ORIGINAL RESEARCH

Synthesis, anti-inflammatory, and cytotoxicity evaluationof 9,10-dihydroanthracene-9,10-a,b-succinimideand bis-succinimide derivatives

Surbhi Arya • Sandeep Kumar • Reshma Rani • Nikhil Kumar •

Partha Roy • Sham M. Sondhi

Received: 24 July 2012 / Accepted: 18 December 2012 / Published online: 5 January 2013

� Springer Science+Business Media New York 2013

Abstract 9,10-Dihydroanthracene-9,10-a,b-succinimide

derivatives 4a–e and bis-succinimide derivatives 6a–e have

been synthesized by grinding 9,10-dihydroanthracene-

9,10-a,b-succinic anhydride 2 with various mono 3a–e and

diamines 5a–e in quantitative yields. All the target com-

pounds were fully characterized by spectrometric and

spectroscopic means. Compounds 4a–e, 6a–e and recently

reported compounds 4f–p were screened for anti-inflam-

matory and for cytotoxicity against five human cancer cell

lines: T47D, NCI H-522, HCT-15, PA1, and HepG-2.

Compounds 4e, 4i, 4j, and 4p exhibited good anti-inflam-

matory activity and compounds with interesting cytotoxic

profile were 4c, 6e (T47D); 4e, 4o (NCI H-522); 4n (HCT-

15); 4e, 4h, 4o (PA1); and 4a, 4e, 4f, 4i, 4o (HepG-2).

Keywords Succinimide derivatives � Green method �Anti-inflammatory � Cytotoxicity

Introduction

Synthesis of new organic molecules and their evaluation

for anti-inflammatory activity and cytotoxicity continues to

be an area of interest and challenge in academics and

pharmaceutical industry. Literature survey reveals that

certain cyclic imides such as amonafide (Fig. 1, Ia)

mitonafide (Fig. 1, Ib) (Brana and Ramos, 2001; Ingrassia

et al., 2009; Tumiatti et al., 2009), bisimides, i.e., 1,4,5,8-

naphthalenetetra carboxylic diimides (NDI) derivatives

(Fig. 1, Ic) (Brana et al., 1996; Tumiatti et al., 2009),

LU79553 (Fig. 1, Id) (Bousquet et al., 1995), DMP 840

(Fig. 1, Ie) (McRipley et al., 1994; Nitiss et al., 1998),

isoquinoline-1,3-diones (Dorr et al., 1996), phenazine-

carboxamides (Gamage et al., 2001), imidoazoacridanones

(Cholody et al., 1995; Hernandez et al., 1995), anthra-

cyclinones (Leng et al., 1998) and acridinecarboxamides

(Atwell et al., 1987) exhibited good in vivo anticancer

activity. Cyclic imides and bis imides are also reported to

exhibit anti-inflammatory (Abdel-Aziz et al., 2011; Amr

et al., 2007; Sondhi et al., 2009a), antibacterial (Khalil

et al., 2010), antimicrobial (Anizon et al., 1997) and

insecticidal (Kennedy et al., 2003) activities. Cyclic imides

and bis imides reported in Fig. 1 contain naphthalene as

core moiety. Since naphthalene and anthracene are similar

in structure but anthracene is more aromatic than naph-

thalene, it was considered worthwhile to synthesize cyclic

imides and bis imides-containing anthracene as core moi-

ety. Various organic molecules mentioned above were

synthesized either by refluxing in a solvent for several h or

involving more than one step of synthesis. Our aim was to

develop a very simple method for synthesis of cyclic imi-

des and bis imides.

In view of the literature reports and our background in

this area (Sondhi et al., 2009b, 2010a, b, 2011, 2012a, b) it

was considered worthwhile to synthesize 9,10-dihydro-

anthracene-9,10-a,b-succinimide and bis succinimide

derivatives by condensation of 9,10-dihydroanthracene-

9,10-a,b-succinic anhydride with various mono-amines and

diamines simply by grinding them together. Mono and bis

imide derivatives synthesized were screened for anti-

inflammatory activity and cytotoxicity, and are reported

herewith.

