), 1.99 (m, 2H); FAB MS m/z 336 (M+H)+.
5.1.18. Methyl 5-(2-(tert-butoxycarbonyl)-2-bromoethyl)-
2-methylbenzoate (24b; R3 = tert-butyl). This compound
was prepared from 23 and tert-butyl acrylate by means
1
of a procedure similar to that used for 18 (35%). H
5.1.25. (E)-tert-Butyl 3-(2-(cyclohexylmethyl)-1-oxoisoin-
dolin-6-yl)acrylate (27c). This compound was prepared
from 26b by means of a procedure similar to that used
NMR (500 MHz, CDCl3) d 7.84 (d, 1H, J = 1.5 Hz),
7.33 (dd, 1H, J = 1.5, 7.6 Hz), 7.25 (d, 1H, J = 7.6 Hz),
4.35 (dd, 1H, J = 7.0, 8.2 Hz), 3.48 (dd, 1H, J = 8.2,
14.3 Hz), 3.25 (dd, 1H, J = 7.0, 14.3 Hz), 3.95 (s, 3H),
2.64 (s, 3H), 1.49 (s, 9H); FAB MS m/z 357, 359 (M+H)+.
1
for 16 (66%). H NMR (500 MHz, CDCl3) d 7.98 (d,
1H, J = 1.5 Hz), 7.65 (d, 1H, J = 6.2), 7.64 (dd, 1H,
J = 1.5, 7.9 Hz), 7.43 (d, 1H, J = 7.9 Hz), 6.45 (d, 1H,
J = 6.2), 4.39 (s, 2H), 3.45 (d, J = 7.4 Hz, 2H), 1.71
(m, 5H), 1.54 (s, 9H), 1.20 (m, 6H); FAB MS m/z 356
(M+H)+.
5.1.19. (E)-Methyl 5-(2-(methoxycarbonyl)vinyl)-2-meth-
ylbenzoate (25a; R3 = Me). This compound was pre-
pared from 24a by means of a procedure similar to
1
that used for 19 (quant.). H NMR (500 MHz, CDCl3)
5.1.26. (E)-tert-butyl 3-(2-((naphthalene-1-yl)methyl)-1-
oxoisoindolin-6-yl)acrylate (27d). This compound was
prepared from 26b by means of a procedure similar to
d 8.06 (d, 1H, J = 1.8 Hz), 7.67 (d, 1H, J = 5.9 Hz),
7.53 (dd, 1H, J = 1.8, 8.0 Hz), 7.26 (d, 1H, J = 8.0 Hz),
6.45 (d, 1H, J = 5.9 Hz), 3.91 (s, 3H), 3.80 (s, 3H),
2.61 (s, 3H); FAB MS m/z 235 (M+H)+.
1
that used for 16 (66%). H NMR (500 MHz, CDCl3) d
8.22 (d, 1H, J = 7.7 Hz), 8.05 (s, 1H), 7.87 (d, 1H,
J = 9.2 Hz), 7.85 (d, 1H, J = 8.8 Hz), 7.61 (d, 1H,
J = 5.8 Hz), 7.59 (d, 1H, J = 7.9 Hz), 7.50 (m, 4H),
7.29 (d, 1H, J = 7.9 Hz), 6.45 (d, 1H, J = 5.8 Hz), 5.26
(s, 2H), 4.15 (s, 2H), 1.54 (s, 9H); FAB MS m/z 400
(M+H)+.
5.1.20. Methyl 5-(2-(tert-butoxycarbonyl)vinyl)-2-meth-
ylbenzoate (25b; R3 = tert-butyl). This compound was
prepared from 24b by means of a procedure similar to
1
that used for 19 (50%). H NMR (500 MHz, CDCl3) d
7.84 (d, 1H, J = 1.5 Hz), 7.33 (dd, 1H, J = 1.5, 7.6 Hz),
7.25 (d, 1H, J = 7.6 Hz), 4.35 (dd, 1H, J = 7.0, 8.2 Hz),
3.48 (dd, 1H, J = 8.2, 14.3 Hz), 3.25 (dd, 1H, J = 7.0,
14.3 Hz), 3.95 (s, 3H), 2.64 (s, 3H), 1.49 (s, 9H); FAB
MS m/z 357, 359 (M+H)+.
5.1.27. (E)-Methyl 3-(2-(2-chlorobenzyl)-1-oxoisoindolin-
6-yl)acrylate (27e). This compound was prepared from
26a by means of a procedure similar to that used for
1
16 (70%). H NMR (500 MHz, CDCl3) d 8.05 (s, 1H),
7.75 (d, 1H, J = 5.9 Hz), 7.66 (d, 1H, J = 7.6 Hz), 7.42
(d, 1H, J = 7.6 Hz), 7.40 (m, 1H), 7.34 (m, 1H), 7.24
(m, 2H), 6.52 (d, 1H, J = 5.9 Hz), 4.96 (s, 2H), 4.36 (s,
2H), 3.82 (s, 3H); FAB MS m/z 342 (M+H)+.
5.1.21. (E)-Methyl 5-(2-(methoxycarbonyl)vinyl)-2-bro-
momethylbenzoate (26a; R3 = Me). This compound was
prepared from 25a by means of a procedure similar to
1
that used for 15 (49%). H NMR (500 MHz, CDCl3) d
8.08 (d, 1H, J = 1.8 Hz), 7.63 (d, 1H, J = 5.9 Hz), 7.59
(dd, 1H, J = 1.8, 8.0 Hz), 7.45 (d, 1H, J = 8.0 Hz), 6.47
(d, 1H, J = 5.9 Hz), 4.92 (s, 2H), 3.94 (s, 3H), 3.79 (s,
3H); FAB MS m/z 313, 315 (M+H)+.
5.1.28. (E)-Methyl 3-(2-(3-chlorobenzyl)-1-oxoisoindolin-
6-yl)acrylate (27f). This compound was prepared from
26a by means of a procedure similar to that used for
1
16 (71%). H NMR (500 MHz, CDCl3) d 8.05 (d, 1H,

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7637
J = 1.5 Hz), 7.75 (d, 1H, J = 6.1 Hz), 7.65 (dd, 1H,
J = 1.5, 7.9 Hz), 7.42 (d, 1H, J = 7.9 Hz), 7.27 (m, 3H),
7.19 (m, 1H), 6.53 (d, 1H, J = 6.1 Hz), 4.77 (s, 2H),
4.30 (s, 2H), 3.82 (s, 3H); FAB MS m/z 342 (M+H)+.
J = 8.3 Hz), 6.50 (d, 1H, J = 5.9 Hz), 4.83 (s, 2H), 4.32
(s, 2H), 3.86 (s, 3H), 3.81 (s, 3H); FAB MS m/z 338
(M+H)+.
5.1.35. (E)-Methyl 3-(2-(4-methoxybenzyl)-1-oxoisoindo-
lin-6-yl)acrylate (27m). This compound was prepared
from 26a by means of a procedure similar to that used
5.1.29. (E)-Methyl 3-(2-(4-chlorobenzyl)-1-oxoisoindolin-
6-yl)acrylate (27g). This compound was prepared from
26a by means of a procedure similar to that used for
1
for 16 (70%). H NMR (500 MHz, CDCl3) d 8.05 (d,
1
16 (87%). H NMR (500 MHz, CDCl3) d 8.05 (d, 1H,
1H, J = 1.3 Hz), 7.74 (d, 1H, J = 5.8 Hz), 7.64 (dd, 1H,
J = 1.3, 7.9 Hz), 7.40 (d, 1H, J = 7.9 Hz), 7.23 (d, 2H,
J = 8.5 Hz), 6.86 (d, 2H, J = 8.5 Hz), 6.53 (d, 1H,
J = 5.8 Hz), 4.74 (s, 2H), 4.26 (s, 2H), 3.82 (s, 3H),
3.79 (s, 3H); FAB MS m/z 338 (M+H)+.
J = 1.2 Hz), 7.75 (d, 1H, J = 5.9 Hz), 7.66 (dd, 1H,
J = 1.2, 8.0 Hz), 7.41 (d, 1H, J = 8.0 Hz), 7.31 (d, 2H,
J = 8.3 Hz), 7.24 (d, 2H, J = 8.3 Hz), 6.53 (d, 1H,
J = 5.9 Hz), 4.78 (s, 2H), 4.28 (s, 2H), 3.82 (s, 3H);
FAB MS m/z 342 (M+H)+.
5.1.36. (E)-tert-Butyl 3-(2-(2-methylbenzyl)-1-oxoisoin-
dolin-6-yl)acrylate (27n). This compound was prepared
from 26b by means of a procedure similar to that used
5.1.30. (E)-tert-Butyl 3-(2-(2-(trifluoromethyl)benzyl)-1-
oxoisoindolin-6-yl)acrylate (27h). This compound was
prepared from 26b by means of a procedure similar to
1
for 16 (43%). H NMR (500 MHz, CDCl3) d 8.03 (d,
1
that used for 16 (69%). H NMR (500 MHz, CDCl3) d
1H, J = 1.2 Hz), 7.65 (d, 1H, J = 6.1 Hz), 7.64 (dd, 1H,
J = 1.2, 7.8 Hz), 7.37 (d, 1H, J = 7.8 Hz), 7.20 (m, 4H),
6.46 (d, 1H, J = 6.1 Hz), 4.83 (s, 2H), 4.20 (s, 2H),
2.34 (s, 3H), 1.54 (s, 9H); FAB MS m/z 364 (M+H)+.
8.04 (s, 1H), 7.67 (d, 1H, J = 7.6 Hz), 7.65 (d, 1H,
J = 7.9 Hz), 7.64 (d, 1H, J = 6.2 Hz), 7.49 (dd, 1H,
J = 7.6, 7.6 Hz), 7.42 (d, 1H, J = 7.9 Hz), 7.40 (d, 1H,
J = 7.6 Hz), 7.37 (dd, 1H, J = 7.6, 7.6 Hz), 6.46 (d, 1H,
J = 6.2 Hz), 5.00 (s, 2H), 4.29 (s, 2H), 1.54 (s, 9H);
FAB MS m/z 418 (M+H)+.
5.1.37. (E)-tert-Butyl 3-(2-(3-methylbenzyl)-1-oxoisoin-
dolin-6-yl)acrylate (27o). This compound was prepared
from 26b by means of a procedure similar to that used
1
5.1.31. (E)-tert-Butyl 3-(2-(3-(trifluoromethyl)benzyl)-1-
oxoisoindolin-6-yl)acrylate (27i). This compound was
prepared from 26b by means of a procedure similar to
for 16 (49%). H NMR (500 MHz, CDCl3) d 8.04 (d,
1H, J = 1.5 Hz), 7.65 (d, 1H, J = 5.9 Hz), 7.63 (dd, 1H,
J = 1.5, 7.9 Hz), 7.38 (d, 1H, J = 7.9 Hz), 7.22 (dd, 1H,
J = 7.3, 7.3 Hz), 7.12 (s, 1H), 7.10 (d, 2H, J = 7.3 Hz),
6.45 (d, 1H, J = 5.9 Hz), 4.77 (s, 2H), 4.27 (s, 2H),
2.33 (s, 3H), 1.54 (s, 9H); FAB MS m/z 364 (M+H)+.
1
that used for 16 (71%). H NMR (500 MHz, CDCl3) d
8.04 (s, 1H), 7.65 (d, 1H, J = 7.9 Hz), 7.65 (d, 1H,
J = 6.2 Hz), 7.56 (s, 1H), 7.55 (d, 1H, J = 7.7 Hz), 7.50
(d, 1H, J = 7.7 Hz), 7.46 (dd, 1H, J = 7.7, 7.7 Hz), 7.40
(d, 1H, J = 7.9 Hz), 6.46 (d, 1H, J = 6.2 Hz), 4.86 (s,
2H), 4.30 (s, 2H), 1.54 (s, 9H); FAB MS m/z 418
(M+H)+.
5.1.38. (E)-tert-Butyl 3-(2-(4-methylbenzyl)-1-oxoisoin-
dolin-6-yl)acrylate (27p). This compound was prepared
from 26b by means of a procedure similar to that used
1
for 16 (69%). H NMR (500 MHz, CDCl3) d 8.02 (s,
5.1.32. (E)-tert-Butyl 3-(2-(4-(trifluoromethyl)benzyl)-1-
oxoisoindolin-6-yl)acrylate (27j). This compound was
prepared from 26b by means of a procedure similar to
1H, J = 1.4 Hz), 7.63 (d, 1H, J = 7.6 Hz), 7.63 (d, 1H,
J = 5.9 Hz), 7.36 (d, 1H, J = 7.6 Hz), 7.19 (d, 2H,
J = 7.6 Hz), 7.14 (d, 2H, J = 7.6 Hz), 6.44 (d, 1H,
J = 5.9 Hz), 4.76 (s, 2H), 4.25 (s, 2H), 2.33 (s, 3H),
1.54 (s, 9H); FAB MS m/z 364 (M+H)+.
1
that used for 16 (52%). H NMR (500 MHz, CDCl3) d
8.04 (s, 1H), 7.65 (d, 1H, J = 7.6 Hz), 7.65 (d, 1H,
J = 6.2 Hz), 7.60 (d, 1H, J = 8.3 Hz), 7.42 (d, 1H,
J = 8.3 Hz), 7.40 (d, 1H, J = 7.6 Hz), 6.46 (d, 1H,
J = 6.2 Hz), 4.86 (s, 2H), 4.30 (s, 2H), 1.54 (s, 9H);
FAB MS m/z 418 (M+H)+.
5.1.39. (E)-Methyl 3-(2-(2-phenylbenzyl)-1-oxoisoindolin-
6-yl)acrylate (27q). This compound was prepared from
26a by means of a procedure similar to that used for
1
16 (51%). H NMR (500 MHz, CDCl3) d 8.00 (d, 1H,
5.1.33. (E)-tert-Butyl 3-(2-(2-methoxybenzyl)-1-oxoisoin-
dolin-6-yl)acrylate (27k). This compound was prepared
from 26b by means of a procedure similar to that used
J = 1.5 Hz), 7.73 (d, 1H, J = 6.2 Hz), 7.62 (dd, 1H,
J = 1.5, 7.9 Hz), 7.43 (m, 2H), 7.31 (m, 8H), 6.51 (d,
1H, J = 6.2 Hz), 4.82 (s, 2H), 4.07 (s, 2H), 3.81 (s,
3H); FAB MS m/z 384 (M+H)+.
1
for 16 (62%). H NMR (500 MHz, CDCl3) d 8.00 (s,
1H), 7.74 (d, 1H, J = 5.9 Hz), 7.61 (d, 1H, J = 7.9 Hz),
7.37 (d, 1H, J = 7.9 Hz), 7.25 (m, 2H), 6.90 (m, 2H),
6.44 (d, 1H, J = 5.9 Hz), 4.83 (s, 2H), 4.31 (s, 2H),
3.86 (s, 3H), 1.54 (s, 9H); FAB MS m/z 380 (M+H)+.
5.1.40. (E)-Methyl 3-(2-(3-phenylbenzyl)-1-oxoisoindolin-
6-yl)acrylate (27r). This compound was prepared from
26a by means of a procedure similar to that used for
1
16 (41%). H NMR (500 MHz, CDCl3) d 8.06 (s, 1H),
5.1.34. (E)-tert-Butyl 3-(2-(3-methoxybenzyl)-1-oxoisoin-
dolin-6-yl)acrylate (27l). This compound was prepared
from 26b by means of a procedure similar to that used
7.76 (d, 1H, J = 5.9 Hz), 7.64 (d, 1H, J = 8.0 Hz), 7.56
(d, 2H, J = 7.4 Hz), 7.52 (s, 1H), 7.51 (d, 1H,
J = 8.0 Hz), 7.43 (dd, 2H, J = 7.3, 7.4 Hz), 7.41 (dd,
1H, J = 7.3, 8.0 Hz), 7.40 (d, 1H, J = 8.0 Hz), 7.34 (t,
1H, J = 7.3 Hz), 7.28 (d, 1H, J = 7.3 Hz), 6.53 (d, 1H,
J = 5.9 Hz), 4.89 (s, 2H), 4.32 (s, 2H), 3.82 (s, 3H);
FAB MS m/z 384 (M+H)+.
1
for 16 (69%). H NMR (500 MHz, CDCl3) d 8.03 (s,
1H), 7.74 (d, 1H, J = 5.9 Hz), 7.62 (d, 1H, J = 8.0 Hz),
7.39 (d, 1H, J = 8.0 Hz), 7.26 (s, 1H), 7.25 (d, 1H,
J = 7.3 Hz), 6.91 (dd, 1H, J = 7.3, 8.3 Hz), 6.89 (d, 1H,

