Synthesis of novel 4,1-benzoxazepine derivatives as squalene synthase inhibitors and their inhibition of cholesterol synthesis

Article abstract of DOI:10.1021/jm020234o

Modification of the carboxyl group at the 3-position and introduction of protective groups to the hydroxy group of the 4,1-benzoxazepine derivative 2 (metabolite of 1) were carried out, and the inhibitory activity for squalene synthase and cholesterol syn

Full text of DOI:10.1021/jm020234o

J . Med. Chem. 2002, 45, 4571-4580
4571
Syn th esis of Novel 4,1-Ben zoxa zep in e Der iva tives a s Squ a len e Syn th a se
In h ibitor s a n d Th eir In h ibition of Ch olester ol Syn th esis
Takashi Miki,*,† Masakuni Kori,† Hiroshi Mabuchi,† Ryu-ichi Tozawa,† Tomoyuki Nishimoto,† Yasuo Sugiyama,‡
Koichiro Teshima,† and Hidefumi Yukimasa†
Pharmaceutical Research Division, Takeda Chemical Industries, Ltd., 2-17-85, J uso-Honmachi, Yodogawa-ku,
Osaka 532-8686, J apan, and Strategic Product Planning Department, Takeda Chemical Industries, Ltd., 4-1-1,
Doshomachi, Chuo-ku, Osaka 540-8645, J apan
Received May 29, 2002
Modification of the carboxyl group at the 3-position and introduction of protective groups to
the hydroxy group of the 4,1-benzoxazepine derivative 2 (metabolite of 1) were carried out,
and the inhibitory activity for squalene synthase and cholesterol synthesis in the liver was
investigated. Among these compounds, the glycine derivative 3a and â-alanine derivative 3f
exhibited the most potent inhibition of squalene synthase prepared from HepG2 cells (IC50
15 nM). On the other hand, the piperidine-4-acetic acid derivative 4a , which was prepared by
acetylation of 3j, was the most effective inhibitor of cholesterol synthesis in rat liver (ED50
)
)
2.9 mg/kg, po). After oral administration, 4a was absorbed and rapidly hydrolyzed to deacylated
3j. Compound 3j was detected mainly in the liver, but the plasma level of 3j was found to be
low. Compounds 3j and 4a were found to be competitive inhibitors with respect to farnesyl
pyrophosphate. Further evaluation of 4a as a cholesterol-lowering and antiatherosclerotic agent
is underway.
In tr od u ction
activity relationship of 2. Modification of the carboxyl
group at the 3-position and introduction of protective
groups to the hydroxy group were carried out, and the
inhibitory activity for squalene synthase and sterol
synthesis in the liver was investigated (Figure 1, 3 and
4). In this paper, we report the synthesis of the 4,1-
benzoxazepine derivatives 3 and 4, which led to the
candidate 4a (Figure 1).
Squalene synthase [EC 2.5.1.21] catalyzes the forma-
tion of squalene from farnesyl pyrophosphate (FPP) in
cholesterol biosynthesis. This enzymatic reaction takes
place after the pathway branches to other isoprene
derivatives such as dolichol, ubiquinones, and isopen-
tenyl t-RNA. Since squalene synthase inhibitors might
not interfere with the biosynthesis of these isoprene
derivatives, inhibition of this step would arrest only
cholesterol biosynthesis and might be useful for the
treatment of hyperlipidemia. Several squalene synthase
inhibitors,1 such as cationic intermediate analogues
(containing ammonium ions or sulfonium ions), sub-
strate analogues (phosphorus-containing compounds
bisphosphonates and R-phosphonosulfonic acids), qui-
nuclidine derivatives, 2,8-dioxabicyclo[3.2.1]octane de-
rivatives2,3 (squalestatins, zaragozic acids, and TAN-
1607A4), and others,5 have been reported.
Previously, we reported the synthesis of a series of
4,1-benzoxazepine derivatives and their inhibitory ac-
tivity for squalene synthase.6a-d Through modification
of the skeleton and substituents, we synthesized a novel
squalene synthase inhibitor, sodium (3R,5S)-7-chloro-
5-(2,3-dimethoxyphenyl)-1-neopentyl-2-oxo-1,2,3,5-tet-
rahydro-4,1-benzoxazepine-3-acetate (Figure 1, 1).
Ch em istr y
Compounds 2 and 3 were synthesized as follows
(Scheme 1). Compound 2 was prepared using the
optically active alcohol 56d as starting material. Com-
pound 5 was treated with 3-hydroxy-2,2-dimethylpro-
pionaldehyde7 and sodium cyanoborohydride (NaBH3-
CN) to obtain alkylated compound 6. Condensation of
6 and a fumaryl chloride monoethyl ester followed by
intramolecular Michael addition of the obtained inter-
mediate 7 afforded the 4,1-benzoxazepine derivative
8.6a,b,8 In this cyclization, a thermodynamically stable
3,5-trans compound was obtained.9 Hydrolysis of the
ester 8 gave the carboxylic acid 2. The enantiomeric
purity of compound 2 was 100% ee by HPLC analysis.
The optically active alcohol 5 was transformed into
compound 2 without racemization. The amide derivative
9 was prepared by condensation of 2 with various amino
acid esters using diethylphosphoryl cyanide (DEPC).
Then compounds 9a -l were hydrolyzed or hydrogenated
to yield acids 3a -l.
Pharmacokinetic studies of compound 1 revealed that
2 was the urinary metabolite of 1 in dogs (Figure 1).
Compound 2 exhibited potent inhibitory activity for
squalene synthase derived from human hepatoma
(HepG2) cells (IC50 ) 18 nM) similar to 1. We focused
our studies on the modification and the structure-
Acid 3j was reacted with acetic anhydride to provide
the acetylated compound 4a (Scheme 2). Compound 3j
was converted to benzyl ester 10, which was treated
with acid chloride or chloromethyl pivalate to afford
acylated compounds 11a -c and alkylated compound
11d . Acids 4b-e were obtained by hydrogenation of
compounds 11a -d (Scheme 3).
* To whom correspondence should be addressed. Phone: +81-6-
6300-6263. Fax: +81-6-6300-6306. E-mail: Miki_Takashi@takeda.co.jp.
† Pharmaceutical Research Division.
‡ Strategic Product Planning Department.
10.1021/jm020234o CCC: $22.00 © 2002 American Chemical Society
Published on Web 08/30/2002

4572 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20
Miki et al.
F igu r e 1. Structures of 4,1-benzoxazepine derivatives.
Sch em e 1a
a
Reagents: (a) 3-hydroxy-2,2-dimethylpropionaldehyde, NaBH3CN, AcOH, MeOH; (b) fumaryl chloride monoethyl ester, NaHCO3,
AcOEt; (c) K2CO3, EtOH; (d) NaOH, EtOH; (e) amino acid esters, DEPC, NEt3, DMF; (f) H2, Pd-C, AcOEt.
Biologica l Resu lts a n d Discu ssion
acetate into sterols in the Wistar rat liver at 1 h after
oral dosing. The results are shown in Tables 1 and 2.
The compounds synthesized were evaluated for their
inhibitory activity for squalene synthase prepared from
HepG2 cells. Inhibitory activity was measured according
to the method of L. H. Cohen et al. with slight
modification.6a,10 The compounds were also evaluated
for their ability to inhibit the incorporation of [14C]-
In a previous paper,6b we described that elongation
of the tether length and the conversion to carboxamide
of the 3-acetic acid moiety decreased the activity. We
then found that the inhibitory activity of glycine deriva-
tive 3a for squalene synthase was as potent as that of

4,1-Benzoxazepine Derivatives
J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20 4573
Sch em e 2a
entries 6-8). Bulky substituents at the 3-position were
found to be undesirable for the inhibition.
Although compounds 3a -f, which have methylene or
ethylene between the carboxyl group and carbamoyl
group, are potent inhibitors of squalene synthase, they
only weakly inhibit the synthesis of sterols in rat liver
(<31% inhibition at 10 mg/kg, po) (Table 1, entries 1-6).
On the other hand, adding methylene groups to 3f
improved the inhibitory activity (butyric acid derivative
3g and pentanoic acid derivative 3h showed 59% and
50% inhibition at 10 mg/kg, po, respectively) (Table 1,
entries 7 and 8). Since we had much success in improv-
ing the oral activity through elongation of the chain,
we added methylene groups to the piperidine-4-carbox-
ylic acid derivative 3i. Although the acetic acid, propi-
onic acid, and butyric acid derivatives 3j-l showed a 3-
to 4-fold reduced activity than 3i at the enzyme level,
their inhibition of sterol synthesis increased signifi-
cantly (Table 1, entries 9-12). In consideration of the
balance between in vitro inhibitory activity for the
human enzyme and de novo synthesis of sterols in rats,
we selected compound 3j for further pharmacological
investigation.
a
Reagents: (a) Ac2O, DMAP, pyridine.
Sch em e 3a
The dose that achieved 50% inhibition of sterol
synthesis in rat liver after oral administration (ED50)
of 3j was 6.8 mg/kg, po (Wistar rats). This compound
was approximately 2-fold less active than 1 (ED50 ) 3.0
mg/kg, po, Wistar rats). Compounds 4a -e having
protective groups added to the hydroxy group were
prepared, and their activity to inhibit squalene synthase
and sterol synthesis was investigated. Their inhibitory
effect for the enzyme was 2- to 10-fold weaker than that
of 3j (Table 2). Acetylated 4a showed the most potent
inhibition of sterol synthesis with an ED50 of 2.9 mg/
kg, po, which was almost equal to that of 1. The
propionyl, butyryl, and isobutyryl derivatives 4b-d
showed 30-40% inhibition at 10 mg/kg, po (p > 0.05).
Although the pivaloyloxymethyl derivative 4e reduced
sterol synthesis by 43% at 10 mg/kg, po (p < 0.01), the
activity was weaker than that of 3j and 4a .
