European Journal of Medicinal Chemistry 46 (2011) 5127e5137

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European Journal of Medicinal Chemistry

journal homepage: http://www.elsevier.com/locate/ejmech

Original article

Synthesis and biological evaluation of 1,9-disubstituted b-carbolines as potent

DNA intercalating and cytotoxic agents
Zhiyong Chen a, Rihui Cao b, *, Buxi Shi b, Liang Guo c, Jie Sun c, Qin Ma c, Wenxi Fan c, Huacan Song b, *
a
Guangdong Provincial Public Laboratory of Analysis and Testing Technology, China National Analytical Center, Building 34, 100 Xianlie Middle Road, Guangzhou, 510070, PR China
b
School of Chemistry and Chemical Engineering, Sun Yat-sen University, 135 Xin Gang West Road, Guangzhou 510275, PR China
c
Xinjiang Huashidan Pharmaceutical Co. Ltd., 45 He Nan East Road, Urumqi 830011, PR China

a r t i c l e i n f o a b s t r a c t

Article history: A series of novel 1,9-disubstituted b-carbolines was designed, synthesized and evaluated as cytotoxic and
Received 13 February 2010 DNA intercalating agents. Compounds 7b, 7c, 8b and 8c exhibited the most potent cytotoxic activities
Received in revised form with IC50 values of lower than 20 mM against ten human tumor cell lines. The results indicated that (1)
15 August 2011
the 3-chlorobenzyl and 3-phenylpropyl substituents in position-9 of b-carboline nucleus were the
Accepted 19 August 2011
Available online 26 August 2011
suitable pharmacophoric group giving rise to significant antitumor agents; (2) the length of the alky-
lamino side chain moiety affected their cytotoxic potencies, and three CH2 units were more favorable. In
addition, these compounds were found to exhibit remarkable DNA intercalating effects.
Keywords:
b-Carboline Ó 2011 Elsevier Masson SAS. All rights reserved.
Synthesis
Cytotoxic
Intercalating
Tm

1. Introduction [7,14,15], CDK [16e18], MK-2 [19,20], PLK1 [21], kinesin Eg5 [22]
and IKK [19].
DNA is one of the most important pharmacological targets of To promote the antitumor activity and DNA intercalating
many drugs currently in clinical use. Small molecules that bind potency of such compounds, previous attempt [5,11] were focused
genomic DNA have proven to be effective antitumor, antivirus and on incorporating substituents into position-1 and 3 of b-carboline
antibacterial therapeutic agents. It is well-known that small nucleus. The introduction of amino group into position-1 or the
molecules can interact with DNA through multiple modes including presence of a flexible amino side chain in position-3 of b-carboline
(i) intercalation; (ii) surface binding; (iii) minor groove binding and nucleus was proved to improve the affinity of the intercalator for
(iv) major groove binding. DNA intercalating agents form an the DNA molecule, consequently to a greater cytotoxic activity. We
important class of drugs in antitumor therapy [1]. To investigate the have ever reported that a series of 9-alkyl or arylated alkyl
interaction of such agents with DNA, in particular, effects of substituted harmine derivatives as efficient DNA intercalating and
structural requirement of them on the interaction, could offer novel potent cytotoxic agents [23]. In addition, our previous investigation
insights into the design of new DNA targeting antitumor drugs [2]. [24e29] on the synthesis of a variety of b-carboline derivatives and
The b-carboline alkaloids are a large group of natural and the evaluation of their antitumor activities unraveled that b-car-
synthetic indole alkaloids associated with a broad spectrum of bolines had potent antitumor activities and the activities was
biochemical effects and pharmaceutical properties [3]. Recent correlated to both the planarity of the molecule and the presence of
reports [4e12] have pointed out b-carbolines as a new class of the ring substituents. Structureeactivity relationships (SARs)
potential antitumor agents, which were discovered to exert their analysis suggested that the introduction of appropriate substitu-
antitumor effects through multiple mechanisms of action, such as ents into position-9 of b-carboline nucleus played a vital role in
intercalating into DNA [5,10,13], inhibiting topoisomerase I and II determining their antitumor potencies, and the n-butyl and phe-
nylpropyl substituents in position-9 was suitable pharmacophoric
group giving rise to some potent antitumor agents.
* Corresponding authors. Tel.: þ86 20 84110918; fax: þ86 20 84112245.
More recently, our group reported that b-carbolines, bearing
E-mail addresses: [email protected] (R. Cao), [email protected] a flexible alkylamino side chain at position-3 and a alkyl or arylated
(H. Song). alkyl substituent at position-9, exhibited significant antitumor

0223-5234/$ e see front matter Ó 2011 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2011.08.027
5128 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137

activities in vitro, and the N9-arylated alkyl substituted b-carbolines corresponding carboxaldehyde by SeO2 in anhydrous dioxane to
represented the most interesting cytotoxic activities [31,32]. In provide compounds 2aef in 62e78% yield. The reaction of
addition, previous literature described that the complex polycyclic compounds 2aef with the corresponding diamines to form schiff
ring system at postion-1 of b-carboline nucleus in manzamine A bases took place readily at room temperature in good yield. The
can be replaced with simpler amino substituents to provide active crude schiff bases without further purification were directly
compounds [11]. reduced with NaBH3CN in anhydrous methanol to give the target
In our continued efforts for simple but efficient antitumor and b-carbolines 3aed, 4aed, 5aed, 6aed, 7aed and 8aed (Table 1) in
DNA intercalating agents, we report here, the molecular design and 35e65% yield. The chemical structures of all the synthesized new
chemical synthesis of a series of novel b-carbolines bearing a flex- compounds were characterized by MS, IR, 1H NMR, and 13C NMR
ible alkylamino side chain at position-1 and a alkyl or arylated alkyl spectra.
substituent at position-9, respectively. These compounds were
expected to intercalate into DNA more strongly and to exhibit
3. Results and discussion
efficient medical and biological use due to the improved water
solubility and the increased flexibility of the aminoalkyl side chains.
3.1. Cytotoxic activity in vitro

2. Chemistry The cytotoxic potencies of novel b-carbolines bearing a N,N-

dialkylaminoalkylamino moiety against a panel of human tumor
The synthetic routes of novel b-carbolines 3aed, 4aed, 5aed, cell lines were investigated and compared with the reference drugs
6aed, 7aed and 8aed are outlined in Scheme 1. The 1-methyl cisplatin, paclitaxel and adriamycin. The tumor cell line panel
b-carboline, prepared by the condensation of L-tryptophan with consisted of renal carcinoma (769-P, 786-0 and OS-RC-2), epider-
acetaldehyde in acid solution and followed by aromatization, moid carcinoma of the nasopharynx (KB), gastric carcinoma (BGC-
oxidation and decarboxylation in a single step through the action of 823), liver carcinoma (HepG2), melanoma (A375), colon carcinoma
potassium dichromate according to the method previously (HT-29), prostate carcinoma (22RV1) and breast carcinoma (MCF-
described by Synder et al. [30], was alkylated or arylated by the 7). As shown in Table 1, compounds 3aed, 4aed, 5aed and 6aed
action of sodium hydride in anhydrous DMF followed by the only demonstrated moderate cytotoxic activities, whereas
addition of the relevant appropriate alkylating and arylating agents compounds 7aed and 8aed, bearing a 3-chlorobenzyl and
to afford intermediates 1aef [24e26] in 65e76% yield. The methyl 3-phenylpropyl substituents in position-9 of b-carboline nucleus,
group in position-1 of compounds 1aef was oxidized to its respectively, exhibited significant cytotoxic potencies. Of all 9-(3-

O O
N DMF/NaH N N
R9Br SeO2
N N N
H CH3 CH3 CHO
R9 R9

1a R9=n-C4H9 2a R9=n-C4H9
1b R9=i-C4H9 2b R9=i-C4H9
1c R9=CH2C6H5 2c R9=CH2C6H5
1d R9=CH2C6H4(4-F) 2d R9=CH2C6H4(4-F)
1e R9=CH2C6H4(3-Cl) 2e R9=CH2C6H4(3-Cl)
1f R9=(CH2)3C6H5 2f R9=(CH2)3C6H5

1) CH3OH/CH2Cl2 (2:1 V:V)

H2N N n
m 2) NaBH3CN/CH3OH
n 3) 4N HCl/CH3CH2OH

.4HCl
N n
H N
N
m n
N
R9
3a R9=n-C4H9 m=0 n=1 6a R9=CH2C6H4(4-F) m=0 n=1
3b R9=n-C4H9 m=1 n=0 6b R9=CH2C6H4(4-F) m=1 n=0
3c R9=n-C4H9 m=1 n=1 6c R9=CH2C6H4(4-F) m=1 n=1
3d R9=n-C4H9 m=2 n=1 6d R9=CH2C6H4(4-F) m=2 n=1
4a R9=i-C4H9 m=0 n=1 7a R9=CH2C6H4(3-Cl) m=0 n=1
4b R9=i-C4H9 m=1 n=0 7b R9=CH2C6H4(3-Cl) m=1 n=0
4c R9=i-C4H9 m=1 n=1 7c R9=CH2C6H4(3-Cl) m=1 n=1
4d R9=i-C4H9 m=2 n=1 7d R9=CH2C6H4(3-Cl) m=2 n=1
5a R9=CH2C6H5 m=0 n=1 8a R9=(CH2)3C6H5 m=0 n=1
5b R9=CH2C6H5 m=1 n=0 8b R9=(CH2)3C6H5 m=1 n=0
5c R9=CH2C6H5 m=1 n=1 8c R9=(CH2)3C6H5 m=1 n=1
5d R9=CH2C6H5 m=2 n=1 8d R9=(CH2)3C6H5 m=2 n=1

