Design, synthesis and anticancer activity of N3,N 11-bis(2-hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthenes-3, 11-dicarboxamide

Article abstract of DOI:10.1248/cpb.c12-00723

A series of novel N³,N¹¹-bis(2-hydroxyethyl)-14-aryl- 14H-dibenzo[a,j]xanthenes-3,11-dicarboxamide, three N³,N ¹¹-bis(2-hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthene-3, 11-dimethanamine derivatives and their intermediates 14-aryl-14H-dibenzo[a,j] xanthenes-3,11-dicarboxylic acid, were synthesized, and the structures of which were characterized by 1H-NMR, 13C-NMR, high resolution (HR)-MS, and IR spectra. The antitumor activities of these molecules were evaluated on five cancer cell lines. The results of in vitro assay against human hepatocellular carcinoma cell lines (SK-HEP-1 and HepG2 and SMMC-7721 cells), acute promyelocytic leukemia NB4 cells and uterine cervix cancer HeLa cells, show several compounds to be endowed with cytotoxicity in micromolar to submicromolar range. The carboxamide derivatives 6c and 6e exhibitted good inhibition on NB4 cancer cells, and the IC₅₀ values of which were 0.82 μm and 0.96 μm, respectively, much lower than 5.01 μm of the positive control As₂O₃. Flow cytometric analysis results revealed that compounds 6e and 6f may induce tumor cell apoptosis.

Full text of DOI:10.1248/cpb.c12-00723

February 2013
Regular Article
Chem. Pharm. Bull. 61(2) 167–175 (2013)
167
Design, Synthesis and Anticancer Activity of N3,N11-Bis(2-hydroxyethyl)-
14-aryl-14H-dibenzo[a,j]xanthenes-3,11-dicarboxamide
,a
Yongbin Song,a Yihui Yang,b Jun You,a Bo Liu,* Lijun Wu,b Yunlong Hou,b Wenji Wang,b and
Jiuxin Zhub
a Key Laboratory of Green Chemical Technology of College of Heilongjiang Province, School of Chemical and
Environmental Engineering, Harbin University of Science and Technology; Harbin 150040, China: and b College of
Pharmacy, Harbin Medical University; Harbin 150081, China.
Received August 16, 2012; accepted October 30, 2012; advance publication released online November 12, 2012
A
series of novel N3,N11-bis(2-hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthenes-3,11-dicarboxamide,
three N3,N11-bis(2-hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthene-3,11-dimethanamine derivatives and
their intermediates 14-aryl-14H-dibenzo[a,j]xanthenes-3,11-dicarboxylic acid, were synthesized, and the
structures of which were characterized by H-NMR, 13C-NMR, high resolution (HR)-MS, and IR spectra.
1
The antitumor activities of these molecules were evaluated on five cancer cell lines. The results of in vitro
assay against human hepatocellular carcinoma cell lines (SK-HEP-1 and HepG2 and SMMC-7721 cells),
acute promyelocytic leukemia NB4 cells and uterine cervix cancer HeLa cells, show several compounds to
be endowed with cytotoxicity in micromolar to submicromolar range. The carboxamide derivatives 6c and
6e exhibitted good inhibition on NB4 cancer cells, and the IC50 values of which were 0.82µm and 0.96µm,
respectively, much lower than 5.01µm of the positive control As2O3. Flow cytometric analysis results revealed
that compounds 6e and 6f may induce tumor cell apoptosis.
Key words dibenzo[a,j]xanthene; synthesis; antitumor activity; cell apoptosis; NMR
The synthesis of xanthenes, especially benzoxanthenes has cytotoxicity against tumor cell lines,10) encouraging us to
received much attention in recent years due to their wide modify the structures of these derivatives. So far the syn-
range of biological and pharmacological activities such as thesis of dibenzo[a,j]xanthene has been mostly focused on
antiviral,1) antibacterial,2) and anti-inflammatory3) activities as modification of 14-position of the heterocyclic ring, with other
well as the activities in photodynamic therapy4) and the activi- positions seldom changed. In order to study the relationship
ties as antagonists for the paralyzing action of zoxazolamine.5) between the structures of mitoxantrone analogues and their
Furthermore, they can be used as dyes,6) pH-sensitive fluores- biological activities, as well as to obtain compounds endowed
cent materials for visualization of biomolecules7) and utilized with antiproliferative activity, we have developed a new series
in laser technologies.8)
of 14-aryl-14H-dibenzo[a,j]xanthene-3,11-dicarboxamide com-
Anthracenes, especially anthraquinones, are the basic pounds bearing a 2-hydroxyethyl group in the nitrogen atom
framework of anthracycline antitumor agents, which contain (6a–i, Fig. 2). In addition, three N3,N11-bis(2-hydroxyethyl)-14-
many clinical drugs such as bisantrene, doxorubicin, dauno- aryl-14H-dibenzo[a,j]xanthene-3,11-dimethanamine
deriva-
rubicin, epirubicin, zorubicin, and aclarubicin. Mitoxantrone tives (9a, 9e, 9f, Fig. 3) and ten intermediates containing
(Fig. 1) is a significant synthesized antineoplastic agent, dicarboxylic acid groups (5a–j, Fig. 2) were prepared. In this
which owns strong clinical activity on patients with breast paper, we reported on the synthesis of target derivatives and
cancer, acute leukemia and lymphoma. In addition, it exhib- theirs intermediates, and tested cytotoxicity on five cancer cell
its antiviral, antibacterial, antiprotozoal, immunomodulating, lines.
and antineoplastic properties.9) Therefore, it is significant to
investigate the analogues of mitoxantrone which maintain its
biological activity.
Results and Discussion
Chemistry The synthetic route for the new dicarbox-
The findings of research indicate that the compounds of amide derivatives 6a–i is outlined in Chart 1. Compounds
14-aryl-14H-dibenzo[a,j]xanthenes (Fig. 1) exhibit in vitro 1–4, which were known compounds, were the materials and
Fig. 1. Structures of Mitoxantrone and 14-Aryl-14H-dibenzo[a,j]xanthenes
The authors declare no conflict of interest.
© 2013 The Pharmaceutical Society of Japan
*To whom correspondence should be addressed. e-mail: songyongbin1981@sina.com

168
Vol. 61, No. 2
Fig. 2. Structures of 5a–j and 6a–i
Fig. 3. Structures of 7a, 7e, 7f and 9a, 9e, 9f
Reagents and conditions: (a) (i) Br2, AcOH, (ii) Sn, AcOH, 3h, reflux; (b) CuCN, DMF, 4h, 160°C; (c) HCl, H2O, reflux, 8h; (d) H2SO4, AcOH, 0.5–2h; (e) (i) SOCl2,
DMF, CHCl3, 4h, 67°C; (ii) CHCl3, 3h, rt.
Chart 1
intermediates for the synthesis of dicarboxamide derivatives
The synthesis of key intermediate dicarboxylic acids (5a–j)
(6a–i). Compounds 2 and 3 were prepared through published is also outlined in Chart 1. 5a–j were prepared through the
methodology,11,12) respectively. Compound 3 was refluxed one-pot condensation of aldehydes with compound 4 in the
for 8h in the presence of 10% hydrochloric acid, the formed present of concentrated sulfuric acid in acetic acid and good
precipitate was filtered and washed with water and EtOAc yields (84–92%) were obtained.
successively, and then dried under vacuum to afford the
6-hydroxy-2-naphthalenecarboxylic acid compound 4 (88–90% responding acyl chlorides, and then turned into dicarbox-
yield) (Chart 1). amides 6a–i (84–92% yield) (Chart 1), by adding excessive
Compounds 5a–i were firstly converted into the cor-

February 2013
169
Reagents and conditions: (f) (i) SOCl2, DMF, CHCl3, 4h, 67°C, (ii) LiAlH4, THF, 15min, 0°C; (g) (i) SOCl2, DMF, CHCl3, 0.5h, r.t.; (ii) 2-aminoethanol, CHCl3, 6h, r.t.