Lineweaver-Burk analysis of the inhibitory effect on
the HepG2 enzyme revealed compounds 3j and 4a to
be competitive inhibitors with respect to FPP (Km ) 5.9
× 10-7 M). Dixon plot analysis showed that the Ki values
for compounds 3j and 4a were 3.4 × 10-9 and 9.4 × 10-9
M, respectively.
a
Reagents: (a) benzyl bromide, K2CO3, DMF; (b) R2Cl, NEt3,
DMAP, THF or R2Cl, NaH, DMF; (c) H2, Pd-C, AcOEt.
2 (Table 1, entry 1). Both the carbamoyl group and
carboxyl group were assumed to play important roles
in exerting the inhibitory activity. The N-methylated
compound 3b also achieved the same extent of inhibition
(Table 1, entry 2). This result indicated that the
hydrogen atom on the nitrogen did not affect the
inhibition.
The L-alanine derivative 3c was found to be as potent
as the D-alanine derivative 3d . The stereochemistry of
the side chain was not essential for the inhibition (Table
1, entries 3 and 4). Conversion of D-alanine to D-leucine
resulted in a slight decrease in activity (Table 1, entry
5). Modification of the tether length between the car-
bamoyl group and carboxyl group gave the following
results. The â-alanine derivative 3f exhibited the same
level of activity as the glycine derivative 3a . Further
elongation resulted in a slight loss of activity (Table 1,
The pharmacokinetic profile of compound 4a in rats
was evaluated. After oral administration, 4a was ab-
sorbed and rapidly hydrolyzed to deacylated 3j. Com-
pound 3j was detected mainly in the liver, but the
plasma level of 3j was found to be low. Thus, it was
assumed that 4a would be distributed mainly to the
liver as deacylated 3j. The ratio of the area under curve
(AUC) (liver/plasma) of compound 3j (injected as 4a )
was 3-fold higher than that of compound 1. Because the
target organ of squalene synthase is the liver, we believe
that 4a has a good profile compared to 1.
Compound 4a significantly reduced plasma non-HDL-
cholesterol and triglyceride levels in marmosets and
Watanabe heritable hyperlipidemic (WHHL) rabbits.11
Thus, 4a (TAK-475) is a promising candidate for an
antihyperlipidemic and antiatherosclerotic drug.

4574 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20
Miki et al.
Ta ble 1. Inhibition of Squalene Synthase and Sterol Synthesis in Rat Liver by 4,1-Benzoxazepine Derivatives 3a -l
sterol synthesisb
% inhibition
(10 mg/kg, po)
compound
R (configuration)
enzyme
IC50 (nM)
a
entry
1
2
3
4
5
6
7
8
9
3a
3b
3c
3d
3e
3f
3g
3h
3i
-NHCH2COOH
15
18
21
18
28
15
32
38
18
45
61
70
14 ( 26
-1 ( 43
14 ( 48
-N(Me)CH2COOH
-NHCH(Me)COOH (S)
-NHCH(Me)COOH (R)
-NHCH(Bui)COOH (R)
-NHCH2CH2COOH
-NHCH2CH2CH2COOH
-NHCH2CH2CH2CH2COOH
4-(carboxy)piperidin-1-yl
4-(carboxymethyl)piperidin-1-yl
4-(2-carboxyethyl)piperidin-1-yl
4-(3-carboxypropyl)piperidin-1-yl
31 ( 16
14 ( 30
19 ( 24
59 ( 13**
50 ( 16*
26 ( 23
77 ( 11**
76 ( 11**
87 ( 5**
10
11
12
3j
3k
3l
a
b
IC50 values were determined by a single experimental run in duplicate. Test compounds (10 mg/kg) as a 0.5% MC solution were
administered to 6-week-old male Wistar rats. Animals were intravenously injected with [14C]acetate (50 µCi/kg) 1 h after the drug
administration. They were sacrificed 1 h after the injection, and the radioactivity of the digitonin-precipitable sterols in the tissues was
measured. Values are the mean ( SD (n ) 6-10). (/) p e 0.05, (//) p e 0.01 (vs control), by Dunnett’s test.
Ta ble 2. Inhibition of Squalene Synthase and Sterol Synthesis in Rat Liver by 4,1-Benzoxazepine Derivatives 3j and 4a -e
sterol synthesisb
compound
% inhibition
3 mg/kg, po
enzyme
IC50 (nM)
ED50
(mg/kg, po)
a
entry
R
1 mg/kg, po
10 mg/kg, po
1
2
3
4
5
6
7
3j
4a
4b
4c
4d
4e
1
H
45
78
87
93
89
6.8
2.9
11 ( 69
31 ( 20*
20 ( 47
42 ( 21**
64 ( 15**
81 ( 9**
40 ( 18
COCH3
COCH2CH3
COCH2CH2CH3
COCH(CH3)2
CH2OCOC(CH3)3
30 ( 32
34 ( 26
43 ( 19**
471
3.0c
a
b
IC50 values were determined by a single experimental run in duplicate. Test compounds (10 mg/kg) as a 0.5% MC solution were
administered to 6-week-old male Wistar rats. Animals were intravenously injected with [14C]acetate (50 µCi/kg) 1 h after the drug
administration. They were sacrificed 1 h after the injection, and the radioactivity of the digitonin-precipitable sterols in the tissues was
measured. Values are the mean ( SD (n ) 6-10). (/) p e 0.05, (//) p e 0.01 (vs control), by Dunnett’s test. c See ref 6b.
Con clu sion
Exp er im en ta l Section
All melting points were determined on
a Yanagimoto
We performed chemical modifications of 4,1-benzox-
azepine derivatives and evaluated the inhibitory activ-
itiy for microsomal squalene synthase and for in vivo
hepatic sterol synthesis. Among these compounds, the
piperidine-4-acetic acid derivative 4a was the most
effective inhibitor of cholesterol synthesis in rat liver
(ED50 ) 2.9 mg/kg, po). A pharmacokinetic study showed
that 4a was deacylated to 3j (major active metabolite)
with distribution to the liver. Pharmacological studies
on this new squalene synthase inhibitor 4a will be
reported in due course.
micromelting point apparatus and are uncorrected. Proton
nuclear magnetic resonance (1H NMR) spectra were recorded
on a Varian GEMINI-200 (200 MHz) spectrometer (with
tetramethylsilane as an internal standard). Infrared absorp-
tion spectra were recorded on a J ASCO IR-810. [R]D values
were determined in the indicated solvents on a J ASCO DIP-
370 polarimeter. TLC analyses were carried out on Merck
Kieselgel 60 F254 plates. Elemental analyses were carried out
by Takeda Analytical Laboratories, Ltd. and are within (0.4%
of the theoretical values. For column chromatography, Merck
Kieselgel 60 (70-230 mesh ASTM) was used. Yields were not

4,1-Benzoxazepine Derivatives
J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20 4575
maximized. The following abbreviations are used: s ) singlet,
d ) doublet, t ) triplet, q ) quartet, m multiplet, br ) broad.
Cl) C, H, N. 100% ee [HPLC analysis; Sumichiral-OA (4.6 i.d.
× 250 mm; Sumitomo Chemical Analysis Center, Ltd.); eluent,
50 mM ammonium acetate methanol solution; flow rate, 1.0
mL/min; retention time, 14.27 min for 2, 14.36 and 19.09 min
for racemic compound].
(S)-5-Ch lor o-r-(2,3-d im et h oxyp h en yl)-2-(3-h yd r oxy-
2,2-d im eth ylp r op yl)a m in oben zyl Alcoh ol (6). 3-Hydroxy-
2,2-dimethylpropionaldehyde (21 g, 0.206 mol) and acetic acid
(12.4 g, 0.206 mol) were added to a solution of (S)-2-amino-5-
chloro-R-(2,3-dimethoxyphenyl)benzyl alcohol (5, 55 g, 0.187
mol) in MeOH (500 mL). After the mixture was stirred for 1 h
at 60 °C, sodium cyanoborohydride (12.8 g, 0.206 mol) was
added to this solution with ice-cooling. The mixture was stirred
for 2 h at 60 °C and diluted with AcOEt (500 mL). The solution
was washed with 1 N NaOH, water, and brine, dried over Na2-
SO4, and then concentrated under reduced pressure. The
residue was subjected to column chromatography [eluent:
hexane-AcOEt (2:1)] to give 6 (62 g, 0.163 mol, 87%) as a
Eth yl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-
(3-h yd r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-
4,1-ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-a ceta te (9j). To
a stirred mixture of 2 (0.6 g, 1.26 mmol) and ethyl piperidine-
4-acetate hydrochloride12 (0.27 g, 1.29 mmol) in DMF (6 mL)
was added diethylphosphoryl cyanide (DEPC) (0.23 g, 1.39
mmol) at room temperature, followed by the addition of NEt3
(0.28 g, 2.97 mmol). The mixture was stirred for 30 min at
room temperature. The mixture was diluted with AcOEt (50
mL) and washed with 5% KHSO4, saturated NaHCO3, and
brine and was then dried over Na2SO4. The solvent was
removed under reduced pressure. The residue was subjected
to column chromatography [eluent: hexane-AcOEt (1:2)] to
give 9j (0.45 g, 0.713 mmol, 57%) as a colorless amorphous
colorless oil. [R]22D -37.9° (c 0.453, MeOH). IR νmax(neat), cm-1
:
3400 (br, OH, NH). 1H NMR (CDCl3): δ 0.91 (3H, s), 0.93
(3H, s), 2.95 (2H, s), 3.37 (2H, s), 3.83 (3H, s), 3.88 (3H, s),
5.99 (1H, s), 6.63 (1H, d, J ) 8.8 Hz), 6.77 (1H, dd, J ) 1.6,
7.6 Hz), 6.90 (1H, dd, J ) 1.6, 7.6 Hz), 7.03 (1H, d, J ) 2.6
Hz), 7.03 (1H, t, J ) 7.6 Hz), 7.13 (1H, dd, J ) 2.6, 8.8 Hz).
powder. [R]22 -163° (c 0.213, MeOH). IR νmax (KBr), cm-1
:
D
3440 (br, OH), 1730, 1640 (CdO). 1H NMR (CDCl3): δ 0.63
(3H, s), 1.05 (3H, s), 1.26 (3H, t, J ) 7.2 Hz), 1.54-1.80 (4H,
m), 2.19-2.28 (2H, m), 2.50-2.75 (2H, m), 2.96-3.23 (3H, m),
3.39 (1H, d, J ) 14.2 Hz), 3.58-3.67 (1H, m), 3.62 (3H, s),
3.89 (3H, s), 3.91-3.98 (1H, m), 4.13 (2H, q, J ) 7.2 Hz), 4.43-
4.53 (3H, m), 6.16 (1H, s), 6.60 (1H, s), 6.96-7.01 (1H, m),
7.18-7.41 (4H, m). Anal. (C33H43ClN2O8) C, H, N.
