Novel seco cyclopropa[c]pyrrolo[3,2-e]indole bisalkylators bearing a 3,3′-arylenebisacryloyl group as a linker

Article abstract of DOI:10.1021/jm000107x

We synthesized the novel seco cyclopropa[c]pyrrolo[3,2-e]indole (CPI) bisalkylators and evaluated their antitumor activity. Among these derivatives, 11a (AT-760), in which the two seco 3-methoxycarbonyl-2-trifluoromethyl CPI (MCTFCPI) moieties are connected with a 3,3′-(1,4-phenylene)bisacryloyl group, was found to exhibit more potent cytotoxicity and antitumor activity against HeLaS3 human uterine cervix carcinoma cells and Colon 26 adenocarcinoma cells, respectively, than 8 (bizelesin, U-77,779). It also appeared that compound 11a exhibits improved in vivo efficacy in the human colon CX-1 model when compared to either compound 8 or mitomycin C (MMC). Efficacious doses for 11a were found to be 2-fold lower than those for 8.

Full text of DOI:10.1021/jm000107x

1396
J . Med. Chem. 2001, 44, 1396-1406
Novel Seco Cyclop r op a [c]p yr r olo[3,2-e]in d ole Bisa lk yla tor s Bea r in g a
3,3′-Ar ylen ebisa cr yloyl Gr ou p a s a Lin k er
Yasumichi Fukuda,* Shigeki Seto, Hirosuke Furuta, Hiroyuki Ebisu, Yasuo Oomori, and Shiro Terashima†
Central Research Laboratories, Kyorin Pharmaceutical Company Ltd., Mitarai, Nogi, Tochigi 329-0114, J apan, and
Sagami Chemical Research Center, Nishi-Ohnuma, Sagamihara, Kanagawa 229-0012, J apan
Received October 11, 2000
We synthesized the novel seco cyclopropa[c]pyrrolo[3,2-e]indole (CPI) bisalkylators and
evaluated their antitumor activity. Among these derivatives, 11a (AT-760), in which the two
seco 3-methoxycarbonyl-2-trifluoromethyl CPI (MCTFCPI) moieties are connected with a 3,3′-
(1,4-phenylene)bisacryloyl group, was found to exhibit more potent cytotoxicity and antitumor
activity against HeLaS3 human uterine cervix carcinoma cells and Colon 26 adenocarcinoma
cells, respectively, than 8 (bizelesin, U-77,779). It also appeared that compound 11a exhibits
improved in vivo efficacy in the human colon CX-1 model when compared to either compound
8 or mitomycin C (MMC). Efficacious doses for 11a were found to be 2-fold lower than those
for 8.
In tr od u ction
ing even more potent antitumor activity than 8 and 9,
we designed and synthesized the seco MCTFCPI bis-
alkylator 11a carrying a 3,3′-(1,4-phenylene)bisacryloyl
group whose length is shorter than that in 9.10 Taking
into account the potent antitumor activity of 11a , we
carried out the synthesis of the various seco CPI
bisalkylators 10 bearing the same 3,3′-(1,4-phenylene)-
bisacryloyl group as a linker (Chart 4).10 On the basis
of the results of these studies, we next investigated the
synthesis and evaluation of the antitumor activity of
novel seco MCTFCPI bisalkylators 11b-q to clarify the
structure-activity relationships (SARs) of the linker
part of the MCTFCPI bisalkylators. Herein, we report
on the synthesis and antitumor activity of 11a -q.
Among them, 11a (AT-760) was found to exhibit more
potent cytotoxicity and antitumor activity against
HeLaS3 human uterine cervix carcinoma cells and
Colon 26 adenocarcinoma cells, respectively, than 8 and
less toxicity than 9. It also appeared that, in the human
colon CX-1 model, 11a exhibits more potent activity
than does mitomycin C and similar activity to 8.
The antitumor antibiotics 1 (CC-1065),1 2 (duocar-
mycin A),2 and 3 (duocarmycin SA),3 carrying a cyclo-
propa[c]pyrrolo[3,2-e]indole (CPI) moiety as the common
pharmacophore, are isolated from Streptomyces sp. The
CPI derivatives have been recognized as monoalkylators
whose CPI systems are responsible for their potent
cytotoxicity through sequence-selective alkylation of
double-strand DNA.4 The seco-type CPI derivatives 5
(carzelesin, U-80,244)5 and 6 (KW-2189)6 derived from
1 and 2, respectively, are presently under clinical trials
(Chart 1). Recently, we reported the synthesis and
antitumor activity of the novel seco 3-methoxycarbonyl-
2-trifluoromethyl CPI (seco MCTFCPI) derivative 7 (AT-
3510) showing antitumor activity against human tumor
xenografts more potent than that of 5 or 6 or the
clinically widely used anticancer agent cisplatin (Chart
2).7 It has been considered that the seco-type CPI de-
rivatives serve as ring-opened prodrugs of corresponding
cytotoxic CPI derivatives.6b,8a,d
It is reported that 8 (bizelesin, U-77,779), in which
two alkylating moieties are connected with a linker, a
1,3-bis(2-carbonyl-1H-indol-5-yl)urea group, behaves as
a bisalkylator.8 The antitumor activity of 8 against
L1210 murine leukemia cells is obviously superior to
that of other CPI mono- and bisalkylators.8 The bis-
alkylator 8 is presently in phase I clinical trials.8d More
recently, we reported the synthesis and antitumor
activity of the novel seco MCTFCPI bisalkylator 9.9 In
this compound, the two seco MCTFCPI groups are con-
nected with a linker, the 5,5′-bis(2-carbonyl-1H-indole)
group. We found that 9 shows more potent antitumor
activity than does 8 (Chart 3). Based on our studies, it
is evident that the length of a linker has a more sig-
nificant influence on cytotoxicity and antitumor activity
rather than does the type of linker.9 Therefore, with the
aim of exploring the novel seco CPI bisalkylators show-
Resu lts a n d Discu ssion
Ch em istr y. We synthesized seco MCTFCPI bisalky-
lators 11b-q following the procedure for synthesizing
11a reported in our earlier paper.10 Deprotection of the
optically pure 12 under acidic conditions gave the
indoline 13 as its hydrochloride. This was immediately
coupled with various 3,3′-arylenebisacrylic acids 14b-q
in the presence of 1-[3-(dimethylamino)propyl]-3-ethyl-
carbodiimide hydrochloride (EDCI) to afford the seco
MCTFCPI bisalkylators 11b-q (Scheme 1).
While 3,3′-(1,4-phenylene)bisacrylic acid (14a ) is com-
mercially available, the synthesis of 14c-e,m -q was
accomplished by employing a Heck reaction of various
diazonium salts (for 14d ), bistriflates (for 14e,m ,n ,p ,q),
or dibromides (for 14c,o) and by employing a Doebner
reaction of various dialdehydes (for 14b,f-l). Thus, the
Heck reaction of 1,2-dibromobenzene 15 with ethyl
acrylate provided diethyl ester 16c. The synthesis of
diethyl ester 16d was achieved from 1,2-diphenylhy-
* To whom correspondence should be addressed. Tel: 81 280 56
2201. Fax: 81 (0) 280 57 1293. E-mail: yasumichi.fukuda@mb2.
kyorin-pharm.co.jp.
† Sagami Chemical Research Center.
10.1021/jm000107x CCC: $20.00 © 2001 American Chemical Society
Published on Web 03/24/2001

Seco Cyclopropa[c]pyrrolo[3,2-e]indole Bisalkylators
J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9 1397
Ch a r t 1
Ch a r t 2
prepared directly from the corresponding 1,4-quinones
24, 25, and 29 without isolation of the unstable hydro-
quinones 26m ,n ,p . The synthesis of bistriflate 23q was
achieved according to the reported method.13 A Heck
reaction of these bistriflates 23e,m ,n ,p ,q with ethyl
acrylate smoothly took place to give diethyl esters
16e,m ,n ,p ,q. Unfortunately, a Heck reaction of bistri-
flate 23o was found to give 9,10-anthraquinone 28 as
the sole product instead of the expected diethyl ester
16o under these conditions. The Heck reaction of 9,10-
dibromoanthrathene (27), however, cleanly provided the
desired 16o. The resulting diethyl esters 16c-e,m -q
were hydrolyzed under basic conditions to afford the
bisacrylic acids 14c-e,m -q.The synthesis of dialde-
hydes 32h -j was achieved by alkylation of dimethyl 2,3-
dihydroxyterephthalate (30)14 in the presence of cesium
carbonate, followed by reduction with Red-Al/N-meth-
ylpiperazine reagent. The syntheses of dialdehydes
32f,g and bisacrylic acid 14k were carried out according
to the reported procedure,15 as was the synthesis of
dialdehyde 32l.16 Thus, treatment of the bisoxazolidine
derivative 34 derived from 2,3-dimethoxyterephthaloyl
dichloride (33)17 with ethylmagnesium bromide cleanly
produced the diethyl derivative 35. This was converted
to dialdehyde 32l by alkylation with methyl iodide and
subsequent reduction with sodium borohydride. The
Doebner reaction of dialdehydes 32b,f-l cleanly pro-
vided the corresponding 3,3′-arylenebisacrylic acids
14b,f-l.
Ch a r t 3
Ch a r t 4
Cytotoxicity. The results of cytotoxicity assay of the
seco MCTFCPI bisalkylator 11a and the newly synthe-
sized derivatives 11b-q against HeLaS3 human uterine
cervix carcinoma cells are summarized in Table 1. The
bisalkylator 11a bearing a 3,3′-(1,4-phenylene)bisacryl-
oyl group exhibited more potent cytotoxicity than did 8
or 9. The strong cytotoxicity observed for 11a disap-
peared in its positional isomers 11b,c.10 Cytotoxicity of
the bisalkylators 11d ,f,g, in which one or more phen-
ylene group(s) are added to the linker of 11a , reduced
dramatically. The bipyridyl analogue 11e showed su-
perior cytotoxicity to the corresponding biphenyl deriva-
tive 11d . Cytotoxicity of 11h -j in which the C-2 and
drazine (17) according to the reported method,11 with
slight modification to avoid the isolation of carcinogenic
benzidine. The homo-coupling reaction of the 2-bro-
mopyridine 2012 in the presence of NiBr2(PPh3)2 cleanly
provided the bipyridyl derivative 21. This was converted
to bistriflate 23e by sequential treatments with 47%
HBr and triflic anhydride. Bistriflates 23m ,n ,p were

1398 J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9
Fukuda et al.
Sch em e 1a
a
(a) (i) 3 M HCl-AcOEt, (ii) EDCI, 14a -q, 46-82% (2 steps).
C-3 positions of the 1,4-phenylene group are substituted
by dimethoxy, methylenedioxy, or ethylenedioxy group-
(s) was comparable to that of 11a . Interestingly, 11k ,
carrying dimethoxy groups at the C-2 and C-5 positions
of 1,4-phenylene group, exhibited cytotoxicity 3500
times less than that of 11a . In contrast to the case of
11h , the additional methoxy group at the C-5 position
of the phenylene ring debilitates cytotoxicity, probably
due to its steric hindrance of the interaction in a minor
groove of duplex DNA. Although the naphthalene
analogues 11m ,n turned out to be 3-4 times less
cytotoxic than 11a , the anthracene analogues 11o-q
exhibited cytotoxicity about 1000 times less than that
of 11a . From these results, it appeared that cytotoxicity
of the bisalkylators depends highly not only on the
length but also on the width of the linker. As for 11a ,
further cytotoxicity assays were also made against the
human cancer cell lines SBC-3 (lung), SBC-3/ADR (lung,
adriamycin-resistant), PC-3 (lung), KATO III (gastric),
ZR-75-1 (breast), A2780 (ovarian), and DLD-1 (colon).