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C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
5.1.41. (E)-Methyl 3-(2-(4-phenylbenzyl)-1-oxoisoindolin-
6-yl)acrylate (27s). This compound was prepared from
26a by means of a procedure similar to that used for
1H), 7.63 (d, 1H, J = 6.1 Hz), 7.59 (d, 1H, J = 7.7 Hz),
7.33 (m, 5H), 7.25 (m, 3H), 7.16 (t, 1H, J = 7.0 Hz),
7.14 (d, 2H, J = 8.2 Hz), 6.43 (d, 1H, J = 6.1 Hz), 5.64
(dd, 1H, J = 6.4, 9.5 Hz), 4.18 (d, 1H, J = 17.4 Hz),
3.95 (d, 1H, J = 17.4 Hz), 2.73 (m, 1H), 2.68 (m, 1H),
2.11 (m, 2H), 1.73 (m, 1H), 1.66 (m, 1H), 1.54 (s, 9H);
FAB MS m/z 468 (M+H)+.
1
16 (quant.). H NMR (500 MHz, DMSO-d6) d 8.06 (s,
1H), 7.76 (d, 1H, J = 5.9 Hz), 7.65 (d, 1H, J = 7.9 Hz),
7.65 (d, 4H, J = 8.2 Hz), 7.38 (m, 6H), 6.52 (d, 1H,
J = 5.9 Hz), 4.84 (s, 2H), 4.33 (s, 2H), 3.82 (s, 3H);
FAB MS m/z 384 (M+H)+.
5.1.48. (E)-(S)-tert-Butyl 3-(2-(2-(benzyloxy)-1-phenyl-
ethyl)-1-oxoisoindolin-6-yl)acrylate (27z). This com-
5.1.42. (E)-tert-Butyl 3-(2-(2-tert-butylbenzyl)-1-oxoiso-
indolin-6-yl)acrylate (27t). This compound was prepared
from 26b by means of a procedure similar to that used
pound was prepared from 26b by means of
a
1
procedure similar to that used for 16 (35%). H NMR
(500 MHz, CDCl3) d 8.00 (d, 1H, J = 1.6 Hz), 7.63 (d,
1H, J = 5.9 Hz), 7.60 (dd, 1H, J = 1.6, 7.6 Hz), 7.31
(m, 11H), 6.43 (d, 1H, J = 5.9 Hz), 5.78 (dd, 1H,
J = 4.9, 7.3 Hz), 4.62 (d, 1H, J = 11.9 Hz), 4.54 (d, 1H,
J = 11.9 Hz), 4.43 (d, 1H, J = 17.4 Hz), (2H), 4.14 (d,
1
for 16 (49%). H NMR (500 MHz, CDCl3) d 8.07 (s,
1H), 7.66 (d, 1H, J = 6.2 Hz), 7.64 (d, 1H, J = 8.0 Hz),
7.43 (d, 1H, J = 8.0 Hz), 7.38 (d, 1H, J = 8.0 Hz), 7.21
(dd, 1H, J = 4.6, 8.0 Hz), 7.15 (m, 2H), 6.46 (d, 1H,
J = 6.2 Hz), 5.13 (s, 2H), 4.24 (s, 2H), 1.55 (s, 9H),
1.48 (s, 9H); FAB MS m/z 406 (M+H)+.
1H, J = 17.4 Hz), 4.12 (dd, 1H, J = 7.3, 10.4 Hz), 4.01
+31.1ꢁ (c
30:4
D
(dd, 1H, J = 4.9, 10.4 Hz), 1.52 (s, 9H); ½aꢀ
0.165, CH3CN); FAB MS m/z 470 (M+H)+.
5.1.43. (E)-tert-Butyl 3-(2-(4-tert-butylbenzyl)-1-oxoiso-
indolin-6-yl)acrylate (27u). This compound was prepared
from 26b by means of a procedure similar to that used for
16 (90%). 1H NMR (500 MHz, CDCl3) d 8.03 (s, 1H), 7.65
(d, 1H, J = 5.8 Hz), 7.63 (d, 1H, J = 8.0 Hz), 7.37 (d, 1H,
J = 8.0 Hz), 7.35 (d, J = 8.3 Hz), 7.23 (d, 1H, J = 8.3 Hz),
6.46 (d, 1H, J = 5.8 Hz), 4.77 (s, 2H), 4.28 (s, 2H), 1.54 (s,
9H), 1.30 (s, 9H); FAB MS m/z 406 (M+H)+.
5.1.49. (E)-(R)-tert-Butyl 3-(2-(2-(benzyloxy)-1-phenyl-
ethyl)-1-oxoisoindolin-6-yl)acrylate (27aa). This com-
pound was prepared from 26b by means of
a
1
procedure similar to that used for 16 (37%). H NMR
(500 MHz, CDCl3) d 8.00 (d, 1H, J = 1.6 Hz), 7.63 (d,
1H, J = 5.9 Hz), 7.60 (dd, 1H, J = 1.6, 7.6 Hz), 7.31
(m, 11H), 6.43 (d, 1H, J = 5.9 Hz), 5.78 (dd, 1H,
J = 4.9, 7.3 Hz), 4.62 (d, 1H, J = 11.9 Hz), 4.54 (d, 1H,
J = 11.9 Hz), 4.43 (d, 1H, J = 17.4 Hz), 4.14 (d, 1H,
5.1.44. (E)-tert-Butyl 3-(2-benzhydryl-1-oxoisoindolin-6-
yl)acrylate (27v). This compound was prepared from 26b
by means of a procedure similar to that used for 16
(72%). 1H NMR (500 MHz, CDCl3) d 8.06 (s, 1H),
7.66 (d, 1H, J = 7.6 Hz), 7.64 (d, 1H, J = 6.7 Hz), 7.26
(m, 11H), 6.46 (d, 1H, J = 7.6 Hz), 5.22 (s, 1H), 4.24
(s, 2H), 1.54 (s, 9H); FAB MS m/z 426 (M+H)+.
J = 17.4 Hz), 4.12 (dd, 1H, J = 7.3, 10.4 Hz), 4.01 (dd,
1H, J = 4.9, 10.4 Hz), 1.52 (s, 9H); ½aꢀ
25:5
D
ꢁ51.0ꢁ (c
0.155, CH3CN); FAB MS m/z 470 (M+H)+.
5.1.50. (E)-3-(1-Oxo-2-phenethylisoindolin-6-yl)acrylic
acid (28a). This compound was prepared from 27a by
means of a procedure similar to that used for 10
(70%). 1H NMR (500 MHz, DMSO-d6) d 12.39 (s,
1H), 7.91 (d, 1H, J = 1.5 Hz), 7.89 (dd, 1H, J = 1.5,
7.8 Hz), 7.68 (d, 1H, J = 6.1 Hz), 7.58 (d, 1H,
J = 7.8 Hz), 7.22 (m, 5H), 6.60 (d, 1H, J = 6.1 Hz),
4.41 (s, 2H), 3.77 (t, 2H, J = 7.3 Hz), 2.92 (t, 2H,
J = 7.3 Hz); FAB MS m/z 308 (M+H)+.
5.1.45. (E)-tert-Butyl 3-(1-oxo-2-(1,2-diphenylethyl)isoin-
dolin-6-yl)acrylate (27w). This compound was prepared
from 26b by means of a procedure similar to that used
1
for 16 (58%). H NMR (500 MHz, CDCl3) d 7.93 (s,
1H), 7.60 (d, 1H, J = 5.9 Hz), 7.58 (d, 1H, J = 7.6 Hz),
7.45 (d, 1H, J = 7.6 Hz), 7.35 (m, 3H), 7.26 (m, 6H),
7.15 (m, 1H), 6.40 (d, 1H, J = 5.9 Hz), 5.97 (dd, 1H,
J = 6.4, 9.7 Hz), 4.39 (d, 1H, J = 17.1 Hz), 4.10 (d, 1H,
J = 17.1 Hz), 3.51 (dd, 1H, J = 6.4, 14.6 Hz), 3.43 (dd,
1H, J = 9.7, 14.6 Hz), 1.53 (s, 9H); FAB MS m/z 440
(M+H)+.
5.1.51.
(E)-3-(1-Oxo-2-(3-phenylpropyl)isoindolin-6-
yl)acrylic acid (28b). This compound was prepared from
27b by means of a procedure similar to that used for 10
1
(76%). H NMR (500 MHz, DMSO-d6) d 12.39 (s, 1H),
7.93 (d, 1H, J = 1.5 Hz), 7.91 (dd, 1H, J = 1.5, 7.8 Hz),
7.68 (d, 1H, J = 5.7 Hz), 7.60 (d, 1H, J = 7.8 Hz), 7.24
(m, 4H), 7.16 (m, 1H), 6.62 (d, 1H, J = 5.7 Hz), 4.51
(s, 2H), 3.54 (t, 2H, J = 7.3 Hz), 2.60 (t, 2H,
J = 7.8 Hz), 1.91 (m, 2H); FAB MS m/z 322 (M+H)+.
5.1.46. (E)-tert-Butyl 3-(1-oxo-2-(1,3-diphenylpropyl)iso-
indolin-6-yl)acrylate (27x). This compound was pre-
pared from 26b by means of a procedure similar to
1
that used for 16 (59%). H NMR (500 MHz, CDCl3) d
8.02 (d, 1H, J = 1.6 Hz), 7.63 (d, 1H, J = 5.8 Hz), 7.61
(dd, 1H, J = 1.6, 8.2 Hz), 7.30 (m, 11H), 6.45 (d, 1H,
J = 5.8 Hz), 5.68 (dd, 1H, J = 6.8, 8.6 Hz), 4.30 (d, 1H,
J = 17.5 Hz), 4.02 (d, 1H, J = 17.5 Hz), 2.72 (m, 1H),
2.67 (m, 1H), 2.43 (m, 2H), 1.53 (s, 9H); FAB MS m/z
454 (M+H)+.
5.1.52. (E)-3-(2-(Cyclohexylmethyl)-1-oxoisoindolin-6-
yl)acrylic acid (28c). A mixture of 27c (109 mg,
0.31 mmol), 8 mL of CH2Cl2, and 8 mL of trifluoroace-
tic acid was stirred overnight at room temperature. The
solvent was evaporated and the residue was recrystal-
lized from n-hexane/AcOEt to afford 92 mg (quant.) of
5.1.47. (E)-tert-Butyl 3-(1-oxo-2-(1,4-diphenylbutyl)isoin-
dolin-6-yl)acrylate (27y). This compound was prepared
from 26b by means of a procedure similar to that used
1
the title compound. H NMR (500 MHz, DMSO-d6) d
2.40 (s, 1H), 7.92 (s, 1H), 7.90 (d, 1H, J = 7.7 Hz),
7.68 (d, 1H, J = 6.0 Hz), 7.60 (d, 1H, J = 7.7 Hz), 6.60
1
for 16 (45%). H NMR (500 MHz, CDCl3) d 8.00 (s,

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7639
(d, 1H, J = 6.0 Hz), 4.48 (s, 2H), 3.35 (d, 2H,
J = 7.1 Hz), 1.61 (m, 6H), 1.19 (m, 3H), 0.94 (m, 2H);
FAB MS m/z 300 (M+H)+.
5.1.59. (E)-3-(2-(4-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl) acrylic acid (28j). This compound was pre-
pared from 27j by means of a procedure similar to
that used for 28c (quant.). 1H NMR (500 MHz,
DMSO-d6) d 12.41 (s, 1H), 8.00 (s, 1H), 7.93 (d, 1H,
J = 7.7 Hz), 7.71 (d, 2H, J = 8.3 Hz), 7.68 (d, 1H,
J = 6.0 Hz), 7.59 (d, 1H, J = 7.7 Hz), 7.49 (d, 2H,
J = 8.3 Hz), 6.63 (d, 1H, J = 6.0 Hz), 4.83 (s, 2H), 4.44
(s, 2H); FAB MS m/z 362 (M+H)+.
5.1.53. (E)-3-(2-((Naphthalene-1-yl)methyl)-1-oxoisoin-
dolin-6-yl)acrylic acid (28d). This compound was pre-
pared from 27d by means of a procedure similar to
that used for 28c (quant.). 1H NMR (500 MHz,
DMSO-d6) d 12.42 (s, 1H), 8.21 (d, 1H, J = 7.7 Hz),
8.02 (s, 1H), 7.95 (d, 1H, J = 7.2 Hz), 7.90 (m, 2H),
7.68 (d, 1H, J = 5.9 Hz), 7.53 (d, 1H, J = 7.2 Hz), 7.52
(m, 4H), 6.63 (d, 1H, J = 5.9 Hz), 5.18 (s, 2H), 4.28 (s,
2H); FAB MS m/z 344 (M+H)+.
5.1.60. (E)-3-(2-(2-Methoxybenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28k). This compound was prepared from
27k by means of a procedure similar to that used for
1
28c (quant.). H NMR (500 MHz, CDCl3) d 12.41 (s,
5.1.54. (E)-3-(2-(2-Chlorobenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28e). This compound was prepared from
27e by means of a procedure similar to that used for
10 (70%). 1H NMR (500 MHz, CD3OD) d 8.00 (d,
1H, J = 1.5 Hz), 7.83 (dd, 1H, J = 1.5, 7.9 Hz), 7.75 (d,
1H, J = 6.2 Hz), 7.66 (m, 1H), 7.56 (d, 1H,
J = 7.9 Hz), 7.44 (m, 1H), 7.30 (m, 2H), 6.58 (d, 1H,
J = 6.2 Hz), 4.94 (s, 2H), 4.42 (s, 2H) ; FAB MS m/z
328 (M+H)+.
1H), 7.97 (d, 1H, J = 1.1 Hz), 7.91 (dd, 1H, J = 1.1,
7.7 Hz), 7.70 (d, 1H, J = 5.9 Hz), 7.59 (d, 1H,
J = 7.7 Hz), 7.27 (dd, 1H, J = 7.2, 8.2 Hz), 7.08 (d,
1H, J = 7.7 Hz), 7.02 (d, 1H, J = 8.2 Hz), 6.90 (dd,
1H, J = 7.2, 7.7 Hz), 6.62 (d, 1H, J = 5.9 Hz), 4.69 (s,
2H), 4.40 (s, 2H), 3.82 (s, 3H); FAB MS m/z 324
(M+H)+.
5.1.61. (E)-3-(2-(3-Methoxybenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28l). This compound was prepared from 27l
by means of a procedure similar to that used for 28c
5.1.55. (E)-3-(2-(3-Chlorobenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28f). This compound was prepared from
27f by means of a procedure similar to that used for
10 (quant.). 1H NMR (500 MHz, CDCl3) d 8.00 (d,
1H, J = 1.2 Hz), 7.83 (dd, 1H, J = 1.2, 7.9 Hz), 7.70 (d,
1H, J = 15.8 Hz), 7.56 (d, 1H, J = 7.9 Hz), 7.33 (m, 3
H), 7.24 (d, 1H, J = 7.3 Hz), 6.58 (d, 1H, J = 15.8 Hz),
4.85 (s, 2H), 4.43 (s, 2H); FAB MS m/z 328.
1
(79%). H NMR (500 MHz, CD3OD) d 7.99 (d, 1H,
J = 1.5 Hz), 7.81 (dd, 1H, J = 1.5, 7.9 Hz), 7.75 (d, 1H,
J = 6.2 Hz), 7.55 (d, 1H, J = 7.9 Hz), 7.25 (dd, 1H,
J = 7.3, 7.3 Hz), 6.85 (m, 3H), 6.57 (d, 1H, J = 6.2 Hz),
4.77 (s, 2H), 4.40 (s, 2H), 3.75 (s, 3H); FAB MS m/z
324 (M+H)+.
5.1.62. (E)-3-(2-(4-Methoxybenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28m). This compound was prepared from
27m by means of a procedure similar to that used for
10 (71%). 1H NMR (500 MHz, CD3OD) d 7.99 (d,
1H, J = 1.5 Hz), 7.81 (dd, 1H, J = 1.5, 7.9 Hz), 7.75 (d,
1H, J = 6.1 Hz), 7.55 (d, 1H, J = 7.9 Hz), 7.24 (d, 2H,
J = 8.7 Hz), 6.90 (d, 2H, J = 8.7 Hz), 6.58 (d, 1H,
J = 6.1 Hz), 4.74 (s, 2H), 3.76 (s, 2H), 3.30 (s, 3H);
FAB MS m/z 324 (M+H)+.
5.1.56. (E)-3-(2-(4-Chlorobenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28g). This compound was prepared from
27g by means of a procedure similar to that used for
10 (71%). 1H NMR (500 MHz, CD3OD) d 8.00 (d,
1H, J = 1.5 Hz), 7.83 (dd, 1H, J = 1.5, 7.8 Hz), 7.75 (d,
1H, J = 6.0 Hz), 7.57 (d, 1H, J = 7.8 Hz), 7.35 (d, 2H,
J = 8.7 Hz), 7.30 (d, 2H, J = 8.7 Hz,), 6.58 (d, 1H,
J = 6.0 Hz), 4.80 (s, 2H), 4.42 (s, 2H); FAB MS m/z
328 (M+H)+.
5.1.63. (E)-3-(2-(2-Methylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28n). This compound was prepared from
28n by means of a procedure similar to that used for
28c (79%). 1H NMR (500 MHz, CDCl3) d 8.03 (d,
1H, J = 1.2 Hz), 7.65 (d, 1H, J = 6.1 Hz), 7.64 (dd,
1H, J = 1.2, 7.8 Hz), 7.37 (d, 1H, J = 7.8 Hz), 7.20
(m, 4H), 6.46 (d, 1H, J = 6.1 Hz), 4.83 (s, 2H), 4.20
(s, 2H), 2.34 (s, 3H), 1.54 (s, 9H); FAB MS m/z 364
(M+H)+.
5.1.57. (E)-3-(2-(2-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl) acrylic acid (28h). This compound was pre-
pared from 27 h by means of a procedure similar to
that used for 28c (quant.). 1H NMR (500 MHz,
DMSO-d6) d 12.43 (s, 1H), 8.03 (s, 1H), 7.95 (d, 1H,
J = 7.8 Hz), 7.78 (d, 1H, J = 7.8 Hz), 7.70 (d, 1H,
J = 6.1 Hz), 7.65 (d, 1H, J = 7.3 Hz), 7.62 (dd, 1H,
J = 7.3, 7.8 Hz), 7.51 (dd, 1H, J = 7.3. 7.3 Hz), 7.35 (d,
1H, J = 7.8 Hz), 6.63 (d, 1H, J = 6.1 Hz), 4.92 (s, 2H),
4.44 (s, 2H); FAB MS m/z 362 (M+H)+.
5.1.64. (E)-3-(2-(3-Methylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28o). This compound was prepared from
28n by means of a procedure similar to that used
for 28c (quant.). 1H NMR (500 MHz, DMSO-d6) d
12.42 (s, 1H), 7.99 (d, J = 1.1 Hz, 1H), 7.91 (dd,
J = 1.1, 7.7 Hz, 1H), 7.70 (d, J = 5.9 Hz, 1H), 7.58
(d, J = 7.7 Hz, 1H), 7.23 (dd, J = 7.2, 7.5 Hz, 1H),
7.09 (d, J = 7.5 Hz, 1H), 7.08 (s, 1H), 7.05 (d,
J = 7.2 Hz, 1H), 6.63 (d, J = 5.9 Hz, 1H), 4.68 (s,
2H), 4.38 (s, 2H), 2.27 (s, 3H); FAB MS m/z 308
(M+H)+.
5.1.58. (E)-3-(2-(3-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl) acrylic acid (28i). This compound was pre-
pared from 27i by means of a procedure similar to
that used for 28c (quant.). 1H NMR (500 MHz,
DMSO-d6) d 12.42 (s, 1H), 8.01 (s, 1H), 7.93 (d, 1H,
J = 7.8 Hz), 7.70 (d, 2H, J = 6.1 Hz), 7.66 (s, 1H), 7.65
(m, 2H), 7.60 (d, 1H, J = 7.8 Hz), 7.59 (m, 1H), 6.63
(d, 1H, J = 6.1 Hz), 4.83 (s, 2H), 4.44 (s, 2H); FAB
MS m/z 362 (M+H)+.