Eth yl (S)-3-[N-[4-Ch lor o-2-(r-h yd r oxy-2,3-d im eth oxy-
b en zyl)p h en yl]-N-(3-h yd r oxy-2,2-d im et h ylp r op yl)ca r -
ba m oyl]a cr yla te (7). A solution of fumaryl chloride mono-
ethyl ester (3, 29 g, 0.179 mol) in AcOEt (50 mL) was added
to a stirred mixture of 6 (62 g, 0.163 mol) and NaHCO3 (21 g,
0.245 mol) in AcOEt (500 mL). After being stirred for 30 min
at room temperature, the reaction mixture was washed with
water, dried over Na2SO4, and then concentrated under
reduced pressure to give 7 (82 g, 0.163 mol, quantitative) as a
colorless oil. IR νmax(neat), cm-1: 3400 (br, OH), 1715, 1650
(CdO), 1620 (CdC). 1H NMR (CDCl3): δ 0.73 (1/2 × 3H, s),
0.74 (1/2 × 3H, s), 0.87 (1/2 × 3H, s), 1.01 (1/2 × 3H, s), 1.18-
1.30 (3H, m), 2.87 (1/2 × 1H, d, J ) 14.6 Hz), 3.05-3.41 (2H
+ 1/2 × 1H, m), 3.70 (1/2 × 3H, s), 3.73 (1/2 × 3H, s), 3.79 (1/2
× 3H, s), 3.89 (1/2 × 3H, s), 4.04-4.22 (2H, m), 4.32-4.55 (2H,
m), 6.01-7.85 (10H, m).
Meth yl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-b en zoxa zep in -3-yl]a cet yl]glycin a t e (9a ). Com-
pound 9a (1.15 g, 2.09 mmol, quantitative) was prepared from
2 (1.0 g, 2.09 mmol) and methyl glycinate hydrochloride (0.28
g, 2.20 mmol) in the same manner as described for the
preparation of 9j. Colorless prisms. Mp 83-84 °C (AcOEt-
hexane). [R]22 -197° (c 0.155, MeOH). IR νmax (KBr), cm-1
:
D
3600-3200 (br, OH, NH), 1749, 1658 (CdO). 1H NMR
(CDCl3): δ 0.64 (3H, s), 1.05 (3H, s), 2.72 (1H, dd, J ) 5.8,
14.6 Hz), 2.96 (1H, dd, J ) 7.2, 14.6 Hz), 3.06-3.22 (1H, br),
3.38 (1H, d, J ) 14.0 Hz), 3.62 (3H, s), 3.62 (1H, d, J ) 11.2
Hz), 3.75 (3H, s), 3.89 (3H, s), 4.01-4.03 (2H, m), 4.40 (1H,
dd, J ) 5.8, 7.2 Hz), 4.47 (1H, d, J ) 14.0 Hz), 6.17 (1H, s),
6.32-6.42 (1H, br), 6.61 (1H, s), 6.97-7.36 (5H, m). Anal.
(C27H33N2O8Cl) C, H, N.
Eth yl (3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-
h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h yd r o-
4,1-ben zoxa zep in e-3-a ceta te (8). A mixture of 7 (82 g, 0.163
mol) and K2CO3 (22.5 g, 0.163 mol) in EtOH (500 mL) was
stirred overnight at room temperature. The reaction mixture
was diluted with AcOEt (1 L). This solution was washed with
water (500 mL) and brine (500 mL), dried over Na2SO4, and
then concentrated under reduced pressure. The solid residue
was recrystallized from EtOH to give 8 (65.2 g, 0.129 mol, 79%)
Ben zyl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
dr o-4,1-ben zoxazepin -3-yl]acetyl]-N-m eth ylglycin ate (9b).
Compound 9b (1.1 g, 1.72 mmol, 82%) was prepared from 2
(1.0 g, 2.09 mmol) and benzyl sarcosinate p-toluenesulfonate
(0.74 g, 2.11 mmol) in the same manner as described for the
preparation of 9j. A colorless amorphous powder. [R]22D -177°
(c 0.136, MeOH). IR νmax (KBr), cm-1: 3600-3200 (br, OH),
1736, 1680, 1651 (CdO). 1H NMR (CDCl3): δ 0.64 (3H, s), 1.02
(3H, s), 2.60 (1/5 × 1H, dd, J ) 4.2, 15.0 Hz), 2.79 (4/5 × 1H,
dd, J ) 4.2, 16.4 Hz), 2.94 (1/5 × 3H, s), 3.12 (4/5 × 3H, s),
3.09 (1/5 × 1H, dd, J ) 8.4, 15.0 Hz), 3.24 (4/5 × 1H, dd, J )
8.4, 16.4 Hz), 3.19 (1H, d, J ) 10.6 Hz), 3.55 (1H, d, J ) 14.2
Hz), 3.61 (3H, s), 3.63 (1H, d, J ) 10.6 Hz), 3.89 (3H, s), 4.41-
4.60 (4H, m), 5.14 (4/5 × 2H, s), 5.19 (1/5 × 2H, s), 6.26 (1/5 ×
1H, s), 6.27 (4/5 × 1H, s), 6.64 (1H, s), 6.96-7.36 (10H, m).
Anal. (C34H39N2O8Cl) C, H, N.
as colorless needles. Mp 188-190 °C (EtOH). [R]22 -205° (c
D
0.318, MeOH). IR νmax(KBr), cm-1: 3430 (br, OH), 1720, 1650
(CdO). 1H NMR (CDCl3): δ 0.64 (3H, s), 1.05 (3H, s), 1.25 (3H,
t, J ) 7.4 Hz), 2.77 (1H, dd, J ) 5.8, 16.8 Hz), 3.07 (1H, dd, J
) 8.0, 16.8 Hz), 3.09-3.20 (1H, m), 3.39 (1H, d, J ) 14.4 Hz),
3.58-3.65 (1H, m), 3.62 (3H, s), 3.89 (3H, s), 4.13 (2H, dq, J )
1.8, 7.4 Hz), 4.12-4.20 (1H, br), 4.38 (1H, dd, J ) 5.8, 8.0 Hz),
4.49 (1H, d, J ) 14.4 Hz), 6.16 (1H, s), 6.61 (1H, s), 6.96-7.36
(5H, m). Anal. (C26H32NO7Cl‚0.5H2O) C, H, N.
(3R,5S)-7-Ch lor o-5-(2,3-dim eth oxyph en yl)-1-(3-h ydr oxy-
2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-ben zox-
a zep in e-3-a cetic Acid (2). An aqueous solution (100 mL) of
NaOH (10.3 g, 0.258 mol) was added to a solution of 8 (65.2 g,
0.129 mmol) in EtOH (400 mL). After being stirred for 2 h at
60 °C, the reaction mixture was diluted with water (500 mL),
acidified, and then extracted with AcOEt (500 mL, twice). The
combined organic extracts were washed with brine, dried over
Na2SO4, and then concentrated under reduced pressure. The
solid residue was recrystallized from AcOEt-hexane (1:2) to
give 2 (58.2 g, 0.122 mol, 94%) as colorless needles. Mp
Eth yl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-
(3-h yd r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-
4,1-ben zoxa zep in -3-yl]a cetyl]-L-a la n in a te (9c). Compound
9c (0.62 g, 1.07 mmol, quantitative) was prepared from 2 (0.5
g, 1.05 mmol) and ethyl L-alaninate hydrochloride (0.18 g, 1.15
mmol) in the same manner as described for the preparation
199-202 °C (AcOEt-hexane). [R]22 -223° (c 0.193, MeOH).
of 9j. Colorless prisms. Mp 130-132 °C (AcOEt-hexane). [R]22
D
D
IR νmax(KBr), cm-1: 3600-2400 (br, COOH, OH), 1710, 1650
(CdO). 1H NMR (CDCl3): δ 0.65 (3H, s), 1.04 (3H, s), 2.76 (1H,
dd, J ) 5.4, 16.8 Hz), 3.08 (1H, dd, J ) 8.0, 16.8 Hz), 3.11-
3.17 (1H, m), 3.39 (1H, d, J ) 14.6 Hz), 3.57-3.64 (1H, m),
3.61 (3H, s), 3.89 (3H, s), 4.37 (1H, dd, J ) 5.4, 8.0 Hz), 4.48
(1H, d, J ) 14.6 Hz), 6.15 (1H, s), 6.61 (1H, s), 6.96-7.01 (1H,
m), 7.18-7.22 (2H, m), 7.36-7.37 (2H, m). Anal. (C24H28NO7-
-191° (c 0.171, MeOH). IR νmax (KBr), cm-1: 3600-3200 (br,
OH, NH), 1739, 1655 (CdO). 1H NMR (CDCl3): δ 0.64 (3H,
s), 1.04 (3H, s), 1.26 (3H, t, J ) 7.4 Hz), 1.40 (3H, d, J ) 7.4
Hz), 2.70 (1H, dd, J ) 5.6, 14.8 Hz), 2.90 (1H, dd, J ) 7.4,
14.8 Hz), 3.14 (1H, d, J ) 11.6 Hz), 3.37 (1H, d, J ) 14.6 Hz),
3.61 (3H, s), 3.61 (1H, d, J ) 11.6 Hz), 3.89 (3H, s), 4.18 (2H,
q, J ) 7.4 Hz), 4.38-4.55 (3H, m), 6.16 (1H, s), 6.27 (1H, d, J

4576 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20
Miki et al.