Chart 2
2-aminoethanol. But the conversion of compound 5j into the show antitumor activity to five cancer cell lines (IC50 >50µm).
corresponding dicarboxamide 6j was unsuccessful, and this As evidenced by the cytotoxicity data, the carboxamide de-
may be due to the formyl group of 5j. It is easy to purify the rivatives 6a–i show better inhibitory activity than carboxylic
compounds type 5 and 6, which have small solubilities in the acid derivatives 5a–j in most cases. Meanwhile, compounds
corresponding solvents, and the dopants are soluble.
with a substituted benzene ring at the position 14 of the mol-
Known compounds 7a, 7e and 7f (Fig. 3) were prepared ecule (6c–i, 5b–i) appear to be more toxic than those with an
through published methodology.13) The synthetic route for unsubstituted benzene ring (6a, 5a); moreover, a comparison
the new dimethanamine derivatives 9a, 9e and 9f (Fig. 3) is of the substitution on the benzene ring suggests that para
outlined in Chart 2. The conversions of compounds, 6a, 6e position-halogen-substituted carboxamide derivatives (e.g.,
and 6f into the corresponding dimethanamines produce very 6c) exhibit stronger inhibitory activity for the cancer cells
low yields using lithium aluminum hydride as reducing agent than those of ortho position-halogen-substituted ones (e.g., 6b,
in tetrahydrofuran (THF), possibly owing to side reactions Table 1).
and the reduction of the halogen atoms in these compounds.
Compounds 6c, 6e, 6f and 6h are proved to be potent cy-
Compounds 5a and 5e can give 8a and 8e, respectively, using totoxic agents towards leukemia cell line NB4. The carbox-
lithium aluminum hydride as reducing agent, but 5f only gives amide derivative 6c demonstrates the best inhibition to NB4
8a, not 8f, owing to the reduction of the bromine atom into cancer cells at a concentration of 0.82µm, lower than 5.01µm
hydrogen atom. LiAlH4 can convert acid chloride of dicar- of As2O3. And the mean IC50 values of 6e, 6f and 6h are 0.96,
boxylic acid 5 into compounds 8 quickly, and the bromine 2.06 and 2.38µm, respectively, which are also lower than that
atom of 5f can be retained in the dimethanol derivative 8f. of arsenic trioxide (Table 1). Compound 6i inhibits the growth
Compounds 8 were treated with thionyl chloride to give corre- of NB4 cells at 6.15µm, which is near the concentration of
sponding dichlorinated compounds, which were not separated As2O3. In addition, carboxylic acid compounds 5b–i exhibit
and reacted with 2-aminoethanol in CHCl3 to afford deriva- a moderate cytotoxicity to NB4 cell line; in contrast, the ana-
tives 9a, 9e and 9f.
logs 5a and 5j are inactive (Table 1). the mean IC50 values of
All the structures of the synthetic intermediates (5a–j), the 9a, 9e and 9f are 3.08, 4.12 and 4.01µm, respectively, which
derivatives (6a–i, 8a, 8e, 8f, 9a, 9e, 9f) were confirmed by are also lower than that of arsenic trioxide (Table 1).
1H-NMR, 13C-NMR, high resolution (HR)-MS, IR spectro-
The derivatives 6c and 6d exhibit respective IC50 values
scopic data which allowed the correct identification and deter- of 5.72 and 5.26µm to human hepatoma cell line (HepG2),
mined the purity of the compounds. which are lower than the positive control. The IC50 value of
Antiproliferation Assay The series of compounds 5a–j, compound 6e to HepG2 cell line is 6.41µm, which is com-
6a–i, 9a, 9e and 9f were tested for cytotoxicity against a panel parable to that of the positive control As2O3. Compounds 6f
of five cancer cell lines of different types of human tumors, and 6g exhibit moderate IC50 values of 11.56 and 16.07µm to
which contained human hepatoma cell (SK-HEP-1, HepG2, HepG2 cell line, respectively, and 6i shows a weak IC50 value
SMMC-7721), acute promyelocytic leukemia cell (NB4), and of 30.47µm (Table 1). The derivatives 9a, 9e, and 9f exhibit
uterine cervix cancer cell (HeLa). The corresponding IC50 IC50 values of 13.24, 14.49, and 16.79µm to HepG2 cell line,
values of all compounds are summarized in Table 1, and ar- respectively (Table 1).
senic trioxide (As2O3) tested in parallel was used as cytotoxic
Compounds 6e and 6f exhibit a good inhibitory effect on
positive control. In addition, dibenzo[a,j]xanthene derivatives human hepatoma cell line (SK-HEP-1), and the IC50 values
(7e, 7e, 7f), which do not possess functional groups on 3- and are 6.46 and 6.41µm, respectively, which are comparable to
11-positions, were used as negative standards, and they don’t that of As2O3 (6.02 µm, Table 1). Compound 6e is also proved

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Vol. 61, No. 2
Table 1. IC50 Values (µm) for Antiproliferative Activity of the Synthesized Compounds
IC50 (µm) for different cell linesa)
Compound
HepG2
SK-HEP-1
SMMC-7721
NB4
>50
HeLa
>50
5a
5b
>50
>50
>50
>50
>50
>50
22.
21.17 0.34
5c
5d
5e
5f
5g
5h
5i
5j
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
18.21 1.43
30.35 1.17
23.24 1.39
17.33 0.14
37.27 0.74
44.41 1.02
47.64 1.38
>50
>50
22.13 1.02
>50
36.39 1.48
>50
>50
>50
>50
6a
>50
>50
>50
>50
>50
6b
>50
>50
>50
>50
>50
6c
6d
6e
6f
6g
6h
6i
9a
5.72 0.54
5.26 0.96
6.41 0.032
11.56 0.69
16.07 0.42
>50
30.47 1.98
13.24 0.52
14.49 0.35
16.79 0.41
>50
9.81 0.38
9.21 1.84
6.46 0.50
6.41 0.52
>50
19.27 1.35
13.98 1.01
7.35 0.29
7.31 0.72
8.44 0.34
>50
12.50 1.16
>50
9.92 1.06
18.71 0.65
>50
0.82 0.042
>50
27.94 2.58
21.51 2.05
17.72 1.83
13.18 1.79
>50
0.96 0.051
2.06 0.21
>50
2.38 0.27
6.15 0.94
3.08 0.25
4.12 0.32
4.01 0.41
>50
>50
>50
20.11 0.24
16.68 0.61
17.25 0.67
16.35 0.68
>50
20.76 1.95
14.27 1.63
15.18 1.79
16.35 2.13
>50
9e
9f
7a, 7e, 7f
As2O3
6.23 0.34
6.02 0.38
9.13 0.82
5.01 0.071
10.92 0.22
a) Values are means standard deviation from three consecutive experiments.
to be more active in inhibition of human hepatoma cell effect study, compounds 6e and 6f were selected to perform
(SMMC-7721) than its analogues. Compound 6f shows a better a biparametric cytofluorimetric analysis using annexin V and
inhibitory activity than its derivatives to uterine cervix cancer PI double-staining as shown in Fig. 4, in order to elucidate
HeLa cells (Table 1).