Scheme 1. Synthesis of b-carbolines 3aed, 4aed, 5aed, 6aed, 7aed and 8aed.
 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137 5129

Table 1
Cytotoxicity of b-carboline derivatives in vitroc (IC50,a mM).
.4HCl
N n
H N
N
m n
N
9
R
Compd R9 m n 769-Pb KB BGC-823 786e0 HepG2 A375 HT-29 OS-RC-2 22RV1 MCF-7
3a n-C4H9 0 1 43.7 >100 >100 >100 >100 31.3 >100 31.9 112 59.6
3b n-C4H9 1 0 75.6 53.3 41.0 22.4 26.2 52.9 10.1 31.3 17.2 66.2
3c n-C4H9 1 1 14.9 61.7 40.0 56.7 27.1 54.4 41.4 26.4 40.3 75.8
3d n-C4H9 2 1 69.2 64.4 32.6 19.9 20.4 56.9 9.3 14.8 12.6 55.6
4a i-C4H9 0 1 25.4 73.0 99.2 28.7 >100 84.6 8.4 50.9 16.5 96.0
4b i-C4H9 1 0 >100 62.6 44.7 20.2 28.3 82.9 12.1 21.3 17.9 72.2
4c i-C4H9 1 1 >100 92.8 72.8 33.5 53.8 68.3 17.0 47.0 23.0 >100
4d i-C4H9 2 1 >100 74.6 56.2 18.6 27.2 75.8 19.5 26.3 19.3 83.1
5a CH2C6H5 0 1 34.9 >100 >100 68.6 59.5 28.0 63.8 32.5 73.2 62.3
5b CH2C6H5 1 0 15.1 44.2 41.0 39.3 23.5 26.9 32.7 12.5 24.5 53.6
5c CH2C6H5 1 1 19.2 60.0 66.9 39.6 22.0 25.8 33.4 22.5 34.3 51.6
5d CH2C6H5 2 1 71.7 39.9 33.8 16.7 27.0 56.2 12.3 20.4 12.8 58.2
6a CH2C6H4(p-F) 0 1 23.9 >100 >100 65.5 61.1 25.2 58.8 34.6 65.9 58.8
6b CH2C6H4(p-F) 1 0 11.7 32.5 28.8 30.1 20.0 28.6 27.8 10.9 29.9 44.2
6c CH2C6H4(p-F) 1 1 19.6 32.9 34.6 36.3 22.8 34.5 29.5 18.1 32.9 53.1
6d CH2C6H4(p-F) 2 1 13.3 30.0 29.7 17.7 20.7 51.8 18.8 12.6 19.1 52.0
7a CH2C6H4(m-Cl) 0 1 26.7 57.9 38.9 28.1 16.4 14.6 27.0 29.8 28.6 17.2
7b CH2C6H4(m-Cl) 1 0 24.5 14.0 12.0 7.5 9.3 13.1 6.1 10.2 6.1 14.7
7c CH2C6H4(m-Cl) 1 1 11.4 12.4 10.8 9.3 18.0 11.9 5.7 10.1 5.7 17.6
7d CH2C6H4(m-Cl) 2 1 28.5 10.3 9.3 8.8 7.2 14.6 5.6 9.9 5.9 20.8
8a (CH2)3C6H5 0 1 14.8 9.2 14.7 18.2 10.6 14.7 16.3 8.1 21.9 25.7
8b (CH2)3C6H5 1 0 1.7 3.2 5.9 4.4 4.6 18.1 3.9 11.1 5.3 17.7
8c (CH2)3C6H5 1 1 2.4 3.1 7.5 5.0 3.5 11.6 3.7 10.0 6.9 16.6
8d (CH2)3C6H5 2 1 12.4 9.7 20.7 10.0 8.5 4.7 3.7 51.1 10.2 27.4
Cisplatin 19.2 4.6 13.4 4.9 16.0 9.4 85.7 3.4 4.6 12.4
Paclitaxel 7.1 0.08 1.5 <0.08 <0.08 0.81 0.38 <0.08 0.08 1.3
Adriamycin 11.5 0.6 <1.3 3.1 <1.3 17.2 4.2 <1.3 <1.3 5.1
a
Cytotoxicity as IC50 for each cell line is the concentration of compound, which reduced by 50% the optical density of treated cells with respect to untreated using the MTT
assay.
b
Cell lines include renal carcinoma (769-P, 786-0 and OS-RC-2), epidermoid carcinoma of the nasopharynx (KB), gastric carcinoma (BGC-823), liver carcinoma (HepG2),
melanoma (A375), colon carcinoma (HT-29), prostate carcinoma (22RV1) and breast carcinoma (MCF-7).
c
The data represent the mean values of three independent determinations.

chloro)benzyl and 9-(3-phenyl)propyl substituted b-carboline from melting curves (not shown) and the results of Tm analysis
derivatives, compounds 7b, 7c, 8b and 8c, having a N,N- performed with CT-DNA were shown in Fig. 2. CT-DNA which melt at
diethylamino propylamino moiety in position-1, were found to be a low temperature (53.2  C in PE buffer) afforded a sensitive deter-
the most potent cytotoxic agents with IC50 value of lower than mination of the DNA binding capacity of the studied molecules. As
20 mM against 10 human tumor cell lines investigated. These results indicated in Fig. 2, compound 3c, bearing a N,N-diethylamino pro-
indicated that (1) the 3-chlorobenzyl and 3-phenylpropyl substit- pylamino group and n-butyl subsituent in position-1 and 9,
uents in position-9 of b-carboline nucleus were the suitable phar- respectively, markedly stabilized CT-DNA against heat denaturation
macophoric group giving rise to potent antitumor agents; (2) the with DTm value (DTm ¼ TdrugeDNA
m
complex
 TDNA
m
alone
) of 19.2  C.
length of the alkylamino side chain moiety also affected their Whereas, 9-isobutyl (compound 4c) and 9-benzyl (compound 5c)
cytotoxic potencies, and three CH2 units were more favorable. substituted b-carboline congeners exhibited weaker effects on CT-
DNA thermal stability with DTm value of 16.1 and 14.8  C, respec-
3.2. UV spectral absorbance tively. Unexpectedly, replacement of the n-butyl substituent in
position-9 of compound 3c with 4-fluorobenzyl group afforded
The interaction of the selected compounds 3e8c with calf compound 6c, which displayed the poorest DNA binding capacity
thymus DNA (CT-DNA) was examined by UV spectroscopy. Fig. 1 with 6Tm value of 8.1  C. Interestingly, compounds 7c and 8c,
illustrates the absorption spectra of compounds 3e8c in the PE bearing 3-chlorobenzyl and 3-phenylpropyl group in position-9,
buffer (1 mM Na2HPO4, 0.1 mM EDTA, pH 7.4) in the presence of respectively, were found to be the most potent compounds with
increasing amounts of CT-DNA. In all cases, the binding of the drugs 6Tm value of 19.9 and 21.5  C, respectively. The results suggested
to CT-DNA results in considerable spectral changes, characterized that (1) such compounds could significantly stabilize the double
by a slight bathochromic shift and a marked hypochromism. These helix of CT-DNA; (2) the introduction of appropriate substituent into
results suggested that these compounds had a significant interac- position-9 of b-carboline nucleus played a vital role in determining
tion with CT-DNA double helix. their DNA binding potency, and 3-chlorobenzyl and 3-phenylpropyl
group were more favorable.

3.3. DNA thermal denaturation studies

3.4. Fluorescence studies
Melting temperature (Tm) measurements were deployed to
evaluate relative affinity for DNA of compounds 3e8c. The Tm of CT- b-Carboline alkaloids present intrinsic fluorescence properties
DNA in the presence and absence of compounds 3e8c were obtained which could be used to evaluate their DNA binding potencies. The
5130 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137

effect of CT-DNA on the fluoresence intensity of the selected decreased with the increasing concentrations of added CT-DNA.
b-carbolines 3e8c was determined by the use of fluoresence When the concentration of added CT-DNA was increased to
spectroscopy. From the titration of compounds 3e8c (Fig. 3), the 90 mM, the fluorescence intensity of all investigated compounds
fluorescence intensity of all investigated compounds gradually was lowered to their minimum. In addition, over the course of the

0.6
0.6
3c
0.5 0 uM 6c
0 uM 30 uM
0.5
30 uM 60 uM
0.4 60 uM 90 uM
90 uM 0.4 120 uM
150 uM
Absorption

120 uM

Absorption
0.3 150 uM 180 uM
180 uM 0.3 210 uM
0.2
0.2

0.1
0.1

0.0
250 300 350 400 0.0
250 300 350 400
Wavelength (nm)
Wavelength (nm)

0.6 0.6 7c
0 uM
0 uM 4c 30 uM
0.5 30 uM
60 uM
60 uM
90 uM
90 uM
0.4 120 uM 0.4 120 uM
Absorption

150 uM
Absorption

150 uM
180 uM 180 uM
0.3 210 uM
210 uM

0.2 0.2

0.1

0.0 0.0
250 300 350 400 250 300 350 400
Wavelength (nm) Wavelength (nm)

0.7 0.7

5c
0.6 0 uM 0.6
0 uM 8c
30 uM 30 uM
0.5
60 uM 60 uM
0.5
90 uM 90 uM
120 uM
120 uM
0.4 150 uM 0.4
Absorption

Absorption

150 uM
180 uM
180 uM
0.3 210 uM
0.3 210 uM

0.2 0.2

0.1 0.1

0.0 0.0
250 300 350 400 250 300 350 400
Wavelength (nm) Wavelength (nm)

Fig. 1. Absorption spectra for compounds 3e8c in 1 ml PE buffer (1 mM Na2HPO4, 0.1 mM EDTA, pH 7.4) at different molarities of CT-DNA: top curve (0.0) and bottom curve
(210 mM) were recorded in quartz cells (10 mm path length) by a UVevisible spectrophotometer at room temperature.
 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137 5131

silica gel (Kieselgel 60 F254, Merck). 9-n-Butyl-1-methyl-b-carbo-

line (1a) [26], 9-benzyl-1-methyl-b-carboline (1c) [26] and 9-(3-
phenyl)propyl-1-methyl-b-carboline (1f) [26] are known
compounds. NMR spectra were recorded on a Varian Mercury-Plus
300 spectrometer, Bruker AVANCE 400 and Varian INOVA500NB in
D2O or DMSO-d6 or D2O þ DMSO-d6 at 25  C. All chemical shifts (d)
are quoted in parts per million downfield from TMS and coupling
constants (J) are given in Hertz. Mass spectra were obtained from
VG ZAB-HS and LCMS-2010A. High-resolution mass spectrometry
(HRMS); was performed on MAT95XP. Infrared (IR) spectra were
measured on VECTOR 22 spectrometer using a potassium bromide
(KBr) disk, scanning from 400 to 4000 cm1.