S. Arya � S. Kumar � R. Rani � S. M. Sondhi (&)

Department of Chemistry, Indian Institute of Technology-

Roorkee, Roorkee 247667, Uttarakhand, India

e-mail: sondifcy@iitr.ernet.in

N. Kumar � P. Roy

Department of Biotechnology, Indian Institute of Technology-

Roorkee, Roorkee 247667, Uttarakhand, India

123

Med Chem Res (2013) 22:4278–4285

DOI 10.1007/s00044-012-0439-6

MEDICINALCHEMISTRYRESEARCH

N

O

O

NMe

HMe

Ia

H2N Amonafide

+

Ib

N

O

O

N

Me

HMe

O2N Mitonafide

+

N

O

O

NH NH

N

O

OLU 79553

Id

N

O

ONH

HN

N

O

ODMP 840

NO2

O2NMe

Me

Ie

Ic

N

O

O

N

Me

HMeN

O

O

N

Me

MeH

++

Fig. 1 Mono-imides Ia, Ib, and di-imdes Ic, Id, Ie possessing potential anticancer activity

R is same for 3a-e and 4a-e R´ is same for 5a-e and 6a-e

4a, R= 6a, R´=

4b, R= 6b, R´=

4c, R= 6c, R´=

4d, R= 6d, R´=

4e, R= 6e, R´=

O H2C

O H2C

HN N CH2CH2

HNH2C

H2C

CH2CH2

CH2CH2CH2

CH2CH2CH2CH2

CHCH3

CH2

N N (CH2)3(CH2)3

N

O

O

R' N

O

O

O

O

O

N

O

O

R

2

R NH2

3a-e

H2N R' NH2

5a-e

4a-e 6a-e

Scheme 1 Synthesis of mono

and bis imide derivatives 4a–e,

6a–e

Med Chem Res (2013) 22:4278–4285 4279

123

Results and discussion

Chemistry

9,10-Dihydroanthracene-9,10-a,b-succinic anhydride (2,

Scheme 1) was synthesized by following the reaction

procedure reported in literature (Vogel, 1968). Condensa-

tion of 9,10-dihydroanthracene-9,10-a,b-succinic anhy-

dride 2 with 2-(amino methyl)tetrahydrofuran 3a was

carried out by grinding both of them in equimolar ratio in a

small mortar with a pestle for 10 min. Thin layer chro-

matography (TLC) of the reaction mixture showed com-

pletion of the reaction. Crude product so obtained was

washed with diethyl ether and then crystallized from

methanol to get pure product 3a,4,9,9a-tetrahydro-2-[(2-

methyl)tetrahydrofuran]-4,9[10,20]-benzeno-1H-benz-[f]-

isoindole-1,3-(2H)-dione (4a; Scheme 1) in 91 % yield.

Spectral (IR, 1H NMR, 13C NMR, and GCMS) and ele-

mental analysis data of 4a reported in the experimental

section fully support the structure assigned to it. Similarly

condensation of 9,10-dihydroanthracene-9,10-a,b-succinic

anhydride 2 with 1-(2-aminoethyl) piperazine 3b, 2-amino

methyl furan 3c, 4-amino methyl piperidine 3d, and

benzylamine 3e (Schumann et al., 1964) gave the

corresponding condensation products 4b–e (Scheme 1) in

excellent yields. Physical constants, spectral, and elemental

analysis data of 4b–e reported in the experimental section

fully support the structures assigned to 4b–e.

This simple method was extended further for the syn-

thesis of bis-imide derivatives (6a–e, Scheme 1). Ethylene

diamine 5a and 9,10-dihydroanthracene-9,10-a,b-succinic

anhydride 2 were mixed in 1:2 molar ratio. This reaction

mixture was grinded in a small mortar with a pestle for

15 min. TLC of the reaction mixture showed completion of

the reaction. Crude product so obtained was washed with

diethyl ether and then crystallized from methanol to get

pure product 9,10-ethanoanthracene-11, 12-dicarboximide,

N,N0-ethylene bis [9,10-dihydro] (6a, Scheme 1) (Kita-

honoki and Kido, 1970) in excellent yield. 1H NMR

(500 MHz, DMSO-d6) of 6a shows signals at d (ppm):

2.678 (s, 4H, CH2 ? CH2), 3.148–3.179 (t, 4H, 49 CH),

4.721–4.750 (d, 4H, 49 CH), 7.045–7.141 (m, 8H, Ar),

7.182–7.231 (m, 4H, Ar), 7.418–7.449 (m, 4H, Ar)

GC–MS m/z 576 (M?; 1.29 %). FT-IR spectra show

absorption bands at 1768 and 1711 (–CO–N–CO–) and 1462

(Ar) cm-1. Spectral data of 6a fully support the structure

assigned to it. Similarly condensation of 1,3-diaminopro-

pane 5b, 1,4-diaminobutane 5c, 1,2-diaminopropane 5d and

1,4 bis (3-aminopropyl) piperazine 5e with 9,10-dihydro-

anthracene-9,10-a,b-succinic anhydride 2 gave the corre-

sponding condensation products 6b–e (Scheme 1) in

excellent yields. Physical constants, spectral and elemental

analysis data of 6b–e reported in the experimental section are

in full agreement with the structures assigned to compounds

6b–e.