whether the compound-induced cell death involved apoptosis
It can be found that halogen substitution at C-4′ (para) or necrosis. The results indicate that the experimental com-
position on the 14-phenyl group of dicarboxamide deriva- pounds could induce the apoptosis of activated HeLa cells. It
tives 6a–i is critical for improving the antitumor activity has been found that compounds 6e and 6f induce more apo-
to five cell lines, especially to eukemia cell line NB4. The ptotic cells than necrotic cells, and this phenomenon is even
ability sequence of the halogen atoms in compounds 6c, 6e more pronounced at a lower concentration (12.5µm) than at a
and 6f to improve the antitumor activity is F>Cl>Br, which higher concentration (25µm).
maybe due to the atomic size difference or the capability of
forming hydrogen bonds with the receptor. Halogen substitu-
tion at C-2' (ortho) position on the 14-phenyl group shows
Conclusion
In conclusion, a series of N3,N11-bis(2-hydroxyethyl)-14-
little effect on antitumor activity, thus we can deduce that aryl-14H-dibenzo[a,j]xanthenes-3,11-dicarboxamide
deriva-
the steric hindrance can block the conjunction between the tives, three N3,N11-bis(2-hydroxyethyl)-14-aryl-14H-dibenzo-
halogen atom and receptor. Derivatives 9a, 9e and 9f show a [a,j]xanthene-3,11-dimethanamine derivatives and their in-
comparative antitumor activity with that of 6f to five cell lines termediate can be synthesized in acceptable overall yields.
(Table 1). An unsubstituted benzene ring at the position 14 of The results of in vitro antitumor activity experiments reveal
the molecule (9a) appears to be more toxic than those with a that the series of compounds exhibit pronounced inhibitory
substituted benzene ring (9e, 9f), and the antitumor activities activities toward a wide range of human tumor cell lines, and
of derivatives 9a, 9e and 9f do not show a similar tendency some of them show a high level of tumor cells selectivity. The
toward halogen substitution with that of compounds 6a–i.
carboxamide derivatives 6c and 6e demonstrate good inhibi-
Several compounds show a high level of tumor cells selec- tion effects on NB4 cancer cells, and the best activities appear
tivity. For instance, Compound 6d shows anti-proliferative at concentrations of 0.82µm and 0.96µm, respectively, much
activity on human hepatoma SK-HEP-1, HepG2 and uterine lower than 5.01µm of the positive control As2O3, which is
cervix cancer HeLa cells, but it does not exhibit cytotoxic believed to be a potential antitumor agent with a potent inhibi-
activity on human hepatoma SMMC-7721 and acute promy- tory activity in tumor growth. Apoptosis assay results by the
elocytic leukemia NB4 cells, the IC50 values of which exceed flow cytometry analysis show that compounds 6e and 6f are
50µm. Compound 6g shows only anti-proliferative activity potential antitumor agents.
to cell line HepG2, and doesn’t exhibit inhibitory activity to
other tumor cell lines (Table 1).
Experimental
Apoptosis Assay On the basis of the antiproliferative
Chemistry All the solvents were of analytic grade. Silica

February 2013
171
Fig. 4. The Apoptosis Induction Effect of 6e and 6f on HeLa Cells, HeLa Cells Were Cultured with Agents for 24h
Cells were stained by Annexin V-FITC/PI and apoptosis was analyzed by flow cytometry.
GF254 plates were purchased from Qingdao Haiyang Chemi- C29H17O5 [M−H]+ 455.1076, Found 455.1071.
cal Co., Ltd., China. The melting points of the compounds
14-(2-Fluorophenyl)-14H-dibenzo[a,j]xanthene-3,11-dicar-
were taken on an X-6 melting piont apparatus and are un- boxylic Acid (5b): White solid. Yield 83.4%. mp >300°C.
corrected. IR spectra were obtained on an Avatar 370 FT-IR 1H-NMR (DMSO-d6, 300MHz) δ: 8.62 (d, J=1.5Hz, 2H, 4,
spectrometer. 1H- and 13C-NMR spectra were recorded at 10-H), 8.50 (d, J=9.0Hz, 2H, 1, 13-H), 8.18 (d, J=8.9Hz, 2H,
300 and 75MHz, respectively in DMSO-d6, using a Bruker 5, 9-H), 8.07 (dd, J=9.0, 1.6Hz, 2H, 2, 12-H), 7.65 (d, J=
Avance 300 spectrometer (tetramethylsilane (TMS) as an 8.9Hz, 2H, 6, 8-H), 7.59–7.62 (m, 1H), 6.70–7.11 (m, 3H), 6.88
internal standard). HR-MS were measured on a Waters LCT (s, 1H, 14-H). 13C-NMR (DMSO-d6, 75MHz) δ: 167.7, 158.9
Premier XE benchtop orthogonal acceleration time-of-flight (1JCF=242.9Hz), 150.0, 133.6, 131.8 (3JCF=7.0Hz), 131.7, 131.5
mass spectrometer.
General Procedure
(2JCF=24.4Hz), 131.4, 130.3, 129.5 (3JCF=8.5Hz), 127.3, 126.9,
of 125.6, 123.1, 118.9, 116.2 (2JCF=22.5Hz), 115.8, 31.4. IR (KBr)
for
the
Preparation
14-Aryl-14H-Dibenzo[a,j]xanthene-3,11-dicarboxylic Acid ν: 1689, 1621, 1466, 1398, 1301, 1242cm−1. HR-MS (ESI)
(5a–j) A mixture of 6-hydroxy-2-naphthalenecarboxylic acid Calcd for C29H16FO5 [M−H]+ 463.0982, Found 463.0987.
4 3.76g (20mmol), appropriate arylaldehyde (10.5mmol), gla-
14-(4-Fluorophenyl)-14H-dibenzo[a,j]xanthene-3,11-dicar-
cial acetic acid (20mL) and concentrated sulfuric acid (1mL) boxylic Acid (5c): White solid. Yield 85.3%. mp >300°C.
was stirred at room temperature for 10min, and then heated 1H-NMR (DMSO-d6, 300MHz) δ: 8.77 (d, J=9.0Hz, 2H, 1,
to reflux for 0.5–2h. TLC was used to monitor the reactions. 13-H), 8.62 (d, J=1.4Hz, 2H, 4, 10-H), 8.16 (d, J=9.0Hz, 2H,
After the reaction had been completed, the system was cooled 5, 9-H), 8.07 (dd, J=9.0, 1.4Hz, 2H, 2, 12-H), 7.66 (d, J=
to room temperature, the formed precipitate was filtered and 9.0Hz, 2H, 6, 8-H), 7.68–7.59 (m, 2H, 2′, 6′-H), 6.99 (t, J=
washed with acetic acid and water successively, then dried 8.8Hz, 2H, 3′, 5′-H), 6.82 (s, 1H, 14-H). 13C-NMR (DMSO-
under vacuum to afford the compounds 5a–j.
d6, 75MHz) δ: 167.7, 161.0 (1JCF=241.9Hz), 149.9, 141.7, 133.5,
14-Phenyl-14H-dibenzo[a,j]xanthene-3,11-dicarboxylic Acid 131.7, 131.3, 130.5, 130.1 (3JCF=8.0Hz), 127.3, 126.7, 124.2,
(5a): White solid. Yield 82.3%. mp >300°C. 1H-NMR 119.1, 117.9, 115.8 (2JCF=21.3Hz), 36.0. IR (KBr) ν: 2925, 1691,
(DMSO-d6, 300MHz) δ: 8.74 (d, J=9.0Hz, 2H, 1, 13-H), 1620, 1466, 1398, 1296, 1245cm−1. HR-MS (ESI) Calcd for
8.60(d, J=1.4Hz, 2H, 4, 10-H), 8.14 (d, J=8.9Hz, 2H, 5, 9-H), C29H16FO5 [M−H]+ 463.0982, Found 463.0989.