5.2. General procedures for the synthesis of 9-alkyl substituted

b-carbolines 1b, 1d and 1e

Fig. 2. Variation of the 6Tm of the complexes between the tested compounds and CT- A mixture of 1-methyl-b-carboline (1.82 g, 10 mmol) and
DNA. Melting temperature measurements were performed in PE buffer (1 mM anhydrous DMF (50 ml) was stirred at room temperature until
Na2HPO4, 0.1 mM EDTA) at pH 7.4 with a drug/DNA ration of 0.2. Adr is abbreviation of clear, and then 60% NaH (0.6 g, 15 mmol) and halogenated alkane
adriamycin (Doxorubicin Hydrochloride). (15 mmol) were added. The mixture was stirred at room temper-
ature. After completion of the reaction as indicated by TLC, the
solution was poured into H2O (50 ml) and extracted with ethyl
fluorescence quenching of all investigated compounds, a slight
acetate. The organic phase was washed with water and brine, then
bathochromic shift was also observed. These results suggested that
dried over anhydrous sodium sulfate, filtered and evaporated. The
such b-carbolines could effectively interact with DNA.
resulting oil was purified by silica column chromatography with
ethyl acetate as the eluent. Upon recrystallization, white solid were
4. Conclusion
obtained.
With the aim of elucidating the structureeactivity relationship
5.2.1. 9-Isobutyl-1-methyl-b-carboline (1b)
studies and probing the structural requirement for the potent
Yield 65%; 1H NMR (300 MHz, CDCl3): d 8.30 (d, J ¼ 5.1 Hz, 1H, H-
antitumor activity of b-carbolines, a series of novel 1-(N,N-dia-
3), 8.08 (d, J ¼ 7.8 Hz, 1H, H-4), 7.82 (d, J ¼ 5.1 Hz, 1H, H-8), 7.22e7.57
lkylaminoalkylamino)- and 9-alkyl substituted b-carbolines has
(m, 3H, H-5, H-6, H-7), 4.33 (d, J ¼ 7.5 Hz, 2H, NCH2CH(CH3)2), 3.03 (s,
been synthesized and evaluated as potent cytotoxic and DNA
3H, CH3), 2.22e2.31(m, 1H, NCH2CH(CH3)2), 0.94 (s, 3H, CH(CH3)2),
intercalating agents. Their cytotoxic potencies and DNA inter-
0.92 (s, 3H, CH(CH3)2). 13C NMR (75 MHz, CDCl3): d 142.0, 141.4, 138.1,
calating capacities evidenced the importance of the length of N,N-
135.4, 129.1, 127.9, 121.4, 121.2, 119.6, 113.0, 110.6, 51.9, 30.9, 24.2(d),
dialkylaminoalkylamino group in position-1 and the nature of alkyl
20.5; FAB-MS m/z (M þ 1) 239.
substituent in position-9 of b-carboline nucleus. The
3-chlorobenzyl and 3-phenylpropy substituents in position-9 were
5.2.2. 9-(4-Fluorobenzyl)-1-methyl-b-carboline (1d)
the suitable pharmacophore giving rise to potent antitumor agents
Yield 72%; 1H NMR (300 MHz, CDCl3): d 8.33 (d, J ¼ 5.1 Hz, 1H, H-
and the optimal length of the alkylamino side chain moiety were
3), 8.13 (d, J ¼ 7.8 Hz, 1H, H-4), 7.86 (d, J ¼ 5.1 Hz, 1H, H-8), 7.50e7.55
three CH2 units.
(m, 1H, H-7), 7.25e7.34 (m, 2H, H-5, H-6), 6.96 (d, 4H, J ¼ 7.2 Hz, Ph-
An overview of the cytotoxic activities data of all new synthe-
H), 5.75 (s, 2H, NCH2-4-fluorophenyl), 2.87 (s, 3H, CH3). 13C NMR
sized b-carbolines bearing a flexible alkylamino side chain at
(75 MHz, CDCl3): d 163.8, 160.5, 141.8, 141.6, 138.7, 135.5, 133.8,
position-1 and of the earlier reported b-carbolines bearing a flex-
129.2, 128.6, 127.2, 127.1, 121.7, 121.5, 120.3, 116.2, 115.9, 113.1, 109.8,
ible alkylamino side chain at position-3 [31,32] clearly indicated
47.7, 23.5(d); FAB-MS m/z (M þ 1) 305.
that (i) the N9-arylated alkyl substituted b-carbolines represented
the most interesting cytotoxic activities; (ii) the optimal length of
5.2.3. 9-(3-Chlorobenzyl)-1-methyl-b-carboline (1e)
the alkylamino side chain moiety were three CH2 units and (iii)
Yield 76%; 1H NMR (300 MHz, CDCl3): d 8.34 (d, J ¼ 5.1 Hz, 1H, H-
their cytotoxic potencies had no difference between b-carbolines
3), 8.14e8.16 (m, 1H, H-4), 7.86 (d, J ¼ 5.1 Hz, 1H, H-8), 7.50e7.56 (m,
bearing a flexible alkylamino side chain at C-1 and C-3.
1H, H-7), 7.15e7.32 (m, 3H, H-5, H-6, Ph-H), 7.03 (s, 1H, Ph-H), 6.81
An attempt to include all of the 1,9-disubstituted b-carbolines
(d, J ¼ 7.2 Hz, 2H, Ph-H), 5.74 (s, 2H, CH2), 2.87 (s, 3H, CH3). 13C NMR
into a correlation of cytotoxic potency with 6Tm values was
(75 MHz, CDCl3): d 141.8, 141.5, 140.3, 138.8, 135.5, 135.1, 130.5,
unsuccessful, but this result has been observed for other DNA
129.3, 128.7, 128.0, 125.8, 123.7, 121.7, 121.6, 120.4, 113.2, 109.8, 47.9,
binding agents. Because the DNA binding process is complicated
23.5(d); FAB-MS m/z (M þ 1) 321.
and dependent on many structural factors, undoubtedly, a more
detailed molecular mechanics study on the DNA binding of these
compounds is needed. 5.3. General procedures for the synthesis of 9-alkyl substituted
b-carboline-1-carboxaldehydes 2aef
5. Experimental protocols
To a solution of 1aef (5 mmol) in dioxane (100 ml) was added
5.1. Reagents and general procedures SeO2 (10 mmol). The suspension was refluxed for 2 h. After
completion of the reaction as indicated by TLC, the mixture was
All solvents were of reagent grade and, when necessary, were cooled and filtered through Celite. The filtrate was evaporated
purified and dried by standard methods. Reactions and products under reduced pressure. The residue was crystallized from acetone
were routinely monitored by thin-layer chromatography (TLC) on or acetoneepetroleum ether to afford white solid.
5132 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137

700

3c 300
600
6c
250
500 0 uM
Fluorescence intensity

15uM

Fluorescence intensity
200 0 uM
400 30uM 15 uM
45uM 30 uM
150
60uM 45 uM
300
75uM 60 uM
100 75 uM
90uM
200 90 uM
50
100
0
0
400 450 500 550 400 450 500 550

/ nm / nm

600 700

4c
600
500 7c

500
400 0 uM
Fluorescence intensity
Fluorescence intensity

15uM
30uM 400 0 uM
300 45uM 15 uM
60uM 300 30 uM
75uM 45 uM
200
90uM 60 uM
200 75 uM
100 90 uM
100

0
0
400 450 500 550
400 450 500 550
/ nm
/ nm
800
500

700 5c

400 8c
600
Fluorescence intensity

Fluorescence intensity

500
0 uM 300
15 uM 0 uM
400
30 uM 15 uM
45 uM 200
30 uM
300 45 uM
60 uM
75 uM 60 uM
200 90 uM 75 uM
100 90 uM
100

0 0
400 450 500 550 400 450 500 550

/ nm / nm

Fig. 3. Fluorescence spectra of the various b-carbolines upon incubation with graded concentration of CT-DNA. A fixed concentration of the various b-carbolines (10 mM) was
incubated with increasing concentration of CT-DNA from 0 to 90 mM in PE buffer (1 mM Na2HPO4, 0.1 mM EDTA, pH 7.4).
 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137 5133