7640
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
5.1.65. (E)-3-(2-(4-Methylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28p). This compound was prepared from
27p by means of a procedure similar to that used for
28c (quant.). 1H NMR (500 MHz, DMSO-d6) 1H
NMR (500 MHz, CDCl3) d 12.41 (s, 1H), 7.98 (s, 1H,
J = 1.4 Hz), 7.91 (dd, 1H J = 1.4, 7.9 Hz), 7.70 (d, 1H
J = 6.1 Hz), 7.57 (d, 1H J = 7.9 Hz), 7.16 (s, 4H), 6.62
(d, 1H J = 6.1 Hz), 4.67 (s, 2H), 4.35 (s, 2H), 2.26 (s,
3H); FAB MS m/z 308 (M+H)+.
5.1.71. (E)-3-(2-Benzhydryl-1-oxoisoindolin-6-yl) acrylic
acid (28v). This compound was prepared from 27v by
means of a procedure similar to that used for 28c
(quant.). 1H NMR (500 MHz, DMSO-d6) d 8.88 (s,
1H), 8.01 (d, 1H, J = 1.5 Hz), 7.94 (dd, 1H, J = 1.5,
8.0 Hz), 7.70 (d, 1H, J = 6.0 Hz), 7.58 (d, 1H,
J = 8.0 Hz), 7.26 (m, 8H), 7.19 (d, 2H, J = 7.5 Hz),
6.64 (d, 1H, J = 6.0 Hz), 5.67 (m, 1H), 4.28 (s, 2H);
FAB MS m/z 370 (M+H)+.
5.1.72. (E)-3-(1-Oxo-2-(1,2-diphenylethyl)isoindolin-6-yl)
acrylic acid (28w). This compound was prepared from
27w by means of a procedure similar to that used for
5.1.66.
(E)-3-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28q). This compound was prepared from
27q by means of a procedure similar to that used for
10 (quant.). H NMR (500 MHz, DMSO-d6) d 12.28
1
1
28c (quant.). H NMR (500 MHz, DMSO-d6) d 12.44
(br, 1H), 7.86 (s, 1H), 7.64 (d, 1H, J = 5.8 Hz), 7.54
(d, 2H, J = 8.6 Hz), 7.45 (d, 2H, J = 7.3 Hz), 7.36 (dd,
2H, J = 7.3, 8.0 Hz), 7.28 (t, 1H, J = 8.0 Hz), 7.28 (d,
2H, J = 7.3 Hz), 7.20 (dd, 2H, J = 7.3, 7.3 Hz), 7.11 (t,
1H, J = 7.3 Hz), 6.58 (d, 1H, J = 5.8 Hz), 5.75 (t, 1H,
J = 8.3 Hz), 4.58 (d, 1H, J = 18.3 Hz), 4.12 (d, 1H,
J = 18.3 Hz), 3.45 (d, 2H, J = 8.3 Hz); FAB MS m/z
384 (M+H)+.
(br, 1H), 7.94 (s, 1H), 7.89 (d, 1H, J = 8.0 Hz), 7.68
(d, 1H, J = 5.9 Hz), 7.54 (d, 1H, J = 8.0 Hz), 7.44 (m,
2H), 7.35 (m, 5H), 7.25 (m, 1H), 7.21 (m, 1H), 6.62 (d,
1H, J = 5.9 Hz), 4.69 (s, 2H), 4.24 (s, 2H); FAB MS
m/z 370 (M+H)+.
5.1.67.
(E)-3-(2-(3-Phenylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28r). This compound was prepared from
27r by means of a procedure similar to that used for
1
5.1.73. (E)-3-(1-Oxo-2-(1,3-diphenylpropyl)isoindolin-6-
yl) acrylic acid (28x). This compound was prepared from
27x by means of a procedure similar to that used for 28c
(quant.). 1H NMR (500 MHz, DMSO-d6) d 8.10 (s, 1H),
7.83 (d, 1H, J = 6.3 Hz), 7.66 (d, 1H, J = 8.2 Hz), 7.41
(d, 1H, J = 8.2 Hz), 7.40 (dd, 2H, J = 7.3, 8.5 Hz), 7.36
(dd, 2H, J = 7.3, 8.5 Hz), 7.29 (t, 1H, J = 7.3 Hz), 7.25
(d, 2H, J = 8.5 Hz), 7.19 (d, 2H, J = 8.5 Hz), 7.16 (t,
1H, J = 7.3 Hz), 6.54 (d, 1H, J = 6.3 Hz), 5.69 (dd, 1H,
J = 6.8, 8.6 Hz), 4.33 (d, 1H, J = 17.5 Hz), 4.06 (d, 1H,
J = 17.5 Hz), 2.73 (m, 1H), 2.67 (m, 1H), 2.43 (m, 2H);
FAB MS m/z 398 (M+H)+.
10 (quant.). H NMR (500 MHz, DMSO-d6) d 12.41
(s, 1H), 8.00 (s, 1H), 7.91 (d, 1H, J = 7.4 Hz), 7.70 (d,
1H, J = 5.9 Hz), 7.63 (d, 2H, J = 7.3 Hz), 7.59 (d, 1H,
J = 7.4 Hz), 7.58 (s, 1H), 7.47 (d, 1H, J = 7.9 Hz), 7.46
(dd, 2H, J = 7.3, 7.3 Hz), 7.44 (dd, 1H, J = 7.9,
7.9 Hz), 7.44 (t, 1H, J = 7.3 Hz), 7.26 (d, 1H,
J = 7.9 Hz), 6.63 (d, 1H, J = 5.5 Hz), 4.80 (s, 2H), 4.44
(s, 2H); FAB MS m/z 370 (M+H)+.
5.1.68.
(E)-3-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28s). This compound was prepared from
27s by means of a procedure similar to that used
for 10 (54%). 1H NMR (500 MHz, DMSO-d6) d
12.41 (s, 1H), 8.01 (d, 1H, J = 1.8 Hz), 7.93 (dd, 1H,
J = 1.8, 7.9 Hz), 7.70 (d, 1H, J = 5.8 Hz), 7.63 (m,
5H), 7.44 (m, 2H), 7.36 (m, 3H), 6.64 (d, 1H,
J = 5.8 Hz), 4.77 (s, 2H), 4.43 (s, 2H); FAB MS m/z
370 (M+H)+.
5.1.74. (E)-3-(1-Oxo-2-(1,4-diphenylbutyl)isoindolin-6-yl)
acrylic acid (28y). This compound was prepared from
27y by means of a procedure similar to that used for
28c (quant.). 1H NMR (500 MHz, DMSO-d6) d 7.95
(s, 1H), 7.90 (d, 1H, J = 8.1 Hz); 7.67 (d, 1H,
J = 6.1 Hz), 7.55 (d, 1H, J = 8.1 Hz), 7.34 (m, 4H),
7.25 (m, 3H), 7.14 (m, 3H), 6.60 (d, 1H, J = 6.1 Hz),
5.41 (dd, 1H, J = 6.1, 10.0 Hz), 4.45 (d, 1H,
J = 18.5 Hz), 4.04 (d, 1H, J = 18.5 Hz), 2.63 (m, 2H),
2.07 (m, 2H), 1.53 (m, 2H); FAB MS m/z 412 (M+H)+.
5.1.69. (E)-3-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28t). This compound was prepared from
27t by means of a procedure similar to that used for
28c (quant.). 1H NMR (500 MHz, DMSO-d6) 1H
NMR (500 MHz, CDCl3) d 8.02 (s, 1H), 7.93 (d, 1H,
J = 7.7 Hz), 7.70 (d, 1H, J = 6.0 Hz), 7.59 (d, 1H,
J = 7.7 Hz), 7.40 (d, 1H, J = 7.7 Hz), 7.20 (dd, 1H,
J = 7.7, 7.7 Hz), 7.15 (dd, 1H, J = 7.7, 7.7 Hz), 7.05
(d, 1H, J = 7.7 Hz), 6.64 (d, 1H, J = 6.0 Hz), 4.99 (s,
2H), 4.35 (s, 2H), 1.43 (s, 9H); FAB MS m/z 350
(M+H)+.
5.1.75.
(E)-(S)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
oxoisoindolin-6-yl) acrylic acid (28z). This compound
was prepared from 28z by means of a procedure similar
to that used for 28c (quant.). 1H NMR (500 MHz,
CDCl3) d 8.01 (s, 1H), 7.80 (d, 1H, J = 6.1 Hz), 7.64
(d, 1H, J = 7.8 Hz), 7.37 (d, 1H, J = 7.8 Hz), 7.31 (m,
10H), 6.50 (d, 1H, J = 6.1 Hz), 5.80 (dd, 1H, J = 4.9,
7.9 Hz), 4.62 (d, 1H, J = 11.9 Hz), 4.54 (d, 1H,
J = 11.9 Hz), 4.47 (d, 1H, J = 17.5 Hz), 4.17 (d, 1H,
5.1.70. (E)-3-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-yl)
acrylic acid (28u). This compound was prepared from
28u by means of a procedure similar to that used for
28c (quant.). 1H NMR (500 MHz, DMSO-d6) d 7.99
(s, 1H), 7.92 (d, 1 H, J = 8.1 Hz), 7.70 (d, 1H,
J = 6.2 Hz), 7.57 (d, 1H, J = 8.1 Hz), 7.36 (d, 1H,
J = 8.5 Hz), 7.19 (d, 1H, J = 8.5 Hz), 6.63 (d, 1H,
J = 6.2 Hz), 4.68 (s, 2H), 4.37 (s, 2H), 1.24 (s, 9H);
FAB MS m/z 350 (M+H)+.
J = 17.5 Hz), 4.12 (dd, 1H, J = 7.9, 10.3 Hz), 4.01 (dd,
1H, J = 4.9, 10.3 Hz); ½aꢀ
30:1
D
+35.3ꢁ (c 0.306, CH3CN);
FAB MS m/z 414 (M+H)+.
5.1.76. (E)-(R)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
oxoisoindolin-6-yl)acrylic acid (28aa). This compound
was prepared from 27aa by means of a procedure similar

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7641
to that used for 28c (quant.). 1H NMR (500 MHz,
CDCl3) d 8.01 (s, 1H), 7.80 (d, 1H, J = 5.9 Hz), 7.64
(d, 1H, J = 7.8 Hz), 7.37 (d, 1H, J = 7.8 Hz), 7.31 (m,
10H), 6.50 (d, 1H, J = 5.9 Hz), 5.80 (dd, 1H, J = 4.9,
7.9 Hz), 4.62 (d, 1H, J = 11.9 Hz), 4.54 (d, 1H,
J = 11.9 Hz), 4.47 (d, 1H, J = 17.5 Hz), 4.17 (d, 1H,
5.1.81. (E)-3-(2-((Naphthalene-1-yl)methyl)-1-oxoisoin-
dolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide
(29d). This compound was prepared from 28d by means
1
of a procedure similar to that used for 11 (49%). H
NMR (500 MHz, CDCl3) d 9.45 (s, 1H), 8.21 (d, 1H,
J = 8.0 Hz), 8.06 (s, 1H), 7.84 (m, 2H), 7.76 (d, 1H,
J = 5.5 Hz), 7.46 (m, 5H), 7.25 (d, 1H, J = 8.0 Hz,),
6.53 (d, 1H, J = 5.5 Hz), 5.25 (s, 2H), 5.06 (m, 1H),
4.13 (s, 2H), 4.01 (m, 1H), 3.65 (m, 1H), 1.84 (m, 3H),
1.53 (m, 3H); FAB MS m/z 443 (M+H)+.
J = 17.5 Hz), 4.12 (dd, 1H, J = 7.9, 10.3 Hz), 4.01 (dd,
ꢁ51.7ꢁ (c 0.130, CH3CN);
34:5
D
1H, J = 4.9, 10.3 Hz); ½aꢀ
FAB MS m/z 414 (M+H)+.
5.1.77. (E)-N-(2-(Trimethylsilyl)ethoxy)-3-(2-benzhydryl-
1-oxoisoindolin-6-yl)acrylamide (29v). To a mixture of
28v (65 mg, 0.18 mmol) and 5 mL of anhydrous THF
were added triethylamine (0.074 mL, 0.53 mmol), and
ethyl chloroformate (0.017 mL, 0.18 mmol) at 0 ꢁC, with
stirring for 1 h at 0 ꢁC. Then 1 equiv of methanolic o-(2-
trimethylsilylethyl)hydroxylamine was added and the
whole was stirred overnight at room temperature. The
reaction mixture was washed with dil NaOH and brine,
dried over anhydrous MgSO4, and evaporated. The res-
idue was purified by silica gel column chromatography
(eluant n-hexane/AcOEt 2:1 v/v) to afford 25 mg (29%)
5.1.82. (E)-3-(2-(2-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29e). This
compound was prepared from 28e by means of a proce-
dure similar to that used for 11 (61%). 1H NMR
(500 MHz, CDCl3) d 8.78 (s, 1H), 8.07 (s, 1H), 7.79
(d, 1H, J = 5.8 Hz), 7.62 (d, 1H, J = 7.9 Hz), 7.41 (d,
1H, J = 7.9 Hz), 7.40 (m, 1H), 7.33 (m, 1H), 7.23 (m,
2H), 6.50 (d, 1H, J = 5.8 Hz), 5.03 (m, 1H), 4.96 (s,
2H), 4.36 (s, 2H), 4.00 (m, 1H), 3.67 (m, 1H), 1.86 (m,
3H), 1.64 (m, 3H); FAB MS m/z 427 (M+H)+.
1
5.1.83. (E)-3-(2-(3-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29f). This
compound was prepared from 28f by means of a proce-
dure similar to that used for 11 (31%). 1H NMR
(500 MHz, CDCl3) d 9.22 (s, 1H), 8.06 (s, 1H), 7.79
(d, 1H, J = 5.3 Hz), 7.61 (d, 1H, J = 7.9 Hz), 7.40 (d,
1H, J = 7.9 Hz), 7.26 (m, 3H), 7.19 (m, 1H), 6.53 (d,
1H, J = 5.3 Hz), 5.05 (m, 1H), 4.78 (s, 2H), 4.30 (s,
2H), 4.01 (m, 1H), 3.67 (m, 1H), 1.85 (m, 3H), 1.61
(m, 3H); FAB MS m/z 427 (M+H)+.
of the title compound as an oil. H NMR (500 MHz,
CDCl3) d 9.21 (br, 1 H), 8.11 (s, 1H), 7.80 (d, 1H,
J = 4.5 Hz), 7.59 (d, 1H, J = 7.5 Hz,), 7.33 (m, 9H),
7.19 (d, 2H, J = 7.0 Hz), 6.90 (s, 1H), 6.50 (d, 1H,
J = 4.5 Hz), 4.23 (s, 2H), 4.01 (m, 2H), 1.05 (t, 2H,
J = 7.8 Hz,), 0.01 (s, 9H); FAB MS m/z 485 (M+H)+.
5.1.78. (E)-3-(1-Oxo-2-phenethylisoindolin-6-yl)-N-(tet-
rahydro-2H-pyran-2-yloxy)acrylamide (29a). This com-
pound was prepared from 28a by means of
a
1
procedure similar to that used for 11 (67%). H NMR
(500 MHz, CDCl3) d 9.27 (s, 1H), 8.00 (s, 1H), 7.76
(d, 1H, J = 4.9 Hz), 7.57 (d, 1H, J = 7.6 Hz), 7.35 (d,
1H, J = 7.6 Hz), 7.23 (m, 5H), 6.51 (d, 1H,
J = 4.9 Hz), 5.05 (m, 1H), 4.21 (s, 2H), 4.01 (m, 1H),
3.88 (t, 2H, J = 7.1 Hz), 3.65 (m, 1H), 3.00 (t, 2H,
J = 7.1 Hz), 1.85 (m, 3H), 1.60 (m, 3H); FAB MS m/z
407 (M+H)+.
5.1.84. (E)-3-(2-(4-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29g). This
compound was prepared from 28g by means of a proce-
dure similar to that used for 11 (38%). 1H NMR
(500 MHz, CDCl3) d 9.42 (s, 1H), 8.03 (s, 1H), 7.76
(d, 1H, J = 4.4 Hz), 7.60 (d, 1H, J = 7.5 Hz), 7.37 (d,
1H, J = 7.5 Hz), 7.29 (d, 2H, J = 8.5 Hz), 7.19 (d, 2H,
J = 8.5 Hz), 6.53 (d, 1H, J = 4.4 Hz), 5.05 (m, 1H),
4.77 (s, 2H), 4.28 (s, 2H), 4.01 (m, 1H), 3.64 (m, 1H),
1.84 (m, 3H), 1.59, (m, 3H); FAB MS m/z 427 (M+H)+.
5.1.79. (E)-3-(1-Oxo-2-(3-phenylpropyl)isoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29b). This
compound was prepared from 28b by means of a proce-
dure similar to that used for 11 (76%). 1H NMR
(500 MHz, CDCl3) d 9.77 (s, 1H), 8.00 (s, 1H), 7.76
(d, 1H, J = 5.2 Hz,), 7.58 (d, 1H, J = 7.6 Hz), 7.40 (d,
1H, J = 7.6 Hz), 7.24 (m, 5H), 6.57 (d, 1H,
J = 5.2 Hz,), 5.08 (m, 1H), 4.37 (s, 2H), 4.04 (m, 1H),
3.68 (t, 2H, J = 7.3 Hz), 3.67 (m, 1H), 2.69 (t, 2H,
J = 7.7 Hz), 2.01 (m, 2H), 1.85 (m, 3H), 1.60 (m, 3H);
FAB MS m/z 421 (M+H)+.
5.1.85. (E)-3-(2-(2-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide
(29h). This compound was prepared from 28h by means
1
of a procedure similar to that used for 11 (84%). H
NMR (500 MHz, CDCl3) d 9.65 (s, 1H), 8.06 (s, 1H),
7.77 (d, 1H, J = 5.2 Hz), 7.67 (d, 1H, J = 7.7 Hz), 7.61
(d, 1H, J = 7.3 Hz), 7.48 (dd, 1H, J = 7.3, 7.3 Hz), 7.38
(m, 3H), 6.56 (d, 1H, J = 5.2 Hz), 5.06 (m, 1H), 5.01
(s, 2H), 4.29 (s, 2H), 4.01 (m, 1H), 3.63 (m, 1H), 1.83
(m, 3H), 1.58 (m, 3H); FAB MS m/z 461 (M+H)+.
5.1.80. (E)-3-(2-(Cyclohexylmethyl)-1-oxoisoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29c).
This compound was prepared from 28c by means of a
5.1.86. (E)-3-(2-(3-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide
(29i). This compound was prepared from 28i by means
1
procedure similar to that used for 11 (52%). H NMR
(500 MHz, CDCl3) d 9.05 (s, 1H), 8.01 (s, 1H), 7.72
(d, 1H, J = 5.6 Hz), 7.60 (d, 1H, J = 7.9 Hz), 7.42 (d,
1H, J = 7.9 Hz), 6.50 (d, 1H, J = 5.6 Hz,), 5.04 (m,
1H), 4.40 (s, 2H), 4.00 (m, 1H), 3.65 (m, 1H), 3.45 (d,
2H, J = 7.3 Hz), 1.74 (m, 12H), 1.20 (m, 3H), 1.05 (m,
2H); FAB MS m/z 399 (M+H)+.
1
of a procedure similar to that used for 11 (72%). H
NMR (500 MHz, CDCl3) d 9.46 (s, 1H), 8.04 (s, 1H),
7.75 (d, 1H, J = 4.4 Hz,), 7.60 (d, 1H, J = 7.7 Hz), 7.55
(s, 1H), 7.54 (d, 1H, J = 7.6 Hz), 7.49 (d, 1H,
J = 7.7 Hz), 7.45 (dd, 1H, J = 7.7, 7.7 Hz), 7.38 (d, 1H,