) 6.6 Hz), 6.61 (1H, s), 6.96-7.35 (5H, m). Anal. (C29H37N2O8-
Cl‚H2O) C, H, N.
ν
max(KBr), cm-1
:
3600-3200 (br, OH, NH), 1738, 1660
1
(CdO). H NMR (CDCl3): δ 0.64 (3H, s), 1.05 (3H, s), 1.45-
1.68 (4H, m), 2.34 (2H, t, J ) 7.0 Hz), 2.63 (1H, dd, J ) 5.6,
14.4 Hz), 2.84 (1H, dd, J ) 7.4, 14.4 Hz), 3.14 (1H, t, J ) 11.2
Hz), 3.24 (2H, q, J ) 6.2 Hz), 3.38 (1H, d, J ) 14.2 Hz), 3.61
(3H, s), 3.61 (1H, dd, J ) 4.4, 11.2 Hz), 3.67 (3H, s), 3.89 (3H,
s), 4.20 (1H, dd, J ) 4.4, 11.2 Hz), 4.40 (1H, dd, J ) 5.6, 7.4
Hz), 4.46 (1H, d, J ) 14.2 Hz), 5.88-5.94 (1H, br), 6.15 (1H,
s), 6.60 (1H, s), 6.96-7.36 (5H, m). Anal. (C30H39N2O8Cl‚H2O)
C, H, N.
Meth yl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-ben zoxa zep in -3-yl]a cetyl]-D-a la n in a te (9d ). Com-
pound 9d (0.61 g, 1.08 mmol, quantitative) was prepared from
2 (0.5 g, 1.05 mmol) and methyl D-alaninate hydrochloride
(0.16 g, 1.15 mmol) in the same manner as described for the
preparation of 9j. A colorless amorphous powder. [R]22D -174°
(c 0.268, MeOH). IR νmax (KBr), cm-1: 3600-3200 (br, OH,
NH), 1743, 1660 (CdO). 1H NMR (CDCl3): δ 0.64 (3H, s), 1.05
(3H, s), 1.41 (3H, t, J ) 7.4 Hz), 2.68 (1H, dd, J ) 6.6, 14.8
Hz), 2.89 (1H, dd, J ) 6.8, 14.8 Hz), 3.15 (1H, d, J ) 10.8 Hz),
3.38 (1H, d, J ) 14.6 Hz), 3.57-3.66 (1H, br), 3.62 (3H, s),
3.74 (3H, s), 3.89 (3H, s), 4.38 (1H, dd, J ) 6.6, 6.8 Hz), 4.48
(1H, d, J ) 14.6 Hz), 4.56 (1H, t, J ) 7.4 Hz), 6.17 (1H, s),
6.43 (1H, d, J ) 7.8 Hz), 6.61 (1H, s), 6.96-7.35 (5H, m). Anal.
(C28H35N2O8Cl‚0.5H2O) C, H, N.
Meth yl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-ca r boxy-
la te (9i). Compound 9i (1.0 g, 1.66 mmol, 79%) was prepared
from 2 (1.0 g, 2.09 mmol) and methyl piperidine-4-carboxylate
hydrochloride (0.38 g, 2.09 mmol) in the same manner as
described for the preparation of 9j. A colorless powder. Mp
121-124 °C (AcOEt-hexane). [R]22 -180° (c 0.418, MeOH).
D
IR νmax (KBr), cm-1: 3600-3300 (br, OH), 1730, 1645 (CdO).
1H NMR (CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 1.56-1.73 (4H,
m), 2.30-2.55 (1H, m), 2.65-2.82 (2H, m), 3.06-3.24 (3H, m),
3.39 (1H, d, J ) 14.2 Hz), 3.61 (3H, s), 3.66-3.72 (4H, m),
3.878 (3H, s), 4.25-4.40 (1H, m), 4.45-4.48 (2H, m), 6.16 (1H,
s), 6.61 (1H, s), 6.96-7.40 (5H, m). Anal. (C31H39N2O8Cl) C,
H, N.
Meth yl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-ben zoxa zep in -3-yl]a cetyl]-D-leu cin e (9e). Com-
pound 9e (1.9 g, 3.14 mmol, quantitative) was prepared from
2 (1.5 g, 3.13 mmol) and methyl D-leucinate hydrochloride (0.63
g, 3.47 mmol) in the same manner as described for the
preparation of 9j. A colorless powder. Mp 110-111 °C (Et2O).
IR νmax (KBr), cm-1: 3600-3200 (br, OH, NH), 1749, 1658
Eth yl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-
(3-h yd r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-
4,1-ben zoxazepin -3-yl]acetyl]piper idin e-4-pr opion ate (9k).
Compound 9k (1.1 g, 1.70 mmol, 81%) was prepared from 2
(1.0 g, 2.09 mmol) and ethyl piperidine-4-propionate hydro-
chloride13 (0.49 g, 2.21 mmol) in the same manner as described
1
(CdO). H NMR (CDCl3): δ 0.64 (3H, s), 0.92 (3H, d, J ) 3.0
Hz), 0.95 (3H, d, J ) 1.6 Hz), 1.05 (3H, s), 1.42-1.85 (3H, s),
2.69 (1H, dd, J ) 6.0, 14.6 Hz), 2.91 (1H, dd, J ) 6.6, 14.6
Hz), 3.28 (1H, d, J ) 14.4 Hz), 3.05-3.22 (1H, m), 3.62 (3H,
s), 3.72 (3H, s), 4.35-4.68 (3H, m), 6.18 (1H, s), 6.28-6.42 (1H,
m), 6.61 (1H, d, J ) 1.6 Hz), 6.94-7.42 (5H, m). Anal.
(C31H41N2O8Cl) C, H, N.
for the preparation of 9j. A colorless amorphous powder. [R]22
D
-189° (c 0.248, MeOH). IR νmax (KBr), cm-1: 3600-3300 (br,
OH), 1732, 1645 (CdO). 1H NMR (CDCl3): δ 0.63 (3H, s), 1.05
(3H, s), 1.26 (3H, t, J ) 7.2 Hz), 0.88-1.20 (2H, m), 1.43-1.80
(4H, m), 2.37-2.40 (2H, m), 2.46-2.58 (1H, m), 2.67-2.77 (1H,
m), 2.90-3.03 (1H, m), 3.11-3.23 (2H, m), 3.39 (1H, d, J )
14.0 Hz), 3.61 (3H, s), 3.65 (1H, d, J ) 12.0 Hz), 3.89 (3H, s),
3.85-3.95 (1H, m), 4.13 (2H, q, J ) 7.2 Hz), 4.30-4.53 (4H,
m), 6.16 (1H, s), 6.60 (1H, s), 6.97-7.36 (5H, m). Anal.
(C34H45N2O8Cl) C, H, N.
Meth yl N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-ben zoxa zep in -3-yl]a cetyl]-â-a la n in a te (9f). Com-
pound 9f (1.80 g, 3.20 mmol, 77%) was prepared from 2 (2.0
g, 4.18 mmol) and methyl â-alaninate hydrochloride (0.64 g,
4.59 mmol) in the same manner as described for the prepara-
tion of 9j. A colorless amorphous powder. [R]22D -208 (c 0.114,
MeOH). IR νmax (KBr), cm-1: 3600-3200 (br, OH, NH), 1738,
1650 (CdO). 1H NMR (CDCl3): δ 0.64 (3H, s), 1.05 (3H, s),
2.51 (2H, t, J ) 6.2 Hz), 2.66 (1H, dd, J ) 5.8, 14.2 Hz), 2.83
(1H, dd, J ) 7.4, 14.2 Hz), 3.14 (1H, t, J ) 11.0 Hz), 3.50 (2H,
q, J ) 6.2 Hz), 3.38 (1H, d, J ) 14.0 Hz), 3.58-3.68 (1H, br),
3.61 (3H, s), 3.67 (3H, s), 3.89 (3H, s), 4.13-4.25 (1H, br), 4.40
(1H, dd, J ) 5.8, 7.4 Hz), 4.46 (1H, d, J ) 14.0 Hz), 6.15 (1H,
s), 6.30-6.40 (1H, br), 6.61 (1H, d, J ) 1.4 Hz), 6.97-7.44 (5H,
m). Anal. (C28H35N2O8Cl) C, H, N.
Eth yl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-
(3-h yd r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-
4,1-ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-bu tyr a te (9l).
Compound 9l (1.0 g, 1.52 mmol, 73%) was prepared from 2
(1.0 g, 2.09 mmol) and ethyl piperidine-4-butyrate hydrochlo-
ride14 (0.52 g, 2.21 mmol) in the same manner as described
for the preparation of 9j. A colorless amorphous powder. [R]22
D
-182° (c 0.152, MeOH). IR νmax (KBr), cm-1: 3600-3300 (br,
OH), 1732, 1645 (CdO). 1H NMR (CDCl3): δ 0.63 (3H, s), 1.04
(3H, s), 0.88-1.80 (9H, m), 1.26 (3H, t, J ) 7.4 Hz), 2.24-2.31
(2H, m), 2.46-2.57 (1H, m), 2.65-2.78 (1H, m), 2.90-3.03 (1H,
m), 3.08-3.24 (2H, m), 3.39 (1H, d, J ) 14.0 Hz), 3.61 (3H, s),
3.64 (1H, dd, J ) 3.0, 11.4 Hz), 3.89 (3H, s), 3.85-3.97 (1H,
m), 4.12 (2H, q, J ) 7.4 Hz), 4.32-4.53 (4H, m), 6.16 (1H, s),
6.59 (1H, s), 6.96-7.36 (5H, m). Anal. (C35H47N2O8Cl) C, H,
N.