Fully characterized and purified compounds 4a–e and 6a–

e along with the earlier reported (Rani et al., 2012) com-

pounds 4f–o (Fig. 2) were screened for anti-inflammatory

activity (Winter et al., 1962) using carrageenan-induced paw

oedema assay and the results are summarized in Table 1.

4 f-p

4f, R= 4g, R=

4h, R= 4i, R=

4j, R= 4k, R=

4l, R= 4m, R=

4n, R= 4o, R=

4p, R=

N

O

O

R

N H2C

NH2C

N HN

HN

O NH2C

N CH2CH2

HN

N CH2CH2

N CH2CH2

O

NN

CH2CH2CH2

Fig. 2 Structures of previously

synthesized monoimide

derivatives 4f–p

4280 Med Chem Res (2013) 22:4278–4285

123

Compounds 4e, 4i, 4j, and 4p exhibited interesting anti-

inflammatory activity 37, 37, 39, and 38 %, respectively,

whereas standard drug ibuprofen exhibited 39 % anti-

inflammatory activity at 50 mg/kg p.o.

In vitro cytotoxicity (Monks et al., 1991; Skehan et al.,

1990) evaluation of compounds 4a–p and 6a–e was carried

out against five human cancer cell lines consisting of breast

(T47D), lung (NCI H-522), colon (HCT-15), ovary (PA1),

and liver (HepG-2). Percentage (%) growth inhibition of

cancer cell lines was determined at a concentration of

10 lM and the results are summarized in Table 1. Com-

pounds with interesting cytotoxic profile are 4c, 6e (T47D);

4e, 4o (NCI H-522); 4n (HCT-15); 4e, 4h, 4o (PA1); and

4a, 4e, 4f, 4i, 4n (HepG-2).

Two series of compounds are reported in this paper, mono

imides, i.e., 4a–p and bis imides, i.e., 6a–e. Data in Table 1

indicate that bis imides possess less biological activities as

compared to mono imides. This may be due to bulky nature

of bis imides which make these difficult to interact with the

biological targets. In the case of mono imide derivatives, i.e.,

4a–p both linker arm and size of the ring attached to linker

arm were changed and the best results were obtained in the

case of 4e and 4o, where linker arm is –CH2– and –NH–,

respectively, and the cyclic ring is phenyl in both the cases.

Compound 4e exhibited very good anti-inflammatory

activity and cytotoxicity against three cancer cell lines

consisting of lung (NCI H-522), ovary (PA1), and liver

(HepG-2), whereas the compound 4o exhibited very good

cytotoxicity against abovementioned three cancer cell lines and

moderate anti-inflammatory activity.

Conclusion

A number of mono and bis imide derivatives, i.e., 4a–o and

6a–e have been synthesized by simple grinding method and

Table 1 Anti-inflammatory activity and in vitro cytotoxicity evaluation of compounds 4a–p and 6a–e

Compound no. Anti-inflammatory

activity (%) at

50 mg/kg p.o.

Cytotoxicity at a concentration of 1 9 10-5 Ma

Breast T47D Lung NCI H-522 Colon HCT-15 Ovary PA1 Liver HepG2

4a 34 23 30 22 31 55

4b 31 NT NT NT NT NT

4c 15 40 25 15 27 38

4d 35 17 25 11 26 32

4e 37 22 62 12 43 52

4f 23 21 24 19 37 53

4g 32 10 15 20 31 40

4h 14 26 20 07 47 35

4i 37 24 27 12 21 61

4j 39 20 24 22 24 34

4k 28 07 13 14 36 47

4l 27 20 23 06 25 38

4m 17 26 15 16 33 41

4n 27 28 32 30 40 48

4o 11 13 52 11 46 52

4p 38 01 27 04 13 37

6a 20 05 11 10 09 29

6b 33 24 17 27 06 45

6c 25 19 23 26 37 25

6d 22 02 20 09 14 41

6e 15 33 17 10 16 33

Ibuprofen 39 – – – – –

CYC-PHO – 12 09 05 15 14

CYC-HEXI – 09 13 12 32 15

5-FU – 10 18 15 18 28

Bold values represent compounds showing good anti-inflammatory activity and cytotoxicity

CYC-PHO cyclophospamide, CYC-HEXI cycloheximide, 5-FU 5-Fluorouracil, NT not testeda Compounds tested in triplicate, data expressed as mean value of three independent experiments

Med Chem Res (2013) 22:4278–4285 4281

123

screened for cytotoxicity and anti-inflammatory activity.