8.06 (dd, J=9.0, 1.5Hz, 2H, 2, 12-H), 7.64 (d, J=8.9Hz, 2H,
14-(2-Chlorophenyl)-14H-dibenzo[a,j]xanthene-3,11-di-
6, 8-H), 7.60 (d, J=7.3Hz, 2H, 2′, 6′-H), 7.15(t, J=7.4Hz, carboxylic Acid (5d): White solid. Yield 84.1%. mp >300°C.
2H, 3′, 5′-H), 6.98 (t, J=7.3Hz, 1H, 4′-H), 6.76(s, 1H, 14-H). 1H-NMR (DMSO, 300MHz) δ: 8.59 (brs, 2H, 4, 10-H), 8.54
13C-NMR (DMSO-d6, 75MHz) δ: 167.6, 149.9, 145.5, 133.7, (d, J=9.1Hz, 2H, 1, 13-H), 8.10 (d, J=9.0Hz, 2H, 5, 9-H),
131.6, 131.2, 130.5, 129.0, 128.3, 127.1, 127.0, 126.7, 124.3, 8.04 (dd, J=9.1, 1.3Hz, 2H, 2, 12-H), 7.54 (d, J=9.0Hz,
119.1, 118.0, 36.9. IR (KBr) ν: 1686, 1620, 1465, 1396, 1300, 2H, 6, 8-H), 7.43 (d, J=7.3Hz, 1H, 6′-H), 7.30 (d, J=7.9Hz,
1242 cm−1. HR-MS (electrospray ionization (ESI)) Calcd for 1H, 3′-H), 7.12–6.96 (m, 2H, 4′, 5′-H), 6.66 (s, 1H, 14-H).

172
Vol. 61, No. 2
13C-NMR (DMSO, 75MHz) δ: 167.7, 150.1, 142.6, 133.7, 132.2, 118.1, 36.5; 20.9. IR (KBr) ν: 1704, 1681, 1621, 1468, 1399,
131.7, 131.5, 130.5, 130.3, 129.2, 128.7, 127.3, 126.7, 126.7, 1289, 1251 cm−1. HR-MS (ESI) Calcd for C30H19O5 [M−H]+
123.8, 119.1, 116.8, 35.1. IR (KBr) ν: 1685, 1620, 1469, 1397, 459.1233, Found 459.1236.
1303, 1250cm−1. HR-MS (ESI) Calcd for C29H16ClO5 [M+H]+
479.0686, Found 479.0679.
14-(4-Formylphenyl)-14H-dibenzo[a,j]xanthene-3,11-
dicarboxylic Acid (5j): White solid. Yield 85.8%. mp >300°C.
14-(4-Chlorophenyl)-14H-dibenzo[a,j]xanthene-3,11- 1H-NMR (DMSO-d6, 300MHz) δ: 9.76 (s, 1H, CHO), 8.78
dicarboxylic Acid (5e): White solid. Yield 87.9%. mp >300°C. (d, J=9.1Hz, 2H, 1, 13-H), 8.62 (d, J=1.7Hz, 2H, 4, 10-H),
1H-NMR (DMSO-d6, 300MHz) δ: 8.75 (d, J=9.0Hz, 2H, 1, 8.19 (d, J=9.1Hz, 2H, 5, 9-H), 8.07 (dd, J=9.0, 1.7Hz, 2H,
13-H), 8.63 (d, J=1.6Hz, 2H, 4, 10-H), 8.17 (d, J=8.9Hz, 2, 12-H), 7.87 (d, J=8.2Hz, 2H, 2′, 6′-H), 7.71 (d, J=8.2Hz,
2H, 5, 9-H), 8.07 (dd, J=9.0, 1.7Hz, 2H, 2, 12-H), 7.66 (d, 2H, 3′, 5′-H), 7.69 (d, J=8.9Hz, 2H, 6, 8-H), 6.93 (s, 1H,
J=8.9Hz, 2H, 6, 8-H), 7.64 (d, J=8.5Hz, 2H, 2′, 6′-H), 7.23 14-H). 13C-NMR (DMSO-d6, 75MHz) δ: 192.7, 167.7, 151.8,
(d, J=8.5Hz, 2H, 3′, 5′-H), 6.82 (s, 1H, 14-H). 13C-NMR 149.9, 135.0, 133.5, 131.6, 131.5, 130.5, 130.3, 129.0, 127.3,
(DMSO-d6, 75MHz) δ: 167.7, 149.9, 144.5, 133.5, 131.6, 126.8, 124.1, 119.1, 117.1, 37.0. IR (KBr) ν: 1708, 1692, 1621,
131.6, 131.4, 130.5, 130.1, 129.0, 127.4, 126.8, 124.2, 119.1, 1468, 1399, 1299, 1251cm−1. HR-MS (ESI) Calcd for C30H17O6
117.6, 36.2. IR (KBr) ν: 1694, 1621, 1466, 1397, 1245, 1157, [M−H]+ 473.1025, Found 473.1029.
1135 cm−1. HR-MS (ESI) Calcd for C29H16ClO5 [M+H]+
479.0686, Found 479.0689.
General Procedure for the Preparation of N3,N11-Bis(2-
hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthene-3,11-
14-(4-Bromophenyl)-14H-dibenzo[a, j]xanthene-3,11- dicarboxamide (6a–i)
A
100mL, three-necked flask,
dicarboxylic Acid (5f): White solid. Yield 85.6%. mp >300°C. equipped with a magnetic stir bar, a condenser, a thermometer
1H-NMR (DMSO-d6, 300MHz) δ: 8.76 (d, J=8.8Hz, 2H, 1, and an addition funnel, was charged with 2mmol dibenzo[a,j]-
13-H), 8.63 (brs, 2H, 4, 10-H), 8.19 (d, J=8.9Hz, 2H, 5, 9-H), xanthenes dicarboxylic acid 5, 20mL of CHCl3 and 1 drop
8.07 (brd, J=8.8Hz, 2H, 2, 12-H), 7.68 (d, J=8.9Hz, 2H, 6, of dimethylformamide. Afterwards a solution of 2.2mL
8-H), 7.59 (d, J=6.9Hz, 2H, 2′, 6′-H), 7.37 (d, J =6.5Hz, 2H, (30mmol) thionyl chloride in CHCl3 (8mL) was added, and
3′, 5′-H), 6.82 (s, 1H, 14-H). 13C-NMR (DMSO-d6, 75MHz) the mixture was heated to reflux at 67°C for 4h. After the ini-
δ: 167.7, 149.9, 144.9, 133.5, 132.0, 131.7, 131.4, 130.5, 130.5, tial suspension turned into a yellow solution, thionyl chloride
127.3, 126.8, 124.2, 120.2, 119.1, 117.5, 36.3. IR (KBr) ν: 1703, was removed by evacuation under reduced pressure to give the
1621, 1467, 1400, 1300, 1252cm−1. HR-MS (ESI) Calcd for acid chloride as a white solid. A CHCl3 (20mL) solution of
C29H16BrO5 [M−H]+ 523.0181, Found 523.0188.
the acid chloride was then added to the solution of 2-amino-
14-(3-Nitrophenyl)-14H-dibenzo[a,j]xanthene-3,11-dicar- ethanol (488mg, 8mmol) in CHCl3(25mL). The mixture was
boxylic Acid (5g): White solid. Yield 84.3%. mp >300°C. stirred for 3h at room temperature. A white solid formed was
1H-NMR (DMSO-d6, 300MHz) δ: 8.84 (d, J=9.0Hz, 2H, 1, filtered and washed with CHCl3 and water, successively, then
13-H), 8.63 (d, J=1.4Hz, 2H, 4, 10-H), 8.52 (brs, 1H, 2′-H), dried under vacuum to afford the compounds 6a–i.