5.3.1. 9-n-Butyl-b-carboline-1-carboxaldehyde (2a) (50 mL) and washed with aqueous Na2CO3 (pH 10, 50 mL). The
Yield 65%; 1H NMR (300 MHz, CDCl3): d 10.31 (s, 1H, CHO); 8.63 organic layer was separated, dried over anhydrous Na2SO4, filtered,
(d, J ¼ 4.8 Hz, 1H, H-3); 8.13e8.17 (m, 2H, H-4, H-8); 7.62e7.68 (m, and concentrated under vacuum. The residue was purified by flash
1H, H-7); 7.53 (d, J ¼ 8.4 Hz, 1H, H-5); 7.24e7.32 (m, 1H, H-6); 4.86 chromatography on silica gel (CH2Cl2/CH3OH/NH4OH, 95:5:1) to
(t, J ¼ 7.5 Hz, 2H, NCH2CH2CH2CH3); 1.70e1.80 (m, 2H, gain yellow oil. The oil was dissolved in 4N HCl/ethanol (20 mL) and
NCH2CH2CH2CH3); 1.26e1.41 (m, 2H, NCH2CH2CH2CH3); 0.93 (t, stirred at room temperature for 30 min, then removed the solvent
J ¼ 7.2 Hz, 3H, NCH2CH2CH2CH3). 13C NMR (75 MHz, CDCl3): d 193.9, under reduced pressure to obtain yellow solid. The solid were dried
142.5, 138.3, 137.7, 135.1, 132.5, 129.3, 121.4, 120.9, 120.8, 118.6, 110.7, in vacuo at 100  C for 3 days to remove the residual ethanol.
46.8, 32.1, 20.3, 14.2; FAB-MS m/z (M þ 1) 253.
5.4.1. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-n-butyl-b-
5.3.2. 9-Isobutyl-b-carboline-1-carboxaldehyde (2b) carboline hydrochloride salt (3a)
Yield 62%; 1H NMR (300 MHz, CDCl3): d 10.31 (s, 1H, CHO), 8.63 Yield 40%; IR (KBr, cm1) v: 2947, 2792, 2581, 1623, 1494, 1443,
(d, J ¼ 4.8 Hz, 1H, H-3), 8.14e8.19 (m, 2H, H-4, H-8), 7.31e7.66 (m, 1389, 1292, 1198, 1134, 1026, 758; 1H NMR (300 MHz, DMSO-
3H, H-5, H-6, H-7), 4.78 (t, J ¼ 7.5 Hz, 2H, NCH2CH(CH3)2), 2.04e2.17 d6 þ D2O): d 8.41 (d, J ¼ 5.1 Hz, 1H, H-3), 8.30e8.33 (m, 2H, H-4, H-
(m, 1H, NCH2CH(CH3)2), 0.83 (s, 3H, CH3), 0.80 (s, 3H, CH3). 13C NMR 8), 7.78 (d, J ¼ 8.1 Hz, 1H, H-5), 7.64e7.69 (m, 1H, H-7), 7.33 (t,
(75 MHz, CDCl3): d 193.9, 142.9, 138.3, 138.0, 135.3, 132.5, 129.2, J ¼ 7.5 Hz, 1H, H-6), 4.99 (s, 2H, CH2NHCH2CH2N), 4.56 (t, J ¼ 7.2 Hz,
121.4, 120.8, 118.7, 111.3, 109.9, 53.4, 30.0, 29.4, 20.1; FAB-MS m/z 2H, NCH2CH2CH2CH3), 3.64e3.66 (m, 4H, NHCH2CH2N), 3.18e3.25
(M þ 1) 253. (m, 4H, N(CH2CH3)2), 1.70e1.80 (m, 2H, NCH2CH2CH2CH3),
1.25e1.37 (m, 8H, NCH2CH2CH2CH3, N(CH2CH3)2), 0.89 (t, J ¼ 7.2 Hz,
5.3.3. 9-Benzyl-b-carboline-1-carboxaldehyde (2c) 3H, NCH2CH2CH2CH3); 13C NMR (75 MHz, D2O): d 144.3, 135.1,
Yield 78%; 1H NMR (300 MHz, CDCl3): d 10.20 (s, 1H, CHO), 8.65 132.6, 132.4, 130.8, 129.7, 122.5, 121.9, 118.6, 116.9, 110.9, 49.2, 48.3,
(d, J ¼ 5.1 Hz, 1H, H-3), 8.18e8.23 (m, 2H, H-4, H-8), 7.34e7.63 (m, 46.7, 45.4, 43.5, 32.2, 19.8, 13.5, 8.6; ESI-MS m/z: 353.4 (M þ 1)þ.
3H, H-5, H-6, H-7), 7.16e7.19 (m, 3H, Ph-H), 6.92e6.95 (m, 2H, Ph-
H), 6.18 (s, 2H, NCH2). 13C NMR (75 MHz, CDCl3): d 193.9, 143.0, 5.4.2. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-n-butyl-b-
138.9, 138.1, 137.7, 135.8, 132.8, 129.7, 128.8, 127.4, 126.3, 121.6, 121.3, carboline hydro-chloride salt (3b)
121.1, 118.8, 111.2, 50.4; FAB-MS m/z (M þ 1) 287. Yield 48%; IR (KBr, cm1) v: 3013, 2956, 2869, 2611, 1625, 1524,
1462, 1382, 1338, 1243, 1170, 1058, 774, 752; 1H NMR (300 MHz,
5.3.4. 9-(4-Fluorobenzyl)-b-carboline-1-carboxaldehyde (2d) DMSO-d6 þ D2O): d 8.49 (d, J ¼ 5.4 Hz, 1H, H-3), 8.43 (d, J ¼ 5.4 Hz,
Yield 72%; 1H NMR (300 MHz, CDCl3): d 10.20 (s, 1H, CHO), 8.66 1H, H-4), 8.37 (d, J ¼ 8.1 Hz, 1H, H-8), 7.82 (d, J ¼ 8.4 Hz, 1H, H-5),
(d, J ¼ 5.1 Hz, 1H, H-3), 8.17e8.23 (m, 2H, H-4, H-8), 7.35e7.64 (m, 7.70 (t, J ¼ 7.5 Hz, 1H H-7,), 7.36 (t, J ¼ 7.2 Hz, 1H, H-6), 4.95 (s, 2H,
3H, H-5, H-6, H-7), 6.84e6.95 (m, 4H, 4-fluorophenyl-H), 6.14 (s, CH2NHCH2CH2CH2N), 4.60 (t, J ¼ 6.6 Hz, 2H, NCH2CH2CH2CH3), 3.32
2H, NCH2). 13C NMR (75 MHz, CDCl3): d 194.0, 163.6, 160.4, 142.8, (t, J ¼ 7.2 Hz, 2H, NHCH2CH2CH2N), 3.22 (t, J ¼ 7.2 Hz, 2H,
139.0, 138.0, 135.6, 133.5, 132.8, 129.8, 128.1, 128.0, 121.6, 121.4, NHCH2CH2CH2N), 2.76 (s, 6H, N(CH3)2), 2.18e2.27 (m, 2H,
121.2, 118.9, 115.8, 115.5, 111.1, 49.8; FAB-MS m/z (M þ 1) 305. NHCH2CH2CH2N), 1.69e1.79 (m, 2H, NCH2CH2CH2CH3), 1.31e1.40
(m, 2H, NCH2CH2CH2CH3), 0.89 (t, J ¼ 7.2 Hz, 3H, NCH2CH2CH2CH3);
13
5.3.5. 9-(3-Chlorobenzyl)-b-carboline-1-carboxaldehyde (2e) C NMR (75 MHz, D2O): d 144.7, 136.3, 133.4, 132.7, 131.1, 126.1,
Yield 76%; 1H NMR (300 MHz, CDCl3): d 10.20 (s, 1H, CHO), 8.67 122.7, 122.0, 118.7, 117.7, 110.9, 54.5, 45.9 (2C), 45.4, 43.3, 31.8, 21.8,
(d, J ¼ 4.8 Hz, 1H, H-3), 8.18e8.23 (m, 2H, H-4, H-8), 7.58e7.64 (m, 19.8, 13.5; ESI-MS m/z: 339.4 (M þ 1)þ.
1H, H-7), 7.36e7.44 (m, 2H, H-5, H-6), 7.09e7.17 (m, 2H, 3-
chlorophenyl-H), 6.81e6.97 (m, 2H, 3-chlorophenyl-H), 6.16 (s, 5.4.3. 1-[N-(3-diethylamino)-propyl]-methylamino-9-n-butyl-b-
2H, NCH2). 13C NMR (75 MHz, CDCl3): d 194.1, 142.7, 140.0, 139.1, carboline hydro-chloride salt (3c)
138.0, 135.7, 134.7, 132.8, 130.1, 129.9, 127.7, 126.5, 124.5, 121.6, 121.5, Yield 46%; IR (KBr, cm1) v: 2958, 2685, 1624, 1496, 1461, 1383,
121.2, 118.9, 111.0, 50.1; FAB-MS m/z (M þ 1) 321. 1337, 1201, 1135, 1044, 758; 1H NMR (300 MHz, DMSO-d6 þ D2O):
d 8.49 (d, J ¼ 5.1 Hz, 1H, H-3), 8.42 (d, J ¼ 5.4 Hz, 1H, H-4), 8.36 (d,
5.3.6. 9-(3-Phenylpropyl)-b-carboline-1-carboxaldehyde (2f) J ¼ 8.1 Hz, 1H, H-8), 7.81 (d, J ¼ 8.4 Hz, 1H, H-5), 7.70 (t, J ¼ 7.5 Hz,
Yield 68%; 1H NMR (300 MHz, CDCl3): d 10.30 (s, 1H, CHO), 8.62 1H, H-7), 7.35 (t, J ¼ 7.2 Hz, 1H, H-6), 4.94 (s, 2H,
(d, J ¼ 4.8 Hz, 1H, H-3), 8.12e8.16 (m, 2H, H-4, H-8), 7.58e7.63 (m, CH2NHCH2CH2CH2N), 4.59 (t, J ¼ 6.9 Hz, 2H, NCH2CH2CH2CH3), 3.31
1H, H-7), 7.13e7.37 (m, 7H, H-5, H-6, Ph-H), 4.89 (t, J ¼ 7.8 Hz, 2H, (t, J ¼ 6.9 Hz, 2H, NHCH2CH2CH2N), 3.20 (t, J ¼ 7.5 Hz, 2H,
NCH2CH2CH2Ph), 2.71 (t, J ¼ 7.5 Hz, 2H, NCH2CH2CH2Ph), 2.07e2.18 NHCH2CH2CH2N), 3.08e3.15 (m, 4H, N(CH2CH3)2), 2.16e2.26 (m,
(m, 2H, NCH2CH2CH2Ph). 13C NMR (75 MHz, CDCl3): d 194.0, 142.3, 2H, NHCH2CH2CH2N), 1.69e1.79 (m, 2H, NCH2CH2CH2CH3),
141.3, 138.4, 137.7, 135.1, 132.5, 129.4, 128.6, 128.5(2C), 126.2, 121.5, 1.30e1.37 (m, 2H, NCH2CH2CH2CH3), 1.23 (t, J ¼ 7.2 Hz, 6H,
120.9, 118.7, 110.6, 46.5, 33.1, 31.5; FAB-MS m/z (M þ 1) 315. N(CH2CH3)2), 0.88 (t, J ¼ 7.2 Hz, 3H, NCH2CH2CH2CH3); 13C NMR
(75 MHz, DMSO-d6 þ D2O): d 144.2, 133.9, 133.9, 133.2, 131.9, 131.5,
5.4. General procedures for the synthesis of b-carbolines 3aed, 123.6, 122.0, 120.0, 117.6, 111.8, 48.6, 47.1, 46.4, 45.6 (2C), 32.6, 21.0,
4aed, 5aed, 6aed, 7aed and 8aed 20.3, 14.6, 9.3; ESI-MS m/z: 367.4 (M þ 1)þ.