Compounds 4e and 4o exhibited very good cytotoxicity

against three cancer cell lines whereas compound 4e also

exhibited good anti-inflammatory activity. From the above

observations it can be concluded that compounds 4e and 4o

meet electronic and stereochemical requirements on the

target site in a better way as compared to other molecules

which failed to act.

Experimental

Melting points (mp) were determined on a JSGW apparatus

and are uncorrected. IR spectra were recorded using a

Perkin Elmer 1600 FT spectrometer. 1H NMR spectra were

recorded on a Bruker WH-500 spectrometer at a ca,

5–15 % (W/V) solution in DMSO-d6 or CDCl3 (TMS as

internal standard). 13C NMR spectra were recorded on a

Bruker WH-125 spectrometer using DMSO-d6 or CDCl3 as

solvent. GC–MS were recorded on Perkin Elmer Clarus

500 gas chromatograph where built in MS detector was

used. Elemental analysis was carried out on a Vario EL III

elementor. Thin layer chromatography (TLC) was per-

formed on silica gel G for TLC (Merck) and spots were

visualized by iodine vapor.

General procedure for synthesis of 9,

10-dihydroanthracene-9,10-a,b-succinimide

derivatives 4a–e

Synthesis of 3a,4,9,9a-tetrahydro-2-[(2-methyl)

tetrahydrofuran)]-4,9[10,20]-benzeno-1H benz-[f]-

isoindole-1,3-(2H)-dione (4a)

9,10-Dihydroanthracene-9,10-a,b-succinic anhydride (2;

552 mg, 2 mmol) and 2-(amino methyl) tetrahydrofuran

(3a; 0.21 mL, 2 mmol) were grinded together in a small

mortar with a pestle for 10 min. TLC of the reaction

mixture on silica gel using ethyl acetate/methanol (19:1) as

mobile phase indicated completion of the reaction. Crude

product so obtained was washed with diethyl ether and then

recrystallized from methanol to get pure condensation

product 3a,4,9,9a-tetrahydro-2-[(2-methyl)tetrahydrofu-

ran)]-4,9[10,20]-benzeno-1H-benz-[f] isoindole-1,3-(2H)-

dione (4a). Yield—650 mg 91 %, mp: 223–224 �C, IR

(KBr) tmax: 1777, 1697 (–CO–N–CO–), 1462 (Ar) cm-1.1H NMR (500 MHz, DMSO-d6) d: 0.961–0.965 (m, 1H,

one H of CH2), 1.143–1.159 (m, 1H, one H of CH2),

1.594–1.652 (m, 2H, CH2), 2.927–2.969 (q, 1H, J = 8 Hz

and 13 Hz, one H of CH2), 3.044–3.081 (dd, 1H, J = 5 and

13.5 Hz, one H of CH2), 3.127–3.140 (m, 1H, [CH–),

3.261–3.269 (t, 2H, J = 2 Hz, CH ? CH), 3.417–3.446 (q,

1H, J = 7.5 and 14.5 Hz, one H of CH2), 3.582–3.601 (q,

1H, J = 2 and 7.5 Hz, one H of CH2), 4.775–4.787 (t, 2H,

J = 3 Hz, CH ? CH), 7.120–7.137 (q, 2H, Ar),

7.155–7.172 (q, 2H, Ar), 7.244–7.270 (q, 2H, Ar),

7.462–7.479 (q, 2H, Ar), 13C NMR (125 MHz, DMSO-d6)

d: 43.30, 44.29, 46.48, 118.69, 122.15, 124.25, 125.00,

126.27, 126.80, 136.74, 139.58, 142.06, 148.86, 154.41 and

176.48 GC–MS: m/z 359 (M?, 15 %), 181 ( N

O

O

CH2

O. +

, 5 %),

178 ( , 90 %), 110 ( N

O

O

CH2

+, 8 %), 85 ( O CH2

+ , 5 %), 84

( O CH2, 100 %), 71 ( O +, 58 %). Anal. Calcd for C23H21NO3:

C 76.86, H 5.89, N 3.90; Found C 76.80, H 5.79, N 3.90.

Similarly compounds 4b–e were synthesized.