As shown in Table 2, cytotoxicity of 11a was superior
to that of adriamycin (ADR) in all human cancer cell
lines tested and 11a showed different cytotoxic proper-
ties compared with those of adriamycin.
shows more potent antitumor activity than 8 and less
toxicity than 9 in the murine Colon 26 model. Next, we
further evaluated in vivo antitumor activity against
human colon CX-1 tumor xenografts for 11a and 8. The
results are shown in Table 3 and Figure 1. Compound
11a exhibits improved in vivo efficacy in the human
colon CX-1 model when compared to either compound
8 or mitomycin C (MMC). Efficacious doses for 11a were
found to be 2-fold lower than those for 8.
Con clu sion
As described above, we have succeeded in the design,
synthesis, and evaluation of the novel seco MCTFCPI
bisalkylators 11a -q carrying various 3,3′-arylenebi-
sacryloyl groups. We found that the bisalkylator 11a ,
bearing a 3,3′-(1,4-phenylene)bisacryloyl group as a
linker, exhibited more potent antitumor activity or less
toxicity than did the other bisalkylators 11b-q in the
Colon 26 model. Moreover, 11a , much like clinical trial
candidate 8, shows more potent antitumor activity in
the human colon CX-1 system xenografts model than
the clinically widely used anticancer agent mitomycin
C. Further investigation of the pharmacological profiles
of 11a is in progress.
In Vivo An titu m or Activity. The seco MCTFCPI
bisalkylators 11a -q were subjected to in vivo antitumor
activity assay against Colon 26 murine adenocarcinoma
cells. The results shown in Table 1 indicate that 11a
Exp er im en ta l Section
All melting points were determined with a Yamato MP-500
melting point apparatus and are uncorrected. Measurements
of optical rotations were carried out using a J ASCODIP-360

Seco Cyclopropa[c]pyrrolo[3,2-e]indole Bisalkylators
J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9 1399
Sch em e 2a
a
(a) (i) Ethyl acrylate, Pd(OAc)2, DPPP, Et3N, 31%, (ii) KOH, 91%; (b) (i) 42% HBF4, (ii) NaNO2, 42% HBF4; (c) (i) ethyl acrylate,
Pd(OAc)2, 29%, (ii) KOH, 83%; (d) (i) NiBr2(PPh3)2, Zn, Et4NI, 87%, (ii) 47% HBr, 84%, (iii) Tf2O, 2,4,6-collidine, 84%; (e) (i) ethyl acrylate,
Pd(OAc)2, DPPP, Et3N, 86%, (ii) KOH, 89%; (f) (i) 10% Pd-C, H2, (ii) Tf2O, 2,4,6-collidine, 84% (for 23m ), Tf2O, DMAP, 67% (for 23n ); (g)
(i) ethyl acrylate, Pd(OAc)2, DPPP, Et3N, 85% (for 16m ), 92% (for 16n ), (ii) KOH, 96% (for 14m ), 93% (for 14n ); (h) (i) ethyl acrylate,
Pd(OAc)2, DPPP, Et3N, 94% (for 16o), (ii) KOH, 96%; (i) ethyl acrylate, Pd(OAc)2, DPPP, Et3N.
automatic digital polarimeter. Infrared (IR) spectra were
recorded on a J ASCO FT/IR-5300 spectrometer. 1H NMR
spectra were measured with a J EOL J NM-EX-400 (400 MHz)
spectrometer. The chemical shifts are expressed in parts per
million (δ value) downfield from tetramethylsilane, using
tetramethylsilane (δ ) 0) and/or residual solvents such as
chloroform (δ ) 7.26) and benzene (δ ) 7.20) as an internal
standard. Splitting patterns are indicated as s, singlet; d,
doublet; t, triplet; q, quartet; m, multiplet; br, broad peak.
Measurements of mass spectra were performed with a Hitachi
M-2000 mass spectrometer. Data for elemental analysis are
within (0.3% of theoretical values and were determined by a
Yanaco CHN corder MT-5. Unless otherwise noted, all the
experiments were carried out using anhydrous solvents under
an atmosphere of dry argon. Tetrahydrofuran and diethyl ether
(ether) were distilled from sodium benzophenone ketyl.
Throughout this work, Merck precoated thin layer chromato-
graphic (TLC) plates (silica gel 60 F254, 0.25 mm, Art 5715)
were used for TLC analyses. Wako Gel C-200 and C-300 were
used as an adsorbent for flash column chromatography. To
minimize the health risks posed by these potent cytotoxic
compounds to analytical service personnel of our laboratory
and to allow preparation of only the very limited quantities
needed for testing, infrared spectra and combustion elemental
analyses were not obtained for the final analogues.4c
Dieth yl 3,3′-(1,2-P h en ylen e)bisa cr yla te (16c). A mixture
of 1,2-dibromobenzene (500 mg, 2.1 mmol), ethyl acrylate (4.6
mL, 42 mmol), triethylamine (1.2 mL, 8.6 mmol), 1,3-(diphen-
ylphosphino)propane (87 mg, 0.21 mmol), and palladium(II)
acetate (48 mg, 0.21 mmol) in DMF (100 mL) was stirred at
100 °C for 18 h. After concentration in vacuo, the residue was
dissolved in CH2Cl2. The dichloromethane solution was washed
with water, dried over anhydrous Na2SO4, filtered, and then
concentrated in vacuo. Flash chromatography (CH2Cl2) of the
residue gave 16c as yellow crystals (181 mg, 31%). Mp: 75-
1
76 °C. H NMR (CDCl3): δ 1.35 (t, J ) 7.3 Hz, 6H), 4.29 (q, J
) 7.3 Hz, 4H), 6.35 (d, J ) 15.6 Hz, 2H), 7.40 (dd, J ) 5.9 Hz,
3.4 Hz, 2H), 7.57 (dd, J ) 5.9 Hz, 3.4 Hz, 2H), 8.04 (d, J )
15.6 Hz, 2H). IR (KBr): 1709, 1637, 1626, 1314, 1186 cm-1
.
MS (EI) m/z: 274 (M+). Anal. (C16H18O4) C, H, N.
3,3′-(1,2-P h en ylen e)bisa cr ylic Acid (14c). A suspension
of 16c (98 mg, 0.36 mmol) and KOH (202 mg, 3.6 mmol) in
EtOH (2 mL) was heated at reflux for 2 h. After cooling, the
mixture was adjusted to pH 1 by the addition of 1 N HCl
solution. The resulting precipitates were collected by filtration,
washed with water, and then dried in vacuo to give 14c as
colorless crystals (71 mg, 91%). Mp: >300 °C. 1H NMR
(DMSO-d6): δ 6.44 (d, J ) 15.7 Hz, 2H), 7.47 (dd, J ) 5.9 Hz,
3.9 Hz, 2H), 7.77 (dd, J ) 5.9 Hz, 3.9 Hz, 2H), 7.90 (d, J )
15.7 Hz, 2H), 12.60 (br, 2H). IR (KBr): 2976, 2828, 1690, 1624,

1400 J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9
Fukuda et al.
Sch em e 3a
a
(a) (i) Na2S2O4, (ii) Tf2O, 2,4,6-collidine, 66%; (b) (i) ethyl acrylate, Pd(OAc)2, DPPP, Et3N, 90%, (ii) KOH, 90%; (c) (i) ethyl acrylate,
Pd(OAc)2, DPPP, Et3N, 90%, (ii) KOH, 90%; (d) Cs2CO3, MeI, 99% (for 31h ), BrCH2Cl, 98% (for 31i), BrCH2CH2Br, 99% (for 31j); (e)
NaAlH2(OCH2CH2OMe)2, N-methylpiperazine, 80% (for 32h ), 66% (for 32i), 68% (for 32j).
Sch em e 4a
a
(a) (i) 2-Amino-2-methyl-1-propanol, 100%, (ii) SOCl2, 47%, (iii) EtMgBr, 100%; (b) (i) MeI, 76%, (ii) NaBH4, 13%; (c) malonic acid,
pyridine, piperidine, 91% (for 14b), 95% (for 14f), 93% (for 14g), 93% (for 14h ), 95% (for 14i), 95% (for 14j), 61% (for 14k ), 74% (for 14l).
1292 cm-1. MS (EI) m/z: 218 (M+). Anal. (C12H10O4‚1/10H2O)
C, H, N.
water. The aqueous solution was adjusted to pH 2-3 by the
addition of 1 N HCl solution. The resulting precipitates were
collected by filtration, washed with water, and then dried in
vacuo to give 14d as colorless crystals (140 mg, 83%). Mp:
>300 °C. 1H NMR (DMSO-d6): δ 6.60 (d, J ) 15.7 Hz, 2H),
7.64 (d, J ) 15.7 Hz, 2H), 7.80 (s, 8H), 12.48 (brs, 2H). IR
(KBr): 2970, 1680, 1624, 1316, 1215 cm-1. MS (EI) m/z: 294
(M+). Anal. (C18H14O4‚1/5H2O) C, H, N.
5,5′-Dim eth oxy-2,2′-bip yr id yl (21). To a suspension of
zinc dust (2.22 g, 34 mmol), bis(triphenylphosphine)nickel(II)
bromide (5.07 g, 6.8 mmol), and tetraethylammonium iodide
(5.82 g, 23 mmol) in THF (45 mL) was added a solution of 20
(4.26 g, 23 mmol) in THF (22 mL) at 50 °C. The mixture was
stirred at the same temperature for 8 h, and then added to 2
M ammonia solution. The resulting solution was extracted
with ether-benzene (1:1). The combined organic extracts were
washed with water and brine, dried over anhydrous Na2SO4,
filtered, and then concentrated in vacuo. Flash chromatogra-
phy (CH2Cl2:AcOEt ) 1:1) of the residue gave 21 as colorless
crystals (2.14 g, 87%). Mp: 130-133 °C (cyclohexane). 1H NMR
(CDCl3): δ 3.91 (s, 6H), 7.30 (dd, J ) 8.8 Hz, 2.9 Hz, 2H), 8.25
(d, J ) 8.8 Hz, 2H), 8.33 (d, J ) 2.9 Hz, 2H). IR (KBr): 1562,
1467, 1289, 1258, 1227 cm-1. MS (EI) m/z: 216 (M+). HRMS
(EI) for C12H12N2O2 (M+): calcd, 216.0899; found, 216.0903.
5,5′-Dih yd r oxy-2,2′-bip yr id yl (22). A mixture of 21 (395
mg, 1.8 mmol) in acetic acid (7 mL) and 47% HBr solution (7
Dieth yl 3,3′-(1,1′-Dip h en yl-4,4′-d iyl)bisa cr yla te (16d ).
To a solution of 42% HBF4 (12.5 g, 58 mmol) was added 17
(2.15 g, 12 mmol) at room temperature, and the mixture was
stirred at the same temperature for 2.5 h. After the addition
of more 42% HBF4 solution (10 mL), NaNO2 (1.64 g, 24 mmol)
in water (4 mL) was added to the mixture at 0 °C, and the
whole mixture was stirred at the same temperature for 1.5 h.
MeOH (0.7 mL), ethyl acrylate (3.5 mL, 32 mmol), and Pd-
(OAc)2 (49 mg, 0.22 mmol) were added to the resulting solution.
The mixture was stirred at 55-60 °C for 1 h and then
extracted with ether. The combined organic extracts were
washed with saturated NaHCO3 solution, dried over anhy-
drous Na2SO4, filtered, and then concentrated in vacuo. Flash
chromatography (CH2Cl2:benzene ) 1:1) of the residue gave
1
16d as colorless crystals (1.17 g, 29%). Mp: 147-149 °C. H
NMR (CDCl3): δ 1.35 (t, J ) 7.3 Hz, 6H), 4.28 (q, J ) 7.3 Hz,
4H), 6.49 (d, J ) 15.6 Hz, 2H), 7.60 (d, J ) 8.3 Hz, 4H), 7.65
(d, J ) 8.3 Hz, 4H), 7.72 (d, J ) 15.6 Hz, 2H). IR (KBr): 1705,
1631, 1306, 1208, 1182 cm-1. MS (EI) m/z: 350 (M+). Anal.