7642
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
J = 7.6 Hz), 6.53 (d, 1H, J = 4.4 Hz), 5.06 (m, 1H), 4.86
(s, 2H), 4.30 (s, 2H), 4.00 (m, 1H), 3.64 (m, 1H), 1.85 (m,
3H), 1.59 (m, 3H) ; FAB MS m/z 461 (M+H)+.
5.1.92. (E)-3-(2-(3-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29o). This
compound was prepared from 28o by means of a proce-
dure similar to that used for 11 (76%). 1H NMR
(500 MHz, CDCl3) d 9.52 (s, 1H), 8.04 (s, 1H), 7.77 (d,
1H, J = 5.6 Hz,), 7.58 (d, 1H, J = 7.7 Hz), 7.35 (d, 1H,
J = 7.7 Hz), 7.21 (dd, 1H, J = 7.5, 7.5 Hz), 7.10 (s, 1H),
7.20 (d, 1H, J = 7.5 Hz), 7.20 (d, 1 H, J = 7.5 Hz), 6.55
(d, 1H, J = 5.6 Hz), 5.06 (m, 1H), 4.76 (s, 2H), 4.27 (s,
2H), 4.02 (m, 1H), 3.66 (m, 1H), 2.31 (s, 3H), 1.84 (m,
3H), 1.59 (m, 3H); FAB MS m/z 407 (M+H)+.
5.1.87. (E)-3-(2-(4-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide
(29j). This compound was prepared from 28j by means
1
of a procedure similar to that used for 11 (57%). H
NMR (500 MHz, CDCl3) d 8.71 (s, 1H), 8.08 (s, 1H),
7.80 (d, 1H, J = 5.6 Hz), 7.63 (d, 1H, J = 7.7 Hz), 7.59
(d, 2H, J = 8.3 Hz), 7.42 (d, 2H, J = 8.3 Hz), 7.41 (d,
1H, J = 7.7 Hz), 6.51 (d, 1H, J = 5.6 Hz), 5.03 (m,
1H), 4.87 (s, 2H), 4.31 (s, 2H), 3.99 (m, 1H), 3.67 (m,
1H), 1.86 (m, 3H), 1.62 (m, 3H); FAB MS m/z 461
(M+H)+.
5.1.93. (E)-3-(2-(4-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29p). This
compound was prepared from 28p by means of a proce-
dure similar to that used for 11 (71%). 1H NMR
(500 MHz, CDCl3) d 9.35 (s, 1H), 8.04 (s, 1H), 7.77
(d, 1H, J = 5.6 Hz), 7.58 (d, 1H, J = 7.6 Hz), 7.34 (d,
1H, J = 7.6 Hz), 7.18 (d, 2H, J = 7.9 Hz), 7.12 (d, 2H,
J = 7.9 Hz), 6.53 (d, 1H, J = 5.6 Hz), 5.05 (m, 1H),
4.76 (s, 2H), 4.26 (s, 2H), 4.01 (m, 1H), 3.65 (m, 1H),
2.32 (s, 3H), 1.84 (m, 3H), 1.59 (m, 3H); FAB MS m/z
407 (M+H)+.
5.1.88.
(E)-3-(2-(2-Methoxybenzyl)-1-oxoisoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29k).
This compound was prepared from 28k by means of a
procedure similar to that used for 11 (47%). H NMR
1
(500 MHz, CDCl3) d 9.11 (s, 1H), 8.04 (s, 1H), 7.78
(d, 1H, J = 5.6 Hz), 7.58 (d, 1H, J = 7.6 Hz), 7.37 (d,
1H, J = 7.6 Hz), 7.26 (dd, 1H, J = 7.3, 8.3 Hz), 7.25 (d,
1H, J = 7.6 Hz), 6.90 (dd, 1H, J = 7.3, 7.6 Hz), 6.88 (d,
1H, J = 8.3 Hz), 6.52 (d, 1H, J = 5.6 Hz), 5.04 (m,
1H), 4.84 (s, 2H), 4.32 (s, 2H), 4.00 (m, 1H), 3.86 (s,
3H), 3.65 (m, 1H), 1.84 (m, 3H), 1.60 (m, 3H); FAB
MS m/z 423 (M+H)+.
5.1.94. (E)-3-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29q). This
compound was prepared from 28q by means of a proce-
dure similar to that used for 11 (62%). 1H NMR
(500 MHz, CDCl3) d 9.45 (s, 1H), 7.97 (s, 1H), 7.72
(d, 1H, J = 5.3 Hz), 7.52 (d, 1H, J = 7.3 Hz), 7.27 (m,
10H), 6.49 (d, 1H, J = 5.3 Hz), 5.01 (m, 1H), 4.77 (s,
2H), 4.03 (s, 2H), 3.95 (m, 1H), 3.59 (m, 1H), 1.79 (m,
3H), 1.54 (m, 3H); FAB MS m/z 469 (M+H)+.
5.1.89.
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide
(E)-3-(2-(3-Methoxybenzyl)-1-oxoisoindolin-6-
(29l).
This compound was prepared from 28l by means of a
procedure similar to that used for 11 (27%). H NMR
1
(500 MHz, CDCl3) d 9.25 (s, 1H), 8.05 (s, 1H), 7.78
(d, 1H, J = 5.6 Hz), 7.59 (d, 1H, J = 7.5 Hz), 7.37 (d,
1H, J = 7.5 Hz), 7.24 (dd, 1H, J = 7.6, 7.9 Hz), 6.87 (d,
1H, J = 7.6 Hz,), 6.83 (s, 1H), 6.82 (d, 1H, J = 7.9 Hz),
6.54 (d, 1H, J = 5.6 Hz), 5.05 (m, 1H), 4.77 (s, 2H),
4.29 (s, 2H), 4.01 (m, 1H), 3.77 (s, 3H), 3.65 (m, 1H),
1.86 (m, 3H), 1.64 (m, 3H); FAB MS m/z 423 (M+H)+.
5.1.95. (E)-3-(2-(3-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29r). This
compound was prepared from 28r by means of a proce-
dure similar to that used for 11 (62%). 1H NMR
(500 MHz, CDCl3) d 9.52 (s, 1H), 8.05 (s, 1H), 7.76
(d, 1H, J = 5.3 Hz), 7.56 (d, 1H, J = 7.6 Hz), 7.54 (d,
2H, J = 7.3 Hz), 7.51 (s, 1H), 7.50 (d, 1H, J = 7.6 Hz),
7.40 (dd, 2 H, J = 7.3, 7.3 Hz), 7.38 (dd, 1H, J = 7.3,
7.6 Hz), 7.33 (d, 1H, J = 7.3 Hz), 7.32 (t, 1H,
J = 7.3 Hz), 7.27 (d, 1H, J = 7.6 Hz), 6.54 (d, 1H,
J = 5.3 Hz), 5.06 (m, 1H), 4.87 (s, 2H), 4.31 (s, 2H),
4.01 (m, 1H), 3.64 (m, 1H), 1.58 (m, 3H), 1.25 (m,
3H); FAB MS m/z 469 (M+H)+.
5.1.90.
(E)-3-(2-(4-Methoxybenzyl)-1-oxoisoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29m).
This compound was prepared from 28m by means of a
procedure similar to that used for 11 (72%). H NMR
1
(500 MHz, CDCl3) d 9.37 (s, 1H), 8.03 (s, 1H), 7.76
(d, 1H, J = 5.6 Hz), 7.57 (d, 1H, J = 7.6 Hz), 7.35 (d,
1H, J = 7.6 Hz), 7.23 (d, 2H, J = 8.7 Hz), 6.85 (d, 2H,
J = 8.7 Hz), 6.53 (d, 1H, J = 5.6 Hz), 5.06 (m, 1H),
4.74 (s, 2H), 4.25 (s, 2H), 4.01 (m, 1H), 3.78 (s, 3H),
3.67 (m, 1H), 1.84 (m, 3H), 1.60 (m, 3H); FAB MS
m/z 423 (M+H)+.
5.1.96. (E)-3-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29s). This
compound was prepared from 28s by means of a proce-
dure similar to that used for 11 (23%). 1H NMR
(500 MHz, CDCl3) d 8.57 (s, 1H), 8.09 (s, 1H), 7.62
(d, 1H, J = 5.8 Hz), 7.62 (d, 1H, J = 7.6 Hz), 7.56 (d,
4H, J = 8.2 Hz), 7.38 (m, 6H), 6.49 (d, 1H,
J = 5.8 Hz), 5.03 (m, 1H), 4.85 (s, 2H), 4.33 (s, 2H),
3.99 (m, 1H), 3.67 (m, 1H), 1.86 (m, 3H), 1.60 (m,
3H); FAB MS m/z 469 (M+H)+.
5.1.91. (E)-3-(2-(2-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29n). This
compound was prepared from 28n by means of a proce-
1
dure similar to that used for 11 (65%). H NMR (500
MHz, CDCl3) d 9.64 (s, 1H), 8.04 (s, 1 H), 7.76 (d,
1H, J = 5.5 Hz), 7.57 (d, 1H, J = 7.6 Hz), 7.34 (d, 1H,
J = 7.6 Hz), 7.18 (m, 4H), 6.55 (d, 1H, J = 5.5 Hz),
5.06 (m, 1H), 4.82 (s, 2H), 4.01 (s, 2H), 4.00 (m, 1H),
3.63 (m, 1H), 2.33 (s, 3H), 1.83 (m, 3H), 1.58 (m, 3H);
FAB MS m/z 407 (M+H)+.
5.1.97. (E)-3-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29t).
This compound was prepared from 28t by means of a
1
procedure similar to that used for 11 (95%). H NMR