8.20 (d, J=9.1Hz, 2H, 5, 9-H), 8.10 (brd, J=7.3Hz, 1H, 6′-H),
N3,N11-Bis(2-hydroxyethyl)-14-phenyl-14H-dibenzo[a,j]-
8.08 (dd, J=9.0, 1.5Hz, 2H, 2, 12-H), 7.87 (dd, J=8.1, 1.5Hz, xanthene-3,11-dicarboxamide (6a): White solid. Yield 88.9%.
1H, 4′-H), 7.70 (d, J=9.0Hz, 2H, 6, 8-H), 7.48 (t, J=8.0Hz, mp 202–204°C. 1H-NMR (DMSO-d6, 300MHz) δ: 8.75
1H, 5′-H), 7.03 (s, 1H, 14-H). 13C-NMR (DMSO-d6, 75MHz) (d, J=9.0Hz, 2H, 1, 13-H), 8.61 (t, J=5.2Hz, 2H, NH×2),
δ: 167.7, 150.1, 148.4, 147.4, 134.7, 133.5, 131.7, 130.7, 130.5, 8.48 (s, 2H, 4, 10-H), 8.05 (m, 4H, 5, 9, 2, 12-H), 7.64 (m,
127.5, 126.9, 124.1, 122.5, 122.3, 119.1, 117.1, 117.1, 36.3. 4H, 6, 8, 2′, 6′-H), 7.15 (t, J=7.5Hz, 2H, 3′, 5′-H), 6.98 (t,
IR (KBr) ν: 1686, 1622, 1534, 1466, 1397, 1349, 1246cm−1. J=7.2Hz, 1H, 4′-H), 6.79 (s, 1H, 14-H), 4.79 (s, 2H, OH×2),
HR-MS (ESI) Calcd for C29H16NO7 [M−H]+ 490.0927, Found 3.62–3.45 (m, 4H, NHCH2CH2OH×2), 3.44–3.31 (m, 4H,
490.0930.
NHCH2CH2OH×2). 13C-NMR (DMSO-d6, 75MHz) δ: 166.6,
14-(4-Nitrophenyl)-14H-dibenzo[a,j]xanthene-3,11-dicar- 149.4, 145.8, 132.7, 131.1, 130.6, 130.4, 128.9, 128.7, 128.4,
boxylic Acid (5h): White solid. Yield 82.3%. mp 284–285°C. 126.9, 125.6, 124.1, 118.9, 117.9, 60.3, 42.8, 36.9. IR (KBr) ν:
1H-NMR (DMSO-d6, 300MHz) δ: 8.78 (d, J=8.9Hz, 2H, 1, 2929, 2876, 1638, 1546, 1462, 1399, 1249cm−1. HR-MS (ESI)
13-H), 8.64 (brs, 2H, 4, 10-H), 8.20 (d, J=9.0Hz, 2H, 5, 9-H), Calcd for C33H29N2O5 [M+H]+ 533.2076, Found 533.2070.
8.08 (brd, J=8.9Hz, 2H, 2, 12-H), 8.04 (d, J=8.4Hz, 2H, 2′,
N3,N11-Bis(2-hydroxyethyl)-14-(2-fluorophenyl)-14H-di-
6′-H), 7.92 (d, J=8.3Hz, 2H, 3′, 5′-H), 7.70 (d, J=8.9Hz, 2H, benzo[a,j]xanthene-3,11-dicarboxamide (6b): White solid.
1
6, 8-H), 7.01 (s, 1H, 14-H). 13C-NMR (DMSO-d6, 75MHz) Yield 90.7%. mp 201–203°C. H-NMR (DMSO-d6, 300MHz)
δ: 167.7, 152.5, 150.0, 146.4, 133.5, 131.7, 131.7, 130.5, 129.5, δ: 8.58 (t, J=5.6Hz, 2H, NH×2), 8.49 (s, 2H, 4, 10-H), 8.47
127.4, 126.9, 124.4, 124.1, 119.1, 116.8, 36.7. IR (KBr) ν: 1703, (d, J=8.9Hz, 2H, 1, 13-H), 8.07 (d, J=9.0Hz, 2H, 5, 9-H),
1621, 1522, 1467, 1400, 1345, 1251cm−1. HR-MS (ESI) Calcd 8.05 (dd, J=8.9, 1.5Hz, 2H, 2, 12-H), 7.63 (d, J=9.0Hz,
for C29H16NO7 [M−H]+ 490.0927, Found 490.0924.
2H, 6, 8-H), 7.64–7.54 (m, 1H, Ar-H), 7.13–6.97 (m, 3H,
14-(4-Methylphenyl)-14H-dibenzo[a,j]xanthene-3,11-di- Ar-H), 6.90 (s, 1H, 14-H), 4.76 (t, J=5.6Hz, 2H, OH×2),
carboxylic Acid (5i): White solid. Yield 84.9%. mp >300°C. 3.54 (q, J=6.0Hz, 4H, NHCH2CH2OH×2), 3.40–3.34 (m, 4H,
1H-NMR (DMSO-d6, 300MHz) δ: 8.74 (d, J=9.0Hz, 2H, 1, NHCH2CH2OH×2). 13C-NMR (DMSO-d6, 75MHz) δ: 166.5,
13-H), 8.61 (s, 2H, 4, 10-H), 8.14 (d, J=9.0Hz, 2H, 5, 9-H), 159.0 (1JCF=242.7Hz), 149.6, 132.6, 131.9 (2JCF=13.1Hz), 131.4,
8.05 (d, J=9.0Hz, 2H, 2, 12-H), 7.64 (d, J=8.9Hz, 2H, 6, 131.1, 131.0, 130.3, 129.5 (3JCF=8.4Hz), 128.9, 125.8, 125.7
8-H), 7.48 (d, J=7.8Hz, 2H, 2′, 6′-H), 6.95 (d, J=7.8Hz, 2H, (2JCF=21.5Hz), 123.0, 118.9, 116.2 (2JCF=22.4Hz), 115.7, 60.3,
3′, 5′-H), 6.72 (s, 1H, 14-H), 2.04 (s, 3H, CH3). 13C-NMR 42.8, 31.6. IR (KBr) ν: 2938, 2880, 1640, 1547, 1463, 1401,
(DMSO-d6, 75MHz) δ: 167.8, 149.8, 142.7, 136.1, 133.6, 1252 cm−1. HR-MS (ESI) Calcd for C33H28FN2O5 [M+H]+
131.6, 131.1, 130.5, 129.5, 128.2, 127.2, 126.6, 124.4, 119.0, 551.1982, Found 551.1989.

February 2013
173
N3,N11-Bis(2-hydroxyethyl)-14-(4-fluorophenyl)-14H- 5, 9, 2, 12, 6′-H), 7.87 (dd, J=8.2, 1.6Hz, 1H, 4′-H), 7.68
dibenzo[a,j]xanthene-3,11-dicarboxamide (6c): White solid. (d, J=8.9Hz, 2H, 6, 8-H), 7.47 (t, J=8.0Hz, 1H, 5′-H),
1
Yield 92.0%. mp 268–269°C. H-NMR (DMSO-d6, 300MHz) 7.04 (s, 1H, 14-H), 4.77 (t, 2H, J=5.6Hz, OH×2), 3.54
δ: 8.75 (d, J=9.0Hz, 2H, 1, 13-H), 8.62 (t, J=5.5Hz, 2H, (q, 4H, J=6.0Hz, NHCH2CH2OH×2), 3.46–3.21 (m, 4H,
NH×2), 8.49 (brs, 2H, 4, 10-H), 8.07–8.03 (m, 4H, 5, 9, 2, NHCH2CH2OH×2). 13C-NMR (DMSO-d6, 75MHz) δ: 166.5,
12-H), 7.69–7.61 (m, 4H, 6, 8, 2′, 6′-H), 6.98 (t, J=8.8Hz, 2H, 149.5, 148.3, 147.7, 134.8,132.5,131.3, 131.3, 130.7, 130.5,
3′, 5′-H), 6.83 (s, 1H, 14-H), 4.79 (t, J=5.1Hz, 2H, OH×2), 128.9, 125.8, 123.8, 122.5, 122.2, 119.0, 116.9, 60.2, 42.8,
3.64–3.50 (m, 4H, NHCH2CH2OH×2), 3.45–3.34 (m, 4H, 36.3. IR (KBr) ν: 2938, 2884, 1647, 1622, 1536, 1462, 1398,
NHCH2CH2OH×2). 13C-NMR (DMSO-d6, 75MHz) δ: 166.6, 1357, 1253 cm−1. HR-MS (ESI) Calcd for C33H28N3O7 [M+H]+
161.0 (1JCF=242.3Hz), 149.4, 142.0, 132.6, 131.1, 130.8, 130.4, 578.1927, Found 578.1929.