Meth yl 4-[[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
d r o-4,1-ben zoxa zep in -3-yl]a cet yl]a m in o]b u t yr a t e (9g).
Compound 9g (1.59 g, 2.76 mmol, 66%) was prepared from 2
(2.0 g, 4.18 mmol) and methyl 4-aminobutyrate hydrochloride
(0.71 g, 4.60 mmol) in the same manner as described for the
preparation of 9j. A colorless powder. Mp 78-80 °C (AcOEt-
hexane). [R]22 -202° (c 0.149, MeOH). IR νmax (KBr), cm-1
:
D
3600-3200 (br, OH, NH), 1738, 1651 (CdO). 1H NMR
(CDCl3): δ 0.64 (3H, s), 1.05 (3H, s), 1.4 (2H, quintet, J ) 7.2
Hz), 2.36 (2H, t, J ) 7.2 Hz), 2.63 (1H, dd, J ) 5.8, 14.2 Hz),
2.83 (1H, dd, J ) 7.4, 14.2 Hz), 3.14 (1H, t, J ) 10.8 Hz), 3.23-
3.34 (2H, m), 3.38 (1H, d, J ) 14.6 Hz), 3.58-3.67 (1H, br),
3.61 (3H, s), 3.67 (3H, s), 3.89 (3H, s), 4.14-4.22 (1H, br), 4.40
(1H, dd, J ) 5.8, 7.4 Hz), 4.46 (1H, d, J ) 14.6 Hz), 5.96-6.03
(1H, br), 6.15 (1H, s), 6.61 (1H, d, J ) 1.4 Hz), 6.97-7.40 (5H,
m). Anal. (C29H37N2O8Cl‚0.5H2O) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-a cetic Acid (3j).
To a solution of 9k (0.45 g, 0.713 mmol) in EtOH (4 mL), 1 N
NaOH (1 mL) was added. The mixture was stirred for 30 min
at 60 °C. The reaction mixture was diluted with water (50 mL),
acidified, and then extracted with AcOEt (50 mL × 3). The
organic extracts were washed with brine, dried over Na2SO4,
and concentrated under reduced pressure. The solid residue
was crystallized with Et2O-hexane (1:1) to give 3k (0.30 g,
0.497 mmol, 70%) as a colorless powder. Mp 135-140 °C
Meth yl 5-[[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-(3-h yd r oxy-2,2-d im et h ylp r op yl)-2-oxo-1,2,3,5-t et r a h y-
dr o-4,1-ben zoxazepin -3-yl]acetyl]am in o]pen tan oate (9h ).
Compound 9h (2.57 g, 4.35 mmol, quantitative) was prepared
from 2 (2.0 g, 4.18 mmol) and methyl 5-aminopentanoate
hydrochloride (0.77 g, 4.60 mmol) in the same manner as
described for the preparation of 9j. Colorless prisms. Mp 84-
(Et2O-hexane). [R]22 -191° (c 0.179, MeOH). IR νmax (KBr),
D
cm-1
:
3600-2400 (br, COOH, OH), 1730, 1640 (CdO). 1H
NMR (CDCl3): δ 0.63 (3H, s), 1.03 (3H, s), 1.70-1.85 (4H, m),
1.96-2.10 (1H, m), 2.24-2.33 (2H, m), 2.50-2.78 (2H, m),
3.01-3.22 (3H, m), 3.39 (1H, d, J ) 15.4 Hz), 3.58-3.67 (1H,
m), 3.61 (3H, s), 3.891 (3H, s), 3.92-3.97 (1H, m), 4.46-4.57
85 °C (AcOEt-hexane). [R]22 -191° (c 0.126, MeOH). IR
D

4,1-Benzoxazepine Derivatives
J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20 4577
(3H, m), 6.149 (1H, s), 6.60 (1H, s), 6.96-7.40 (5H, m). Anal.
Hz), 4.07 (3H, s), 4.36 (1H, dd, J ) 5.4, 7.2 Hz), 4.45 (1H, d, J
) 14.2 Hz), 4.52-4.66 (1H, m), 6.16 (1H, s), 6.57-6.66 (2H,
m). Anal. (C30H39N2O8Cl) C, H, N.
(C31H39N2O8Cl) C, H, N.
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]glycin e (3a ). Compound 3a (0.94
g, 1.76 mmol, 97%) was prepared from 9a (1.0 g, 1.82 mmol)
in the same manner as described for the preparation of 3j.
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]-â-a la n in e (3f). Compound 3f (1.1
g, 2.00 mmol, 76%) was prepared from 9f (1.49 g, 2.65 mmol)
in the same manner as described for the preparation of 3j. A
Colorless prisms. Mp 122-123 °C (AcOEt-hexane). [R]22
D
-191° (c 0.332, MeOH). IR νmax (KBr), cm-1: 3600-2400 (br,
COOH, OH, NH), 1736, 1657 (CdO). 1H NMR (CDCl3): δ 0.65
(3H, s), 1.03 (3H, s), 2.72 (1H, dd, J ) 5.2, 14.6 Hz), 2.97 (1H,
dd, J ) 7.8, 14.6 Hz), 3.18 (1H, d, J ) 11.6 Hz), 3.39 (1H, d, J
) 14.2 Hz), 3.61 (3H, s), 3.61 (1H, d, J ) 11.6 Hz), 3.89 (3H,
s), 4.03 (2H, t, J ) 3.6 Hz), 4.38 (1H, dd, J ) 5.2, 7.8 Hz), 4.45
(1H, d, J ) 14.2 Hz), 6.16 (1H, s), 6.61 (1H, s), 6.64-6.74 (1H,
br), 6.96-7.35 (5H, m). Anal. (C26H31N2O8Cl) C, H, N.
colorless amorphous powder. [R]22 -232° (c 0.133, MeOH).
D
IR νmax (KBr), cm-1: 3600-2200 (br, COOH, OH, NH), 1716,
1651 (CdO). 1H NMR (CDCl3): δ 0.64 (3H, s), 1.04 (3H, s),
2.53 (2H, t, J ) 5.4 Hz), 2.65 (1H, dd, J ) 5.2, 14.2 Hz), 2.84
(1H, dd, J ) 7.4, 14.2 Hz), 3.16 (1H, t, J ) 11.2 Hz), 3.43-
3.56 (2H, m), 3.38 (1H, d, J ) 14.2 Hz), 3.60 (3H, s), 3.60 (1H,
d, J ) 11.2 Hz), 3.89 (3H, s), 4.43 (1H, dd, J ) 5.2, 7.4 Hz),
4.47 (1H, d, J ) 14.2 Hz), 6.15 (1H, s), 6.60 (1H, d, J ) 1.8
Hz), 6.82-6.92 (1H, br), 6.96-7.36 (5H, m). Anal. (C27H33N2O8-
Cl) C, H, N.
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxazepin -3-yl]acetyl]-N-m eth ylglycin e (3b). 10% Pd-C
catalyst (100 mg) was added to a stirred solution of 9b (0.95
g, 1.49 mmol) in AcOEt (20 mL) at room temperature. The
reaction mixture was stirred under H2 atmosphere for 30 min
at room temperature. The catalyst was removed by filtration
and the solvent was removed under reduced pressure to give
3b (0.86 g, 1.57 mmol, quantitative) as a colorless amorphous
4-[[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]a m in o]bu tyr ic Acid (3g). Com-
pound 3g (1.1 g, 1.95 mmol, 76%) was prepared from 9g (1.49
g, 2.58 mmol) in the same manner as described for the
preparation of 3j. Colorless prisms. Mp 111-113 °C (AcOEt-
hexane). [R]22 -203° (c 0.106, MeOH). IR νmax(KBr), cm-1
:
D
powder. [R]22 -194° (c 0.201, MeOH). IR νmax (KBr), cm-1
:
3600-2200 (br, COOH, OH, NH), 1716, 1651 (CdO). 1H NMR
(CDCl3): δ 0.64 (3H, s), 1.04 (3H, s), 1.84 (2H, quintet, J )
6.8 Hz), 2.37 (2H, t, J ) 6.8 Hz), 2.65 (1H, dd, J ) 5.4, 14.2
Hz), 2.84 (1H, dd, J ) 7.6, 14.2 Hz), 3.16 (1H, t, J ) 10.8 Hz),
3.25-3.33 (2H, m), 3.39 (1H, d, J ) 14.6 Hz), 3.60 (3H, s),
3.60 (1H, d, J ) 10.8 Hz), 3.89 (3H, s), 4.37-4.48 (2H, m),
6.15 (1H, s), 6.22-6.30 (1H, br), 6.61 (1H, d, J ) 1.4 Hz), 6.96-
7.36 (5H, m). Anal. (C28H35N2O8Cl‚0.5H2O) C, H, N.
D
3600-2400 (br, COOH, OH), 1734, 1653 (CdO). 1H NMR
(CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 2.63 (1/5 × 1H, dd, J )
4.0, 15.2 Hz), 2.80 (4/5 × 1H, dd, J ) 4.4, 16.4 Hz), 2.95 (1/5
× 3H, s), 3.13 (4/5 × 3H, s), 3.17 (1H, d, J ) 11.0 Hz), 3.25
(1H, dd, J ) 9.2, 16.4 Hz), 3.40 (1H, d, J ) 14.4 Hz), 3.61 (3H,
s), 3.63 (1H, d, J ) 11.0 Hz), 3.79 (1/5 × 3H, s), 3.89 (4/5 ×
3H, s), 4.31-4.52 (4H, m), 6.13 (1/5 × 1H, s), 6.16 (4/5 × 1H,
s), 6.61 (1H, s), 6.95-7.37 (5H, m). Anal. (C27H33N2O8Cl‚
0.3H2O) C, H, N.