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7643
1
(500 MHz, CDCl3) d 8.49 (br, 1H), 8.11 (s, 1H), 7.82 (d,
1H, J = 5.2 Hz), 7.62 (d, 1H, J = 7.6 Hz), 7.43 (d, 1H,
J = 7.6 Hz), 7.38 (d, 1H, J = 8.0 Hz), 7.22 (dd, 1H,
J = 4.0, 8.6 Hz), 7.14 (m, 2H), 6.47 (d, 1H, J = 5.2 Hz),
5.13 (s, 2H), 5.03 (m, 1H), 4.25 (s, 2H), 3.99 (m, 1H),
3.69 (m, 1H), 1.86 (m, 3H), 1.58 (m, 3H), 1.48 (s, 9H);
FAB MS m/z 449 (M+H)+.
(65%). H NMR (500 MHz, CDCl3) d 9.37 (br, 1H),
8.03 (s, 1H), 7.75 (d, 1H, J = 5.0 Hz), 7.56 (d, 1H,
J = 7.6 Hz), 7.31 (m, 11H), 6.50 (d, 1H, J = 5.0 Hz),
5.79 (dd, 1H, J = 5.2, 7.3 Hz), 5.05 (m, 1H), 4.62 (d,
1H, J = 12.2 Hz), 4.54 (d, 1H, J = 12.2 Hz), 4.45 (d,
1H, J = 17.7 Hz), 4.17 (d, 1H, J = 17.7 Hz,), 4.12 (dd,
1 H, J = 7.3, 10.4 Hz), 2H, 4.03 (dd, 1H, J = 5.2,
10.4 Hz), 4.00 (m, 1H), 3.64 (m, 1H), 1.83 (m, 3H),
+50.4ꢁ (c 0.163, CH3CN); FAB
30:8
D
5.1.98. (E)-3-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29u).
This compound was prepared from 28u by means of a
1.59 (m, 3H); ½aꢀ
MS m/z 513 (M+H)+.
1
procedure similar to that used for 11 (95%). H NMR
5.1.103. (E)-(R)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
(500 MHz, CDCl3) d 8.56 (br, 1H), 8.07 (s, 1H), 7.80
(d, 1H, J = 5.4 Hz), 7.60 (d, 1H, J = 8.0 Hz), 7.38 (d,
1H, J = 8.0 Hz), 7.35 (d, 2H, J = 8.2 Hz), 7.23 (d, 2H,
J = 8.2 Hz), 6.46 (d, 1H, J = 5.4 Hz), 5.03 (m, 1H),
4.78 (s, 2H), 4.29 (s, 2H), 3.99 (m, 1H), 3.67 (m, 1H),
1.86 (m, 3H), 1.62 (m, 3H), 1.30 (s, 9H); FAB MS m/z
449 (M+H)+.
oxoisoindolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)ac-
rylamide (29aa). This compound was prepared from
28aa by means of a procedure similar to that used for
11 (77%). 1H NMR (500 MHz, CDCl3) d 9.37 (br,
1H), 8.03 (s, 1H), 7.75 (d, 1H, J = 5.0 Hz), 7.56 (d,
1H, J = 7.6 Hz), 7.31 (m, 11H), 6.50 (d, 1H,
J = 5.0 Hz), 5.79 (dd, 1H, J = 5.2, 7.3 Hz), 5.05 (m,
1H), 4.62 (d, 1H, J = 12.2 Hz), 4.54 (d, 1H,
J = 12.2 Hz), 4.45 (d, 1H, J = 17.7 Hz), 4.17 (d, 1H,
J = 17.7 Hz), 4.12 (dd, 1H, J = 7.3, 10.4 Hz), 4.03 (dd,
5.1.99.
(E)-3-(1-Oxo-2-(1,2-diphenylethyl)isoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29w).
This compound was prepared from 28w by means of a
procedure similar to that used for 11 (99%). H NMR
1H, J = 5.2, 10.4 Hz), 4.00 (m, 1H), 3.64 (m, 1H), 1.83
ꢁ52.3ꢁ (c 0.146, CH3CN);
34:5
D
1
(m, 3H), 1.59 (m, 3H); ½aꢀ
(500 MHz, CDCl3) d 8.79 (s, 1H), 7.92 (s, 1H), 7.72
(d, 1H, J = 5.0 Hz), 7.53 (d, 1H, J = 7.2 Hz), 7.45 (d,
2H, J = 7.3 Hz), 7.26 (m, 8H), 7.14 (t, 1H, J = 7.3 Hz),
6.38 (d, 1H, J = 5.0 Hz), 5.96 (dd, 1H, J = 6.4, 9.8 Hz),
5.02 (m, 1H), 4.38 (d, 1H, J = 17.1 Hz), 4.08 (d, 1H,
J = 17.1 Hz), 3.99 (m, 1H), 3.67 (m, 1H), 3.50 (dd, 1H,
J = 6.4, 14.5 Hz), 3.42 (dd, 1H, J = 9.8, 14.5 Hz), 1.86
(m, 3H), 1.61 (m, 3H); FAB MS m/z 483 (M+H)+.
FAB MS m/z 513 (M+H)+.
5.1.104. (E)-3-(1-Oxo-2-phenethylisoindolin-6-yl)-N-
hydroxyacrylamide (30a). This compound was prepared
from 29a by means of a procedure similar to that used
for 12 (80%). Mp 180–182 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 7.70 (s, 1H), 7.69 (d, 1 H, J = 5.9 Hz),
7.52 (d, 1H, J = 7.9 Hz), 7.25 (m, 6H), 6.51 (d, 1H,
J = 5.9 Hz), 4.38 (s, 2H), 3.78 (t, 2H, J = 7.3 Hz), 2.92
(t, 2H, J = 7.3 Hz); HR FAB MS: (M+H)+ calcd for
C19H19N2O3, 323.1396. Found: 323.1421.
5.1.100. (E)-3-(1-Oxo-2-(1,3-diphenylpropyl)isoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29x).
This compound was prepared from 28x by means of a
1
procedure similar to that used for 11 (61%). H NMR
5.1.105. (E)-3-(1-Oxo-2-(3-phenylpropyl)isoindolin-6-yl)-
N-hydroxyacrylamide (30b). This compound was pre-
pared from 29b by means of a procedure similar to that
used for 12 (49%). Mp 117–119 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 7.81 (s, 1H), 7.76 (d, 1H, J = 7.9 Hz), 7.60
(d, 1H, J = 7.9 Hz), 7.53 (d, 1H, J = 5.9 Hz), 7.21 (m,
5H), 6.55 (d, 1H, J = 5.9 Hz), 4.50 (s, 2H), 3.54 (t, 2H,
J = 7.3 Hz), 2.60 (t, 2H, J = 7.9 Hz), 1.91 (m, 2H); HR
FAB MS: (M+H)+ calcd for C20H21N2O3, 337.1552.
Found: 337.1552.
(500 MHz, CDCl3) d 9.20 (br, 1H), 8.04 (s, 1H), 7.78
(d, 1H, J = 5.3 Hz), 7.57 (d, 1H, J = 7.7 Hz), 7.40 (m,
2H), 7.34 (m, 3 H), 7.25 (m, 3H), 7.15 (m, 3H), 6.50
(d, 1H, J = 5.3 Hz), 5.67 (dd, 1H, J = 6.9, 8.6 Hz), 5.05
(b. 1H), 4.30 (d, 1H, J = 17.3 Hz), 4.02 (m, 1H,
J = 17.3 Hz), 4.01 (m, 1H), 3.67 (m, 1H), 2.72 (m, 1H),
2.66 (m, 1H), 2.42 (m, 2H), 1.85 (m, 3H), 1.60 (m,
3H); FAB MS m/z 497 (M+H)+.
5.1.101. (E)-3-(1-Oxo-2-(1,4-diphenylbutyl)isoindolin-6-
yl)-N-(tetrahydro-2H-pyran-2-yloxy)acrylamide (29y).
This compound was prepared from 28y by means of a
5.1.106. (E)-3-(2-(Cyclohexylbenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30c). This compound was
prepared from 29c by means of a procedure similar
to that used for 12 (95%). Mp >300 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.77 (s, 1H), 9.09 (s, 1H),
7.78 (s, 1H), 7.74 (d, 1H, J = 7.3 Hz), 7.58 (d, 1H,
J = 7.3 Hz), 7.48 (d, 1H, J = 5.9 Hz), 6.56 (d, 1H,
J = 5.9 Hz), 4.47 (s, 2H), 3.35 (d, 2H, J = 7.0 Hz,),
1.61 (m, 6H), 1.17 (m, 3H), 0.95 (m, 2H); HR FAB
MS: (M+H)+ calcd for C18H23N2O3, 315.1709. Found:
315.1742.
1
procedure similar to that used for 11 (81%). H NMR
(500 MHz, CDCl3) d 9.55 (br, 1H), 8.02 (s, 1H), 7.77
(d, 1H, J = 5.8 Hz), 7.54 (d, 1H, J = 6.4 Hz), 7.31 (m,
5H), 7.23 (m, 3H), 7.15 (t, 1H, J = 7.3 Hz), 7.14 (d, 2H,
J = 7.3 Hz), 6.54 (d, 1H, J = 5.8 Hz), 5.63 (dd, 1H,
J = 6.6, 9.2 Hz), 5.07 (m, 1H), 4.18 (d, 1H,
J = 17.1 Hz), 4.01 (m, 1H), 3.95 (d, 1H, J = 17.1 Hz),
3.65 (m, 1H), 2.69 (m, 1H), 2.67 (m, 1H), 2.09 (m, 2H),
1.85 (m, 3H), 1.65 (m, 5H); FAB MS m/z 511 (M+H)+.
5.1.102.
(E)-(S)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
5.1.107. (E)-3-(2-((Naphthalene-1-yl)methyl)-1-oxoisoin-
dolin-6-yl)-N-hydroxyacrylamide (30d). This compound
was prepared from 29d by means of a procedure similar
to that used for 12 (29%). Mp >300 ꢁC; 1H NMR
oxoisoindolin-6-yl)-N-(tetrahydro-2H-pyran-2-yloxy)ac-
rylamide (29z). This compound was prepared from 28z
by means of a procedure similar to that used for 11

7644
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
(500 MHz, DMSO-d6) d 8.21 (d, 1H, J = 9.1 Hz), 7,95
(d, 1H, J = 7,4 Hz), 7.91 (dd, 1H, J = 4.9, 4.9 Hz), 7.81
(s, 1H), 7.68 (d, 1H, J = 7.4 Hz), 7.52 (m, 5H), 7.29 (d,
1H, J = 7.1 Hz), 6.53 (d, 1H, J = 7.1 Hz), 5.18 (s, 2H),
4.25 (s, 2H); HR FAB MS: (M+H)+ calcd for
C22H19N2O3, 359.1396. Found: 359.1370.
5.1.113. (E)-3-(2-(4-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-hydroxyacrylamide (30j). This compound
was prepared from 29j by means of a procedure similar
to that used for 12 (35%). Mp >300 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.83 (s, 1H), 9.12 (s, 1H),
7.81 (s, 1H), 7.72 (d, 1H, J = 7.3 Hz, 1H), 7.71 (d, 2H,
J = 8.0 Hz), 7.54 (d, 1H, J = 7.3 Hz), 7.48 (d, 2H,
J = 8.0 Hz), 7.35 (d, 1H, J = 5.6 Hz), 6.58 (d, 1H,
J = 5.6 Hz), 4.82 (s, 2H), 4.41 (s, 2H); HR FAB MS:
(M+H)+ calcd for C19H16F3N2O3, 377.1113. Found:
377.1094.
5.1.108. (E)-3-(2-(2-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30e). This compound was pre-
pared from 29e by means of a procedure similar to that
1
used for 12 (67%). Mp >300 ꢁC; H NMR (500 MHz,
DMSO-d6) d 10.74 (s, 1H), 9.07 (s, 1H), 7.87 (s, 1H),
7.78 (d, 1H, J = 7.9 Hz), 7.60 (d, 1H, J = 7.9 Hz), 7.55
(d, 1H, J = 5.9 Hz), 7.49 (m, 1H), 7.33 (m, 2H), 7.26
(m, 1H), 6.57 (d, 1H, J = 5.9 Hz), 4.82 (s, 2H), 4.42 (s,
2H); HR FAB MS: (M+H)+ calcd for C18H16ClN2O3,
3 43.0849. Found: 343.0898.
5.1.114. (E)-3-(2-(2-Methoxybenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30k). This compound was
prepared from 29k by means of a procedure similar to
that used for 12 (78%). Mp >300 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.95 (s, 1H), 9.03 (s, 1H),
7.86 (s, 1H), 7.70 (d, 1H, J = 7.3 Hz), 7.53 (d, 1H,
J = 7.3 Hz), 7.28 (d, 1H, J = 7.7 Hz), 7.27 (dd, 1H,
J = 7.3, 7.3 Hz,), 7.07 (d, 1H, J = 7.3 Hz),7.03 (d, 1H,
J = 7.9 Hz), 6.89 (dd, 1H, J = 7.3, 7.9 Hz), 6.56 (d, 1H,
J = 7.7 Hz), 4.68 (s, 2H), 4.37 (s, 2H), 3.82 (s, 3H) HR
FAB MS: (M+H)+ calcd for C19H19F3N2O4, 339.1345.
Found: 339.1367.
5.1.109. (E)-3-(2-(3-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30f). This compound was pre-
pared from 29f by means of a procedure similar to that
used for 12 (35%). Mp 147–149 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.76 (s, 1H), 9.09 (s, 1H), 7.86 (s, 1H),
7.77 (d, 1H, J = 7.3 Hz), 7.58 (d, 2H, J = 7.9 Hz), 7.51
(d, 1H, J = 5.9 Hz), 7.36 (m, 3H), 7.23 (d, 1H,
J = 7.3 Hz), 6.58 (d, 1H, J = 5.9 Hz), 4.73 (s, 2H), 4.41
(s, 2H); HR FAB MS: (M+H)+ calcd for
C18H16ClN2O3, 343.0849. Found: 343.0865.
5.1.115. (E)-3-(2-(3-Methoxybenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30l). This compound was pre-
pared from 29l by means of a procedure similar to that
used for 12 (46%). Mp 177–179 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.73 (s, 1H), 9.07 (s, 1H), 7.86 (s, 1H),
7.77 (d, 1H, J = 7.9 Hz), 7.57 (d, 1H, J = 7.9 Hz), 7.54
(d, 1H, J = 5.9 Hz), 7.26 (dd, 1H, J = 7.9, 7.9 Hz), 6.85
(d, 1H, J = 7.9 Hz), 6.84 (s, 1H), 6.83 (d, 1H,
J = 7.9 Hz), 6.56 (d, 1H, J = 5.9 Hz), 4.69 (s, 2H), 4.38
(s, 2H), 3.72 (s, 3H); HR FAB MS: (M+H)+ calcd for
C19H19F3N2O4, 339.1345. Found: 339.1312.
5.1.110. (E)-3-(2-(4-Chlorobenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30g). This compound was pre-
pared from 29g by means of a procedure similar to that
1
used for 12 (67%). Mp >300 ꢁC; H NMR (500 MHz,
DMSO-d6) d 7.82 (s, 1H), 7.74 (d, 1H, J = 7.3 Hz,),
7.56 (d, 1H, J = 7.3 Hz), 7.40 (d, 2H, J = 8.2 Hz), 7.39
(d, 1H, J = 5.9 Hz), 7.29 (d, 2H, J = 8.2 Hz), 6.56 (d,
1H, J = 5.9 Hz), 4.72 (s, 2 H), 4.38 (s, 2H); HR FAB
MS: (M+H)+ calcd for C18H16ClN2O3, 343.0849.
Found: 343.0889.
5.1.116. (E)-3-(2-(4-Methoxybenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30m). This compound was
prepared from 29m by means of a procedure similar to
that used for 12 (22%). Mp 145–147 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.72 (s, 1H), 9.06 (s, 1H),
7.85 (s, 1H), 7.76 (d, 1H, J = 7.9 Hz), 7.57 (d, 1H,
J = 7.9 Hz), 7.54 (d, 1H, J = 5.9 Hz), 7.21 (d, 2H,
J = 8.5 Hz), 6.90 (d, 2H, J = 8.5 Hz), 6.56 (d 1H,
J = 5.9 Hz), 4.65 (s, 2H), 4.34 (s, 2H), 3.72 (s, 3H);
HR FAB MS: (M+H)+ calcd for C19H19F3N2O4,
339.1345. Found: 339.1368.
5.1.111. (E)-3-(2-(2-(trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-hydroxyacrylamide (30h). This compound
was prepared from 29h by means of a procedure similar
1
to that used for 12 (61%). Mp 188–190 ꢁC; H NMR
(500 MHz, DMSO-d6) d 10.71 (s, 1H), 9.15 (s, 1H),
7.88 (s, 1H), 7.79 (d, 1H, J = 8.0 Hz), 7.77 (d, 1H,
J = 7.3 Hz), 7.65 (d, 1H, J = 7.3 Hz), 7.61 (dd, 1H,
J = 7.3, 7.3 Hz), 7.51 (dd, 1H, J = 7.3, 7.3 Hz), 7.51 (d,
1H, J = 5.9 Hz), 7.34 (d, 1H, J = 8.0 Hz), 6.56 (d, 1H,
J = 5.9 Hz), 4.92 (s, 2H), 4.43 (s, 2H); HR FAB MS:
(M+H)+ calcd for C19H16F3N2O3, 377.1113. Found:
377.1089.
5.1.117. (E)-3-(2-(2-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30n). This compound was pre-
pared from 29n by means of a procedure similar to that
1
used for 12 (80%). Mp >300 ꢁC; H NMR (500 MHz,
5.1.112. (E)-3-(2-(3-(Trifluoromethyl)benzyl)-1-oxoisoin-
dolin-6-yl)-N-hydroxyacrylamide (30i). This compound
was prepared from 29i by means of a procedure similar
DMSO-d6) d 7.75 (s, 1H), 7.70 (d, 1H, J = 7.3 Hz),
7.52 (d, 1H, J = 7.3 Hz), 7.18 (d, 1H, J = 5.9 Hz), 7.15
(m, 4H), 6.52 (d, 1H, J = 5.9 Hz), 4.72 (s, 2H), 4.28 (s,
2H), 2.28 (s, 3H); HR FAB MS: (M+H)+ calcd for
C19H19N2O3, 323.1396. Found: 323.1386.
1
to that used for 12 (73%). Mp 179–181 ꢁC; H NMR
(500 MHz, DMSO-d6) d 10.75 (s, 1H), 9.07 (s, 1H),
7.87 (s, 1H), 7.78 (d, 1H, J = 8.0 Hz), 7.65 (m, 2H),
7.58 (m, 3H), 7.51 (d, 1H, J = 5.8 Hz), 6.56 (d, 1H,
J = 5.8 Hz), 4.83 (s, 2H), 4.43 (s, 2H); HR FAB MS:
(M+H)+ calcd for C19H16F3N2O3, 377.1113. Found:
377.1084.
5.1.118. (E)-3-(2-(3-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30o). This compound was pre-
pared from 29o by means of a procedure similar to that
1
used for 12 (80%). Mp >300 ꢁC; H NMR (500 MHz,