A mixture of b-carboline-1-carboxaldehydes (1 mmol), diamine 5.4.4. 1-[N-(4-diethylamino)-butyl]-methylamino-9-n-butyl-b-

(1.2 mmol), anhydrous methanol (6 mL) and anhydrous CH2Cl2 carboline hydro-chloride salt (3d)
(3 mL) was stirred at room temperature overnight. The solvent was Yield 50%; IR (KBr, cm1) v: 2958, 2693, 1625, 1545, 1496, 1462,
evaporated under vacuum to give the crude schiff base, which was 1337, 1296, 1201, 1136, 1040, 759; 1H NMR (300 MHz, DMSO-
used directly in the next step without further purification. d6 þ D2O): d 8.63 (d, J ¼ 5.4 Hz, 1H, H-3), 8.57 (d, J ¼ 5.4 Hz, 1H, H-4),
NaBH3CN (10 mmol) was added to a solution of the above- 8.39 (d, J ¼ 8.1 Hz, 1H, H-8), 7.91 (t, J ¼ 7.5 Hz, 1H, H-7), 7.84 (d,
mentioned crude schiff base in anhydrous CH3OH (10 mL) at 0  C. J ¼ 8.4 Hz, 1H, H-5), 7.53 (t, J ¼ 7.2 Hz, 1H, H-6), 5.09 (s, 2H,
The mixture was stirred at room temperature for 24 h and then CH2NHCH2CH2CH2CH2N), 4.62 (t, J ¼ 7.5 Hz, 2H, NCH2CH2CH2CH3),
concentrated under vacuum. The residue was dissolved in CH2Cl2 3.47 (t, J ¼ 7.5 Hz, 2H, NHCH2CH2CH2CH2N), 3.23e3.31 (m, 6H,
5134 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137