(C22H22O4) C, H, N.
3,3′-(1,1′-Dip h en yl-4,4′-d iyl)bisa cr ylic Acid (14d ). A sus-
pension of 16d (200 mg, 0.57 mmol) and KOH (377 mg, 6.7
mmol) in EtOH (3 mL) was heated at reflux for 1 h. After
concentration in vacuo, the resulting residue was dissolved in

Seco Cyclopropa[c]pyrrolo[3,2-e]indole Bisalkylators
J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9 1401
Ta ble 1. Cytotoxicity against HeLaS3 Human Uterine Cervix
Carcinoma Cells and in Vivo Antitumor Activity against Colon
26 Murine Adenocarcinoma Cells of the MCTFCPI
Bisalkylators Bearing 3,3′-Aryldiacryloyl Linkers
IC50
max; TGI%
body wt
compd
(ng/mL)a
(µg/kg)b
change (%) MTDc/TGI50
11a
11b
11c
11d
11e
11f
11g
11h
11i
11j
11k
11l
11m
11n
11o
11p
11q
8
0.00274
54.6
87.6
89 (1.95)
91 (4000)
70 (4000)
83 (15.6)
88 (3.91)
nt
84 (4000)
89 (0.977)
90 (1.95)
91 (0.488)
81 (2000)
nt
-5
-5
-6
4
4
nt
-11
-1
-2
3
3
nt
30.7
nt
nt
3.7
3.6
nt
0.141
0.0198
>100
48.3
0.00158
nt
4.6
13.3
9.0
>6.5
nt
7.4
nt
nt
3.5
6.6
8.4
3.3
0.00595
0.00276
9.71
0.00318
0.011
0.00934
1.51
3.25
85 (0.977)
nt
3
nt
75 (4000)
82 (15.6)
84 (15.6)
90 (15.6)
84 (0.977)
-18
-6
-4
-3
1
F igu r e 1. In vivo antitumor activity against human colon
CX-1 tumor xenografts.
0.523
0.060
0.0049
9
filtered, and then concentrated in vacuo. Flash chromatogra-
a
phy (CH2Cl2) of the residue gave 23e as colorless crystals (567
Drug concentration required to inhibit the growth of HeLaS3
1
b
mg, 84%). Mp: 158-162 °C. H NMR (CDCl3): δ 7.78 (dd, J
human uterine cervix carcinoma cells by 50%. The percentage
) 8.9 Hz, 2.4 Hz, 2H), 8.56 (d, J ) 8.8 Hz, 2H), 8.65 (d, J )
2.4 Hz, 2H). IR (KBr): 1468, 1422, 1408, 1376, 1254, 1216,
1162, 1139 cm-1. MS (EI) m/z: 452 (M+). Anal. (C12H6F6N2O6S2)
C, H, N.
tumor growth inhibition as compared with the untreated group.
c Maximum dose within 10% body weight loss. nt, not tested.
Ta ble 2. Cytotoxicity of 11a and Adriamycin against Human
Cancer Cell Lines
Dieth yl 3,3′-(2,2′-Bip yr id yl-5,5′-d iyl)bisa cr yla te (16e).
A mixture of 23e (83.2 mg, 0.18 mmol), ethyl acrylate (0.4 mL,
3.6 mmol), triethylamine (0.1 mL, 0.72 mmol), 1,3-(diphen-
ylphosphino)propane (7.4 mg, 18 µmol), and palladium(II)
acetate (4.0 mg, 18 µmol) in DMF (14 mL) was stirred at 100
°C for 10 h. After concentration in vacuo, the residue was
dissolved in CH2Cl2. The dichloromethane solution was washed
with water, dried over anhydrous Na2SO4, filtered, and then
concentrated in vacuo. Flash chromatography (CH2Cl2:AcOEt
) 10:1) of the residue gave 16e as yellow crystals (54.3 mg,
86%). Mp: 173.5-175 °C. 1H NMR (CDCl3): δ 1.36 (t, J ) 7.3
Hz, 6H), 4.30 (q, J ) 7.3 Hz, 4H), 6.58 (d, J ) 16.1 Hz, 2H),
7.73 (d, J ) 16.1 Hz, 2H), 7.99 (dd, J ) 8.3 Hz, 2.0 Hz, 2H),
8.48 (d, J ) 8.3 Hz, 2H), 8.80 (d, J ) 2.0 Hz, 2H). IR (KBr):
1705, 1637, 1472, 1369, 1308, 1266, 1208, 1175 cm-1. MS (EI)
m/z: 352 (M+). Anal. (C20H20N2O4) C, H, N.
IC50 (ng/mL)a
SBC-3/
ADR
KATO
III ZR-75-1 A2780 DLD-1
compd SBC-3
11a
PC-3
0.00122 0.00349 0.148 0.816 0.00463 0.000397 0.0149
ADR 11.2
129
275
905
6.39
12.1
136
a
Drug concentration required to inhibit the growth of HeLaS3
human uterine cervix carcinoma cells by 50%.
Ta ble 3. Antitumor Activity of the Bisalkylators 8 and 11a
Evaluated Using Athymic Mice on Which Human Colon CX-1
Tumor Xenografts Were Implanted
compd
dose (µg/kg)a
TGI%b
11a
3.91
7.81
7.81
77
83
53
56
65
3,3′-(2,2′-Bip yr id yl-5,5′-d iyl)b isa cr ylic Acid (14e). A
suspension of 16e (100 mg, 0.28 mmol) and KOH (157 mg, 2.8
mmol) in EtOH (3 mL) was heated at reflux for 2 h and then
concentrated in vacuo. The resulting residue was dissolved in
water, and the pH of the aqueous solution was adjusted to 7
by the addition of 1 N HCl solution. The resulting precipitates
were collected by filtration, washed with water and ethanol,
and then dried in vacuo to give 14e as colorless crystals (73.8
8
15.6
9180
MMC
a
Tumor fragments of human colon CX-1 were implanted sc on
day 0. Drugs were administered iv on day 19. The percentage
tumor growth inhibition as compared with the untreated group.
b
1
mL) was heated at reflux for 37 h. After cooling, the resulting
precipitates were collected by filtration and dried in air. The
obtained crystals were dissolved in water, and the aqueous
solution was adjusted to pH 7 by the addition of 10% NaOH
solution. The resulting precipitates were collected by filtration
and dried in vacuo to give 22 as a pale yellow powder (290
mg, 89%). Mp: >300 °C. H NMR (DMSO-d6): δ 6.78 (d, J )
15.6 Hz, 2H), 7.71 (d, J ) 15.6 Hz, 2H), 8.35 (dd, J ) 8.8 Hz,
2.4 Hz, 2H), 8.45 (d, J ) 8.8 Hz, 2H), 9.00 (d, J ) 2.4 Hz, 2H).
IR (KBr): 3435, 1692, 1593, 1547, 1468 cm-1. MS (FAB+)
m/z: 297 (M+ + 1). HRMS (FAB+) for C16H13N2O4 (M+ + 1):
calcd, 297.0875; found, 297.0875. Anal. (C16H12N2O4‚1/4H2O)
C, H, N.
1
mg, 84%). Mp: 273-276 °C. H NMR (CDCl3): δ 7.74 (d, J )
8.8 Hz, 2H), 8.29 (d, J ) 2.4 Hz, 2H), 8.37 (dd, J ) 8.8 Hz, 2.4
1,4-Bis[(t r iflu or om et h a n esu lfon yl)oxy]n a p h t h a len e
(23m ). A suspension of 24 (500 mg, 3.2 mmol) and 10% Pd-C
(50 mg) in THF (10 mL) was stirred at room temperature for
2 h under H2 atmosphere (1 atm). After being ventilated with
Ar gas, 2,4,6-collidine (3.3 mL, 25 mmol) and trifluoromethane-
sulfonic anhydride (1.4 mL, 8.3 mmol) were added to the
resulting mixture at 0 °C, and the whole was stirred at room
temperature for 18 h. After insoluble materials were filtered
off, the filtrate was concentrated in vacuo, and the residue was
dissolved in ether. The ethereal solution was washed with
water, saturated CuSO4 solution, and brine, dried over anhy-
drous Na2SO4, filtered, and then concentrated in vacuo. Flash
chromatography (Hex:AcOEt ) 15:1) of the residue gave 23m
Hz, 2H), 11.28 (brs, 2H). IR (KBr): 2992, 1480, 1292 cm-1
.
MS (EI) m/z: 188 (M+). HRMS (EI) for C10H8N2O2 (M+): calcd,
188.0586; found, 188.0587.
5,5′-B is[(t r iflu o r o m e t h a n e su lfo n yl)ox y]-2,2′-b ip y -
r id yl (23e). Trifluoromethanesulfonic anhydride (0.6 mL, 3.7
mmol) was added to a suspension of 22 (280 mg, 1.5 mmol)
and 2,4,6-colidine (2.0 mL, 15 mmol) in CH2Cl2 (7 mL) at 0
°C, and the mixture was stirred at room temperature for 3 h.
After adding more trifluoromethanesulfonic anhydride (0.6
mL, 3.7 mmol), the mixture was stirred at room temperature
for 3 h and then diluted with ether. The resulting solution was
washed with water and brine, dried over anhydrous Na2SO4,

1402 J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9
Fukuda et al.
as a pale yellow solid (1.11 g, 83%). Mp: 35.5-37 °C. 1H NMR
(CDCl3): δ 7.51 (s, 2H), 7.80 (dd, J ) 6.3 Hz, 3.4 Hz, 2H), 8.15
(dd, J ) 6.3 Hz, 3.4 Hz, 2H). IR (KBr): 1607, 1424, 1387, 1221,
1142 cm-1. MS (EI) m/z: 424 (M+). HRMS (EI) for C12H6F6O6S2
(M+): calcd, 423.9510; found, 423.9512. Anal. (C12H6F6O6S2)
C, H, N.
MS (EI) m/z: 318 (M+). HRMS (EI) for C20H14O4 (M+): calcd,
318.0892; found, 318.0915. Anal. (C20H14O4‚1/5H2O) C, H, N.
1,4-Bis[(t r iflu or om e t h a n e su lfon yl)oxy]a n t h r a ce n e
(23p ). The compound 23p (526 mg, 23%) was prepared from
29 (1.00 g, 4.8 mmol), using 10% Pd-C (50 mg), 2,4,6-colidine
(5.0 mL, 38 mmol), and trifluoromethanesulfonic anhydride
(2.0 mL, 12 mmol), in the same manner as described for 23m .
Mp: 152.5-153.5 °C. 1H NMR (CDCl3): δ 7.47 (s, 2H), 7.67
(dd, J ) 6.3 Hz, 3.4 Hz, 2H), 8.14 (dd, J ) 6.3 Hz, 3.4 Hz, 2H),
8.71 (s, 2H). IR (KBr): 1423, 1213, 1139 cm-1. MS (EI) m/z:
474 (M+). Anal. (C16H8F6O6S2) C, H, N.
Dieth yl 3,3′-(1,4-An th r a cen ed iyl)bisa cr yla te (16p ). The
compound 16p (71.0 mg, 90%) was prepared from 23p (100
mg, 0.21 mmol) in the same manner as described for 16e. Mp:
98.5-99.5 °C. 1H NMR (CDCl3): δ 1.41 (t, J ) 6.8 Hz, 6H),
4.37 (q, J ) 6.8 Hz, 4H), 6.63 (d, J ) 16.1 Hz, 2H), 7.56 (dd,
J ) 6.3 Hz, 3.4 Hz, 2H), 7.75 (s, 2H), 8.08 (dd, J ) 6.3 Hz, 3.4
Hz, 2H), 8.67 (d, J ) 16.1 Hz, 2H), 8.78 (s, 2H). IR (KBr): 1706,
1629, 1255, 1178 cm-1. MS (EI) m/z: 374 (M+). Anal. (C24H22O4)
C, H, N.