130.2 (3JCF=8.1Hz), 128.8, 125.6, 124.0, 119.0, 117.8, 115.7
N3,N11-Bis(2-hydroxyethyl)-14-(4-nitrophenyl)-14H-
(2JCF=21.0Hz), 60.3, 42.8, 36.0. IR (KBr) ν: 2933, 2876, 1624, dibenzo[a,j]xanthene-3,11-dicarboxamide (6h): White solid.
1549, 1506, 1463, 1400, 1250cm−1. HR-MS (ESI) Calcd for Yield 90.5%. mp 228–230°C. H-NMR (DMSO-d6, 300MHz)
1
C33H28FN2O5 [M+H]+ 551.1982, Found 551.1974.
δ: 8.75 (d, J=8.4Hz, 2H, 1, 13-H), 8.63 (brs, 2H, NH×2),
N3,N11-Bis(2-hydroxyethyl)-14-(2-chlorophenyl)-14H- 8.49 (s, 2H, 4, 10-H), 8.15–7.98 (m, 6H, 5, 9, 2, 12, 2′, 6′-H),
dibenzo[a,j]xanthene-3,11-dicarboxamide (6d): White solid. 7.94 (d, J=7.2Hz, 2H, 3′, 5′-H), 7.66 (d, J=8.7Hz, 2H, 6,
1
Yield 85.5%. mp 295–297°C. H-NMR (DMSO-d6, 300MHz) 8-H), 7.00 (s, 1H, 14-H), 4.79 (brs, 2H, OH×2), 3.54 (s,
δ: 8.64–8.56 (m, 4H, 1, 13-H, NH×2), 8.47 (d, J=1.3Hz, 2H, 4H, NHCH2CH2OH×2), 3.38 (s, 4H, NHCH2CH2OH×2).
4, 10-H), 8.07–8.04 (m, 4H, 5, 9, 2, 12-H), 7.61 (d, J=8.9Hz, 13C-NMR (DMSO-d6, 75MHz) δ: 166.5, 152.8, 149.4, 146.4,
2H, 6, 8-H), 7.51 (brd, J=7.8Hz, 1H, 6′-H), 7.32 (dd, J=7.9, 132.5, 131.3, 131.3, 130.4, 129.5, 128.8, 125.8, 124.3, 123.8,
1.2Hz, 1H, 3′-H), 7.16–7.00 (m, 2H, 4′, 5′-H), 6.84 (s, 1H, 119.0, 116.7, 60.3, 42.8, 36.7. IR (KBr) ν: 2938, 2880, 1640,
14-H), 4.75 (t, J=5.6Hz, 2H, OH×2), 3.54 (q, J =6.0Hz, 4H, 1546, 1516, 1462, 1343, 1253cm−1. HR-MS (ESI) Calcd for
NHCH2CH2OH×2), 3.38–3.34 (m, 4H, NHCH2CH2OH×2). C33H28N3O7 [M+H]+ 578.1927, Found 578.1933.
13C-NMR (DMSO-d6, 75MHz) δ: 166.5, 149.7, 142.8, 132.8,
N3,N11-Bis(2-hydroxyethyl)-14-(4-methylphenyl)-14H-
132.3, 131.1, 131.1, 130.6, 130.5, 130.3, 129.2, 128.9, 128.7, dibenzo[a,j]xanthene-3,11-dicarboxamide (6i): White solid.
125.6, 123.6, 119.0, 116.7, 60.3, 42.8, 35.2. IR (KBr) ν: 2929, Yield 89.8%. mp >300°C. 1H-NMR (DMSO-d6, 300MHz)
2868, 1645, 1548, 1464, 1399, 1254cm−1. HR-MS (ESI) Calcd δ: 8.72 (d, J=8.8Hz, 2H, 1, 13-H), 8.60 (brs, 2H, NH×2),
for C33H28ClN2O5 [M+H]+ 567.1687, Found 567.1690.
8.47 (s, 2H, 4, 10-H), 8.12–7.99 (m, 4H, 5, 9, 2, 12-H), 7.62
N3,N11-Bis(2-hydroxyethyl)-14-(4-chlorophenyl)-14H- (d, J=8.8Hz, 2H, 6, 8-H), 7.49 (d, J=7.6Hz, 2H, 2′, 6′-H),
dibenzo[a,j]xanthene-3,11-dicarboxamide (6e): White solid. 6.93 (d, J=7.5Hz, 2H, 3′, 5′-H), 6.74 (s, 1H, 14-H), 4.78 (t,
1
Yield 91.5%. mp 202–203°C. H-NMR (DMSO-d6, 300MHz) J=5.1Hz, 2H, OH×2), 3.62–3.50 (m, 4H, NHCH2CH2OH×2),
δ: 8.72 (d, J=9.0Hz, 2H, 1, 13-H), 8.60 (t, J=5.5Hz, 2H, 3.43–3.32 (m, 4H, NHCH2CH2OH×2), 2.04 (s, 3H, CH3).
NH×2), 8.47 (s, 2H, 4, 10-H), 8.06–8.03 (m, 4H, 5, 9, 2, 13C-NMR (DMSO-d6, 75MHz) δ: 166.6, 149.3, 142.9, 136.0,
12-H), 7.65–7.61 (m, 4H, 6, 8, 2′, 6′-H), 7.21(d, J=8.5Hz, 2H, 132.7, 131.0, 130.5, 130.4, 129.4, 128.7, 128.3, 125.5, 124.1,
3′, 5′-H), 6.81 (s, 1H, 14-H), 4.76 (t, J=5.5Hz, 2H, OH×2), 118.9, 118.0, 60.3, 42.8, 36.5, 20.8. IR (KBr) ν: 2921, 2848,
3.56–3.50 (m, 4H, NHCH2CH2OH×2), 3.40–3.34 (m, 4H, 1644, 1556, 1536, 1401, 1252cm−1. HR-MS (ESI) Calcd for
NHCH2CH2OH×2). 13C-NMR (DMSO-d6, 75MHz) δ: 166.6, C34H31N2O5 [M+H]+ 547.2233, Found 547.2228.