5-[[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
b en zoxa zep in -3-yl]a cet yl]a m in o]p en t a n oic Acid (3h ).
Compound 3h (2.1 g, 3.64 mmol, 94%) was prepared from 9h
(2.3 g, 3.89 mmol) in the same manner as described for the
preparation of 3j. A colorless amorphous powder. [R]22D -191°
(c 0.237, MeOH). IR νmax(KBr), cm-1: 3600-2200 (br, COOH,
OH, NH), 1714, 1651 (CdO). 1H NMR (CDCl3): δ 0.64 (3H,
s), 1.04 (3H, s), 1.45-1.75 (4H, m), 2.36 (2H, t, J ) 7.0 Hz),
2.63 (1H, dd, J ) 5.6, 14.4 Hz), 2.85 (1H, dd, J ) 7.6, 14.4
Hz), 3.16 (1H, t, J ) 12.0 Hz), 3.23-3.28 (2H, m), 3.38 (1H, d,
J ) 14.4 Hz), 3.60 (3H, s), 3.60 (1H, dd, J ) 12.0 Hz), 3.89
(3H, s), 4.40 (1H, dd, J ) 5.6, 7.6 Hz), 4.45 (1H, d, J ) 14.4
Hz), 6.02-6.14 (1H, br), 6.14 (1H, s), 6.60 (1H, s), 6.96-7.36
(5H, m). Anal. (C29H37N2O8Cl‚H2O) C, H, N.
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]-L-a la n in e (3c). Compound 3c
(0.44 g, 0.801 mmol, 89%) was prepared from 9c (0.52 g, 0.901
mmol) in the same manner as described for the preparation
of 3j. A colorless powder. Mp 133-135 °C (AcOEt-hexane).
[R]22D -189° (c 0.226, MeOH). IR νmax (KBr), cm-1: 3600-2400
1
(br, COOH, OH, NH), 1732, 1651 (CdO). H NMR (CDCl3): δ
0.65 (3H, s), 1.04 (3H, s), 1.43 (3H, t, J ) 7.4 Hz), 2.73 (1H,
dd, J ) 6.2, 14.6 Hz), 2.89 (1H, dd, J ) 6.6, 14.6 Hz), 3.16
(1H, d, J ) 12.0 Hz), 3.39 (1H, d, J ) 14.2 Hz), 3.60 (1H, d, J
) 12.0 Hz), 3.60 (3H, s), 3.88 (3H, s), 4.37-4.55 (3H, m), 6.16
(1H, s), 6.48 (1H, d, J ) 6.6 Hz), 6.62 (1H, d, J ) 1.6 Hz),
6.96-7.36 (5H, m). Anal. (C27H33N2O8Cl) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cet yl]p ip er id in e-4-ca r boxylic Acid
(3i). Compound 3i (0.54 g, 0.917 mmol, 79%) was prepared
from 9i (0.7 g, 1.16 mmol) in the same manner as described
for the preparation of 3j. A colorless powder. Mp 162-165 °C
(AcOEt-hexane). [R]22D -193° (c 0.378, MeOH). IR νmax (KBr),
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]-D-a la n in e (3d ). Compound 3d
(0.37 g, 0.674 mmol, 74%) was prepared from 9d (0.51 g, 0.906
mmol) in the same manner as described for the preparation
of 3j. A colorless powder. Mp 130-132 °C (AcOEt-hexane).
[R]22D -174° (c 0.358, MeOH). IR νmax (KBr), cm-1: 3600-2400
cm-1
:
3600-2400 (br, OH, COOH), 1720, 1640 (CdO). 1H
1
(br, COOH, OH, NH), 1732, 1658 (CdO). H NMR (CDCl3): δ
NMR (CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 1.60-1.90 (4H, m),
2.40-2.59 (1H, m), 2.60-2.83 (2H, m), 3.0-3.3 (2H, m), 3.15
(1H, d, J ) 13.0 Hz), 3.39 (1H, d, J ) 15.2 Hz), 3.61 (3H, s),
3.65 (1H, d, J ) 13.0 Hz), 3.77-3.97 (1H, m), 3.89 (3H, s),
4.43-4.51 (3H, m), 6.16 (1H, s), 6.61 (1H, s), 6.96-7.42 (5H,
m). Anal. (C30H37N2O8Cl‚0.3H2O) C, H, N.
0.65 (3H, s), 1.04 (3H, s), 1.44 (3H, t, J ) 7.4 Hz), 2.69 (1H,
dd, J ) 5.8, 14.6 Hz), 2.93 (1H, dd, J ) 7.0, 14.6 Hz), 3.18
(1H, d, J ) 12.2 Hz), 3.39 (1H, d, J ) 14.2 Hz), 3.61 (1H, d, J
) 12.2 Hz), 3.61 (3H, s), 3.89 (3H, s), 4.33-4.58 (3H, m), 6.16
(1H, s), 6.61 (1H, s), 6.66 (1H, d, J ) 7.0 Hz), 6.96-7.35 (5H,
m). Anal. (C27H33N2O8Cl) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
b en zoxa zep in -3-yl]a cet yl]p ip er id in e-4-p r op ion ic Acid
(3k ). Compound 3k (0.60 g, 0.972 mmol, 90%) was prepared
from 9k (0.7 g, 1.08 mmol) in the same manner as described
for the preparation of 3j. A colorless powder. Mp 152-154 °C
(AcOEt-hexane). [R]22D -186° (c 0.185, MeOH). IR νmax (KBr),
N-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]-D-leu cin e (3e). Compound 3e
(1.2 g, 2.03 mmol, 94%) was prepared from 9e (1.3 g, 2.15
mmol) in the same manner as described for the preparation
of 3j. A colorless amorphous powder. IR νmax (KBr), cm-1
:
3600-2400 (br, COOH, OH, NH), 1736, 1657 (CdO). 1H NMR
(CDCl3): δ 0.65 (3H, s), 0.93 (6H, d, J ) 5.6 Hz), 1.03 (3H, s),
1.45-1.82 (3H, m), 2.69 (1H, dd, J ) 5.4, 14.5 Hz), 2.95 (1H,
dd, J ) 7.4, 14.5 Hz), 3.18 (1H, d, J ) 11.8 Hz), 3.40 (1H, d, J
) 14.2 Hz), 3.61 (3H, s), 3.61 (3H, s), 3.61 (1H, d, J ) 11.8
cm-1 3600-2400 (br, OH, COOH), 1732, 1651 (CdO). 1H
:
NMR (CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 0.95-1.14 (2H, m),
1.54-1.78 (4H, m), 2.34-2.40 (2H, m), 2.48-2.56 (1H, m),
2.67-2.77 (1H, m), 2.89-3.08 (1H, m), 3.12-3.21 (2H, m), 3.39
(1H, d, J ) 14.0 Hz), 3.61 (3H, s), 3.65 (1H, d, J ) 12.0 Hz),

4578 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20
Miki et al.
3.89 (3H, s), 3.85-3.97 (1H, m), 4.47-4.55 (3H, m), 6.15, 6.16
(total 1H, each s), 6.59 (1H, s), 6.97-7.39 (5H, m). Anal.
(C32H41N2O8Cl) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-1-(3-h y-
d r oxy-2,2-d im eth ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-4,1-
ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-bu tyr ic Acid (3l).
Compound 3l (0.50 g, 0.792 mmol, 75%) was prepared from 9l
(0.7 g, 1.06 mmol) in the same manner as described for the
preparation of 3j. A colorless powder. Mp 133-135 °C (AcOEt-
(1H, d, J ) 14.0 Hz), 3.61 (3H, s), 3.71 (1H, d, J ) 11.4 Hz),
3.86 (1H, d, J ) 11.4 Hz), 3.88 (3H, s), 3.88-4.00 (1H, m),
4.45-4.51 (2H, m), 4.56 (1H, d, J ) 14.0 Hz), 5.11 (2H, s),
6.24 (1H, s), 6.62 (1H, s), 6.98-7.35 (10H, m). Anal. (C41H49N2O9-
Cl) C, H, N.
Ben zyl 1-[[(3R,5S)-1-[3-(Bu tyr yloxy)-2,2-d im eth ylp r o-
p yl]-7-ch lor o-5-(2,3-d im et h oxyp h en yl)-2-oxo-1,2,3,5-t et -
r a h yd r o-4,1-ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-a ce-
ta te (11b). Compound 11b (0.12 g, 0.157 mmol, 84%) was
prepared from 10 (0.13 g, 0.188 mmol) and butyryl chloride
(48 mg, 0.442 mmol) in the same manner as described for the
hexane). [R]22 -187° (c 0.108, MeOH). IR νmax (KBr), cm-1
:
D
3600-2400 (br, OH, COOH), 1730, 1651 (CdO). 1H NMR
(CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 0.95-1.80 (9H, m), 2.29-
2.36 (2H, m), 2.46-2.58 (1H, m), 2.65-2.78 (1H, m), 2.91-
3.03 (1H, m), 3.11-3.23 (2H, m), 3.39 (1H, d, J ) 14.0 Hz),
3.61 (3H, s), 3.64 (1H, d, J ) 11.4 Hz), 3.89 (3H, s), 3.85-3.97
(1H, m), 4.45-4.52 (3H, m), 6.15 (1H, s), 6.59 (1H, s), 6.96-
7.36 (5H, m). Anal. (C33H43N2O8Cl) C, H, N.
preparation of 11a . A colorless amorphous powder. [R]22
D
-165° (c 0.185, MeOH). IR νmax(KBr), cm-1: 1734, 1682, 1639
(CdO). 1H NMR (CDCl3): δ 0.93 (3H, t, J ) 7.4 Hz), 0.97 (3H,
s), 1.02 (3H, s), 1.10-1.30 (2H, m), 1.55-1.72 (4H, m), 1.95-
2.10 (1H, m), 2.23-2.32 (4H, m), 2.48-2.61 (1H, m), 2.72 (1H,
dd, J ) 4.4, 16.0 Hz), 2.92-3.05 (1H, m), 3.11 (1H, dd, J )
8.8, 16.0 Hz), 3.56 (1H, d, J ) 13.8 Hz), 3.61 (3H, s), 3.70 (1H,
d, J ) 11.4 Hz), 3.87 (1H, d, J ) 11.4 Hz), 3.89 (3H, s), 3.92-
4.00 (1H, m), 4.45-4.60 (3H, m), 5.11 (2H, s), 6.24 (1H, s), 6.62
(1H, s), 6.95-7.35 (10H, m). Anal. (C42H51N2O9Cl) C, H, N.