3,3′-(1,4-An th r a cen ed iyl)bisa cr ylic Acid (14p ). The com-
pound 14p (152 mg, 90%) was prepared from 16p (200 mg,
0.53 mmol) similarly to the preparation of 14d . Mp: >300 °C.
1H NMR (DMSO-d6): δ 6.74 (d, J ) 15.7 Hz, 2H), 7.60 (dd, J
) 5.9 Hz, 2.9 Hz, 2H), 7.99 (s, 2H), 8.28 (dd, J ) 5.9 Hz, 2.9
Hz, 2H), 8.58 (d, J ) 15.7 Hz, 2H), 8.96 (s, 2H), 12.71 (br, 2H).
IR (KBr): 1684, 1621, 1419, 1286, 1259, 1210 cm-1. MS (EI)
m/z: 318 (M+). HRMS (EI) for C20H14O4 (M+): calcd, 318.0892;
found, 318.0894. Anal. (C20H14O4‚1/4H2O) C, H, N.
Dieth yl 3,3′-(1,4-Na p h th a len e)bisa cr yla te (16m ). The
compound 16m (19.5 mg, 85%) was prepared from 23m (30.0
mg, 71 µmol) in the same manner as described for 16e. Mp:
1
84-87 °C. H NMR (CDCl3): δ 1.38 (t, J ) 7.3 Hz, 6H), 4.33
(q, J ) 7.3 Hz, 4H), 6.55 (d, J ) 15.6 Hz, 2H), 7.63 (dd, J )
6.3 Hz, 3.4 Hz, 2H), 7.76 (s, 2H), 8.24 (dd, J ) 6.3 Hz, 3.4 Hz,
2H), 8.52 (d, J ) 15.6 Hz, 2H). IR (KBr): 1705, 1638, 1628,
1616, 1393, 1371, 1312, 1259 cm-1. MS (EI) m/z: 324 (M+).
Anal. (C20H20O4) C, H, N.
3,3′-(1,4-Na p h th a len e)bisa cr ylic Acid (14m ). The com-
pound 14m (90.2 mg, 96%) was prepared from 16m (115 mg,
0.35 mmol) in a manner similar to that described for the
preparation of 14d . Mp: >300 °C. 1H NMR (DMSO-d6): δ 6.66
(d, J ) 15.7 Hz, 2H), 7.70 (dd, J ) 6.9 Hz, 2.9 Hz, 2H), 7.98 (s,
2H), 8.28 (dd, J ) 6.9 Hz, 2.9 Hz, 2H), 8.40 (d, J ) 15.7 Hz,
2H), 12.66 (s, 2H). MS (EI) m/z: 268 (M+). HRMS (EI) for
C
16H12O4 (M+): calcd, 268.0736; found, 268.0743. Anal.
(C16H12O4) C, H, N.
1,4-Bis[(tr iflu or om eth a n esu lfon yl)oxy]-5,8-d im eth oxy-
n a p h th a len e (23n ). A suspension of 25 (200 mg, 0.92 mmol)
and 10% Pd-C (20 mg) in THF (3 mL) was stirred at room
temperature for 1 h under H2 atmosphere (1 atm). After
insoluble materials were filtered off, the filtrate was concen-
trated in vacuo. To a solution of the residue in CH2Cl2 (3 mL)
were added 4-(dimethylamino)pyridine (DMAP; 450 mg, 3.7
mmol) and trifluoromethanesulfonic anhydride (0.4 mL, 2.3
mmol) at 0 °C, and the mixture was stirred at room temper-
ature for 2 h. The mixture was washed with water, 1 N HCl
solution, and brine, dried over anhydrous Na2SO4, filtered, and
then concentrated in vacuo. Flash chromatography (Hex:CH2-
Cl2 ) 1:1) of the residue gave 23n as a colorless solid (299 mg,
Dieth yl 3,3′-(1,4-An th r a qu in oyl)bisa cr yla te (16q). The
compound 16q (49.0 mg, 60%) was prepared from 23q (100
mg, 0.20 mmol) in the same manner as described for 16e. Mp:
1
222.5-224.5 °C. H NMR (CDCl3): δ 1.39 (t, J ) 7.3 Hz, 6H),
4.34 (q, J ) 7.3 Hz, 4H), 6.28 (d, J ) 15.6 Hz, 2H), 7.77 (s,
2H), 7.81 (dd, J ) 5.9 Hz, 3.4 Hz, 2H), 8.24 (dd, J ) 5.9 Hz,
3.4 Hz, 2H), 8.58 (d, J ) 15.6 Hz, 2H). IR (KBr): 1700, 1661,
1631, 1592, 1331, 1281 cm-1. MS (EI) m/z: 404 (M+). Anal.
(C24H20O6) C, H, N.
1
67%). Mp: 145.5-146.5 °C. H NMR (CDCl3): δ 3.97 (s, 6H),
3,3′-(1,4-An th r a qu in oyl)bisa cr ylic Acid (14q). The com-
pound 14q (215 mg, 100%) was prepared from 16q (250 mg,
0.62 mmol) in a manner similar to that described for the
preparation of 14d . Mp: >300 °C. 1H NMR (DMSO-d6): δ 6.40
(d, J ) 15.7 Hz, 2H), 7.93 (dd, J ) 5.9 Hz, 2.9 Hz, 2H), 8.01 (s,
2H), 8.13 (dd, J ) 5.9 Hz, 2.9 Hz, 2H), 8.41 (d, J ) 15.7 Hz,
2H), 12.61 (br, 2H). IR (KBr): 1694, 1667, 1629, 1589, 1419,
1326, 1278 cm-1. MS (EI) m/z: 258 (M+ - 2CO2H). Anal.
(C20H12O6) C, H, N.
3,3′-(1,3-P h en ylen e)bisa cr ylic Acid (14b). A suspension
of 1,3-benzenedicarboxaldehyde (500 mg, 3.7 mmol), malonic
acid (1.72 g, 16.5 mmol), and pyridine-piperidine (70:1 v/v, 7
mL) was heated at reflux for 24 h. After cooling, the mixture
was adjusted to pH 1 by the addition of 1 N HCl solution. The
resulting precipitates were collected by filtration, washed with
water, and then dried in vacuo to give 14b as colorless crystals
(741 mg, 91%). Mp: 287-290 °C. 1H NMR (DMSO-d6): δ 6.67
(d, J ) 16.6 Hz, 2H), 7.46 (t, J ) 7.8 Hz, 1H), 7.61 (d, J )16.6
Hz, 2H), 7.72 (d, J ) 7.8 Hz, 2H), 8.08 (s, 1H), 12.46 (s, 2H).
IR (KBr): 3005, 1680, 1631, 1440, 1302 cm-1. MS (EI) m/z:
218 (M+). Anal. (C12H10O4‚1/10H2O) C, H, N.
7.00 (s, 2H), 7.29 (s, 2H). IR (KBr): 1609, 1431, 1421, 1379,
1272, 1248, 1217, 1202 cm-1. MS (EI) m/z: 484 (M+). Anal.
(C14H10F6O8S2) C, H, N.
Dieth yl 3,3′-(5,8-Dim eth oxy-1,4-n a p h th a len e)bisa cr y-
la te (16n ). The compound 16n (219 mg, 92%) was prepared
from 23n (300 mg, 0.62 mmol) in the same manner as
described for 16e. Mp: 119-122 °C. 1H NMR (CDCl3): δ 1.36
(t, J ) 7.3 Hz, 6H), 3.90 (s, 6H), 4.29 (q, J ) 7.3 Hz, 4H), 6.09
(d, J ) 15.6 Hz, 2H), 6.86 (s, 2H), 7.42 (s, 2H), 8.81 (d, J )
15.6 Hz, 2H). IR (KBr): 1724, 1710, 1633, 1457, 1394 cm-1
.
MS (EI) m/z: 384 (M+). Anal. (C22H24O6) C, H, N.
3,3′-(5,8-Dim et h oxy-1,4-n a p h t h a len e)b isa cr ylic Acid
(14n ). The compound 14n (79.5 mg, 93%) was prepared from
16n (100 mg, 0.26 mmol) similarly to the preparation of 14d .
1
Mp: 290-299.5 °C dec. H NMR (DMSO-d6): δ 3.84 (s, 6H),
6.07 (d, J ) 15.7 Hz, 2H), 7.07 (s, 2H), 7.54 (s, 2H), 8.63 (d, J
) 15.7 Hz, 2H), 12.33 (s, 2H). IR (KBr): 2959, 2681, 2587,
1688, 1624, 1420, 1314, 1289, 1238 cm-1. MS (FAB-) m/z: 327
(M+ - 1). HRMS (FAB-) for C18H15O6 (M+ - 1): calcd,
327.0869; found, 327.0934. Anal. (C18H16O6‚1/2H2O) C, H, N.
Dieth yl 3,3′-(9,10-An th r acen ediyl)bisacr ylate (16o). The
compound 16o (314 mg, 94%) was prepared from 27 (300 mg,
0.89 mmol) in the same manner as described for 16e. 1H NMR
(CDCl3): δ 1.42 (t, J ) 7.3 Hz, 6H), 4.39 (q, J ) 7.3 Hz, 4H),
6.40 (d, J ) 16.1 Hz, 2H), 7.53 (dd, J ) 6.8 Hz, 3.4 Hz, 4H),
8.25 (dd, J ) 6.8 Hz, 3.4 Hz, 4H), 8.61 (d, J ) 16.1 Hz, 2H).
IR (KBr): 1713, 1281, 1046 cm-1. MS (EI) m/z: 374 (M+).
HRMS (EI) for C24H22O4 (M+): calcd, 374.1518; found, 374.1521.
3,3′-[3,3′′-(1,1′:4′,1′′-Ter p h en yl)]b isa cr ylic Acid (14f).
The compound 14f (1.06 g, 95%) was prepared from [3,3′′-(1,1′:
4′,1′′-terphenyl)]dicarboxaldehyde (32f) (80 mg, 0.28 mmol) in
the same manner as described for 14b. Mp: >300 °C. 1H NMR
(DMSO-d6): δ 6.70 (d, J ) 16.1 Hz, 2H), 7.52 (t, J ) 7.8 Hz,
2H), 7.69-7.81 (m, 6H), 7.88 (s, 4H), 8.01 (s, 2H), 12.43 (br,
2H). MS (FAB-) m/z: 369 (M+ - 1). HRMS (FAB-) for C24H18O4
(M+ - 1): calcd, 369.1127; found, 369.1214. Anal. (C24H18O4)
C, H, N.
3,3′-(9,10-An th r a cen ed iyl)bisa cr ylic Acid (14o). The
compound 14o (161 mg, 96%) was prepared from 16o (200 mg,
0.53 mmol) in a manner similar to that described for the
preparation of 14d . Mp: >300 °C. 1H NMR (DMSO-d6): δ 6.32
(d, J ) 15.7 Hz, 2H), 7.64 (dd, J ) 6.8 Hz, 2.9 Hz, 4H), 8.22
(dd, J ) 6.8 Hz, 2.9 Hz, 4H), 8.48 (d, J ) 15.7 Hz, 4H), 12.87
3,3′-[4,4′′-(1,1′:4′,1′′-Ter p h en yl)]b isa cr ylic Acid (14g).
The compound 14g (96.4 mg, 93%) was prepared from [4,4′′-
(1,1′:4′,1′′-terphenyl)]dicarboxaldehyde (32g) (80 mg, 0.28
mmol) in the same manner as described for 14b. Mp: >300
1
°C. H NMR (DMSO-d6): δ 6.60 (d, J ) 15.7 Hz, 2H), 7.77 (d,
(s, 2H). IR (KBr): 1694, 1630, 1424, 1308, 1260, 1200 cm-1
.
J ) 15.7 Hz, 2H), 7.81 (s, 8H), 7.85 (s, 4H), 12.43 (br, 2H). IR

Seco Cyclopropa[c]pyrrolo[3,2-e]indole Bisalkylators
J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9 1403
(KBr): 1676, 1624, 1601, 1429, 1312, 1280, 1214 cm-1. Anal.