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7645
DMSO-d6) d 10.88 (s, 1H), 9.08 (s, 1H), 7.78 (s, 1H),
7.71 (d, 1H, J = 6.1 Hz), 7.52 (d, 1H, J = 6.1 Hz), 7.31
(d, 1H, J = 5.7 Hz), 7.22 (dd, 1H, J = 7.9, 7.9 Hz), 7.09
(d, 1H, J = 7.9 Hz), 7.08 (s, 1H), 7.05 (d, 1H,
J = 7.9 Hz), 6.57 (d, 1H, J = 5.7 Hz), 4.67 (s, 2H), 4.34
(s, 2H), 2.27 (s, 3H); HR FAB MS: (M+H)+ calcd for
C19H19N2O3, 323.1396. Found: 323.1416.
(M+H)+ calcd for C22H25N2O3, 365.1865. Found:
365.1891.
5.1.124. (E)-3-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30u). This compound was pre-
pared from 29u by means of a procedure similar to that
used for 12 (51%). Mp 169–170 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.69 (br, 1H), 9.13 (br, 1H), 7.85 (s, 1H),
7.76 (d, 1H, J = 8.0 Hz), 7.56 (d, 1H, J = 8.0 Hz), 7.51
(d, 1H, J = 5.8 Hz), 7.36 (d, 2H, J = 7.9 Hz), 7.19 (d,
2H, J = 7.9 Hz), 6.54 (d, 1H, J = 5.8 Hz), 4.68 (s, 2H),
4.37 (s, 2H), 1.25 (s, 9H); HR FAB MS: (M+H)+ calcd
for C22H25N2O3, 365.1865. Found: 365.1842.
5.1.119. (E)-3-(2-(4-Methylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30p). This compound was pre-
pared from 29p by means of a procedure similar to
that used for 12 (69%). Mp >300 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 7.77 (s, 1H), 7.71 (d, 1H,
J = 6.7 Hz), 7.52 (d, 1H, J = 6.7 Hz), 7.29 (d, 1H,
J = 8.3 Hz,), 7.15 (s, 4H), 6.53 (d, 1H, J = 8.3 Hz),
4.67 (s, 2H), 4.32 (s, 2H), 2.26 (s, 3H); HR FAB
MS: (M+H)+ calcd for C19H19N2O3, 323.1396. Found:
323.1362.
5.1.125.
(E)-3-(2-Benzhydryl-1-oxoisoindolin-6-yl)-N-
hydroxyacrylamide (30v). This compound was prepared
from 29v by means of a procedure similar to that used
for 12 (73%). Mp 122–124 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.76 (s, 1H), 9.09 (s, 1H), 7.89 (s, 1H),
7.78 (d, 1H, J = 8.0 Hz), 7.58 (d, 1H, J = 8.0 Hz), 7.56
(d, 1H, J = 5.8 Hz), 7.39 (d, 2H, J = 7.3 Hz), 7.37 (d,
2H, J = 7.3 Hz), 7.33 (t, 2H, J = 7.3 Hz), 7.18 (dd,
4 H, J = 7.3, 7.3 Hz), 6.65 (s, 1H), 6.57 (d, 1H,
J = 5.8 Hz), 4.28 (s, 2H); HR FAB MS: (M+H)+ calcd
for C24H21N2O3, 385.1552. Found: 385.1528.
5.1.120. (E)-3-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30q). This compound was pre-
pared from 29q by means of a procedure similar to that
used for 12 (88%). Mp 187–189 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 7.77 (s, 1H), 7.71 (d, 1H, J = 7.4 Hz), 7.50
(d, 1H, J = 7.4 Hz), 7.43 (m, 3H), 7.36 (m, 5H), 7.25 (m,
1H), 7.21 (m, 1H), 6.53 (d, 1H, J = 5.9 Hz), 4.69 (s, 2H),
4.22 (s, 2H); HR FAB MS: (M+H)+ calcd for
C24H21N2O3, 385.1552. Found: 385.1528.
5.1.126. (E)-3-(1-Oxo-2-(1,2-diphenylethyl)isoindolin-6-
yl)-N-hydroxyacrylamide (30w). This compound was
prepared from 29w by means of a procedure similar to
that used for 12 (68%). Mp 136–137 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.73 (s, 1H), 9.07 (s, 1H),
7.73 (s, 1H), 7.72 (d, 1H, J = 7.4 Hz), 7.53 (d, 1H,
J = 7.4 Hz), 7.48 (d, 1H, J = 5.8 Hz), 7.45 (d, 2H,
J = 7.3 Hz), 7.36 (dd, 2H, J = 7.3, 7.9 Hz), 7.28 (d, 2H,
J = 7.3 Hz), 7.27 (t, 1 H, J = 7.9 Hz), 7.20 (dd, 2H,
J = 7.3, 7.9 Hz), 7.11 (t, 1 H, J = 7.9 Hz), 6.50 (d, 1H,
J = 5.8 Hz), 5.74 (t, 1H, J = 8.2 Hz), 4.58 (d, 1H,
J = 18.3 Hz), 4.12 (d, 1H, J = 18.3 Hz), 3.44 (d, 2H,
J = 8.2 Hz); HR FAB MS: (M+H)+ calcd for
C25H23N2O3, 399.1709. Found: 399.1744.
5.1.121. (E)-3-(2-(3-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30r). This compound was pre-
pared from 29r by means of a procedure similar to that
1
used for 12 (93%). Mp >300 ꢁC; H NMR (500 MHz,
DMSO-d6) d 7.79 (s, 1H), 7.71 (d, 1H, J = 7.5 Hz),
7.63 (d, 2H, J = 7.3 Hz), 7.58 (s, 1H), 7.57 (d, 1H,
J = 5.5 Hz), 7.53 (d, 1H, J = 7.5 Hz), 7.45 (dd, 2H,
J = 7.3, 7.3 Hz), 7.44 (dd, 1H, J = 7.5, 7.9 Hz), 7.44 (d,
1H, J = 7.9 Hz), 7.35 (t, 1H, J = 7.3 Hz), 7.26 (d, 1H,
J = 7.5 Hz), 6.53 (d, 1H, J = 5.5 Hz), 4.80 (s, 2H), 4.41
(s, 2H); HR FAB MS: (M+H)+ calcd for C24H21N2O3,
385.1552. Found: 385.1555.
5.1.127. (E)-3-(1-Oxo-2-(1,3-diphenylpropyl)isoindolin-6-
yl)-N-hydroxyacrylamide (30x). This compound was
prepared from 29x by means of a procedure similar to
that used for 12 (69%). Mp 133–134 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 9.12 (br, 1H), 7.84 (s, 1H),
7.75 (d, 1H, J = 8.0 Hz), 7.56 (d, 2H, J = 8.0 Hz), 7.51
(d, 1H, J = 5.8 Hz), 7.36 (m, 3H), 7.23 (m, 5 H), 7.16
(t, 1H, J = 6.8 Hz), 6.54 (d, 1H, J = 5.8 Hz), 5.40 (dd,
1H, J = 7.7, 9.8 Hz), 4.58 (d, 1H, J = 18.2 Hz), 4.11 (d,
1H, J = 18.2 Hz), 2.57 (m, 2H), 2.38 (m, 2H) ; HR
FAB MS: (M+H)+ calcd for C26H25N2O3, 413.1865.
Found: 413.1848.
5.1.122. (E)-3-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl)-
N-hydroxyacrylamide (30s). This compound was pre-
pared from 29s by means of a procedure similar to that
used for 12 (66%). Mp 176–178 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 7.84 (s, 1H), 7.75 (d, 1H, J = 7.4 Hz), 7.63
(d, 1H, J = 5.9 Hz), 7.63 (m, 4H), 7.56 (d, 1H,
J = 7.4 Hz), 7.44 (m, 2H), 7.36 (m, 3H), 6.57 (d, 1H,
J = 5.9 Hz), 4.77 (s, 2H), 4.41 (s, 2H); HR FAB MS:
(M+H)+ calcd for C24H21N2O3, 385.1552. Found:
385.1542.
5.1.123. (E)-3-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-
yl)-N-hydroxyacrylamide (30t). This compound was pre-
pared from 29t by means of a procedure similar to that
used for 12 (60%). Mp 173–174 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.71 (s, 1H), 9.21 (s, 1H), 7.88 (s, 1H),
7.77 (d, 1H, J = 7.9 Hz), 7.57 (d, 1H, J = 7.9 Hz), 7.50
(d, 1H, J = 5.3 Hz), 7.40 (d, 1H, J = 7.0 Hz), 7.20 (dd,
1H, J = 7.0, 7.0 Hz), 7.15 (dd, 1H, J = 7.0, 7.0 Hz),
7.05 (d, 1H, J = 7.0 Hz), 6.57 (d, 1H, J = 5.3 Hz), 4.99
(s, 2H), 4.34 (s, 2H), 1.99 (s, 9H); HR FAB MS:
5.1.128. (E)-3-(1-Oxo-2-(1,4-diphenylbutyl)isoindolin-6-
yl)-N-hydroxyacrylamide (30y). This compound was pre-
pared from 29y by means of a procedure similar to that
used for 12 (27%). Mp 153–154 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 7.68 (s, 1H), 7.67 (d, 1H, J = 7.0 Hz), 7.47
(d, 1H, J = 7.9 Hz), 7.33 (m, 5H), 7.23 (m, 3H), 7.15 (d,
3H), 6.48 (d, 1H, J = 7.0 Hz), 5.41 (dd, 1H, J = 6.1,
9.2 Hz), 4.39 (d, 1H, J = 17.8 Hz), 4.01 (d, 1H,
J = 17.8 Hz), 2.63 (m, 2H), 2.06 (m, 2H), 1.53 (m, 2H);

7646
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
HR FAB MS: (M+H)+ calcd for C27H27N2O3, 427.2022.
Found: 427.2071.
HR FAB MS: (M+H)+ calcd for C24H19N2O4,
399.1345. Found: 399.1352.
5.1.129.
(E)-(S)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
5.1.135. (E)-3-(2-(2,2-Diphenylethyl)-1,3-dioxoisoindolin-
5-yl)-N-hydroxyacrylamide (36e). Mp 199–201 ꢁC;
1HNMR (500 MHz, DMSO) d 10.81 (s, 1H), 9.16 (s,
1H), 7.95 (s, 1H), 7.93 (d, 1H, J = 7.9 Hz), 7.80 (d,
1H, J = 7.9 Hz), 7.57 (d, 1H, J = 5.9 Hz), 7.33 (d, 4H,
J = 7.3 Hz), 7.25 (dd, 4H, J = 7.3 Hz), 7.17 (d, 2H,
J = 7.3 Hz), 6.66 (d, 1H, J = 5.9 Hz), 4.57 (t, 1H,
J = 8.0 Hz), 4.23 (d, 2H, J = 8.0 Hz); HR FAB MS:
(M+H)+ calcd for C25H21N2O4, 413.1501. Found:
413.1541.
oxoisoindolin-6-yl)-N-hydroxyacrylamide (30z). This
compound was prepared from 29z by means of a proce-
dure similar to that used for 12 (85%). Mp 102–104 ꢁC;
1H NMR (500 MHz, DMSO-d6) d 10.72 (s, 1H), 9.03
(s, 1H), 7.80 (s, 1H), 7.74 (d, 1H, J = 7.9 Hz), 7.56 (d,
1H, J = 7.9 Hz), 7.41 (d, 1H, J = 6.4 Hz), 7.30 (m,
10H), 6.54 (d, 1H, J = 6.4 Hz), 5.59 (dd, 1H, J = 5.5,
8.5 Hz), 4.59 (d, 1H, J = 12.2 Hz), 4.53 (d, 1H,
J = 18.0 Hz), 4.52 (d, 1H, J = 12.2 Hz), 4.30 (d, 1H,
J = 18.0 Hz), 4.11 (dd, 1H, J = 8.5, 10.3 Hz), 3.99 (dd,
1H, J = 5.5, 10.3 Hz); ½aꢀ
28:5
D
+63.5ꢁ (c 0.158, CHCl3);
5.1.136.
(E)-(S)-Hydroxy-3-(1,3-dioxo-2-(1-phenyleth-
HR FAB MS: (M+H)+ calcd for C26H25N2O4,
429.1814. Found: 429.1792.
1
yl)isoindolin-5-yl)acrylamide (36f). Mp 137–138 ꢁC; H
NMR (500 MHz, DMSO-d6) d 10.84 (s, 1H), 9.17 (s,
1H), 8.01 (s, 1H), 8.00 (d, 1H, J = 7.9 Hz), 7.86 (d,
1H, J = 7.9 Hz), 7.60 (d, 1H, J = 5.8 Hz), 7.32 (m,
5H), 6.68 (d, 1H, J = 5.8 Hz), 5.45 (q, 1H, J = 7.3 Hz),
1.81 (d, 3H, J = 7.3 Hz): HR FAB MS: (M+H)+ calcd
for C19H17N2O4 337.1188. Found: 337.1196.
5.1.130.
(E)-(R)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1-
oxoisoindolin-6-yl)-N-hydroxyacrylamide (30aa). This
compound was prepared from 29aa by means of a pro-
cedure similar to that used for 12 (42%). Mp 102–
104 ꢁC; 1H NMR (500 MHz, DMSO-d6) d 10.72 (s,
1H), 9.03 (s, 1H), 7.80 (s, 1H), 7.74 (d, 1H,
J = 7.9 Hz), 7.56 (d, 1H, J = 7.9 Hz), 7.41 (d, 1H,
J = 6.4 Hz), 7.30 (m, 10H), 6.54 (d, 1H, J = 6.4 Hz),
5.59 (dd, 1H, J = 5.5, 8.5 Hz), 4.59 (d, 1H,
J = 12.2 Hz), 4.53 (d, 1H, J = 18.0 Hz), 4.52 (d, 1H,
5.1.137. (E)-(R)-Hydroxy-3-(1,3-dioxo-2-(1-phenyleth-
yl)isoindolin-5-yl)acrylamide (36g). Mp 136–138 ꢁC;
1HNMR (500 MHz, DMSO-d6) d 10.82 (s, 1H), 9.17
(s, 1H), 8.01 (s, 1H), 8.00 (d, 1H, J = 7.9 Hz), 7.86 (d,
1H, J = 7.9 Hz), 7.60 (d, 1H, J = 5.8 Hz), 7.32 (m,
5H), 6.68 (d, 1H, J = 5.8 Hz), 5.45 (q, 1H, J = 7.3 Hz),
1.81 (d, 3H, J = 7.3 Hz): HR FAB MS : (M+H)+ calcd
for C19H17N2O4 337.1188. Found: 337.1190.
J = 12.2 Hz), 4.30 (d, 1H, J = 18.0 Hz), 4.11 (dd, 1H,
J = 8.5, 10.3 Hz), 3.99 (dd, 1H, J = 5.5, 10.3 Hz); ½aꢀ
28:5
D
ꢁ86.3ꢁ (c 0.143, CHCl3); HR FAB MS: (M+H)+ calcd
for C26H25N2O4, 429.1814. Found: 429.1822.
5.1.138. (E)-(S)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1,3-
dioxoisoindolin-5-yl)-N-hydroxyacrylamide (36h). Mp
84–86 ꢁC; HNMR (500 MHz, CDCl3) d 9.33 (s, 1H),
7.94 (s, 1H), 7.79 (d, 1H, J = 8.0 Hz), 7.74 (d, 1H,
J = 5.5 Hz), 7.72 (d, 1H, J = 8.0 Hz), 7.48 (m, 2H),
7.26 (m, 8H), 6.46 (d, 1H, J = 5.5 Hz), 5.61 (dd, 1H,
5.1.131.
(E)-3-(2-Phenyl-1,3-dioxoisoindolin-5-yl)-N-
1
1
hydroxyacrylamide (36a). Mp 199–201 ꢁC; HNMR(500
MHz, DMSO-d6) d 10.85 (s, 1H), 9.19 (s, 1H), 8.13 (d,
1H, J = 0.6 Hz), 8.06 (dd, 1H, J = 0.6, 7.4 Hz), 7.97 (d,
1H, J = 7.4 Hz), 7.65 (d, 1H, J = 5.9 Hz), 7.52 (m,
2H), 7.44 (m, 3H), 6.74 (d, 1H, J = 5.9 Hz); HR FAB
MS: (M+H)+ calcd for C17H13N2O4, 309.0875. Found:
309.0865.
J = 5.5, 10.3 Hz), 4.65 (dd, 1H, J = 10.3, 10.3 Hz) 4.57
+5.47ꢁ (c
25:5
D
(s, 2H), 4.02 (dd, 1H, J = 5.5, 10.3 Hz); ½aꢀ
0.150, CH3CN); HR FAB MS: (M+H)+ calcd for
C26H23N2O5, 443.1607. Found: 443.1643.
5.1.132.
(E)-3-(2-Benzyl-1,3-dioxoisoindolin-5-yl)-N-
hydroxyacrylamide (36b). Mp 163–166 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 10.84 (s, 1H), 9.17 (s, 1H), 8.05
(s, 1H), 8.00 (d, 1H, J = 8.0 Hz), 7.90 (d, 1H,
J = 8.0 Hz), 7.62 (d, 1H, J = 5.9 Hz), 7.30 (m, 5H), 6.70
(d, 1H, J = 5.9 Hz), 4.77 (s, 2H); HR FAB MS: (M+H)+
calcd for C18H15N2O4, 323.1032. Found: 323.1038.
5.1.139. (E)-(R)-3-(2-(2-(Benzyloxy)-1-phenylethyl)-1,3-
dioxoisoindolin-5-yl)-N-hydroxyacrylamide (36i). Mp
84–86 ꢁC; H NMR (500 MHz, CDCl3) d 9.33 (s, 1H),
7.94 (s, 1H), 7.79 (d, 1H, J = 8.0 Hz), 7.74 (d, 1H,
J = 5.5 Hz), 7.72 (d, 1H, J = 8.0 Hz), 7.48 (m, 2H),
7.26 (m, 8H), 6.46 (d, 1H, J = 5.5 Hz), 5.61 (dd, 1H,
1
J = 5.5, 10.3 Hz), 4.65 (dd, 1H, J = 10.3, 10.3 Hz) 4.57
(s, 2H), 4.02 (dd, 1H, J = 5.5, 10.3 Hz); ½aꢀ
5.1.133. (E)-3-(2-Phenethyl-1,3-dioxoisoindolin-5-yl)-N-
hydroxyacrylamide (36c). Mp 187–189 ꢁC; 1HNMR (500
MHz, DMSO-d6) d 10.78 (s, 1H), 9.28 (s, 1H), 7.99 (s,
1H), 7.96 (d, 1H, J = 7.9 Hz), 7.84 (d, 1H, J = 7.9 Hz),
7.57 (d, 1H, J = 5.9 Hz), 7.24 (m, 2H), 7.18 (m, 3H), 6.68
(d, 1H, J = 5.9 Hz), 3.81 (t, 1H, J = 7.3 Hz), 2.91 (t, 1H,
J = 7.3 Hz); HR FAB MS: (M+H)+ calcd for
C19H17N2O4, 337.1188. Found: 337.1209.
25:5
D
ꢁ5.37ꢁ
(c 0.149, CH3CN); HR FAB MS: (M+H)+ calcd for
C26H23N2O5, 443.1607. Found: 443.1638.
5.1.140. Methyl 5-hydroxy-2-methylbenzoate (37). To a
mixture of methyl 5-amino-2-methylbenzoate (626 mg,
3.79 mmol), 1 mL of concd H2SO4, and 16 mL of water,
cooled to below 0 ꢁC, was added an aqueous solution of
NaNO2 (262 mg, 3.79 mmol in mL of water), and the
mixture was stirred for a further 5 min. The solution
was added to a refluxing solution of copper (II) sulfate
(1.6 g in 28 mL water) and refluxed for 30 min. After
5.1.134. (E)-3-(2-Benzhydryl-1,3-dioxoisoindolin-5-yl)-N-
hydroxyacrylamide (36d). Mp 142–144 ꢁC; 1HNMR
(500 MHz, CDCl3), d 8.48(s, 1H), 7.96 (s, 1H), 7.80
(m, 3H), 7.36 (m, 10H), 6.72 (s, 1H), 6.47 (m, 1H);