1-[[(3R,5S)-1-(3-Acetoxy-2,2-d im eth ylp r op yl)-7-ch lor o-
5-(2,3-dim eth oxyph en yl)-2-oxo-1,2,3,5-tetr ah ydr o-4,1-ben -
zoxa zep in -3-yl]a cetyl]p ip er id in e-4-a cetic Acid (4a ). A
mixture of 3j (7.2 g, 11.9 mmol), 4-(dimethylamino)pyridine
(DMAP) (1.0 mg, 8.19 µmol), acetic anhydride (1.4 g, 14.0
mmol), and pyridine (70 mL) was stiired for 1 h at room
temperature. The mixture was concentrated under reduced
pressure, and the residue was dissolved in AcOEt (100 mL).
The solution was washed with 1 N HCl, saturated NaHCO3,
and brine, dried over Na2SO4, and then concentrated under
reduced pressure. The residue was recrystallized from EtOH-
H2O (1:1) to give 4a (5.5 g, 9.15 mmol, 77%) as a colorless
Ben zyl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-[2,2-d im et h yl-3-(isob u t yr yloxy)p r op yl]-2-oxo-1,2,3,5-
t et r a h yd r o-4,1-b en zoxa zep in -3-yl]a cet yl]p ip er id in e-4-
a ceta te (11c). Compound 11c (0.40 g, 0.524 mmol, 73%) was
prepared from 10 (0.5 g, 0.721 mmol) and isobutyryl chloride
(113 mg, 1.06 mmol) in the same manner as described for the
preparation of 11a . A colorless amorphous powder. [R]22
D
-164° (c 0.150, MeOH). IR νmax(KBr), cm-1: 1732, 1682, 1639
(CdO). 1H NMR (CDCl3): δ 0.94 (3H, s), 1.03 (3H, s), 1.15 (3H,
d, J ) 7.0 Hz), 1.17 (3H, d, J ) 7.0 Hz), 0.88-1.22 (2H, m),
1.55-1.80 (2H, m), 1.90-2.10 (1H, m), 2.24-2.32 (2H, m),
2.47-2.77 (3H, m), 2.98-3.18 (2H, m), 3.57 (1H, d, J ) 14.0
Hz), 3.61 (3H, s), 3.69 (1H, d, J ) 11.2 Hz), 3.87 (1H, d, J )
11.2 Hz), 3.88 (3H, s), 3.88-3.97 (1H, m), 4.45-4.51 (2H, m),
4.57 (1H, d, J ) 14.0 Hz), 5.11, 5.12 (total 2H, each s), 6.23
(1H, s), 6.62 (1H, s), 6.96-7.35 (10H, m). Anal. (C42H51N2O9-
Cl‚0.5H2O) C, H, N.
powder. Mp 194-196 °C. [R]20 -199° (c 1.00, MeOH). IR
D
ν
max(KBr), cm-1: 3600-2400 (br, COOH), 1732, 1682 (CdO).
1H NMR (CDCl3): δ 0.94 (3H, s), 1.02 (3H, s), 1.09-1.39 (2H,
m), 1.70-1.85 (2H, m), 1.90-2.10 (1H, m), 2.02 (3H, s), 2.22-
2.31 (2H, m), 2.50-2.77 (2H, m), 2.94-3.17 (2H, m), 3.55 (1H,
d, J ) 14.0 Hz), 3.61 (3H, s), 3.72 (1H, d, J ) 11.0 Hz), 3.85
(1H, d, J ) 11.0 Hz), 3.89 (3H, s), 3.90-3.98 (1H, m), 4.44-
4.60 (3H, m), 6.25 (1/2 × 1H, s), 6.26 (1/2 × 1H, s), 6.62 (1H,
s), 6.95-7.33 (5H, m). Anal. (C33H41N2O9Cl) C, H, N.
Ben zyl 1-[[(3R,5S)-7-Ch lor o-1-(3-h yd r oxy-2,2-d im eth -
ylp r op yl)-2-oxo-1,2,3,5-tetr a h yd r o-5-(2,3-d im eth oxyp h e-
n yl)-4,1-b en zoxa zep in -3-yl]a cet yl]p ip er id in e-4-a cet a t e
(10). A mixture of 3j (0.3 g, 0.497 mmol), K2CO3 (0.20 g, 1.45
mmol), and benzyl bromide (83 mg, 0.488 mmol) in DMF (3
mL) was stirred for 30 min at 60 °C. The mixture was diluted
with AcOEt (50 mL), washed with water, 5% KHSO4, satu-
rated NaHCO3, and brine, dried over Na2SO4, and then
concentrated under reduced pressure. The residue was recrys-
tallized from AcOEt-hexane (1:3) to give 10 (0.29 g, 0.418
mmol, 84%) as colorless prisms. Mp 115-117 °C. IR νmax(KBr),
Ben zyl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-[2,2-d im eth yl-3-(p iva loyloxym eth yloxy)p r op yl]-2-oxo-
1,2,3,5-t et r a h yd r o-4,1-b en zoxa zep in -3-yl]a cet yl]p ip er i-
d in e-4-a ceta te (11d ). To a solution of 10 (0.5 g, 0.721 mmol)
and chloromethyl pivalate (0.16 g, 1.06 mmol) in DMF (5 mL)
was added NaH (22 mg, 0.922 mmol) at 0 °C. The mixture
was stirred overnight at room temperature, diluted with
AcOEt, washed with water, 5% KHSO4, saturated NaHCO3,
and brine, dried over Na2SO4, and then concentrated under
reduced pressure. The residue was subjected to column chro-
matography [eluent: hexane-AcOEt (1:1)] to give 11d (0.35
cm-1
:
3600-3200 (br, OH), 1732, 1639 (CdO). 1H NMR
g, 0.434 mmol, 60%) as a colorless amorphous powder. [R]22
D
(CDCl3): δ 0.63 (3H, s), 1.04 (3H, s), 1.15-1.30 (2H, m), 1.65-
1.80 (2H, m), 1.95-2.10 (1H, m), 2.25-2.34 (2H, m), 2.50-
2.78 (2H, m), 3.00-3.23 (4H, m), 3.39 (1H, d, J ) 13.8 Hz),
3.61 (3H, s), 3.62-3.67 (1H, m), 3.89 (3H, s), 3.85-4.00 (1H,
m), 4.30-4.53 (3H, m), 5.11, 5.12 (total 2H, each s), 6.16 (1H,
s), 6.60 (1H, s), 6.96-7.39 (10H, m). Anal. (C38H45N2O8Cl) C,
H, N.
-138° (c 0.293, MeOH). IR νmax(KBr), cm-1: 1738, 1682, 1643
(CdO). 1H NMR (CDCl3): δ 0.93 (3H, s), 1.00 (3H, s), 1.17 (9H,
s), 1.06-1.26 (2H, m), 1.65-1.80 (2H, m), 1.90-2.10 (1H, m),
2.24-2.32 (2H, m), 2.47-2.60 (1H, m), 2.72 (1H, dd, J ) 4.4,
15.4 Hz), 2.98-3.35 (4H, m), 3.64 (3H, s), 3.64 (1H, d, J )
13.4 Hz), 3.89 (3H, s), 3.89-4.00 (1H, m), 4.41-4.71 (3H, m),
4.69 (1H, d, J ) 6.2 Hz), 5.11 (2H, s), 5.19 (1H, d, J ) 6.2 Hz),
6.23 (1H, s), 6.59 (1H, s), 6.95-7.40 (10H, m). Anal. (C45H55N2O10-
Cl‚0.5H2O) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im et h oxyp h en yl)-1-[2,2-
d im eth yl-3-(p r op ion yloxy)p r op yl]-2-oxo-1,2,3,5-tetr a h y-
dr o-4,1-ben zoxazepin -3-yl]acetyl]piper idin e-4-acetic Acid
(4b). 10% Pd-C catalyst (50 mg) was added to a solution of
11a (0.36 g, 0.480 mmol) in AcOEt (10 mL). The reaction
mixture was stirred under H2 atmosphere for 30 min at room
temperature. The catalyst was removed by filtration, and the
filtrate was concentrated under reduced pressure. The residue
was recrystallized from AcOEt-hexane (1:1) to give 4b (0.27
g, 0.410 mmol, 85%) as a colorless powder. Mp 191-192 °C.