(C24H18O4) C, H, N.
Na2SO4, filtered, and then concentrated in vacuo to give 31j
as a colorless solid (5.54 g, 98%). Mp: 107.5-110 °C. 1H NMR
(CDCl3): δ 3.90 (s, 6H), 4.41 (s, 4H), 7.35 (s, 2H). IR (KBr):
1730, 1442, 1278, 1238 cm-1. MS (EI) m/z: 252 (M+). HRMS
(EI) for C12H12O6 (M+): calcd, 252.0634; found, 252.0641.
Dim eth yl 2,3-Dim eth oxyter ep h th a la te (31h ). A suspen-
sion of 30 (5.00 g, 22 mmol) and cesium carbonate (17.3 g, 53
mmol) in DMF (50 mL) was stirred at room temperature for
0.5 h. Methyl iodide (3.3 mL, 53 mmol) was added to the
resulting mixture, and the whole was stirred at room temper-
ature for 2 h. After concentration in vacuo, the residue was
dissolved in CH2Cl2. The dichloromethane solution was washed
with water and brine, dried over anhydrous Na2SO4, filtered,
and then concentrated in vacuo to give 31h as a pale brown
oil (5.55 g, 99%). 1H NMR (CDCl3): δ 3.93 (s, 6H), 3.95 (s, 6H),
7.50 (s, 2H). IR (Neat): 1734, 1290 cm-1. MS (EI) m/z: 254
(M+). HRMS (EI) for C12H14O6 (M+): calcd, 254.0790; found,
254.0776.
2,3-Dim eth oxyter ep h th a la ld eh yd e (32h ). A solution of
N-methylpiperazine (5.20 g, 52 mmol) in toluene (16 mL) was
added to a solution of sodium bis(methoxyethoxy)aluminum
hydride (13.6 g, 47 mmol, 70% toluene solution) at 0 °C for
0.5 h. The obtained toluene solution was added to a solution
of 31h (3.00 g, 12 mmol) in toluene (120 mL) at -20 °C over
50 min, and the mixture was stirred at the same temperature
for 10 min. After the reaction was quenched by adding water
(20 mL), the insoluble materials formed were filtered off. The
filtrate was washed with 1 N HCl solution, water, and brine,
dried over anhydrous Na2SO4, filtered, and then concentrated
in vacuo. Flash chromatography (CH2Cl2) of the residue gave
32h as colorless crystals (1.84 g, 80%). Mp: 99.5-100.5 °C.
1H NMR (CDCl3): δ 4.06 (s, 6H), 7.64 (s, 2H), 10.45 (s, 2H).
IR (KBr): 1688, 1570, 1464, 1420, 1395, 1385, 1250 cm-1. MS
(EI) m/z: 194 (M+). Anal. (C10H10O4) C, H, N.
2,3-(Eth ylen ed ioxy)ter ep h th a la d eh yd e (32j). The com-
pound 32j (1.01 g, 66%) was prepared from 31j (2.00 g, 7.9
mmol) in a manner similar to that described for the prepara-
1
tion of 32h . Mp: 139-140.5 °C. H NMR (CDCl3): δ 4.48 (s,
4H), 7.44 (s, 2H), 10.43 (s, 2H). IR (KBr): 1688, 1578, 1466,
1451, 1404, 1379, 1280, 1256, 1240 cm-1. MS (EI) m/z: 192
(M+). Anal. (C10H8O4) C, H, N.
3,3′-(2,3-Eth ylen ed ioxy-1,4-p h en ylen e)bisa cr ylic Acid
(14j). The compound 14j (1.39 g, 95%) was prepared from 32j
(1.01 g, 5.3 mmol) in the same manner as that described for
1
14b. Mp: >300 °C. H NMR (DMSO-d6): δ 4.40 (s, 4H), 6.57
(d, J ) 15.6 Hz, 2H), 7.29 (s, 2H), 7.74 (d, J ) 15.6 Hz, 2H),
12.43 (s, 2H). IR (KBr): 1687, 1618, 1442, 1419, 1322, 1286,
1229 cm-1. MS (EI) m/z: 276 (M+). HRMS (EI) for C14H12O6
(M+): calcd, 276.0634; found, 276.0634. Anal. (C14H12O6) C,
H, N.
2,2′-(2,3-Dim eth oxy-1,4-p h en ylen e)bis(4,4-d im eth yl-2-
oxa zolin e) (34). (a ) P r ep a r a tion of N,N′-bis(2-h yd r oxy-
1,1-d im eth yleth yl)-2,3-d im eth oxyter ep h th a la m id e: To a
solution of 2-amino-2-methyl-1-propanol (6.06 g, 68 mmol) in
CH2Cl2 (8 mL) was added a solution of 33 (4.47 g, 17 mmol) in
CH2Cl2 (8 mL) at 5-10 °C, and the mixture was stirred at
room temperature for 2 h. After insoluble materials were
filtered off, the filtrate was kept standing at room temperature.
The precipitates formed were collected by filtration and dried
in vacuo to give N,N′-bis(2-hydroxy-1,1-dimethylethyl)-2,3-
dimethoxyterephthalamide as colorless crystals (6.27 g, 100%).
3,3′-(2,3-Dim eth oxy-1,4-ph en ylen e)bisacr ylic Acid (14h ).
The compound 14h (1.34 g, 93%) was prepared from 32h (1.00
g, 5.2 mmol) in the same manner as described for 14b. Mp:
>300 °C. 1H NMR (DMSO-d6): δ 3.84 (s, 6H), 6.62 (d, J )
16.6 Hz, 2H), 7.57 (s, 2H), 7.76 (d, J ) 16.6 Hz, 2H), 12.53 (s,
2H). IR (KBr): 1686, 1629, 1460, 1413, 1308, 1282, 1263, 1223
cm-1. MS (EI) m/z: 278 (M+). HRMS (EI) for C14H14O6 (M+):
calcd, 278.0790; found, 278.0781. Anal. (C14H14O6) C, H, N.
Dim eth yl 2,3-(Meth ylen ed ioxy)ter ep h th a la te (31i). A
suspension of 30 (5.00 g, 22 mmol) and cesium carbonate (17.3
g, 53 mmol) in DMF (50 mL) was stirred at room temperature
for 0.5 h. Bromochloromethane (3.5 mL, 53 mmol) was added
to the DMF solution, and the mixture was stirred at 50 °C for
10 h and then diluted with CH2Cl2. The dichloromethane
solution was washed with water and brine, dried over anhy-
drous Na2SO4, filtered, and then concentrated in vacuo to give
31i as a colorless solid (5.14 g, 98%). Mp: 208-210 °C. 1H
NMR (CDCl3): δ 3.94 (s, 6H), 6.25 (s, 2H), 7.43 (s, 2H). IR
(KBr): 1723, 1439, 1298 cm-1. MS (EI) m/z: 238 (M+). HRMS
(EI) for C11H10O6 (M+): calcd, 238.0477; found, 238.0494.
2,3-(Meth ylen edioxy)ter eph th aladeh yde (32i). The com-
pound 32i (1.52 g, 68%) was prepared from 31i (3.00 g, 13
mmol) in a manner similar to that described for the prepara-
1
Mp: 151-153 °C. H NMR (CDCl3): δ 1.43 (s, 12H), 3.71 (s,
4H), 3.96 (s, 6H), 4.55 (brs, 2H), 7.89 (s, 2H), 8.06 (brs, 2H).
IR (KBr): 1651, 1631, 1546, 1453, 1402, 1311, 1274, 1246 cm-1
.
MS (EI) m/z: 368 (M+). Anal. (C18H28N2O6) C, H, N.
(b) P r ep a r a tion of 2,2′-(2,3-d im eth oxy-1,4-p h en ylen e)-
bis(4,4-d im eth yl-2-oxa zolin e) (34): A mixture of N,N′-bis-
(2-hydroxy-1,1-dimethylethyl)-2,3-dimethoxyterephthala-
mide (6.00 g, 16 mmol) and thionyl chloride (7 mL) was stirred
at room temperature for 3 h. After an additional thionyl
chloride (7 mL) was added, the mixture was stirred at room
temperature for 1 h. After the addition of ether (50 mL), the
supernatant was removed by decantation. The lower residue
was dissolved in water (50 mL), and the aqueous solution was
adjusted to pH 8 by the addition of 10% NaOH solution. The
aqueous solution was extracted with ether, dried over anhy-
drous Na2SO4, filtered, and then concentrated in vacuo. Flash
chromatography (CH2Cl2:EtOH ) 20:1) of the residue gave 34
as colorless crystals (2.52 g, 47%). Mp: 84.5-85.5 °C. 1H NMR
(CDCl3): δ 1.40 (s, 12H), 3.90 (s, 6H), 4.11 (s, 4H), 7.50 (s,
2H). IR (KBr): 1643, 1489, 1460, 1401, 1349, 1307, 1237, 1189
cm-1. MS (EI) m/z: 332 (M+). Anal. (C18H24N2O4) C, H, N.
2,2′-(2,3-Diet h yl-1,4-p h en ylen e)bis(4,4-d im et h yl-2-ox-
a zolin e) (35). Ethylmagnesium bromide (16.4 mL, 1.0 M
solution in THF) was added to a solution of 34 (2.00 g, 6.0
mmol) in THF (20 mL) at 0 °C over 0.5 h, and the mixture
was stirred at room temperature for 2 h. After the reaction
was quenched by the addition of saturated NH4Cl solution and
water, the mixture was extracted with ether. The combined
ethereal extracts washed with brine, dried over anhydrous
Na2SO4, filtered, and then concentrated in vacuo gave 35 as
1
tion of 32h . Mp: 151.5-152 °C. H NMR (CDCl3): δ 6.34 (s,
2H), 7.39 (s, 2H), 10.21 (s, 2H). IR (KBr): 1696, 1472, 1454,
1400, 1364, 1254, 1219 cm-1. MS (EI) m/z: 178 (M+). Anal.
(C9H6O4) C, H, N.
3,3′-(2,3-Meth ylen edioxy-1,4-ph en ylen e)bisacr ylic Acid
(14i). The compound 14i (1.39 g, 95%) was prepared from 31i
(1.00 g, 5.6 mmol) in a manner similar to that described for
the preparation of 14b. Mp: >300 °C. 1H NMR (DMSO-d6): δ
6.29 (s, 2H), 6.61 (d, J ) 16.6 Hz, 2H), 7.18 (s, 2H), 7.51 (d, J
) 16.6 Hz, 2H), 12.53 (s, 2H). IR (KBr): 1689, 1617, 1437,
1320, 1267, 1233 cm-1. MS (EI) m/z: 262 (M+). HRMS (EI)
for C13H10O6 (M+): calcd, 262.0477; found, 262.0462. Anal.
(C13H10O6) C, H, N.
1
pale yellow crystals (1.98 g, 100%). Mp: 49-50 °C. H NMR
(CDCl3): δ 1.16 (t, J ) 7.8 Hz, 6H), 1.39 (s, 12H), 2.98 (q, J )
7.8 Hz, 4H), 4.08 (s, 4H), 7.44 (s, 2H). IR (KBr): 2967, 2932,
1653, 1460, 1346, 1287, 1188, 1132, 1049 cm-1. MS (EI) m/z:
328 (M+). Anal. (C20H28N2O2) C, H, N.