149.4, 144.7, 132.6, 131.5, 131.1, 130.9, 130.4, 130.1, 128.9,
General Procedure for the Preparation of N3,N11-Bis(2-
128.8, 125.7, 123.9, 118.9, 117.4, 60.3, 42.8, 36.2. IR (KBr) ν: hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthene-3,11-
2921, 2864, 1638, 1624, 1547, 1462, 1399, 1251cm−1. HR-MS dimethanol (8a, 8e, 8f) The acid chloride of 4mmol di-
(ESI) Calcd for C33H28ClN2O5 [M+H]+ 567.1687, Found carboxylic acid 5 was prepared with the former method, and
567.1693.
it was added in portions to a solution of 0.3g (8mmol) of
N3,N11-Bis(2-hydroxyethyl)-14-(4-bromophenyl)-14H- lithium aluminum hydride in 20mL of anhydrous THF. The
dibenzo[a,j]xanthene-3,11-dicarboxamide (6f): White solid. mixture was stirred at 0°C for 15min. Decomposition was
1
Yield 88.8%. mp 252–254°C. H-NMR (DMSO-d6, 300MHz) carried out with water and then with 10mL of 10% NaOH to
δ: 8.73 (d, J=9.0Hz, 2H, 1, 13-H), 8.62 (t, J=5.5Hz, 2H, dissovle the precipitation and the mixed solution was extracted
NH×2), 8.49 (s, 2H, 4, 10-H), 8.07–8.04 (m, 4H, 5, 9, 2, three times with EtOAc (3×20mL) and the collected EtOAc
12-H), 7.63 (d, J=9.0Hz, 2H, 6, 8-H), 7.59 (d, J=8.4Hz, layers were washed with brine and water successively and
2H, 2′, 6′-H), 7.35 (d, J=8.2Hz, 2H, 3′, 5′-H), 6.81 (s, 1H, then dried (MgSO4), filtered and concentrated to give com-
14-H), 4.78 (t, J=5.5Hz, 2H, OH×2), 3.58–3.52 (m, 4H, pounds 8a, 8e, and 8f.
NHCH2CH2OH×2), 3.42–3.36 (m, 4H, NHCH2CH2OH×2).
N3,N11-Bis(2-hydroxyethyl)-14-phenyl-14H-dibenzo[a,j]-
13C-NMR (DMSO-d6, 75MHz) δ: 166.6, 149.3, 145.1, 132.6, xanthene-3,11-dimethanol (8a): White solid. Yield 91.2%.
131.9, 131.1, 130.9, 130.5, 130.4, 128.8, 125.7, 123.9, 120.1, mp 216–218°C. 1H-NMR (DMSO-d6, 300MHz) δ: 8.62 (d,
118.9, 117.4, 60.3, 42.8, 36.3. IR (KBr) ν: 2929, 2884, 1642, J=8.8Hz, 2H, 1, 13-H), 7.89 (d, J=8.9Hz, 2H, 5, 9-H), 7.83
1622, 1539, 1464, 1400, 1256cm−1. HR-MS (ESI) Calcd for (brs, 2H, 4, 10-H), 7.60–7.55 (m, 4H, 2, 12, 2′, 6′-H), 7.52 (d,
C33H28BrN2O5 [M+H]+ 611.1182, Found 611.1184.
J=8.9Hz, 2H, 6, 8-H), 7.12 (t, J=7.7Hz, 2H, 3′, 5′-H), 6.95
N3,N11-Bis(2-hydroxyethyl)-14-(3-nitrophenyl)-14H- (t, J=7.7Hz, 1H, 4′-H), 6.68 (s, 1H, 14-H), 5.26 (t, J=5.7Hz,
dibenzo[a,j]xanthene-3,11-dicarboxamide (6g): White solid. 2H, CH2OH×2), 4.64 (d, J=5.6Hz, 4H, CH2OH×2). 13C-NMR
1
Yield 88.9%. mp 299–300°C. H-NMR (DMSO-d6, 300MHz) (DMSO-d6, 75MHz) δ: 148.2, 146.1, 139.2, 131.0, 130.4, 129.3,
δ: 8.82 (d, J=9.0Hz, 2H, 1, 13-H), 8.71–8.54 (m, 3H, 2′-H, 128.8, 128.4, 126.7, 126.7, 125.8, 123.8, 118.1, 117.9, 63.3, 37.1.
NH×2), 8.49 (d, J=1.5Hz, 2H, 4, 10-H), 8.10–8.05 (m, 5H, IR (KBr) ν: 1600, 1467, 1401, 1253, 1243, 1039cm−1. HR-MS

174
Vol. 61, No. 2
(ESI) Calcd for C29H23O3 [M+H]+ 419.1647, Found 419.1658.
5, 9-H), 7.72 (brs, 2H, 4, 10-H), 7.54 (dd, J=8.7, 1.6Hz,
N3,N11-Bis(2-hydroxyethyl)-14-(4-chlorophenyl)-14H- 2H, 2, 12-H), 7.46 (d, J=9.0Hz, 2H, 6, 8-H), 7.42 (d,
dibenzo[a,j]xanthene-3,11-dimethanol (8e): White solid. Yield J=8.4Hz, 2H, 2′, 6′-H), 7.08 (d, J=8.4Hz, 2H, 3′, 5′-H),
1
92.1%. mp 222–224°C. H-NMR (DMSO-d6, 300MHz) δ: 8.59 6.39 (s, 1H, 14-H), 3.93 (s, 4H, CH2NHCH2CH2OH×2),
(d, J=8.7Hz, 2H, 1, 13-H), 7.89 (d, J=9.0Hz, 2H, 5, 9-H), 7.83 3.68 (t, J=5.1Hz, 4H, CH2NHCH2CH2OH×2), 2.83 (t,
(brs, 2H, 4, 10-H), 7.60 (d, J=8.5Hz, 2H, 2′, 6′-H), 7.56 (brd, J=5.1Hz, 4H, CH2NHCH2CH2OH×2), 2.26 (brs, 4H,
J=8.7Hz, 2H, 2, 12-H), 7.51 (d, J=9.0Hz, 2H, 6, 8-H), 7.18 CH2NHCH2CH2OH×2). 13C-NMR (CDCl3, 75MHz) δ: 148.5,
(d, J=8.4Hz, 3′, 5′-H), 6.70 (s, 1H, 14-H), 5.25 (t, J=5.7Hz, 143.5, 135.9, 132.1, 131.0, 130.4, 129.5, 128.9, 128.6, 127.7,
2H, CH2OH×2), 4.63 (d, J=5.6Hz, 4H, CH2OH×2). 13C-NMR 127.5, 122.8, 118.2, 116.6, 61.0, 53.3, 50.3, 37.5. IR (KBr) ν:
(DMSO-d6, 75MHz) δ: 148.2, 145.0, 139.3, 131.3, 131.0, 130.3, 2924, 2852, 1597, 1489, 1466, 1400, 1243cm−1. HR-MS (ESI)
130.1, 129.5, 128.8, 126.8, 125.9, 123.6, 118.1, 117.4, 63.2, Calcd for C33H32ClN2O3 [M+H]+ 539.2101, Found 539.2106.
36.4. IR (KBr) ν: 1598, 1489, 1466, 1400, 1252, 1015cm−1.
N3,N11-Bis(2-hydroxyethyl)-14-(4-bromophenyl)-14H-
HR-MS (ESI) Calcd for C29H22ClO3 [M+H]+ 453.1257, Found dibenzo[a,j]xanthene-3,11-dimethanamine (9f): White solid.
453.1250.