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7647
cooling, the reaction mixture was extracted with CHCl3.
The product was re-extracted with 1 mol/L NaOH solu-
tion, acidified with 2 mol/L HCL, and extracted with
CHCl3. The extract was dried over anhydrous MgSO4
and evaporated. The residue was purified by silica gel
column chromatography (eluant n-hexane/AcOEt 2:1
1H, J = 8.3 Hz), 7.33 (d, 1H, J = 2.4 Hz), 7.27 (d, 1H,
J = 8.2 Hz), 7.20 (m, 1H), 7.17 (dd, 1H, J = 2.4,
8.2 Hz), 7.14 (m, 2H), 5.11 (s, 2H), 4.60 (s, 2H), 4.16
(s, 2H), 1.51 (s, 9H), 1.47 (s, 9H); FAB MS m/z 410
(M+H)+.
1
v/v) to afford 369 mg (59%) of the title compound. H
5.1.147. tert-Butyl 2-(2-(4-tert-butylbenzyl)-1-oxoisoindo-
lin-6-yloxy)acetate (40e). This compound was prepared
from 39 by means of a procedure similar to that used
NMR (500 MHz, CDCl3) d 7.41 (d, 1H, J = 2.8 Hz),
7.11 (d, 1H, J = 8.2 Hz), 6.91 (dd, 1H, J = 2.8, 8.2 Hz),
5.01 (m, 1H), 3.89 (s, 3H), 2.51 (s, 3H); FAB MS m/z
167 (M+H)+.
1
for 16 (43%). H NMR (500 MHz, CDCl3) d 7.34 (d,
2H, J = 8.3 Hz), 7.29 (d, 1H, J = 2.4 Hz), 7.27 (d, 1H,
J = 8.2 Hz), 7.22 (d, 2H, J = 8.3 Hz), 7.15 (dd, 1H,
J = 2.4, 8.2 Hz), 4.76 (s, 2H), 4.58 (s, 2H), 4.20 (s,
2H), 1.50 (s, 9H), 1.30 (s, 9H); FAB MS m/z 410
(M+H)+.
5.1.141. Methyl 5-((tert-butoxycarbonyl)methoxy)-2-
methylbenzoate (38). To a mixture of 37 (369 mg,
2.22 mmol) and potassium carbonate (460 mg,
3.33 mmol) in 10 mL of DMF was added t-butyl bro-
moacetate (360 ll, 2.44 mmol). The reaction mixture
was stirred for 1 day at 100 ꢁC. After cooling, the reac-
tion mixture was poured into water, extracted with
AcOEt, washed with dil HCl, dil NaOH, and brine,
dried over anhydrous MgSO4, and evaporated. The res-
idue was purified by silica gel column chromatography
(eluant n-hexane/AcOEt 8:1 to 5:1 v/v) to afford
573 mg (92%) of the title compound. 1H NMR
(500 MHz, CDCl3) d 7.43 (d, 1H, J = 2.4 Hz), 7.14 (d,
1H, J = 8.3 Hz), 6.98 (dd, 1H, J = 2.4, 8.3 Hz), 4.51 (s,
2H), 3.87 (s, 3H), 2.51 (s, 3H), 1.48 (s, 9H); FAB MS
m/z 281 (M+H)+.
5.1.148. 2-(2-Benzyl-1-oxoisoindolin-6-yl-oxy)acetic acid
(41a). This compound was prepared from 40a by means
1
of a procedure similar to that used for 28c (quant.). H
NMR (500 MHz, DMSO-d6) d 13.01 (s, 1H), 7.44 (d,
1H, J = 8.2 Hz), 7.34 (dd, 2H, J = 7.2, 7.2 Hz), 7.27 (t,
1H, J = 7.2 Hz), 7.25 (d, 2H, J = 7.2 Hz), 7.16 (dd, 1H,
J = 2.2, 8.2 Hz), 7.15 (d, 1H, J = 2.2 Hz), 4.77 (s, 2H),
4.71 (s, 2H), 4.28 (s, 2H); FAB MS m/z 298 (M+H)+.
5.1.149. 2-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
acetic acid (41b). This compound was prepared from 40b
by means of a procedure similar to that used for 28c
1
(quant.). H NMR (500 MHz, DMSO-d6) d 13.01 (s,
5.1.142. Methyl 5-((tert-butoxycarbonyl)methoxy)-2-
(bromomethyl)benzoate (39). This compound was pre-
pared from 39 by means of a procedure similar to that
1H), 7.44 (d, 1H, J = 8.2 Hz), 7.34 (dd, 2H, J = 7.2,
7.2 Hz), 7.27 (t, 1H, J = 7.2 Hz), 7.25 (d, 2H,
J = 7.2 Hz), 7.16 (dd, 1H, J = 2.2, 8.2 Hz), 7.15 (d, 1H,
J = 2.2 Hz), 4.77 (s, 2H), 4.71 (s, 2H), 4.28 (s, 2H);
FAB MS m/z 298 (M+H)+.
1
used for 15 (57%). H NMR (500 MHz, CDCl3) d 7.47
(d, 1H, J = 2.7 Hz), 7.37 (d, 1H, J = 8.6 Hz), 7.02 (dd,
1H, J = 2.7, 8.6 Hz), 4.92 (s, 2H), 4.54 (s, 2H), 3.93 (s,
3H), 1.49 (s, 9H); FAB MS m/z 358, 360 (M)+.
5.1.150. 2-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
acetic acid (41c). This compound was prepared from 40c
by means of a procedure similar to that used for 28c
5.1.143. tert-Butyl 2-(2-benzyl-1-oxoisoindolin-6-yloxy)-
acetate (40a). This compound was prepared from 39 by
means of a procedure similar to that used for 16 (55%).
1H NMR (500 MHz, CDCl3) d 7.26 (m, 7H), 7.11 (dd,
1H, J = 2.4, 8.2 Hz), 4.76 (s, 2H), 4.55 (s, 2H), 4.16 (s,
2H), 1.47 (s, 9H); FAB MS m/z 354 (M+H)+.
1
(quant.). H NMR (500 MHz, CDCl3) d 7.55 (d, 2H,
J = 7.8 Hz), 7.55 (d, 2H, J = 7.8 Hz), 7.50 (d, 1H,
J = 2.0 Hz), 7.43 (dd, 2H, J = 7.8, 7.8 Hz), 7.35 (d, 2H,
J = 7.8 Hz), 7.31 (d, 1H, J = 8.3 Hz), 7.23 (dd, 1H,
J = 2.0, 8.3 Hz), 7.17 (t, 1H, J = 7.8 Hz), 4.83 (s, 2H),
4.82 (s, 2H), 4.28 (s, 2H); FAB MS m/z 374 (M+H)+.
5.1.144. tert-Butyl 2-(2-(2-phenylbenzyl)-1-oxoisoindolin-
6-yloxy)acetate (40b). This compound was prepared
from 39 by means of a procedure similar to that used
5.1.151. 2-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)acetic acid (41d). This compound was prepared
from 40d by means of a procedure similar to that used
1
for 16 (80%). H NMR (500 MHz, CDCl3) d 7.22–7.44
(m, 11H), 7.12 (m, 1H), 4.79 (s, 2H), 4.57 (s, 2H), 4.00
1
for 28c (quant.). H NMR (500 MHz, CDCl3) d 11.00
(s, 2H), 1.50 (s, 9H); FAB MS m/z 430 (M+H)+.
(br, 1H), 7.52 (s, 1H), 7.40 (d, 1H, J = 8.0 Hz), 7.27
(d, 1H, J = 8.6 Hz), 7.20 (m, 2H), 7.13 (d, 1H,
J = 7.3 Hz), 7.08 (d, 1H, J = 8.0 Hz), 5.10 (s, 2H), 4.80
(s, 2H), 4.18 (s, 2H), 1.45 (s, 9H); FAB MS m/z 354
(M+H)+.
5.1.145. tert-Butyl 2-(2-(4-phenylbenzyl)-1-oxoisoindolin-
6-yloxy)acetate (40c). This compound was prepared
from 39 by means of a procedure similar to that used
1
for 16 (22%). H NMR (500 MHz, CDCl3) d 7.55 (m,
4H), 7.26–7.44 (m, 7H), 7.16 (m, 1H), 4.83 (s, 2H),
4.59 (s, 2H), 4.25 (s, 2H), 1.50 (s, 9H); FAB MS m/z
430 (M+H)+.
5.1.152. 2-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)acetic acid (41e). This compound was prepared
from 40e by means of a procedure similar to that used
1
for 28c (quant.). H NMR (500 MHz, CDCl3) d 12.02
5.1.146. tert-Butyl 2-(2-(2-tert-butylbenzyl)-1-oxoisoindo-
lin-6-yloxy)acetate (40d). This compound was prepared
from 39 by means of a procedure similar to that used
(br, 1H), 7.56 (d, 1H, J = 1.8 Hz), 7.38 (d, 2H,
J = 8.0 Hz), 7.31 (d, 1H, J = 8.6 Hz), 7.25 (m, 3H),
4.86 (s, 2H), 4.80 (s, 2H), 4.26 (s, 2H), 1.34 (s, 9H);
FAB MS m/z 354 (M+H)+.
1
for 16 (22%). H NMR (500 MHz, CDCl3) d 7.42 (d,

7648
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
5.1.153. 2-(2-Benzyl-1-oxoisoindolin-6-yloxy)-N-(tetra-
hydro-2H-pyran-2-yloxy)acetamide (42a). This com-
pound was prepared from 41a by means of a procedure
DMSO-d6) d 10.92 (br, 1H), 8.86 (br, 1H), 7.37 (m,
8H), 7.25 (m, 1H), 7.16 (m, 3H), 4.67 (s, 2H), 4.48 (s,
2H), 4.13 (s, 2H); HR FAB MS: (M+H)+ calcd for
C23H21N2O4, 389.1501. Found: 389.1548.
1
similar to that used for 11 (58%). H NMR (500 MHz,
CDCl3) d 9.17 (s, 1H), 7.40 (s, 1H), 7.30 (m, 6H), 7.10
(d, 1H, J = 8.0 Hz), 5.03 (m, 1H), 4.79 (s, 2H), 4.64 (s,
2H), 4.21 (s, 2H), 4.01 (m, 1H), 3.67 (m, 1H), 1.84 (m,
3H), 1.61 (m, 3H); FAB MS m/z 397 (M+H)+.
5.1.160. 2-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
N-hydroxyacetamide (43c). This compound was pre-
pared from 42c by means of a procedure similar to that
used for 12 (9%). Mp 157–158 ꢁC; H NMR (500 MHz,
1
5.1.154. 2-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
N-(tetrahydro-2H-pyran-2-yloxy)acetamide (42b). This
compound was prepared from 41b by means of a proce-
dure similar to that used for 11 (45%). 1H NMR
(500 MHz, CDCl3) d 9.12 (br, 1H), 7.35 (m, 11H),
7.08 (d, 1H, J = 6.4 Hz), 5.03 (m, 1H), 4.81 (s, 2H),
4.62 (s, 2H), 4.01 (s, 2H), 4.00 (m, 1H), 3.66 (m, 1H),
1.84 (m, 3H), 1.60 (m, 3H); FAB MS m/z 473 (M+H)+.
DMSO-d6) d 10.92 (br, 1H), 8.89 (br, 1H), 7.63 (m, 4H),
7.44 (m, 3H), 7.33 (m, 4H), 7.16 (m, 1H), 4.75 (s, 2H),
4.50 (s, 2H), 4.30 (s, 2H); HR FAB MS: (M+H)+ calcd
for C23H21N2O4, 389.1501. Found: 389.1529.
5.1.161. 2-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)-N-hydroxyacetamide (43d). This compound was
prepared from 42d by means of a procedure similar to
that used for 12 (41%). Mp 157–158 ꢁC; 1H NMR
(500 MHz, CD3OD) d 7.45 (d, 1H, J = 7.7 Hz), 7.44
(d, 1H, J = 8.4 Hz), 7.38 (d, 1H, J = 2.2 Hz), 7.26 (dd,
1H, J = 2.2, 8.4 Hz), 7.20 (dd, 1H, J = 7.7, 7.7 Hz),
7.14 (dd, 1H, J = 7.7, 7.7 Hz), 7.07 (d, 1H, J = 7.7 Hz),
5.10 (s, 2H), 4.64 (s, 2H), 4.26 (s, 2H), 1.48 (s, 9H);
HR FAB MS: (M+H)+ calcd for C21H25N2O4,
369.1814. Found: 369.1824.
5.1.155. 2-(2-(4-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
N-(tetrahydro-2H-pyran-2-yloxy)acetamide (42c). This
compound was prepared from 41c by means of a proce-
dure similar to that used for 11 (33%). 1H NMR
(500 MHz, CDCl3) d 8.57 (br, 1H), 7.33-7.57 (m,
11H), 7.11 (m, 1H), 4.84 (s, 2H), 4.65 (s, 2H), 4.27 (s,
2H), 1.29 (s, 9H); FAB MS m/z 445 (M+H)+.
5.1.156. 2-(2-(2-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)-N-(tetrahydro-2H-pyran-2-yloxy)acetamide (42d).
This compound was prepared from 41d by means of a
5.1.162. 2-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)-N-hydroxyacetamide (43e). This compound was
prepared from 42e by means of a procedure similar to
that used for 12 (61%). Mp 155–157 ꢁC; 1H NMR
(500 MHz, DMSO-d6) d 7.43 (d, 1H, J = 8.2 Hz), 7.38
(d, 2H, J = 8.3 Hz,), 7.35 (d, 1H, J = 2.6 Hz), 7.24 (dd,
1H, J = 2.6, 8.2 Hz), 7.21 (d, 2H, J = 8.3 Hz), 4.75 (s,
2H), 4.62 (s, 2H), 4.29 (s, 2H), 1.29 (s, 9H); HR FAB
MS: (M+H)+ calcd for C21H25N2O4, 369.1814. Found:
369.1807.
1
procedure similar to that used for 11 (31%). H NMR
(500 MHz, CDCl3) d 9.18 (s, 1H), 7.43 (m, 1H), 7.42
(s, 1H), 7.30 (d, 1H, J = 8.2 Hz), 7.20 (m, 1H), 7.13
(m, 2H), 7.11 (d, 1H, J = 8.2 Hz), 5.11 (s, 2H), 5.03
(m, 1H), 4.65 (s, 2H), 4.17 (s, 2H), 4.01 (m, 1H), 3.66
(m, 1H), 1.84 (m, 3H), 1.60 (m, 3H), 1.47 (s, 9H);
FAB MS m/z 453 (M+H)+.
5.1.157. 2-(2-(4-tert-Butylbenzyl)-1-oxoisoindolin-6-yl-
oxy)-N-(tetrahydro-2H-pyran-2-yloxy)acetamide (42e).
This compound was prepared from 41e by means of a
5.1.163. N-Benzyl-2-methyl-5-nitrobenzamide (45). To a
solution of 2-methyl-5-nitrobenzoic acid (500 mg,
2.76 mmol), triethylamine (0.846 mL, 6.07 mmol), and
5 mL of anhydrous THF, cooled to 0 ꢁC, was added eth-
yl chloroformate (0.264 mL, 2.76 mmol). The mixture
was stirred for 1 h, then benzylamine (0.362 mL, 3.31
mmol) was added and the whole was stirred overnight.
The reaction mixture was evaporated and the residue
was redissolved in AcOEt. This solution was washed
with brine, dried over anhydrous MgSO4, and evaporat-
ed. The residue was purified by silica gel column chro-
matography (eluant n-hexane/AcOEt 2:1 v/v) to afford
520 mg (70%) of the title compound. 1H NMR
(500 MHz, CDCl3) d 8.24 (d, 1H, J = 2.5 Hz), 8.17
(dd, 1H, J = 2.5, 8.6 Hz), 7.40 (d, 1H, J = 8.6 Hz), 7.38
(m, 5H), 6.12 (m, 1H), 4.65 (d, 2H, J = 5.8 Hz), 2.58
(s, 3H); FAB MS m/z 271 (M+H)+.
1
procedure similar to that used for 11 (25%). H NMR
(500 MHz, CDCl3) d 9.24 (s, 1H), 7.38 (s, 1H), 7.34
(d, 2H, J = 8.3 Hz), 7.28 (d, 1H, J = 8.2 Hz), 7.22 (d,
2H, J = 8.3 Hz), 7.09 (d, 1H, J = 8.2 Hz), 5.03 (m,
1H), 4.75 (s, 2H), 4.63 (s, 2H), 4.21 (s, 2H), 4.01 (m,
1H), 3.65 (m, 1H), 1.83 (m, 3H), 1.60 (m, 3H), 1.29 (s,
9H); FAB MS m/z 453 (M+H)+.
5.1.158. 2-(2-Benzyl-1-oxoisoindolin-6-yloxy)-N-hydroxyac-
etamide (43a). This compound was prepared from 42a
by means of a procedure similar to that used for 12
1
(29%). Mp 126–128 ꢁC; H NMR (500 MHz, DMSO-
d6) d 10.83 (s, 1H), 8.96 (s, 1H), 7.45 (d, 1H,
J = 8.5 Hz), 7.34 (dd, 2H, J = 7.3, 7.3 Hz), 7.27 (t, 1H,
J = 7.3 Hz), 7.25 (d, 2H, J = 7.3 Hz), 7.23 (d, 1H,
J = 2.5 Hz), 7.18 (dd, 1H, J = 2.5, 8.5 Hz), 4.71 (s,
2H), 4.54 (s, 2H), 4.28 (s, 2H); HR FAB MS: (M+H)+
calcd for C17H17N2O4, 313.1188. Found: 313.1218.
5.1.164. 5-Amino-N-benzyl-2-methylbenzamide (46). A
mixture of 45 (520 mg, 1.92 mmol), 48 mg of 10% Pd–
C 48 mg (0.846 mL, 6.07 mmol), and 6 mL of AcOEt
was hydrogenated at room temperature for 2 h. The
reaction mixture was filtered through Celite, and the fil-
trate was evaporated to afford 420 mg (91%) of the title
compound. 1H NMR (500 MHz, CDCl3) d 7.30 (m,
5H), 6.93(d, 1H, J = 7.9 Hz), 6.62 (d, 1H, J = 2.5 Hz),
5.1.159. 2-(2-(2-Phenylbenzyl)-1-oxoisoindolin-6-yl-oxy)-
N-hydroxyacetamide (43b). This compound was pre-
pared from 42b by means of a procedure similar to that
1
used for 12 (9%). Mp 126–128 ꢁC; HNMR (500 MHz,