Dim eth yl 2,3-(Eth ylen ed ioxy)ter ep h th a la te (31j). A
suspension of 30 (5.00 g, 22 mmol) and cesium carbonate (17.3
g, 53 mmol) in DMF (50 mL) was stirred at room temperature
for 0.5 h. 1,2-Dibromoethane (2.5 mL, 29 mmol) was added to
the DMF solution, and the mixture was stirred at 80 °C for 2
h and then diluted with CH2Cl2. The dichloromethane solu-
tion was washed with water and brine, dried over anhydrous
2,3-Dieth ylter ep h th a la ld eh yd e (32l). (a ) P r ep a r a tion
of 2,2′-(2,3-d ieth yl-1,4-p h en ylen e)bis(4,4-d im eth yl-2-ox-
a zolin iu m iod id e): A mixture of 35 (1.86 g, 5.7 mmol) and
methyl iodide (7 mL) in nitromethane (5 mL) was stirred at
80 °C for 4 h. After dilution with ether, the resulting
precipitates were collected by filtration and dried in vacuo to
give 2,2′-(2,3-diethyl-1,4-phenylene)bis(4,4-dimethyl-2-oxazo-

1404 J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9
Fukuda et al.
linium iodide) as colorless crystals (2.62 g, 76%). Mp: 281-
285 °C (EtOH). 1H NMR (DMSO-d6): δ 1.15 (t, J ) 7.8 Hz,
6H), 1.66 (s, 12H), 2.70 (q, J ) 7.8 Hz, 4H), 3.24 (s, 6H), 5.09
(s, 4H), 8.00 (s, 2H). IR (KBr): 3441, 2969, 2936, 2882, 1730,
1657, 1495, 1427, 1416 cm-1. MS (EI) m/z: 358 (M+-2I). Anal.
(C22H34I2N2O2) C, H, N.
7.97 (m, 2H), 8.05 (d, J ) 15 Hz, 2H), 8.11 (s, 2H), 10.57 (s,
2H), 13.06 (s, 2H). MS (FAB+) m/z: 879 (M+ + 1). HRMS
(FAB+) for C40H31Cl2F6N4O8 (M+ + 1): calcd, 879.1423; found,
879.1401.
(S,S)-Dim et h yl
6,6′-[3,3′-(1,1′-Dip h en yl-4,4′-d iyl)d i-
acr yloyl]bis[4-ch lor om eth yl-8-h ydr oxy-2-tr iflu or om eth yl-
1,4,5,6-tetr ah ydr opyr r olo[3,2-e]in dole-3-car boxylate](11d).
The compound 11d (1.7 mg, 12%) was prepared from 12 (13.5
(b ) P r ep a r a t ion of 2,3-d iet h ylt er ep h t h a la ld eh yd e
(32l): To a suspension of 2,2′-(2,3-diethyl-1,4-phenylene)bis-
(4,4-dimethyl-2-oxazolinium iodide) (2.45 g, 5.1 mmol) in EtOH
(50 mL) was added sodium borohydride (0.77 g, 20 mmol) at 0
°C for 1 h, and the mixture was stirred at 5 °C for 3 h. After
the reaction was quenched by the addition of 2 N HCl solution,
the mixture was extracted with ether. The ethereal organic
extracts were washed with brine. After treatment with acti-
vated carbon, the organic layer was dried over anhydrous
Na2SO4, filtered, and then concentrated in vacuo to give 32l
32
mg, 30 µmol) and 14d (4.4 mg, 15 µmol). [R]D ) -36° (c )
0.05, THF). 1H NMR (DMSO-d6): δ 3.50 (dd, J ) 11 Hz, 9 Hz,
2H), 3.78-3.90 (m, 2H), 3.88 (s, 6H), 4.23-4.32 (m, 2H), 4.38-
4.52 (m, 4H), 7.29 (d, J ) 15 Hz, 2H), 7.71 (d, J ) 15 Hz, 2H),
7.84 (d, J ) 8 Hz, 4H), 7.93 (d, J ) 8 Hz, 4H), 8.11 (s, 2H),
10.56 (s, 2H), 13.07 (s, 2H). MS (FAB+) m/z: 955 (M+ + 1).
HRMS (FAB+) for C46H35Cl2F6N4O8 (M+ + 1): calcd, 955.1736;
found, 955.1703.
1
as a pale yellow oil (102 mg, 13%). Mp: 32-34.5 °C. H NMR
(S,S)-Dim et h yl
6,6′-[3,3′-(2,2′-Bip yr id yl-5,5′-d iyl)d i-
(CDCl3): δ 1.28 (t, J ) 7.3 Hz, 6H), 3.14 (q, J ) 7.3 Hz, 4H),
7.84 (s, 2H), 10.41 (s, 2H). IR (KBr): 3443, 2973, 2932, 1690,
1458, 1393, 1229 cm-1. MS (EI) m/z: 190 (M+). HRMS (EI)
for C12H14O2 (M+): calcd, 190.0994; found, 190.0990. Anal.
(C12H14O2‚1/4H2O) C, H, N.
acr yloyl]bis[4-ch lor om eth yl-8-h ydr oxy-2-tr iflu or om eth yl-
1,4,5,6-tetr ah ydr opyr r olo[3,2-e]in dole-3-car boxylate] (11e).
The compound 11e (2.4 mg, 16%) was prepared from 12 (13.5
32
mg, 30 µmol) and 14e (4.4 mg, 15 µmol). [R]D ) -35° (c )
0.05, THF). 1H NMR (DMSO-d6): δ 3.51 (dd, J ) 10 Hz, 9 Hz,
2H), 3.77-3.92 (m, 2H), 3.88 (s, 6H), 4.25-4.33 (m, 2H), 4.39-
4.55 (m, 4H), 7.47 (d, J ) 15 Hz, 2H), 7.78 (d, J ) 15 Hz, 2H),
8.13 (s, 2H), 8.51 (s, 4H), 9.08 (s, 2H), 10.60 (s, 2H), 13.09 (s,
3,3′-(2,3-Diet h yl-1,4-p h en ylen e)b isa cr ylic Acid (14l).
The compound 14l (62.6 mg, 74%) was prepared from 31l (59.8
mg, 0.31 mmol) in the same manner as that described for 14b.
1
Mp: 264-268 °C dec. H NMR (DMSO-d6): δ 1.11 (t, J ) 7.8
2H). MS (FAB+) m/z: 957 (M+ + 1). HRMS (FAB+) for C44H33
-
Hz, 6H), 2.77 (q, J ) 7.8 Hz, 4H), 6.44 (d, J ) 15.7 Hz, 2H),
7.57 (s, 2H), 7.88 (d, J ) 15.7 Hz, 2H), 12.50 (br, 2H). IR
(KBr): 3432, 2971, 1688, 1628, 1416, 1287 cm-1. MS (EI) m/z:
274 (M+). HRMS (EI) for C16H18O4 (M+): calcd, 274.1205;
found, 274.1160. Anal. (C16H18O4) C, H, N.
Cl2F6N6O8 (M+ + 1): calcd, 957.1641; found, 957.1594.
(S,S)-Dim eth yl 6,6′-[3,3′-(3,3′′-(1,1′:4′,1′′-Ter p h en yl))d i-
acr yloyl]bis[4-ch lor om eth yl-8-h ydr oxy-2-tr iflu or om eth yl-
1,4,5,6-tetr ah ydr opyr r olo[3,2-e]in dole-3-car boxylate] (11f).
The compound 11f (3.1 mg, 10%) was prepared from 12 (27.0
28
(S,S)-Dim eth yl 6,6′-[3,3′-(1,4-P h en ylen e)d ia cr yloyl]bis-
[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-tet-
r a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxyla te] (11a ). A so-
lution of 12 (13.5 mg, 30 µmol) in 3 M HCl-AcOEt (0.6 mL)
was stirred at room temperature for 1 h. Concentration of the
mixture in vacuo gave the crude hydrochloride 13 as a pale
yellow powder, which was directly added to a solution of 14a
(3.3 mg, 15 µmol) and EDCI (17.3 mg, 90 µmol) in DMF (0.3
mL). The mixture was stirred at room temperature overnight.
After dilution with water, the mixture was extracted with
CHCl3-MeOH (5:1), dried over anhydrous Na2SO4, filtered,
then concentrated in vacuo. Flash chromatography (CHCl3:
MeOH:acetone ) 5:1:1) of the residue gave 11a as pale yellow
crystals (3.3 mg, 25%). [R]D24 ) -21° (c ) 0.20, THF). 1H NMR
(DMSO-d6): δ 3.48 (t, J ) 8.3 Hz, 2H), 3.83 (d, J ) 8.3 Hz,
2H), 3.88 (s, 6H), 4.28 (br, 2H), 4.40-4.49 (m, 4H), 7.30 (d, J
) 15.1 Hz, 2H), 7.70 (d, J ) 15.6 Hz, 2H), 7.87 (s, 4H), 8.11
(brs, 2H), 10.52 (br, 2H), 13.02 (br, 2H). MS (FAB+) m/z: 879
(M+ + 1). HRMS (FAB+) for C40H31Cl2F6N4O8 (M+ + 1): calcd,
879.1423; found, 879.1442.
mg, 60 µmol) and 14f (11.2 mg, 30 µmol). [R]D ) -19° (c )
0.05, THF). 1H NMR (DMSO-d6): δ 3.51 (dd, J ) 10 Hz, 9 Hz,
2H), 3.78-3.94 (m, 2H), 3.88 (m, 6H), 4.24-4.34 (m, 2H), 4.39-
4.54 (m, 4H), 7.37 (d, J ) 15 Hz, 2H), 7.58 (t, J ) 8 Hz, 2H),
7.78 (d, J ) 15 Hz, 2H), 7.81 (d, J ) 8 Hz, 2H), 7.83 (d, J ) 8
Hz, 2H), 7.92 (s, 4H), 8.13 (s, 2H), 8.18 (s, 2H), 10.56 (s, 2H),
13.07 (s, 2H). MS (FAB+) m/z: 1031 (M+ + 1). HRMS (FAB+)
for
C
52H39Cl2F6N4O8 (M+ + 1): calcd, 1031.2049; found,
1031.2085.
(S,S)-Dim eth yl 6,6′-[3,3′-(4,4′′-(1,1′:4′,1′′-Ter p h en yl))d i-
acr yloyl]bis[4-ch lor om eth yl-8-h ydr oxy-2-tr iflu or om eth yl-
1,4,5,6-tetr ah ydr opyr r olo[3,2-e]in dole-3-car boxylate] (11g).
The compound 11g (4.9 mg, 31%) was prepared from 12 (13.5
28
mg, 30 µmol) and 14g (5.6 mg, 15 µmol). [R]D ) -17° (c )
0.05, THF). 1H NMR (DMSO-d6): δ 3.50 (t, J ) 10 Hz, 2H),
3.79-3.93 (m, 2H), 3.88 (s, 6H), 4.23-4.33 (m, 2H), 4.38-4.53
(m, 4H), 7.29 (d, J ) 16 Hz, 2H). 7.72 (d, J ) 16 Hz, 2H), 7.83
(d, J ) 8 Hz, 4H), 7.88 (s, 4H), 7.93 (d, J ) 8 Hz, 4H), 8.13 (s,
2H), 10.56 (s, 2H), 13.06 (s, 2H). MS (FAB+) m/z: 1031 (M+
+
1). HRMS (FAB+) for C52H39Cl2F6N4O8 (M+ + 1): calcd,
The other seco MCTFCPI bisalkylators 11b-q were pre-
pared in a manner similar to that described for the preparation
of 11a .
1031.2049; found, 1031.1991.
(S,S)-Dim eth yl 6,6′-[3,3′-(2,3-Dim eth oxy-1,4-ph en ylen e)-
d ia cr yloyl]b is[4-ch lor om e t h yl-8-h yd r oxy-2-t r iflu or o-
m eth yl-1,4,5,6-tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxy-
la te] (11h ). The compound 11h (1.1 mg, 8%) was prepared
(S,S)-Dim eth yl 6,6′-[3,3′-(1,3-P h en ylen e)d ia cr yloyl]bis-
[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-tet-
r a h yd r op yr r olo[3,2-e]in d ole-3-ca r b oxyla t e] (11b ). The
compound 11b (3.5 mg, 27%) was prepared from 12 (13.5 mg,
32
from 12 (13.5 mg, 30 µmol) and 14h (4.2 mg, 15 µmol). [R]D
1
) -26° (c ) 0.05, THF). H NMR (DMSO-d6): δ 3.51 (dd, J )
27
30 µmol) and 14b (3.3 mg, 15 µmol). [R]D ) -28° (c ) 0.05,
10 Hz, 9 Hz, 2H), 3.79-3.94 (m, 2H), 3.88 (s, 6H), 3.90 (s, 6H),
4.24-4.33 (m, 2H), 4.39-4.49 (m, 4H), 7.31 (d, J ) 16 Hz, 2H),
7.78 (s, 2H), 7.88 (d, J ) 16 Hz, 2H), 8.11 (s, 2H), 10.57 (s,
2H), 13.08 (s, 2H). MS (FAB+) m/z: 939 (M+ + 1). HRMS
(FAB+) for C42H35Cl2F6N4O10 (M+ + 1): calcd, 939.1634; found,
939.1617.