Yield 82.1%. mp 120–122°C. 1H-NMR (CDCl3, 300MHz)
N3,N11-Bis(2-hydroxyethyl)-14-(4-bromophenyl)-14H- δ: 8.22 (d, J=8.7Hz, 2H, 1, 13-H), 7.72 (d, J=8.9Hz, 2H,
dibenzo[a,j]xanthene-3,11-dimethanol (8f): White solid. Yield 5, 9-H), 7.69 (d, J=1.1Hz, 2H, 4, 10-H), 7.52 (dd, J=8.7,
89.1%. mp 230–232°C. 1H-NMR (CDCl3, 300MHz) δ: 8.28 1.7Hz, 2H, 2, 12-H), 7.44 (d, J=8.9Hz, 2H, 6, 8-H), 7.34
(d, J=8.7Hz, 2H, 1, 13-H), 7.78 (s, 2H, 4, 10-H), 7.77 (d, (d, J=8.5Hz, 2H, 2′, 6′-H), 7.20 (d, J=8.5Hz, 2H, 3′, 5′-H),
J=8.8Hz, 2H, 5, 9-H), 7.57 (dd, J=8.7, 1.7Hz, 2H, 2, 12-H), 6.34 (s, 1H, 14-H), 3.90 (s, 4H, CH2NHCH2CH2OH×2),
7.47 (d, J=8.9Hz, 2H, 6, 8-H), 7.36 (d, J=8.5Hz, 2H, 2′, 3.66 (t, J=5.3Hz, 4H, CH2NHCH2CH2OH×2), 2.80 (t,
6′-H), 7.23 (d, J=8.5Hz, 3′, 5′-H), 6.40 (s, 1H, 14-H), 4.82 (s, J=5.3Hz, 4H, CH2NHCH2CH2OH×2), 2.24 (brs, 4H,
4H, CH2OH×2) 1.96 (s, 2H, CH2OH×2). 13C-NMR (CDCl3, CH2NHCH2CH2OH×2). 13C-NMR (CDCl3, 75MHz) δ: 148.5,
75MHz) δ: 148.6, 143.9, 136.9, 131.6, 131.0, 130.7, 129.8, 129.1, 144.0, 136.0, 131.6, 131.0, 130.4, 129.9, 128.9, 127.7, 127.5,
126.6, 126.3, 122.9, 120.3, 118.3, 116.6, 65.2, 37.6. IR (KBr) ν: 122.7, 120.2, 118.2, 116.5, 61.0, 53.3, 50.6, 37.6. IR (KBr) ν:
1597, 1485, 1466, 1400, 1253, 1242, 1010cm−1. HR-MS (ESI) 2920, 2848, 1597, 1485, 1466, 1400, 1252cm−1. HR-MS (ESI)
Calcd for C29H22BrO3 [M+H]+ 497.0752, Found 497.0760.
Calcd for C33H32BrN2O3 [M+H]+ 583.1596, Found 583.1590.
Cell Culture The antitumor activity testing was per-
General Procedure for the Preparation of N3,N11-Bis(2-
hydroxyethyl)-14-aryl-14H-dibenzo[a,j]xanthene-3,11- formed on 5 human tumor cell lines from three cancer types
dimethanamine (9a, 9e, 9f) A 100mL, three-necked flask, (leucemia, liver, cervix) using the standard 3-(4,5-dimethylthi-
equipped with a magnetic stir bar, a condenser, a thermometer azol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-dye reduc-
and an addition funnel, was charged with 2mmol compounds tion assay.
8, 20mL of CHCl3 and 1 drop of dimethylformamide. Af-
The tumor cells were cultured in RPMI1640 medium
terwards a solution of 1mL (13.6mmol) thionyl chloride in supplemented with 10% fetal bovine serum and 1% penicillin-
CHCl3 (4mL) was added, and the mixture was stirred at streptomycin under a humidified atmosphere of 5% CO2 at
room temperature for 0.5h. After the solvent was removed 37°C. The density of inoculum depended on the type of tumor
by evacuation under reduced pressure, a yellow solid can be cell, and 400–2000 cells were seeded in 100µL of medium
obtained. A CHCl3 (10mL) solution of the yellow solid was per well of 96-well flat-bottomed microplates.
then added to the solution of 2-aminoethanol (488mg, 8mmol)
Cell Proliferation The compounds for test were dissolved
in CHCl3 (25mL). The mixture was then stirred for 6h at in dimethylsulfoxide (DMSO) and further diluted in medium,
room temperature. The CHCl3 solution was washed with brine and seven different concentrations were set. The final concen-
and water successively and then dried (MgSO4), filtered and trations of these compounds were 50, 25, 12.5, 6.2, 3.1, 0.3,
concentrated. The crude products were recrystallized from pe- 0µm in medium of 96-well microplate, respectively, and the
troleum ether–EtOAc (3:1) to give compounds 9a, 9e, and 9f. volume ratio of DMSO was 0.2% in medium each well.
N3,N11-Bis(2-hydroxyethyl)-14-phenyl-14H-dibenzo[a,j]-
After the cells were allowed to adhere for 24h, the com-
xanthene-3,11-dimethanamine (9a): White solid. Yield 81.2%. pounds for test were incubated for 48h with the tumor cells.
mp 118–120°C. 1H-NMR (CDCl3, 300MHz) δ: 8.31 (d, J= The relative amount of live cells was determined using
9.0Hz, 2H, 1, 13-H), 7.72 (d, J=8.7Hz, 2H, 5, 9-H), 7.68 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
(brs, 2H, 4, 10-H), 7.54–7.44 (m, 6H, 6, 8, 2, 12, 2′, 6′-H), (a standard colorimetric MTT-test).14) Absorbance was mea-
7.11 (t, J=7.5Hz, 2H, 3′, 5′-H), 6.95 (t, J=7.5Hz, 1H, 4′-H), sured at 490nm or 570nm. The drug concentration that
6.38 (s, 1H, 14-H), 3.90 (s, 4H, CH2NHCH2CH2OH×2), caused 50% cell growth inhibition (IC50) was calculated by
3.66 (t, J=5.1Hz, 4H, CH2NHCH2CH2OH×2), 2.80 (t, SPSS program. The results are expressed as mean standard
J=5.1Hz, 4H, CH2NHCH2CH2OH×2), 2.62 (brs, 4H, deviation (S.D.) of at least three independent experiments.
CH2NHCH2CH2OH×2). 13C-NMR (CDCl3, 75MHz) δ: 148.6,
Apoptosis Assay HeLa cells were treated with com-
145.0, 135.3, 131.0, 130.7, 128.6, 128.5, 128.2, 127.7, 127.4, pounds 6e and 6f at the concentrations of 12.5µm and 25µm
126.4, 123.1, 118.2, 117.2, 60.8, 53.2, 50.5, 38.2. IR (KBr) ν: for 24h, respectively, and then the cells were stained with
2923, 2852, 1597, 1466, 1400, 1252, 1242cm−1. HR-MS (ESI) both Annexin V fluorescein isothiocyanate (FITC) and prop-
Calcd for C33H33N2O3 [M+H]+ 505.2491, Found 505.2483.
idium iodide (PI), and measured by FACSAria flow cytometer
N3,N11-Bis(2-hydroxyethyl)-14-(4-chlorophenyl)-14H- (Becton Dickinson, CA, U.S.A.). Flow cytometry data were
dibenzo[a,j]xanthene-3,11-dimethanamine (9e): White solid. analyzed by FCS express V3 (De Novo Software, Los Ange-
Yield 79.2%. mp 119–121°C. 1H-NMR (CDCl3, 300MHz) les, CA, U.S.A.).
δ: 8.25 (d, J=8.7Hz, 2H, 1, 13-H), 7.75 (d, J=9.0Hz, 2H,

February 2013
175
7, 83–86 (1972).
Acknowledgments The authors thank Dr. Yan Gao and
1
M.E. Xiang Li for testing the H- and 13C-NMR spectra, M.E.
6) Banerjee A., Mukherjee A. K., Stain Technol., 56, 83–85 (1981).
7) Knight C. G., Stephens T., Biochem. J., 258, 683–687 (1989).
8) Ahmad M., King T. A., Ko D.-K., Cha B. H., Lee J., J. Phys. D
Appl. Phys., 35, 1473–1476 (2002).
Yanchao Yu and Xuelian Liu for help with the experiment,
and Dr. Yi Wang for the checking language.
9) Shenkenberg T. D., Von Hoff D. D., Ann. Intern. Med., 105, 67–81
(1986).
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