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7649
6.56 (dd, 1H, J = 2.5, 7.9 Hz), 6.17 (m, 1H), 4.54 (d, 2H,
J = 5.8 Hz), 3.57 (br, 2H), 2.28 (s, 3H); FAB MS m/z 241
(M+H)+.
1H), 7.93 (m, 1H), 7.55 (m, 1H), 7.37 (m, 5H), 7.15(m,
1H), 4.50 (s, 2H), 3.07 (s, 3H); FAB MS m/z 254
(M+H)+.
5.1.165. tert-Butyl 3-(3-(benzylcarbamoyl)-4-methylphe-
nyl)-2-bromopropanoate (47). This compound was pre-
pared from 47 by means of a procedure similar to that
5.1.171. (E)-Methyl 3-(3-(N-benzyl-N-methylcarba-
moyl)phenyl)acrylate (54). To a mixture of diethy-
lphosphonoacetate (0.176 mL, 0.97 mmol), potassium
tert-butoxide (109 mg, 0.97 mmol), and 10 mL of
anhydrous THF was added a THF solution of 53
((223 mg, 0.88 mmol)/5 mL) under an Ar atmosphere,
and the whole was stirred overnight at room
temperature. The reaction mixture was evaporated and
the residue was redissolved in AcOEt. This solution
was washed with brine, dried over anhydrous MgSO4,
and evaporated. The residue was purified by silica gel
column chromatography (eluant n-hexane/AcOEt 1:1
v/v) to afford 214 mg (79%) of the title compound (mix-
1
used for 18 (85%). H NMR (500 MHz, CDCl3) d 7.36
(m, 5H), 7.23 (s, 1H), 7.16 (m, 2H), 6.00 (m, 1H), 4,62
(d, 2H, J = 5.5 Hz), 4.23 (dd, 1H, J = 7.0, 8.3 Hz), 3.39
(dd, 1H, J = 8.3, 14.1 Hz), 3.14 (dd, 1H, J = 7.0,
14.1 Hz,), 2.44 (s, 3H), 1.42 (s, 9H); FAB MS m/z 432,
434 (M+H)+.
5.1.166. (E)-tert-Butyl 3-(3-(benzylcarbamoyl)-4-methyl-
phenyl)acrylate (48). This compound was prepared
from 47 by means of a procedure similar to that used
for 19 (quant.). 1H NMR (500 MHz, CDCl3) d 7.50
(d, 1H, J = 5.8 Hz), 7.47 (s, 1H), 7.42 (d, 1H,
J = 7.9 Hz), 7.36 (m, 5H), 7.20 (d, 1H, J = 7.9 Hz),
6.30 (d, 1H, J = 5.8 Hz), 6.20 (m, 1H), 4.61 (d, 2H,
J = 5.5 Hz), 2.45 (s, 3H), 1.52 (s, 9H); FAB MS m/z
352 (M+H)+.
1
ture of regioisomers). H NMR (500 MHz, CDCl3) d
7.68 (d, 1H, J = 5.9 Hz), 7.60 (s, 1H), 7.56 (d, 1H,
J = 6.7 Hz), 7.37 (m, 6H), 7.16 (d, 1H, J = 6.0 Hz),
6.48 (d, 1H, J = 5.9 Hz), 4.76 (s, 2H), 3.80 (s, 3H),
2.86 (s, 3H), minor; d 7.65 (d, 1H, J = 15.5 HzH), 7.60
(s, 1H), 7.52 (d, 1H, J = 6.7 Hz), 7.37 (m, 6H), 7.17 (d,
1H, J = 6.3 Hz), 6.33 (d, 1H, J = 15.5 Hz), 4.50 (s,
2H), 3.80 (s, 3H), 3.06 (s, 3H); FAB MS m/z 310
(M+H)+.
5.1.167.
(E)-3-(3-(Benzylcarbamoyl)-4-methylphenyl)-
acrylic acid (49). This compound was prepared from
47 by means of a procedure similar to that used for
1
28c (quant.). H NMR (500 MHz, DMSO-d6) d 12.36
5.1.172.
(E)-3-(3-(N-Benzyl-N-methylcarbamoyl)phen-
(br, 1H), 8.86 (t, 1H, J = 6.3 Hz), 7.67 (s, 1H), 7.64 (d,
1H, J = 7.8 Hz), 7.58 (d, 1H, J = 5.6 Hz), 7.34 (m,
5H), 7.28 (d, 1H, J = 7.8 Hz), 6.52 (d, 1H, J = 5.6 Hz),
4.44 (d, 2H, J = 6.3 Hz), 2.34 (s, 3H); FAB MS m/z
296 (M+H)+.
yl)acrylic acid (55). This compound was prepared from
54 by means of a procedure similar to that used for 10
(75%). 1H NMR (500 MHz, CDCl3) d major; d 7.77
(d, 1H, J = 5.9 Hz), 7.64 (s, 1H), 7.59 (d, 1H,
J = 6.7 Hz), 7.37 (m, 6H), 7.17 (d, 1H, J = 6.3 Hz),
6.48 (d, 1H, J = 5.9 Hz), 4.78 (s, 2H), 2.88 (s, 3H),
minor; d 7.70 (d, 1H, J = 5.9 Hz), 7.64 (s, 1H), 7.56 (d,
1H, J = 6.7 Hz,), 7.37 (m, 6H), 7.17 (d, 1H,
J = 6.3 Hz), 6.33 (d, 1H, J = 5.9 Hz), 4.52 (s, 2H), 3.08
(s, 3H) FAB MS m/z 296 (M+H)+.
5.1.168. 5-((E)-2-(Tetrahydro-2H-pyran-2-yloxycarba-
moyl)vinyl)-N-benzyl-2-methylbenzamide (50). This com-
pound was prepared from 49 by means of a procedure
1
similar to that used for 11 (52%). H NMR (500 MHz,
CDCl3) d 8.96 (br, 1H), 7.58 (d, 1H, J = 5.5 Hz), 7.44
(s, 1H), 7.32 (m, 6H), 7.18 (d, 1H, J = 7.7 Hz), 6.41 (d,
1H, J = 5.5 Hz), 6.28 (br, 1H), 4.97 (m, 1H), 4.59 (d,
2H, J = 5.6 Hz), 3.96 (m, 1H), 3.63 (m, 1H), 2.43 (s,
3H), 1.81 (m, 3H), 1.63 (m, 3H); FAB MS m/z 395
(M+H)+.
5.1.173. 3-((E)-2-(Tetrahydro-2H-pyran-2-yloxycarba-
moyl)vinyl)-N-benzyl-N-methylbenzamide (56). This
compound was prepared from 55 by means of a
procedure similar to that used for 11 (37%). 1H
NMR (500 MHz, CDCl3) d major; d 8.99 (br, 1H),
7.38 (m, 8H), 7.16 (d, 1H, J = 6.4 Hz), 6.33 (br,
1H), 5.01 (m, 1H), 4.76 (s, 2H), 3.99 (m, 1H),
3.65 (m, 1H), 2.85 (s, 3H), 1.85 (m, 3H), 1.60 (m,
3H), minor; d 9.18 (br, 1H), 7.38 (m, 8H), 7.16
(d, 1H, J = 6.4 Hz), 6.08 (br, 1H), 5.01 (m, 1H),
4.50 (s, 2H), 3.99 (m, 1H), 3.65 (m, 1H), 3.07 (s,
3H), 1.85 (m, 3H), 1.60 (m, 3H); FAB MS m/z
395 (M+H)+.
5.1.169. 5-((E)-2-(Hydroxycarbamoyl)vinyl)-N-benzyl-2-
methylbenzamide (51). This compound was prepared
from 49 by means of a procedure similar to that used
for 12 (51%). Mp 172–173 ꢁC; 1H NMR (500 MHz,
DMSO-d6) d 10.69 (s, 1H), 9.04 (s, 1H), 8.89 (t, 1H,
J = 6.1 Hz), 7.53 (s, 1H), 7.50 (d, 1H, J = 8.1 Hz), 7.42
(d, 1H, J = 6.2 Hz), 7.35 (m, 3H), 7.27 (d, 2H,
J = 8.1 Hz), 7.25 (m, 1H), 6.45 (d, 1H, J = 6.2 Hz),
4.43 (d, 2H, J = 6.1 Hz), 2.33 (s, 3H); HR FAB MS:
(M+H)+ calcd for C18H19N2O3, 311.1396. Found:
311.1382.
5.1.174. 3-((E)-2-Hydroxyvinyl)-N-benzyl-N-methylben-
zamide (57). This compound was prepared from 56 by
means of a procedure similar to that used for 12
1
(78%). H NMR (500 MHz, DMSO-d6) major; d 10.75
5.1.170. N-Benzyl-3-formyl-N-methylbenzamide (53).
This compound was prepared from 52 by means of a
procedure similar to that used for 45 (34%). H NMR
(500 MHz, CDCl3) d 10.04 (s, 1H), 9.97 (s, 1H), 7.73
(m, 1H), 7.60 (m, 1H), 7.37 (m, 5H), 7.31(m, 1H), 4.77
(s, 2H), 2.88 (s, 3H), minor; d 9.98 (s, 1H), 9.97 (s,
(s, 1H), 9.08 (s, 1H), 7.17–7.63 (m, 10H), 6.52 (d, 1H,
J = 6.6 Hz), 4.68 (s, 2H), 2.82 (s, 3H), minor; d 10.75
(s, 1H), 9.08 (s, 1H), 7.17–7.63 (m, 10H), 6.46 (d, 1H,
J = 6.6 Hz), 4.46 (s, 2H), 2.89 (s, 3H); HR FAB MS:
(M+H)+ calcd for C18H19N2O3, 311.1396. Found:
311.1440.
1

7650
C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
5.2. HDAC activity assay kit
5.7. Molecular modeling
The assay of HDAC-inhibitory activity was performed
using an HDAC fluorescence activity assay/drug discov-
ery kit (AK-500, BIOMOL Research Laboratories),
according to the supplier’s protocol. The assay was per-
formed in triplicate and repeated at least two times. The
EC50 of SAHA (7) determined with this assay system
was reported to be 280 nM.20
5.7.1. Hardware & software. Molecular modeling studies
were carried out using QUANTA97 and CHARMm
(ver. 23.2) on a Silicon Graphics octane.
5.7.2. Reference PDB data. Construction of protein–
ligand complexes was based on an X-ray structure of
the HDAC 8 (PDB entry 1W22).
5.3. Other enzyme assays
5.7.3. Molecular modeling of human PPARa-ligand
complexes. Ligands were manually docked into the
active site based on the X-ray structure (PDB 1W22).
Minimizations were performed with CHARMm (energy
minimization condition; gradient convergence condition
is 0.01 kcal/molA, amino acid residues 5 A far from the
binding site fix, peptide main chain and Zn ion fix,
180His, 178Asp, 267Asp fix).
Assay of inhibitory activities against partially purified
HDAC1, 4, and 6 was done according to the reported
methods.19,21 HDAC1, 4, and 6 were used as represen-
tative class I, class IIa, and class IIb enzymes,
respectively.
˚
˚
In this study, kit assay was performed as an initial
screening method to evaluate the importance of the cyc-
lic amide/imide structure. The individual enzyme assays
were performed as a second screen to evaluate the rela-
tionship between structure and class-selective HDAC
inhibitory activity.
Acknowledgment
The work described in this paper was partially supported
by a Grant-in-aid for Scientific Research from The
Ministry of Education, Culture, Sports, Science and
Technology, Japan.
5.4. P21 promoter assay
P21 promoter assay was done according to the reported
methods.6,20
References and notes
5.5. Assay of growth-inhibitory activity toward human
prostate cancer cell line LNCap
1. Grunstein, M. Nature 1997, 389, 349.
2. Cheung, W. L.; Briggs, S. D.; Allis, C. D. Curr. Opin. Cell
Biol. 2000, 12, 326.
3. Roth, S.; Denu, J. M.; Allis, C. D. Annu. Rev. Biochem.
2001, 70, 81.
4. Cress, W. D.; Seto, E. J. Cell Physiol. 2000, 184, 1.
5. Rosato, R. R.; Wang, Z.; Gopalkrishnan, R. V.; Fisher,
P. B.; Grant, S. Int. J. Oncol. 2001, 19, 181.
6. Weidle, U. H.; Grossmann, A. Anticancer Res. 2000, 20,
1471.
7. Hashimoto, Y. Curr. Med. Chem. 1998, 5, 163.
8. Hashimoto, Y. Bioorg. Med. Chem. 2002, 10, 461.
9. Miyachi, H.; Azuma, A.; Ogasawara, A.; Uchimura, E.;
Watanabe, N.; Kobayashi, Y.; Kato, F.; Kato, M.;
Hashimoto, Y. J. Med. Chem. 1997, 40, 2858.
10. Miyachi, H.; Ogasawara, A.; Azuma, A.; Hashimoto,
Y. Bioorg. Med. Chem. 1997, 5, 2095.
11. Miyachi, H.; Azuma, A.; Kitamoto, T.; Hayashi, K.;
Kato, S.; Koga, M.; Sato, B.; Hashimoto, Y. Bioorg. Med.
Chem. Lett. 1997, 7, 1483.
12. Miyachi, H.; Kato, M.; Kato, F.; Hashimoto, Y. J. Med.
Chem. 1998, 41, 263.
13. Shinji, C.; Nakamura, T.; Maeda, S.; Yoshida, M.;
Hashimoto, Y.; Miyachi, H. Bioorg. Med. Chem. Lett.
2005, 15, 4427–4431.
Exponentially growing cells in RPMI1640 medium sup-
plemented with 10% fetal bovine serum were adjusted to
2 · 104 cells/mL, then 100 lL aliquots were plated in
96-well plates and incubated for 24 h at 37 ꢁC under
an atmosphere of 5% CO2 in air. After incubation, var-
ious concentrations of test compounds were added and
incubation was continued for a further 4 days. Viable
cells were counted with a cell counting kit (Dojindo).
Experiments were repeated at least three times.
5.6. Western blot analysis
Aliquots of 3 mL each of LNCaP cells (1 · 105 cells/
mL) in RPMI1640 medium supplemented with 10% fe-
tal bovine serum were plated in 6-well plates and incu-
bated for 24 at 37 ꢁC under an atmosphere of 5% CO2
in air. After incubation, various concentrations of test
compounds were added and incubation was continued
for the indicated periods. The supernatant was discard-
ed, then the cells were rinsed twice with PBS, and
lysed.
14. Finnin, M. S.; Donigian, J. R.; Cohen, A.; Richon, V. M.;
Rifkind, R. A.; Marks, P. A.; Breslow, R.; Pavletich, N. P.
Nature 1999, 401, 188.
15. Vannini, A.; Volpari, C.; Filocamo, G.; Casavola, E.
C.; Brunetti, M.; Renzoni, D.; Chakravarty, P.; Paolini,
C.; Francesco, R.; Gallinari, P.; Steinkuhler, C. D.;
Marco, S. Proc. Natl. Acad. Sci. U.S.A. 2004, 101,
15064.
Expression of the target genes (p21, b-actin, and
androgen receptor) was quantified with appropriate
monoclonal antibodies (p21 monoclonal antibody
(NeOMarkers), b-actin monoclonal antibody (SIGMA),
and androgen receptor monoclonal antibody (NeOMar-
kers)) by using a chemiluminescence assay system
(Amersham Biosciences) according to the supplier’s
protocol.
16. Yoshida, M.; Kijima, M.; Akita, M.; Beppu, T. J. Biol.
Chem. 1990, 265, 17174.

C. Shinji et al. / Bioorg. Med. Chem. 14 (2006) 7625–7651
7651
17. Richon, V. M.; Emiliani, S.; Verdin, E.; Webb, Y.;
Breslow, R.; Rifkind, R. A.; Marks, P. A. Proc. Natl.
Acad. Sci. U.S.A. 1998, 95, 3003.
19. Furumai, R.; Komatsu, Y.; Nishino, N.; Khochbin, S.;
Yoshida, M.; Horinouchi, S. Proc. Natl. Acad. Sci. U.S.A.
2001, 98, 87.
18. Somoza, J. R.; Skene, R. J.; Katz, B. A.; Joseph, C. M.; Ho,
D.; Jennings, A. J.; Andrew, C. L.; Buggy, A. J. J.; Chi, E.;
Tang, J.; Sang, B.; Verner, E.; Wynands, R.; Leahy, E. M.;
Dougan, D. R.; Snell, G.; Navre, M.; Knuth, M. W.;
Swanson, R. V.; E, D.; Leslie, M.; Tari, W. Structure 2004,
12, 1325.
20. Suzuki, T.; Matsuura, A.; Kouketsu, A.; Nakagawa, H.;
Miyata, N. Bioorg. Med. Chem. Lett. 2005, 7, 331.
21. Furumai, R.; Matsuyama, A.; Kobashi, N.; Lee, K.-H.;
Nishiyama, M.; Nakajima, H.; Tanaka, A.; Komatsu, Y.;
Nishino, N.; Yoshida, M.; Horinouchi, S. Cancer Res.
2002, 62, 4916.