1
THF). H NMR (DMSO-d6): δ 3.51 (dd, J ) 10 Hz, 9 Hz, 2H),
3.75-3.93 (m, 2H), 3.88 (s, 6H), 4.24-4.33 (m, 2H), 4.38-4.53
(m, 4H), 7.33 (d, J ) 16 Hz, 2H), 7.52 (t, J ) 8 Hz, 1H), 7.73
(d, J ) 16 Hz, 2H), 7.88 (d, J ) 8 Hz, 2H), 8.12 (s, 2H), 8.25
(s, 1H), 10.56 (s, 2H), 13.06 (s, 2H). MS (FAB+) m/z: 879 (M+
+ 1). HRMS (FAB+) for C40H31Cl2F6N4O8 (M+ + 1): calcd,
879.1423; found, 879.1410.
(S,S)-Dim eth yl 6,6′-[3,3′-(2,3-(Meth ylen edioxy)-1,4-ph en -
ylen e)d ia cr yloyl]bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu o-
r om et h yl-1,4,5,6-t et r a h yd r op yr r olo[3,2-e]in d ole-3-ca r -
boxyla te] (11i). The compound 11i (2.4 mg, 17%) was
prepared from 12 (13.5 mg, 30 µmol) and 14i (3.9 mg, 15 µmol).
(S,S)-Dim eth yl 6,6′-[3,3′-(1,2-P h en ylen e)d ia cr yloyl]bis-
[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-tet-
r ah ydr opyr r olo[3,2-e]in dole-3-car boxylate] (11c). The com-
pound 11c (3.9 mg, 29%) was prepared from 12 (13.5 mg, 30
[R]D ) -12° (c ) 0.05, THF). 1H NMR (DMSO-d6): δ 3.52
30
28
µmol) and 14c (3.3 mg, 15 µmol). [R]D ) -107° (c ) 0.05,
(dd, J ) 11 Hz, 9 Hz, 2H), 3.78-3.85 (m, 2H), 3.88 (s, 6H),
4.24-4.32 (m, 2H), 4.32-4.45 (m, 4H), 6.38 (s, 2H), 7.28 (d, J
) 16 Hz, 2H), 7.37 (s, 2H), 7.65 (d, J ) 16 Hz, 2H), 8.10 (s,
THF). 1H NMR (DMSO-d6): δ 3.51 (t, J ) 9 Hz, 2H), 3.75-
3.92 (m, 2H), 3.87 (s, 6H), 4.22-4.32 (m, 2H), 4.38-4.50 (m,
4H), 7.16 (d, J ) 15 Hz, 2H), 7.52 (dd, J ) 6 Hz, 4 Hz, 2H),
2H), 10.58 (s, 2H), 13.17 (s, 2H). MS (FAB+) m/z: 923 (M+
+

Seco Cyclopropa[c]pyrrolo[3,2-e]indole Bisalkylators
J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9 1405
1). HRMS (FAB+) for C41H31Cl2F6N4O10 (M+ + 1): calcd,
(S,S)-Dim eth yl 6,6′-[3,3′-(1,4-An th r a cen e)d ia cr yloyl]-
bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-
tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxyla te] (11p ). The
compound 11p (2.3 mg, 15%) was prepared from 12 (13.5 mg,
923.1321; found, 923.1298.
(S,S)-Dim eth yl 6,6′-[3,3′-(2,3-(Eth ylen ed ioxy)-1,4-p h en -
ylen e)d ia cr yloyl]bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu o-
r om et h yl-1,4,5,6-t et r a h yd r op yr r olo[3,2-e]in d ole-3-ca r -
boxyla te] (11j). The compound 11j (4.1 mg, 15%) was
prepared from 12 (26.9 mg, 60 µmol) and 14j (8.3 mg, 30 µmol).
32
30 µmol) and 14p (4.8 mg, 15 µmol). [R]D ) -36° (c ) 0.05,
1
THF). H NMR (DMSO-d6): δ 3.54 (t, J ) 10 Hz, 2H), 3.78-
3.93 (m, 2H), 3.89 (s, 6H), 4.27-4.36 (m, 2H), 4.46-4.57 (m,
4H), 7.43 (d, J ) 15 Hz, 2H), 7.62 (dd, J ) 6 Hz, 3 Hz, 2H),
8.19 (s, 4H), 8.30 (dd, J ) 6 Hz, 3 Hz, 2H), 8.67 (d, J ) 15 Hz,
2H), 9.04 (s, 2H), 10.60 (s, 2H), 13.07 (s, 2H). MS (FAB+) m/z:
979 (M+ + 1). HRMS (FAB+) for C48H35Cl2F6N4O8 (M+ + 1):
calcd, 979.1736; found, 979.1699.
30
1
[R]D ) -12° (c ) 0.05, THF). H NMR (DMSO-d6): δ 3.51 (t,
J ) 10 Hz, 2H), 3.79-3.85 (m, 2H), 3.88 (s, 6H), 4.23-4.32
(m, 2H), 4.38-4.44 (m, 4H), 4.46 (s, 4H), 7.25 (d, J ) 16 Hz,
2H), 7.52 (s, 2H), 7.89 (d, J ) 16 Hz, 2H), 8.10 (s, 2H), 10.56
(s, 2H), 13.07 (s, 2H). MS (FAB+) m/z: 937 (M+ + 1). HRMS
(FAB+) for C42H33Cl2F6N4O10 (M+ + 1): calcd, 937.1478; found,
937.1411.
(S,S)-Dim eth yl 6,6′-[3,3′-(1,4-(9,10-An th r a qu in oyl))d i-
acr yloyl]bis[4-ch lor om eth yl-8-h ydr oxy-2-tr iflu or om eth yl-
1,4,5,6-t e t r a h yd r op yr r olo[3,2-e]in d ole -3-ca r b oxyla t e ]
(11q). The compound 11q (1.6 mg, 4%) was prepared from 12
(S,S)-Dim eth yl 6,6′-[3,3′-(2,5-Dim eth oxy-1,4-ph en ylen e)-
d ia cr yloyl]b is[4-ch lor om e t h yl-8-h yd r oxy-2-t r iflu or o-
m eth yl-1,4,5,6-tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxy-
la te] (11k ). The compound 11k (2.1 mg, 15%) was prepared
32
(40.4 mg, 90 µmol) and 14q (15.7 mg, 45 µmol). [R]D ) -48°
1
(c ) 0.05, THF). H NMR (DMSO-d6): δ 3.53 (dd, J ) 10 Hz,
32
9 Hz, 2H), 3.79-3.91 (m, 2H), 3.88 (s, 6H), 4.24-4.34 (m, 2H),
4.41-4.51 (m, 4H), 7.11 (d, J ) 16 Hz, 2H), 7.95 (dd, J ) 6
Hz, 3 Hz, 2H), 8.10-8.19 (m, 4H), 8.21 (s, 2H), 8.53 (d, J ) 16
Hz, 2H), 10.61 (s, 2H), 13.10 (s, 2H). MS (FAB+) m/z: 1009
(M+ + 1). HRMS (FAB+) for C48H33Cl2F6N4O10 (M+ + 1): calcd,
1009.1478; found, 1009.1534.
from 12 (13.5 mg, 30 µmol) and 14k (4.2 mg, 15 µmol). [R]D
1
) -56° (c ) 0.05, THF). H NMR (DMSO-d6): δ 3.52 (dd, J )
11 Hz, 8 Hz, 2H), 3.78-3.90 (m, 2H), 3.88 (s, 6H), 3.98 (s, 6H),
4.23-4.33 (m, 2H), 4.38-4.48 (m, 4H), 7.30 (d, J ) 16 Hz, 2H),
7.53 (s, 2H), 7.96 (d, J ) 16 Hz, 2H), 8.10 (s, 2H), 10.56 (s,
2H), 13.07 (s, 2H). MS (FAB+) m/z: 939 (M+ + 1). HRMS
(FAB+) for C42H35Cl2F6N4O10 (M+ + 1): calcd, 939.1634; found,
939.1699.
(S,S)-Dim eth yl 6,6′-[3,3′-(2,3-Dieth yl-1,4-p h en ylen e)-
d ia cr yloyl]b is[4-ch lor om e t h yl-8-h yd r oxy-2-t r iflu or o-
m eth yl-1,4,5,6-tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxy-
Ack n ow led gm en t. We are grateful to Dr. S. Suzue,
Kyorin Pharmaceutical Co. Ltd., for many valuable
suggestions and encouragement.
la te] (11l). The compound 11l (5.5 mg, 19%) was prepared
31
Refer en ces
from 12 (26.9 mg, 60 µmol) and 14l (8.2 mg, 30 µmol). [R]D
)
1
-16° (c ) 0.05, THF). H NMR (DMSO-d6): δ 1.18 (t, J ) 8
Hz, 6H), 2.85 (q, J ) 8 Hz, 4H), 3.51 (t, J ) 10 Hz, 2H), 3.72-
3.85 (m, 2H), 3.88 (s, 6H), 4.22-4.32 (m, 2H), 4.38-4.48 (m,
4H), 7.12 (d, J ) 16 Hz, 2H), 7.78 (s, 2H), 7.99 (d, J ) 16 Hz,
2H), 8.11 (s, 2H), 10.54 (s, 2H), 13.06 (s, 2H). MS (FAB+) m/z:
935 (M+ + 1). HRMS (FAB+) for C44H39Cl2F6N4O8 (M+ + 1):
calcd, 935.2049; found, 935.2008.
(1) Martin, D. G.; Chidester, C. G.; Duchamp, D. J .; Mizsak, S. A.
Structure of CC-1065 (NSC 298223), a new antitumor antibiotic.
J . Antibiot. 1980, 33, 902.
(2) (a) Ichimura, M.; Muroi, K.; Asano, K.; Kawamoto, I.; Tomita,
F.; Morimoto, M.; Nakano, H. DC89-A1, a new antitumor
antibiotic from streptomyces. J . Antibiot. 1988, 41, 1285. (b)
Yasuzawa, T.; Iida, T.; Muroi, K.; Ichimura, M.; Takahashi, K.;
Sano, H. Structures of duocarmycins, novel antitumor antibiotics
produced by streptomyces sp. Chem. Pharm. Bull. 1988, 36, 3728.
(c) Takahashi, I.; Takahashi, K.; Ichimura, M.; Morimoto, M.;
Asano, K.; Kawamoto, I.; Tomita, F.; Nakano, H. Duocarmycin
A, a new antitumor antibiotic from streptomyces. J . Antibiot.
1988, 41, 1915. (d) Ogawa, T.; Ichimura, M.; Katsumata, S.;
Morimoto, M.; Takahashi, K. New antitumor antibiotics, duo-
carmycins B1 and B2. J . Antibiot. 1989, 42, 1299.
(3) (a) Ichimura, M.; Ogawa, T.; Takahashi, K.; Kobayashi, E.;
Kawamoto, I.; Yasuzawa, T.; Takahashi, I.; Nakano, H. Duo-
carmycin SA, a new antitumor antibiotic from streptomyces sp.
J . Antibiot. 1990, 43, 1037. (b) Yasuzawa, T.; Saito, Y.; Ichimura,
M.; Takahashi, I.; Sano, H. Structure of duocarmycin SA, a
potent antitumor antibiotic. J . Antibiot. 1991, 42, 445.