NHCH2CH2CH2CH2N), 1.91e1.97 (m, 4H, NHCH2CH2CH2CH2N), 134.0, 132.4, 132.1, 127.4, 122.9, 122.2, 119.2, 117.7, 111.9, 52.2, 51.2,
1.81e1.87 (m, 2H, NCH2CH2CH2CH3), 1.38e1.42 (m, 2H, 48.3, 47.8, 46.5, 30.1, 23.4, 21.2, 19.5, 8.7; ESI-MS m/z: 381.4 (M þ 1)þ.
NCH2CH2CH2CH3), 1.34 (t, J ¼ 7.5 Hz, 6H, N(CH2CH3)2), 0.94 (t,
J ¼ 7.5 Hz, 3H, NCH2CH2CH2CH3); 13C NMR (75 MHz, D2O): d 144.9, 5.4.9. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-benzyl-b-
136.4, 133.6, 132.8, 131.3, 126.4, 122.9, 122.2, 118.9, 117.8, 111.1, 51.2, carboline hydro-chloride salt (5a)
48.4, 47.7, 45.9, 45.5, 31.8, 23.3, 21.1, 19.9, 13.5, 8.7; ESI-MS m/z: Yield 45%; IR (KBr, cm1) v: 2977, 2660, 1624, 1495, 1460, 1389,
381.4 (M þ 1)þ. 1336, 1297, 1134, 1025, 753; 1H NMR (300 MHz, DMSO-d6 þ D2O):
d 8.46 (d, J ¼ 5.1 Hz, 1H, H-3), 8.39 (br s, 1H, H-4), 8.37 (br s, 1H, H-8),
5.4.5. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-isobutyl-b- 7.72 (d, J ¼ 8.4 Hz, 1H, H-5), 7.63 (t, J ¼ 7.2 Hz, 1H, H-7), 7.35 (t,
carboline hydro-chloride salt (4a) J ¼ 7.2 Hz,1H, H-6), 7.22e7.29 (m, 3H, Ph-H), 6.96e6.99 (m, 2H, Ph-H),
Yield 55%; CH2Cl2/CH3OH, 95:5; 2963, 2672, 1623, 1497, 1464, 5.93 (s, 2H, NCH2-Ph), 4.79 (s, 2H, CH2NHCH2CH2N), 3.45e3.49 (m,
1384, 1335, 1135, 1042, 777; 1H NMR (300 MHz, DMSO-d6 þ D2O): 4H, NHCH2CH2N), 3.12e3.19 (m, 4H, N(CH2CH3)2), 1.23 (t, J ¼ 7.2 Hz,
d 8.45 (d, J ¼ 5.4 Hz, 1H, H-3), 8.32e8.36 (m, 2H, H-4, H-8), 7.83 (d, 6H, CH3); 13C NMR (75 MHz, DMSO-d6 þ D2O): d 143.6, 138.1, 135.6,
J ¼ 8.7 Hz, 1H, H-5), 7.63e7.69 (m, 1H, H-7), 7.33 (t, J ¼ 7.8 Hz, 1H, H- 134.2, 133.3, 132.4, 131.1, 129.7, 128.3, 126.3, 123.2, 121.9, 120.6, 117.0,
6), 4.96 (s, 2H, CH2NHCH2CH2N), 4.42 (d, J ¼ 7.5 Hz, 2H, 111.7, 48.7, 47.8, 47.5, 46.9, 42.1, 9.4; ESI-MS m/z: 387.4 (M þ 1)þ.
NCH2CH(CH3)2), 3.66e3.68 (m, 4H, NHCH2CH2N), 3.18e3.25 (m, 4H,
N(CH2CH3)2), 2.11e2.20 (m, 1H, NCH2CH(CH3)2), 1.28 (t, J ¼ 7.2 Hz, 5.4.10. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-benzyl-b-
6H, N(CH2CH3)2), 0.86 (d, J ¼ 6.6 Hz, 6H, NCH2CH(CH3)2); 13C NMR carboline hydro-chloride salt (5b)
(75 MHz, D2O,): d 145.0, 135.8, 133.1, 132.4, 130.0, 129.4, 122.6, 122.0, Yield 65%; IR (KBr, cm1) v: 2976, 2661, 1624, 1494, 1458, 1336,
118.6, 117.3, 111.6, 52.0, 48.5, 48.3, 46.6, 43.3, 30.1, 19.4, 8.6; ESI-MS 1297, 1199, 1134, 1027, 749; 1H NMR (300 MHz, DMSO-d6 þ D2O):
m/z: 353.4 (M þ 1)þ. d 8.46 (d, J ¼ 5.4 Hz, 1H, H-3), 8.36e8.39 (m, 2H, H-4, H-8), 7.75 (d,
J ¼ 8.4 Hz, 1H, H-5), 7.61e7.67 (m, 1H, H-7), 7.36 (t, J ¼ 7.5 Hz, 1H, H-
5.4.6. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-isobutyl-b- 6), 7.20e7.31 (m, 3H, Ph-H), 6.94e6.97 (m, 2H, Ph-H), 5.92 (s, 2H,
carboline hydro-chloride salt (4b) NCH2-Ph), 4.69 (s, 2H, CH2NHCH2CH2CH2N), 3.05e3.15 (m, 4H,
Yield 36%; IR (KBr, cm1) v: 3049, 2957, 2673, 2485, 1623, 1459, NHCH2CH2CH2N), 2.79 (s, 6H, CH3), 2.02e2.12 (m, 2H,
1338, 1289, 1207, 1139, 1045, 749; 1H NMR (300 MHz, DMSO- NHCH2CH2CH2N); 13C NMR (75 MHz, D2O): d 144.8, 136.3, 135.5,
d6 þ D2O): d 8.50 (d, J ¼ 5.4 Hz, 1H, H-3), 8.42 (d, J ¼ 5.4 Hz, 1H, H-4), 134.0, 132.7, 132.5, 129.6, 128.5, 127.1, 125.6, 123.1, 122.5, 119.1, 117.9,
8.37 (d, J ¼ 7.8 Hz, 1H, H-8), 7.86 (d, J ¼ 8.7 Hz, 1H, H-5), 7.69 (t, 110.4, 54.3, 48.4, 45.6 (2C), 43.3, 21.5; ESI-MS m/z: 373.4 (M þ 1)þ.
J ¼ 7.5 Hz, 1H, H-7), 7.35 (t, J ¼ 7.2 Hz, 1H, H-6), 4.91 (s, 2H,
CH2NHCH2CH2CH2N), 4.44 (d, J ¼ 7.2 Hz, 2H, NCH2CH(CH3)2), 3.32 5.4.11. 1-[N-(3-diethylamino)-propyl]-methylamino-9-benzyl-b-
(t, J ¼ 7.2 Hz, 2H, NHCH2CH2CH2N), 3.23 (t, J ¼ 7.5 Hz, 2H, carboline hydro-chloride salt (5c)
NHCH2CH2CH2N), 2.77 (s, 6H, N(CH3)2), 2.20e2.28 (m, 2H, Yield 42%; IR (KBr, cm1) v: 2977, 2661, 1624, 1495, 1459, 1336,
NHCH2CH2CH2N), 2.11e2.18 (m, 1H, NCH2CH(CH3)2), 0.87 (d, 1297, 1199, 1134, 1026, 751; 1H NMR (300 MHz, DMSO-d6 þ D2O):
J ¼ 6.6 Hz, 6H, NCH2CH(CH3)2); 13C NMR (75 MHz, D2O): d 145.3, d 8.45 (d, J ¼ 5.1 Hz, 1H, H-3), 8.35 (m, 2H, H-4, H-8), 7.73 (d,
136.3, 133.8, 132.5, 131.6, 126.7, 122.9, 122.1, 118.9, 117.8, 111.8, 54.5, J ¼ 8.4 Hz, 1H, H-5), 7.63 (t, J ¼ 7.5 Hz, 1H, H-7), 7.35 (t, J ¼ 7.5 Hz, 1H,
52.1, 46.3, 45.9, 43.3, 30.0, 21.8, 19.6; ESI-MS m/z: 339.4 (M þ 1)þ. H-6), 7.22e7.30 (m, 3H, Ph-H), 6.93e6.95 (m, 2H, Ph-H), 5.91 (s, 2H,
NCH2-Ph), 4.68 (s, 2H, CH2NHCH2CH2CH2N), 3.04e3.13 (m, 8H,
5.4.7. 1-[N-(3-diethylamino)-propyl]-methylamino-9-isobutyl-b- NHCH2CH2CH2N(CH2CH3)2), 2.00e2.10 (m, 2H, NHCH2CH2CH2N),
carboline hydro-chloride salt (4c) 1.20 (t, J ¼ 7.5 Hz, 6H, CH3); 13C NMR (75 MHz, D2O): d 144.0, 136.2,
Yield 39%; IR (KBr, cm1) v: 2926, 2685, 1624, 1495, 1463, 1385, 134.6, 134.1, 133.9, 131.8, 129.6, 129.1, 128.4, 125.65, 122.7, 122.0,
1335, 1289, 1137, 1044, 758; 1H NMR (300 MHz, DMSO-d6 þ D2O): 119.5, 117.1, 110.3, 48.6, 48.2, 47.9, 46.6, 45.4, 21.0, 8.7; ESI-MS m/z:
d 8.49 (d, J ¼ 5.4 Hz, 1H, H-3), 8.42 (d, J ¼ 5.4 Hz, 1H, H-4), 8.36 (d, 401.4 (M þ 1)þ.
J ¼ 7.5 Hz, 1H, H-8), 7.85 (d, J ¼ 8.4 Hz, 1H, H-5), 7.65e7.71 (m, 1H, H-
7), 7.34 (t, J ¼ 7.2 Hz, 1H, H-6), 4.90 (s, 2H, CH2NHCH2CH2CH2N), 4.43 5.4.12. 1-[N-(4-diethylamino)-butyl]-methylamino-9-benzyl-b-
(d, J ¼ 7.5 Hz, 2H, NCH2CH(CH3)2), 3.31 (t, J ¼ 7.2 Hz, 2H, carboline hydro-chloride salt (5d)
NHCH2CH2CH2N), 3.20 (t, J ¼ 7.5 Hz, 2H, NHCH2CH2CH2N), 3.07e3.15 Yield 61%; IR (KBr, cm1) v: 2941, 2677, 1624, 1459, 1338, 1199,
(m, 4H, N(CH2CH3)2), 2.19e2.27 (m, 2H, NHCH2CH2CH2N), 2.10e2.17 1038, 746; 1H NMR (C300 MHz, DMSO-d6 þ D2O): d 8.55e8.60 (m,
(m, 1H, NCH2CH(CH3)2), 1.24 (t, J ¼ 7.2 Hz, 6H, N(CH2CH3)2), 0.86 (d, 2H, H-3, H-4), 8.47 (d, J ¼ 7.8 Hz, 1H, H-8), 7.80 (d, J ¼ 8.4 Hz, 1H, H-
J ¼ 6.3 Hz, 6H, NCH2CH(CH3)2); 13C NMR (75 MHz, DMSO-d6 þ D2O): 5), 7.70 (t, J ¼ 7.5 Hz, 1H, H-7), 7.40 (t, J ¼ 7.5 Hz, 1H, H-6), 7.23e7.29
d 144.5,134.1,133.9,133.5,132.1,131.4,123.4,121.8,120.0, 117.3,112.4, (m, 3H, Ph-H), 7.01 (d, J ¼ 6.3 Hz, 2H, Ph-H), 6.01 (s, 2H, NCH2-Ph),
52.1, 48.6, 47.1, 46.7, 45.6, 30.7, 21.0, 20.5, 9.3; ESI-MS m/z: 367.4 4.73 (s, 2H, CH2NHCH2CH2CH2CH2N), 3.00e3.10 (m, 8H,
(M þ 1)þ. NHCH2CH2CH2CH2N(CH2CH3)2), 1.72 (br s, 4H, NHCH2CH2C
H2CH2N), 1.21 (t, J ¼ 7.2 Hz, 6H, CH3); 13C NMR (75 MHz, DMSO-
5.4.8. 1-[N-(4-diethylamino)-butyl]-methylamino-9-isobutyl-b- d6 þ D2O): d 143.9, 138.0, 135.3, 134.3, 132.9, 132.8, 131.5, 129.7,
carboline hydro-chloride salt (4d) 128.3, 126.2, 123.4, 122.0, 120.4, 117.1, 111.6, 50.6, 48.7, 47.3, 46.9,
Yield 40%; IR (KBr, cm1) v: 2928, 2677, 1623, 1550, 1461, 1335, 46.7, 23.4, 21.1, 9.3; ESI-MS m/z: 415.4 (M þ 1)þ.
1211, 1138, 1041, 767; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.48 (d,
J ¼ 5.4 Hz,1H, H-3), 8.39 (d, J ¼ 5.4 Hz,1H, H-4), 8.35 (d, J ¼ 7.8 Hz,1H, 5.4.13. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-(4-
H-8), 7.85 (d, J ¼ 8.1 Hz, 1H, H-5), 7.67 (t, J ¼ 8.1 Hz, 1H, H-7), 7.34 (t, fluorobenzyl)-b-carboline hydrochloride salt (6a)
J ¼ 7.2 Hz, 1H, H-6), 4.87 (s, 2H, CH2NHCH2CH2CH2CH2N), 4.42 (d, Yield 60%; IR (KBr, cm1) v: 2976, 2662, 1624, 1494, 1457, 1337,
J ¼ 7.5 Hz, 2H, NCH2CH(CH3)2), 3.22 (t, J ¼ 6 Hz, 2H, 1297, 1246, 1199, 1134, 1029, 749; 1H NMR (300 MHz, DMSO-
NHCH2CH2CH2CH2N), 3.05e3.12 (m, 6H, NHCH2CH2CH2CH2N), d6 þ D2O): d 8.45 (d, J ¼ 5.1 Hz, 1H, H-3), 8.35e8.39 (m, 2H, H-4, H-
2.07e2.20(m, 1H, NCH2CH(CH3)2), 1.81 (br s, 4H, 8), 7.72 (d, J ¼ 8.4 Hz, 1H, H-5), 7.63 (t, J ¼ 7.5 Hz, 1H, H-7), 7.36 (t,
NHCH2CH2CH2CH2N), 1.21 (t, J ¼ 7.2 Hz, 6H, N(CH2CH3)2), 0.86 (d, J ¼ 7.5 Hz, 1H, H-6), 7.08e7.14 (m, 2H, 4-fluorophenyl-H), 6.99e7.04
J ¼ 6.3 Hz, 6H, NCH2CH(CH3)2); 13C NMR (75 MHz, D2O): d 145.3,136.1, (m, 2H, 4-fluorophenyl-H), 5.91 (s, 2H, NCH2-4-fluorophenyl), 4.79
 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137 5135