[R]22D -191° (c 0.143, MeOH). IR νmax(KBr), cm-1: 3600-2400
(br, COOH), 1732, 1680, 1639 (CdO). 1H NMR (CDCl3): δ 0.95
(3H, s), 1.02 (3H, s), 1.12 (3H, t, J ) 7.6 Hz), 0.95-1.26 (2H,
m), 1.70-1.85 (2H, m), 1.90-2.10 (1H, m), 2.23-2.37 (4H, m),
2.50-2.77 (2H, m), 2.95-3.20 (2H, m), 3.56 (1H, d, J ) 13.6
Ben zyl 1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im eth oxyp h en yl)-
1-[2,2-d im eth yl-3-(p r op ion yloxy)p r op yl]-2-oxo-1,2,3,5-tet-
r a h yd r o-4,1-ben zoxa zep in -3-yl]a cetyl]p ip er id in e-4-a ce-
ta te (11a ). To a solution of 10 (0.5 g, 0.721 mmol), NEt3 (0.14
g, 1.42 mmol), and DMAP (9 mg, 0.0709 mmol) in THF (5 mL)
was added propionyl chloride (98 mg, 1.06 mmol) at 0 °C. The
mixture was stirred for 1 h at room temperature, diluted with
AcOEt (50 mL), washed with 5% KHSO4, saturated NaHCO3,
and brine, dried over Na2SO4, and then concentrated under
reduced pressure. The residue was subjected to column chro-
matography on silica gel [eluent: hexane-AcOEt (3:2)] to give
11a (0.46 g, 0.614 mmol, 85%) as a colorless amorphous
powder. [R]22 -167 (c 0.221, MeOH). IR νmax(KBr), cm-1
:
D
1736, 1682, 1639 (CdO). 1H NMR (CDCl3): δ 0.94 (3H, s), 1.02
(3H, s), 1.12 (3H, t, J ) 7.8 Hz), 0.88-1.26 (2H, m), 1.65-1.80
(2H, m), 1.90-2.10 (1H, m), 2.24-2.36 (4H, m), 2.48-2.61 (1H,
m), 2.71 (1H, dd, J ) 4.0, 15.0 Hz), 2.97-3.17 (2H, m), 3.56

4,1-Benzoxazepine Derivatives
J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 20 4579
Hz), 3.61 (3H, s), 3.71 (1H, d, J ) 11.0 Hz), 3.86 (1H, d, J )
11.0 Hz), 3.89 (3H, s), 3.89-3.99 (1H, m), 4.47-4.53 (2H, m),
4.56 (1H, d, J ) 13.6 Hz), 6.24 (1H, s), 6.62 (1H, s), 6.95-7.33
(5H, m). Anal. (C34H43N2O9Cl) C, H, N.
(for 30 s, twice). From the sonicate thus obtained, the mi-
crosome fraction was obtained by the same procedure as in
the preparation of rat-derived enzyme, which was suspended
in an ice-cooled 100 mM potassium phosphate buffer (pH 7.4)
(about 4 mg/mL protein concentration). This suspension was
used as the enzyme preparation.
1-[[(3R,5S)-1-[3-(Bu t yr yloxy)-2,2-d im et h ylp r op yl]-7-
ch lor o-5-(2,3-d im eth oxyp h en yl)-2-oxo-1,2,3,5-tetr a h yd r o-
4,1-ben zoxazepin -3-yl]acetyl]piper idin e-4-acetic Acid (4c).
Compound 4c (89 mg, 0.132 mmol, 84%) was prepared from
11b (0.12 g, 0.157 mmol) in the same manner as described for
Assa y of Sq u a len e Syn t h a se In h ib it or y Act ivit y.
Squalene synthase activity was monitored by the formation
of [3H]squalene from [1-3H]FPP. Fifty microliters of assay
mixture included 5 µM [1-3H]FPP (25 µCi/mol), 1 mM NADPH,
5 mM MgCl2, 6 mM glutathione, 100 mM buffer solution of
potassium phosphate (pH 7.4), the test compound dissolved
in DMSO (a final concentration of DMSO was 2%), and enzyme
solution prepared from rat or HepG2 cells (protein content of
0.8 µg). The assay ran for 45 min at 37 °C and was stopped by
adding 150 µL of CHCl3-MeOH (1:2) containing 0.2% cold
squalene as carrier. An aqueous solution of 3 N NaOH (50 µM)
and CHCl3 (50 µM) was added to the mixture. The chloroform
layer containing the reaction mixture having squalene as the
principal component and 3 mL of toluene-based liquid scin-
tillator were mixed, and the mixture’s radioactivity was
determined by means of a liquid scintillation counter. The
squalene synthase inhibitory activity was expressed in terms
of the concentration of the test compound inhibiting by 50%
the radioactivity taken into the chloroform layer (IC50, molar
concentration (M)).
Ch olester ogen esis in th e Liver in Wista r Ra ts. Six-
week-old male Wistar rats were orally administered test
compounds (0.5%)-methylcellulose emulsion and were intra-
venously injected with [14C]acetate (50 µCi/kg) 1 h after
administration. Animals were killed 1 h after the injection.
The livers were removed, saponified, extracted with petroleum
ether, and then dried under nitrogen blowing. The residue was
dissolved in the ethanol-acetone (1:1) (3 mL). The 0.5%
solution of digitonin in 50% ethanol (2 mL) was added to the
solution. After the mixture stood for 4 h at room temperature,
the cholesterol fraction was obtained as the digitonin precipi-
tate. The radioactivity taken into the digitonin precipitate was
measured.
the preparation of 4b. A colorless amorphous powder. [R]22
D
-182° (c 0.171, MeOH). IR νmax(KBr), cm-1: 3600-2400 (br,
1
COOH), 1732, 1680, 1635 (CdO). H NMR (CDCl3): δ 0.89-
1.02 (9H, s), 1.10-1.40 (2H, m), 1.57-1.85 (4H, m), 1.90-2.10
(1H, m), 2.22-2.30 (4H, m), 2.50-2.78 (2H, m), 3.00-3.20 (2H,
m), 3.57 (1H, d, J ) 14.0 Hz), 3.61 (3H, s), 3.70 (1H, d, J )
11.0 Hz), 3.80-3.98 (2H, m), 3.89 (3H, s), 4.45-4.60 (3H, m),
6.24 (1H, s), 6.62 (1H, s), 6.95-7.33 (5H, m). Anal. (C35H45N2O9-
Cl) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im et h oxyp h en yl)-1-[2,2-
d im eth yl-3-(isobu tyr yloxy)p r op yl]-2-oxo-1,2,3,5-tetr a h y-
dr o-4,1-ben zoxazepin -3-yl]acetyl]piper idin e-4-acetic Acid
(4d ). Compound 4d (0.21 g, 0.312 mmol, 79%) was prepared
from 11c (0.3 g, 0.393 mmol) in the same manner as described
for the preparation of 4b. A colorless powder. Mp 200-202 °C
(AcOEt-hexane). [R]22D -181° (c 0.120, MeOH). IR νmax(KBr),
cm-1: 3600-2400 (br, COOH), 1732, 1680, 1635 (CdO). 1H
NMR (CDCl3): δ 0.95 (3H, s), 1.03 (3H, s), 1.15 (3H, d, J )
7.0 Hz), 1.17 (3H, d, J ) 7.0 Hz), 0.90-1.35 (2H, m), 1.70-
1.85 (2H, m), 1.90-2.10 (1H, m), 2.22-2.31 (2H, m), 2.47-
2.77 (3H, m), 2.95-3.20 (2H, m), 3.57 (1H, d, J ) 14.0 Hz),
3.61 (3H, s), 3.69 (1H, d, J ) 11.0 Hz), 3.87 (1H, d, J ) 11.0
Hz), 3.89 (3H, s), 3.89-3.99 (1H, m), 4.48-4.53 (2H, m), 4.57
(1H, d, J ) 14.0 Hz), 6.23 (1H, s), 6.62 (1H, s), 6.99-7.33 (5H,
m). Anal. (C35H45N2O9Cl) C, H, N.
1-[[(3R,5S)-7-Ch lor o-5-(2,3-d im et h oxyp h en yl)-1-[2,2-
d im eth yl-3-(p iva loyloxym eth yloxy)p r op yl]-2-oxo-1,2,3,5-
t et r a h yd r o-4,1-b en zoxa zep in -3-yl]a cet yl]p ip er id in e-4-
a cetic Acid (4e). 10% Pd-C catalyst (50 mg) was added to a
solution of 11d (0.25 g, 0.310 mmol) in AcOEt (10 mL). The
reaction mixture was stirred under H2 atmosphere for 30 min
at room temperature. The catalyst was removed by filtration,
and the filtrate was concentrated under reduced pressure to
give 4e (0.19 g, 0.265 mmol, 85%) as a colorless amorphous
Ack n ow led gm en t. The authors thank Dr. Teruaki
Okuda for the pharmacokinetic analysis.
Su p p or tin g In for m a tion Ava ila ble: Graphs from Line-
weaver-Burk and Dixon analyses of squalene synthase inhibi-
tion by compounds 3j and 4a . This material is available free
of charge via the Internet at http://pubs.acs.org.
powder. [R]22 -147 (c 0.447, MeOH). IR νmax(KBr), cm-1
:
D
3600-2400 (br, COOH), 1732, 1680, 1641 (CdO). 1H NMR
(CDCl3): δ 0.94 (3H, s), 1.00 (3H, s), 1.17 (9H, s), 1.10-1.60
(2H, m), 1.70-1.90 (2H, m), 1.90-2.10 (1H, m), 2.23-2.31 (2H,
m), 2.55-2.80 (2H, m), 3.00-3.35 (4H, m), 3.64 (3H, s), 3.64
(1H, d, J ) 11.2 Hz), 3.90 (3H, s), 3.90-3.99 (1H, m), 4.33-
4.59 (3H, m), 4.70 (1H, d, J ) 6.2 Hz), 5.19 (1H, d, J ) 6.2
Hz), 6.23 (1H, s), 6.59 (1H, s), 6.97-7.35 (5H, m). Anal.
(C38H49N2O10Cl‚0.5H2O) C, H, N.
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An im a ls a n d Ma ter ia ls. Animals were purchased from
Clea, J apan, Inc. unless otherwise mentioned. Male Wistar
rats were allowed access to standard rodent chow (CE-2 in
pellet form, Clea J apan). RS-[2-14C]Mevalonolactone and [1-3H]-
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