(S,S)-Dim eth yl 6,6′-[3,3′-(1,4-Na p h th a len e)d ia cr yloyl]-
bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-
tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxyla te] (11m ). The
compound 11m (3.0 mg, 21%) was prepared from 12 (13.5 mg,
32
30 µmol) and 14m (4.0 mg, 15 µmol). [R]D ) -41° (c ) 0.05,
1
THF). H NMR (DMSO-d6): δ 3.53 (t, J ) 10 Hz, 2H), 3.78-
3.92 (m, 2H), 3.88 (s, 6H), 4.25-4.34 (m, 2H), 4.45-4.54 (m,
4H), 7.36 (d, J ) 15 Hz, 2H), 7.74 (dd, J ) 6 Hz, 3 Hz, 2H),
8.16 (s, 2H), 8.20 (s, 2H), 8.36 (dd, J ) 6 Hz, 3 Hz, 2H), 8.51
(d, J ) 15 Hz, 2H), 10.60 (s, 2H), 13.09 (s, 2H). MS (FAB+)
m/z: 929 (M+ + 1). HRMS (FAB+) for C44H33Cl2F6N4O8 (M+
1): calcd, 929.1580; found, 929.1530.
+
(4) (a) Boger, D. L.; Coleman, R. S.; Invergo, B. J .; Sakya, S. M.;
Ishizaki, T.; Munk, S. A.; Zarrinmayeh, H.; Kitos, P. A.;
Thompson, S. C. Synthesis and evaluation of aborted and
extended CC-1065 functional analogues: (+)- and (-)-CPI-
CDPI3. Preparation of key partial structures and definition of
an additional functional role of the CC-1065 central and right-
hand subunits. J . Am. Chem. Soc. 1990, 112, 4623. (b) Hurley,
L. H.; Warpehoski, M. A.; Lee, C.-S.; McGovren, J . P.; Scahill,
T. A.; Kelly, R. C.; Mitchell, M. A.; Wicnienski, N. A.; Gebhard,
I.; J ohnson, P. D.; Bradford, V. S. Sequence specificity of DNA
alkylation by the unnatural enantiomer of CC-1065 and its
synthetic analogues. J . Am. Chem. Soc. 1990, 112, 4633. (c)
Warpehoski, M. A.; Gebhard, I.; Kelly, R. C.; Krueger, W. C.;
Li, L. H.; McGovren, J . P.; Prairie, M. D.; Wicnienski, N.;
Wierenga, W. Stereoelectronic factors influencing the biological
activity and DNA interaction of synthetic antitumor agents
modeled on CC-1065. J . Med. Chem. 1988, 31, 590. (d) Chin, H.
L.; Dinshaw, J . P. Site-specific covalent duocarmycin A-Intramo-
lecular DNA triplex complex. J . Am. Chem. Soc. 1992, 114,
10658. (e) Boger, D. L.; Ishizaki, T.; Zarrinmayeh, H.; Munk, S.
A.; Kitos, P. A.; Suntornwat, O. Duocarmycin-pyrindamycin
DNA alkylation properties and identification, synthesis, and
evaluation of agents incorporating the pharmacophore of the
duocarmycin-pyrindamycin alkylation subunit. Identification of
the CC-1065-duocarmycin common pharmacophore. J . Am.
Chem. Soc. 1990, 112, 8961. (f) Boger, D. L. Chemtracts: Org.
Chem. 1991, 4, 329. (g) Sugiyama, H.; Hosoda, M.; Saito, I.; Asai,
A.; Saito, H. Covalent alkylation of DNA with duocarmycin A.
Identification of abasic site structure. Tetrahedron Lett. 1990,
31, 7197 and references therein.
(S,S)-Dim eth yl 6,6′-[3,3′-(5,8-Dim eth oxy-1,4-n a p h th a -
len e)d ia cr yloyl]bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu o-
r om et h yl-1,4,5,6-t et r a h yd r op yr r olo[3,2-e]in d ole-3-ca r -
boxyla te] (11n ). The compound 11n (2.2 mg, 15%) was
prepared from 12 (13.5 mg, 30 µmol) and 14n (4.9 mg, 15
µmol). [R]D ) -56° (c ) 0.05, THF). H NMR (DMSO-d6): δ
3.52 (dd, J ) 9 Hz, 11 Hz, 2H), 3.80-3.87 (m, 2H), 3.88 (s,
6H), 3.89 (s, 6H), 4.23-4.31 (m, 2H), 4.40-4.47 (m, 4H), 6.77
(d, J ) 15 Hz, 2H), 7.10 (s, 2H), 7.75 (s, 2H), 8.16 (s, 2H), 8.77
(d, J ) 15 Hz, 2H), 10.54 (s, 2H), 13.06 (s, 2H). MS (FAB+)
m/z: 989 (M+ + 1). HRMS (FAB+) for C46H37Cl2F6N4O10 (M+
+ 1): calcd, 989.1791; found, 989.1769.
30
1
(S,S)-Dim eth yl 6,6′-[3,3′-(9,10-An th r a cen e)d ia cr yloyl]-
bis[4-ch lor om eth yl-8-h yd r oxy-2-tr iflu or om eth yl-1,4,5,6-
tetr a h yd r op yr r olo[3,2-e]in d ole-3-ca r boxyla te] (11o). The
compound 11o (3.7 mg, 25%) was prepared from 12 (13.5 mg,
29
30 µmol) and 14o (4.8 mg, 15 µmol). [R]D ) -144° (c ) 0.05,
1
THF). H NMR (DMSO-d6): δ 3.56 (t, J ) 10 Hz, 2H), 3.79-
3.85 (m, 2H), 3.87 (s, 6H), 4.22-4.29 (m, 2H), 4.33-4.47 (m,
4H), 7.05 (d, J ) 16 Hz, 2H), 7.66 (dd, J ) 4 Hz, 7 Hz, 4H),
8.22 (s, 2H), 8.37 (dd, J ) 7 Hz, 4 Hz, 4H), 8.55 (d, J ) 16 Hz,
2H), 10.64 (s, 2H), 13.11 (s, 2H). MS (FAB+) m/z: 979 (M+
+
1). HRMS (FAB+) for C48H35Cl2F6N4O8 (M+ + 1): calcd,
979.1736; found, 979.1787.

1406 J ournal of Medicinal Chemistry, 2001, Vol. 44, No. 9
Fukuda et al.
(5) Li, L. H.; DeKoning, T. F.; Kelly, R. C.; Krueger, W. C.;
McGovren, J . P.; Padbury, G. E.; Petzold, G. L.; Wallace, T. L.;
Ouding, R. J .; Prairie, M. D.; Gebhard, I. Cytotoxicity and
antitumor activity of carzelesin, a prodrug cyclopropapylpyrro-
loindole analogue. Cancer Res. 1992, 52, 4904.
(6) (a) Asai, A.; Nagamura, S.; Saito, H. A novel property of
duocarmycin and its analogues for covalent reaction with DNA.
J . Am. Chem. Soc. 1994, 116, 4171. (b) Nagamura, S.; Asai, A.;
Amishiro, N.; Kobayashi, E.; Gomi, K.; Saito, H. Synthesis and
antitumor activity of duocarmycin derivatives: A-ring pyrrole
compounds bearing cinnamoyl groups. J . Med. Chem. 1997, 40,
972.
(11) Sengupta, S.; Bhattacharya, S. Heck reaction of arenediazonium
salts: a palladium-catalysed reaction in an aqueous medium.
J . Chem. Soc., Perkin Trans. 1 1993, 1943.
(12) Leeson, P. D.; Emmett, J . C. Synthesis of thyroid hormone ana-
logues. Part 1. Preparation of 3′-heteroarylmethyl-3,5-di-iodo-
L-thyronines via phenol-dinitrophenol condensation and rela-
tionships between structure and selective thyromimetic activity.
J . Chem. Soc., Perkin Trans. 1 1988, 3085.
(13) Showalter, H. D. H.; Berman, E. M.; J ohnson, J . L.; Atwood, J .
L.; Hunter, W. E. A facile synthesis of functionalized 9,10-
anthracenediones via tosylate and triflate phenolic activation.
Tetrahedron Lett. 1985, 26, 157.
(14) Weitl, F. L.; Raymond, K. N.; Durbin, P. W. Synthetic entero-
bactin analogues. Carboxamido-2,3-dihydroxyterephthalate con-
jugates of spermine and spermidine. J . Med. Chem. 1981, 24,
203.
(15) (a) Cristofaro, M. F.; Chamberlin, A. R. Enantioselective and
diastereoselective molecular recognition of cyclic dipeptides by
a C2 macrolactam Host. J . Am. Chem. Soc. 1994, 116, 5089. (b)
Helms, A.; Heiler, D.; McLendon, G. Electron transfer in bis-
porphyrin doner-acceptor compounds with polyphenylene spac-
ers shows a weak distance dependence. J . Am. Chem. Soc. 1992,
114, 6227. (c) Leighton, P.; Sanders, J . K. M. Synthesis of
bipyridyl-, viologen-, quinone-bridged porphyrins. J . Chem. Soc.,
Perkin Trans. 1 1987, 2385.
(16) Meyers, A. I.; Mihelich, E. D. Oxazolines. XXII. Nucleophilic
aromatic substitution on aryl oxazolines. An efficient approach
to unsymmetrically substituted biphenyls and o-alkyl benzoic
acids. J . Am. Chem. Soc. 1975, 97, 7383.
(7) Fukuda, Y.; Furuta, H.; Kusama, Y.; Ebisu, H.; Oomori, Y.;
Terashima, S. Novel cyclopropapyrroloindole derivative (AT-
3510) bearing methoxycarbonyl and trifluoromethyl groups. J .
Med. Chem. 1999, 42, 1448.
(8) (a) Mitchell, M. A.; Kelly, R. C.; Wicnienski, N. A.; Hatzenbuhler,
N. T.; Williams, M. G.; Petzold, G. L.; Slightom, J . L.; Siemien-
iak, D. R. Synthesis and DNA cross-linking by a rigid CPI dimer.
J . Am. Chem. Soc. 1991, 113, 8994. (b) Sun, D.; Hurley, L. H.
Analysis of the monoalkylation and cross-linking sequence
specificity of bizelesin, a bifunctional alkylation agent related
to (+)-CC-1065. J . Am. Chem. Soc. 1993, 115, 5925. (c) Mitchell,
M. A.; J ohnson, P. D.; Williams, M. G.; Aristoff, P. A. Interstrand
DNA cross-linking with dimers of the spirocyclopropyl alkylating
moiety of CC-1065. J . Am. Chem. Soc. 1989, 111, 6428. (d)
Seaman, F. C.; Hurley, L. H. Manipulative interplay of the
interstrand cross-linker bizelesin with d(TAATTA)2 to achieve
sequence recognition of DNA. J . Am. Chem. Soc. 1996, 118,
10052. (e) Kelly, R. C.; Aristoff, P. A. Novel CC-1065 analogues
having two CPI subunits. EP0359454.
(9) Fukuda, Y.; Furuta, H.; Kusama, Y.; Ebisu, H.; Oomori, Y.;
Terashima, S. The novel cyclopropapyrroloindole(CPI) bisalky-
lators bearing methoxycarbonyl and trifluoromethyl group.
Bioorg. Med. Chem. Lett. 1998, 8, 1387.
(10) Fukuda, Y.; Seto, S.; Furuta, H.; Ebisu, H.; Oomori, Y.; Terash-
ima, S. The novel cyclopropapyrroloindole (CPI) bisalkylators
bearing 3,3′-(1,4-phenylene)diacryloyl group as a linker. Bioorg.
Med. Chem. Lett. 1998, 8, 2003.
(17) Kiggen, W.; Vo¨gtle, F.; Franken, S.; Puff, H. Large oligocyclic
cavities for strong cation complexation. Tetrahedron 1986, 42,
1859.
J M000107X