(s, 2H, CH2NHCH2CH2N), 3.49 (m, 4H, NHCH2CH2N) 3.13e3.20 (m, J ¼ 5.4 Hz, 1H, H-3), 8.37e8.39 (m, 2H, H-4, H-8), 7.70 (d, J ¼ 8.4 Hz,
4H, N(CH2CH3)2), 1.24 (t, J ¼ 7.2 Hz, 6H, CH3); 13C NMR (75 MHz, 1H, H-5), 7.64 (t, J ¼ 8.1 Hz, 1H, H-7), 7.36 (t, J ¼ 6.9 Hz, 1H, H-6),
D2O): d 163.5, 160.2, 144.1, 134.8, 133.3, 133.1, 132.1, 131.0, 127.5, 7.28e7.29 (m, 2H, 3-chlorophenyl-H), 6.98 (s, 1H, 3-chlorophenyl-
127.4, 122.8, 122.2, 119.2, 116.9, 116.1, 115.8, 110.4, 49.4, 48.2, 47.9, H), 6.83e6.87 (m, 1H, 3-chlorophenyl-H), 5.92 (s, 2H, NCH2-
46.8, 43.1, 8.6; ESI-MS m/z: 405.4 (M þ 1)þ. 3-chlorophenyl), 4.68 (s, 2H, CH2NHCH2CH2CH2N), 3.07e3.14 (m,
4H, NHCH2CH2CH2N), 2.73 (s, 6H, CH3), 2.01e2.11 (m, 2H,
5.4.14. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-(4- NHCH2CH2CH2N); 13C NMR (75 MHz, DMSO-d6 þ D2O): d 144.2,
fluorobenzyl)-b-carboline hydrochloride salt (6b) 140.1, 134.4, 134.3, 134.2, 133.9, 132.2, 131.6, 131.1, 128.4, 126.3, 125.0,
Yield 35%; IR (KBr, cm1) v: 2950, 2692, 1625, 1547, 1505, 1466, 123.8, 122.5, 120.1, 118.0, 111.7, 54.1, 48.4, 45.9, 45.4, 42.8, 21.5; ESI-
1384, 1332, 1290, 1222, 1158, 1130, 1047, 1019, 751; 1H NMR MS m/z: 407.4 (M þ 1)þ.
(300 MHz, DMSO-d6 þ D2O): d 8.49 (d, J ¼ 5.1 Hz, 1H, H-3),
8.38e8.42 (m, 2H, H-4, H-8), 7.77 (d, J ¼ 8.4 Hz, 1H, H-5), 7.63e7.69 5.4.19. 1-[N-(3-diethylamino)-propyl]-methylamino-9-(3-
(m, 1H, H-7), 7.37 (t, J ¼ 7.2 Hz, 1H, H-6), 7.08e7.14 (m, 2H, chlorobenzyl)-b-carboline hydrochloride salt (7c)
4-fluorophenyl-H), 6.99e7.04 (m, 2H, 4-fluorophenyl-H), 5.93 (s, Yield 55%; IR (KBr, cm1) v: 2971, 2927, 2881, 2752, 2534, 2447,
2H, NCH2-4-fluorophenyl), 4.70 (s, 2H, CH2NHCH2CH2CH2N), 1623, 1562, 1468, 1432, 1341, 1288, 1195, 1137, 1092, 1040, 772; 1H
3.11e3.18 (m, 4H, NHCH2CH2CH2N), 2.74 (s, 6H, CH3), 2.06e2.16 (m, NMR (300 MHz, DMSO-d6 þ D2O): d 8.66e8.67 (d, J ¼ 5.7 Hz, 1H, H-
2H, NHCH2CH2CH2N); 13C NMR (75 MHz, DMSO-d6 þ D2O): d 163.7, 3), 8.60 (d, J ¼ 5.7 Hz, 1H, H-4), 8.49 (d, J ¼ 7.8 Hz, 1H, H-8), 7.80 (d,
160.4, 144.1, 134.5, 134.3, 134.0, 133.7, 132.0, 131.9, 128.6, 128.5, J ¼ 8.4 Hz, 1H, H-5), 7.71 (t, J ¼ 7.8 Hz, 1H, H-7), 7.41 (t, J ¼ 7.5 Hz, 1H,
123.6, 122.3, 120.4, 117.6, 116.7, 116.4, 111.7, 54.2, 48.2, 46.3, 45.4, H-6), 7.28e7.29 (m, 2H, 3-chlorophenyl-H), 7.14 (s, 1H, 3-
42.8, 21.5; ESI-MS m/z: 391.4 (M þ 1)þ. chlorophenyl-H), 6.93 (t, J ¼ 3.9 Hz, 1H, 3-chlorophenyl-H), 6.06
(s, 2H, NCH2-3-chlorophenyl), 4.77 (s, 2H, CH2NHCH2CH2CH2N),
5.4.15. 1-[N-(3-diethylamino)-propyl]-methylamino-9-(4- 3.16e3.27 (m, 4H, NHCH2CH2CH2N), 3.05e3.13 (m, 4H,
fluorobenzyl)-b-carboline hydrochloride salt (6c) NHCH2CH2CH2N(CH2CH3)2), 2.13e2.23 (m, 2H, NHCH2CH2CH2N),
Yield 40%; IR (KBr, cm1) v: 2944, 2683, 1625, 1506, 1465, 1385, 1.22 (t, J ¼ 7.2 Hz, 6H, CH3); 13C NMR (75 MHz, DMSO-d6 þ D2O):
1336, 1290, 1224, 1159, 1134, 1045, 754; 1H NMR (300 MHz, DMSO- d 144.0, 140.4, 134.8, 134.4, 134.3, 133.5, 132.3, 131.8, 131.6, 128.3,
d6 þ D2O): d 8.44 (d, J ¼ 5.1 Hz, 1H, H-3), 8.31e8.37 (m, 2H, H-4, H-8), 126.3, 125.0, 123.6, 122.3, 120.4, 117.5, 111.7, 48.5, 48.3, 47.1, 46.4,
7.72 (d, J ¼ 8.4 Hz,1H, H-5), 7.63 (t, J ¼ 7.5 Hz,1H, H-7), 7.35 (t, J ¼ 7.5 Hz, 45.5, 20.9, 9.3; ESI-MS m/z: 435.4 (M þ 1)þ.
1H, H-6), 7.09e7.13 (m, 2H, 4-fluorophenyl-H), 6.96e7.00 (m, 2H, 4-
fluorophenyl-H), 5.89 (s, 2H, NCH2-4-fluorophenyl), 4.68 (s, 2H, 5.4.20. 1-[N-(4-diethylamino)-butyl]-methylamino-9-(3-
CH2NHCH2CH2CH2N), 3.05e3.14 (m, 8H, NHCH2CH2CH2N(CH2CH3)2), chlorobenzyl)-b-carboline hydrochloride salt (7d)
2.03e2.09 (m, 2H, NHCH2CH2CH2N), 1.21 (t, J ¼ 7.2 Hz, 6H, CH3); 13C Yield 50%; IR (KBr, cm1) v: 2939, 2671,1624,1464,1337,1197,1041,
NMR (75 MHz, D2O): d 163.5, 160.3, 144.3, 135.6, 134.0, 133.3, 132.3, 770; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.55 (br s, 2H, H-3, H-4),
131.7, 128.0, 127.6, 127.5, 122.9, 122.3, 119.4, 117.6, 116.3, 116.0, 110.4, 8.43 (d, J ¼ 7.8 Hz, 1H, H-8), 7.75 (d, J ¼ 8.4 Hz, 1H, H-5), 7.69 (t,
48.7, 47.9, 47.6, 46.1, 45.7, 21.1, 8.7; ESI-MS m/z: 419.4 (M þ 1)þ. J ¼ 6.9 Hz, 1H, H-7), 7.39 (t, J ¼ 7.2 Hz, 1H, H-6), 7.28e7.30 (m, 2H, 3-
chlorophenyl-H), 7.09 (s, 1H, 3-chlorophenyl-H), 6.90e6.91 (m, 1H,
5.4.16. 1-[N-(4-diethylamino)-butyl]-methylamino-9-(4- 3-chlorophenyl-H), 6.00 (s, 2H, NCH2-3-chlorophenyl), 4.72 (s, 2H,
fluorobenzyl)-b-carboline hydrochloride salt (6d) CH2NHCH2CH2CH2CH2N), 3.01e3.09 (m, 8H, NHCH2CH2CH2CH2
Yield 38%; IR (KBr, cm1) v: 2946, 2683, 1625, 1504, 1462, 1337, N(CH2CH3)2), 1.73 (br s, 4H, NHCH2CH2CH2CH2N), 1.20 (t, J ¼ 6.9 Hz,
1221, 752; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.51 (br s, 2H, H-3, 6H, CH3); 13C NMR (75 MHz, DMSO-d6 þ D2O): d 143.5, 140.6, 135.7,
H-4), 8.37 (d, J ¼ 7.8 Hz, 1H, H-8), 7.63e7.74 (m, 2H, H-5, H-7), 7.36 (t, 134.3, 134.2, 133.1, 132.7, 131.7, 131.4, 128.3, 126.2, 124.9, 123.3, 122.1,
J ¼ 7.5 Hz, 1H, H-6), 7.00e7.10 (m, 4H, 4-fluorophenyl-H), 5.92 (s, 2H, 120.5,117.0, 111.5, 50.7, 48.2, 47.4, 46.9 (2C), 23.4, 21.1, 9.3; ESI-MS m/z:
NCH2-4-fluorophenyl), 4.71 (s, 2H, CH2NHCH2CH2CH2CH2N), 449.4 (M þ 1)þ.
3.00e3.10 (m, 8H, NHCH2CH2CH2CH2N(CH2CH3)2), 1.72 (br s, 4H,
NHCH2CH2CH2CH2N), 1.81 (t, J ¼ 6.9 Hz, 6H, CH3); 13C NMR (75 MHz, 5.4.21. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-(3-
DMSO-d6 þ D2O): d 163.6, 160.4, 143.6, 135.6, 134.1, 132.8, 131.4, phenylpropyl)-b-carboline hydrochloride salt (8a)
128.4, 128.3, 123.3, 122.1, 120.5, 117.0, 116.7, 116.4, 111.4, 50.8, 48.0, Yield 56%; IR (KBr, cm1) v: 2932, 2615, 1622, 1571, 1490, 1451,
47.4, 47.0 (2C), 23.4, 21.1, 9.3; ESI-MS m/z: 433.4 (M þ 1)þ. 1386, 1336, 1208, 1135, 1042, 773; 1H NMR (500 MHz, D2O): d 8.18
(d, J ¼ 7.5 Hz, 1H, H-3), 8.12 (d, J ¼ 7.5 Hz, 1H, H-4), 7.89 (d, J ¼ 10 Hz,
5.4.17. 1-[N-(2-diethylamino)-ethyl]-methylamino-9-(3- 1H, H-8), 7.52 (t, J ¼ 9.5 Hz, 1H, H-7), 7.13e7.19 (m, 5H, H-5, H-6, Ph-
chlorobenzyl)-b-carboline hydrochloride salt (7a) H), 6.95 (d, J ¼ 8.5 Hz, 2H, Ph-H), 4.13 (s, 2H, CH2NHCH2CH2N), 4.03
Yield 47%; IR (KBr, cm1) v: 2971, 2649,1623,1465,1434,1336,1195, (t, J ¼ 9.5 Hz, 2H, NCH2CH2CH2Ph), 3.37 (t, J ¼ 8.5 Hz, 2H,
772; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.43e8.49 (m, 2H, H-3, H- NHCH2CH2N), 3.24e3.27 (m, 4H, N(CH2CH3)2), 3.10 (t, J ¼ 8.5 Hz,
4), 8.29 (d, J ¼ 7.8 Hz, 1H, H-8), 7.55e7.63 (m, 2H, H-5, H-7), 7.29e7.34 2H, NHCH2CH2N), 2.48 (t, J ¼ 8 Hz, 2H, NCH2CH2CH2Ph), 1.74e1.79
(m, 1H, H-6), 7.17e7.25 (m, 2H, 3-chlorophenyl-H), 6.89e6.93 (m, 2H, (m, 2H, NCH2CH2CH2Ph), 1.30 (t, J ¼ 9 Hz, 6H, CH3); 13C NMR
3-chlorophenyl-H), 5.88 (s, 2H, NCH2-3-chlorophenyl), 4.77 (s, 2H, (125 MHz, D2O): d 143.8, 140.7, 134.5, 134.1, 132.1, 131.9, 129.0, 128.7,
CH2NHCH2CH2N), 3.47e3.54 (m, 4H, NHCH2CH2N), 3.13e3.20 (m, 4H, 128.5, 126.4, 122.6, 121.9, 119.1, 116.3, 110.7, 50.1, 47.8, 46.6, 44.3,
NHCH2CH2N(CH2CH3)2), 1.21 (d, J ¼ 7.2 Hz, 6H, CH3); 13C NMR 43.2, 31.7, 30.6, 8.1; ESI-MS m/z: 415.4 (M þ 1)þ.
(75 MHz, DMSO-d6 þ D2O): d 143.4, 140.1, 135.3, 134.3, 133.9, 132.9
(2C), 131.6 (2C),128.4, 125.9, 124.9,123.2,122.3,120.4,117.0,111.3, 48.1, 5.4.22. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-(3-
47.6, 47.5, 42.3, 9.3; ESI-MS m/z: 421.1 (M þ 1)þ. phenylpropyl)-b-carboline hydrochloride salt (8b)
Yield 40%; IR (KBr, cm1) v: 2924, 2684, 1625, 1458, 1383, 1338,
5.4.18. 1-[N-(3-dimethylamino)-propyl]-methylamino-9-(3- 1052, 752; 1H NMR (500 MHz, DMSO-d6 þ D2O): d 8.47 (d,
chlorobenzyl)-b-carboline hydrochloride salt (7b) J ¼ 5.4 Hz, 1H, H-3), 8.34e8.40 (m, 2H, H4, H-8), 7.65e7.74 (m, 2H,
Yield 41%; IR (KBr, cm1) v: 2969, 2648, 1622, 1463, 1336, 1196, H-5, H-7), 7.32e7.38 (m, 1H, H-6), 7.16e7.27 (m, 5H, Ph-H), 4.86 (s,
1038, 771; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.47 (d, 2H, CH2NHCH2CH2CH2N), 4.62 (t, J ¼ 7.5 Hz, 2H, NCH2CH2CH2Ph),
5136 Z. Chen et al. / European Journal of Medicinal Chemistry 46 (2011) 5127e5137

3.19e3.25 (m, 4H, NHCH2CH2CH2N), 2.69e2.77 (m, 8H, CH3, excess of DNA by means of a dispenser equipped with a 25ul
NCH2CH2CH2Ph), 2.16e2.26 (m, 2H, NCH2CH2CH2Ph), 2.02e2.10 (m, syringe and adequate Teflon tubing.
2H, NCH2CH2CH2Ph); 13C NMR (100 MHz, D2O): d 144.5, 140.5,
136.2, 133.5, 132.5, 131.7, 128.7, 128.3, 126.8, 126.4, 123.0, 122.2,
5.7. Determination of 6Tm
119.4, 117.7, 111.0, 54.2, 46.0, 45.2, 44.4, 42.9, 31.8, 29.8, 21.5; ESI-MS
m/z: 401.4 (M þ 1)þ
Tm measurements were performed using a Shimadzu UV
2501PC Spectrometer and following the methods described by Xiao
5.4.23. 1-[N-(3-diethylamino)-propyl]-methylamino-9-(3-
et al. [5] with slight modification. Experiments were carried out in
phenylpropyl)-b-carboline hydrochloride salt (8c)
PE buffer (1 mM Na2HPO4, 0.1 mM EDTA, pH 7.4) in a thermostati-
Yield 55%; IR (KBr, cm1) v: 2933, 2669, 1624, 1493, 1457, 1385,
cally controlled cell hold, and the quartz cuvette (1 cm path length)
1337,1042, 753; 1H NMR (500 MHz, D2O): d 8.45e8.49 (m, 2H H-3, H-
was heated by circulating water at a heating rate of 0.5  C/min from
4), 8.30 (d, J ¼ 8 Hz, 1H, H-8), 7.84 (t, J ¼ 7 Hz, 1H, H-7), 7.65 (d,
25 to 95  C. Doxorubicin Hydrochloride was used as standards. In
J ¼ 8.5 Hz, 1H, H-5), 7.49 (t, J ¼ 8 Hz, 1H, H-6), 7.25e7.32 (m, 3H, Ph-
all cases, the ratio of compound to CT-DNA is 0.2.
H), 7.12 (d, J ¼ 6.5 Hz, 2H, Ph-H), 4.65 (s, 2H, CH2NHCH2CH2CH2N),
4.49 (t, J ¼ 7.5 Hz, 2H, NCH2CH2CH2Ph), 3.27e3.33 (m, 6H,
NHCH2CH2CH2N(CH2CH3)2), 3.13 (t, J ¼ 7.5 Hz, 2H, NHCH2CH2CH2N), 5.8. Fluorescence spectroscopy
2.71 (t, J ¼ 7 Hz, 2H, NCH2CH2CH2Ph), 2.13e2.24 (m, 4H,
NCH2CH2CH2Ph, NHCH2CH2CH2N), 1.35 (t, J ¼ 7 Hz, 6H, CH3); 13C Fluorescence spectral measurement was recorded using 10 mM
NMR (125 MHz, D2O): d 144.4, 140.6, 135.8, 133.5, 132.3, 131.9, 128.7, of the fluorescent drugs incubated in 1 ml of PE buffer in the
128.4, 127.2, 126.4, 122.9, 122.1, 119.5, 117.5, 110.9, 48.4, 47.7, 46.2, presence or absence of increasing concentrations of CT-DNA (0, 15,
45.3, 44.4, 31.8, 29.8, 21.0, 8.3; ESI-MS m/z: 429.4 (M þ 1)þ. 30, 45, 60, 75 and 90 mM) in a quartz cuvette of 10 mm path length.
The corresponding changes in the fluorescence intensity of the
5.4.24. 1-[N-(4-diethylamino)-butyl]-methylamino-9-(3- selected compounds were observed on a Shimadzu RF-5310PC
phenylpropyl)-b-carboline hydrochloride salt (8d) spectrofluorometer at a fluorescence excitation wavelength of
Yield 59%; IR (KBr, cm1) v: 2938, 2667, 1625, 1455, 1337, 1292, 372 nm.
1143, 1041, 762; 1H NMR (300 MHz, DMSO-d6 þ D2O): d 8.43e8.48
(m, 2H, H-3, H-4), 8.31 (d, J ¼ 7.8 Hz, 1H, H-8), 7.63e7.70 (m, 2H, Acknowledgment
H-5, H-7), 7.31e7.36 (m, 1H, H-6), 7.11e7.22 (m, 5H, Ph-H), 4.77 (s,
2H, CH2NHCH2CH2CH2CH2N), 4.56 (t, J ¼ 6.9 Hz, 2H, This work was supported by MEGA-Project (2009ZX09102-004)
NHCH2CH2CH2Ph), 3.05e3.12 (m, 8H, NHCH2CH2CH2CH2N and the Fundamental Research Funds for the Central Universities.
(CH2CH3)2), 2.65 (t, J ¼ 7.2 Hz, 2H, NCH2CH2CH2Ph), 1.97e2.06 (m,
2H, NCH2CH2CH2Ph), 1.77 (br s, 4H, NHCH2CH2CH2CH2N), 1.19 (t,
J ¼ 7.2 Hz, 6H, CH3); 13C NMR (75 MHz, DMSO-d6 þ D2O): d 143.5, References
141.4, 134.5, 133.6, 133.0, 132.0, 131.4, 129.1, 129.0, 126.7, 123.2, 121.9,
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