3a,4,9,9a-Tetrahydro-2-[2-(1-piperazinyl)ethyl]-4,9[10,20]-

benzeno-1H-benz[f]isoindole-1,3-(2H)-dione (4b)

Yield—90 %, mp: 228–230 �C, IR (KBr) tmax: 3408 (NH),

1780, 1705, 1662 (–CO–N–CO–), 1603 and 1542 (Ar)

cm-1. 1H NMR (500 MHz, DMSO-d6) d: 1.343–1.374 (m,

2H, CH2), 2.072 (bs, 4H, CH2 ? CH2), 2.568–2.587 (t, 4H,

J = 4.5 Hz, CH2 ? CH2), 3.039–3.070 (q, 2H, J = 5.5

and 7.5 Hz, CH2), 3.225–3.260 (m, 2H, CH ? CH), 4.769

(s, 2H, CH ? CH), 7.110–7.128 (m, 2H, Ar), 7.158–7.175

(m, 2H, Ar), 7.234–7.271 (m, 2H, Ar), 7.454–7.487 (m,

2H, Ar), 13C NMR (125 MHz, DMSO-d6) d: 34.76, 44.54,

45.28, 46.19, 53.81, 54.56, 99.49, 124.22, 124.71, 126.25,

126.44, 139.15, 141.75 and 176.45 GC–MS: m/z 387 (M?,

1 %), 386 (M?-H, 10 %), 178 (. +

, 19 %), 99 (H2C N NH+ ,

100 %). Anal. Calcd for C24H25N3O2: C 74.39, H 6.50, N

10.84; Found C 74.30, H 6.45, N 10.83.

3a,4,9,9a-Tetrahydro-2-[furan-2-yl-methyl]-4,9[10,20]-

benzeno-1H-benz-[f]-isoindole-1,3-(2H)-dione (4c)

Yield—98 %, mp: 238–240 �C, IR (KBr) tmax: 1772,

1706, 1662 (–CO–N–CO–), 1603 and 1541 (Ar) cm-1. 1H

NMR (500 MHz, DMSO-d6) d: 2.812–2.837 (q, 1H, J =

2 Hz and 11 Hz, one H of CH2), 3.171–3.198 (q, 1H,

J = 2.5, 11 Hz, one H of CH2), 4.417–4.422 (d, 2H,

J = 2.5 Hz, CH ? CH), 4.536–4.540 (d, 2H, J = 2 Hz,

CH ? CH), 6.241–6.246 (d, 1H, J = 2.5 Hz, Ar),

6.413–6.423 (q, 1H, J = 2, 3 Hz, Ar), 6.975–7.098 (m, 5H,

Ar), 7.290–7.343 (m, 3H, Ar), 7.601–7.603 (d, 1H,

J = 1.0 Hz, Ar), 13C NMR (125 MHz, DMSO-d6) d:

34.42, 44.32, 46.22, 48.58, 99.49, 107.13, 110.07, 124.19,

124.61, 126.22, 126.53, 139.01, 142.00, 142.25, 148.07 and

175.89 GC–MS: m/z 355 (M?, 12 %), 178(. +

, 100 %),

177 ( N

O

O

CH2

O. +

, 10 %). Anal. Calcd for C23H17NO3: C 77.73,

H 4.82, N 3.94; Found C 77.70, H 4.79, N 3.95.

4282 Med Chem Res (2013) 22:4278–4285

123

3a,4,9,9a-Tetrahydro-2-[(1-piperadin)-methyl]-4,9[10,20]-

benzeno-1H-benz-[f]-isoindole-1,3-(2H)-dione (4d)

Yield—95 %, mp: 235 �C, IR (KBr) tmax: 3593 (NH),

1772, 1697, 1665 (–CO–N–CO–), 1598 and 1544 (Ar)

cm-1. 1H NMR (500 MHz, DMSO-d6) d: 1.301–1.403 (m,

8H, 4 9 CH2), 2.108 (bs, 1H,[CH–), 3.034–3.065 (t, 2H,

J = 8 Hz, –CH2–), 3.239 (s, 2H, CH ? CH), 4.767

(s, 2H, CH ? CH), 7.116–7.174 (m, 4H, Ar), 7.231–7.248

(q, 2H, J = 3, 5 Hz, Ar), 7.461–7.478 (q, 2H, J = 3, 5 Hz,

Ar), 13C NMR (125 MHz, DMSO-d6) d: 29.90, 36.39,

44.30, 45.43, 46.40, 47.90, 124.46, 124.84, 126.52, 127.05,

139.06, 141.08, and 171.55 GC–MS: m/z 372 (M?, 8 %),

178. (. +

, 20 %), 84 ( NH+ , 100 %). Anal. Calcd for

C24H24N2O2: C 77.41, H 6.45, N 7.52; Found C 77.39,

H 6.44, N 7.50.

3a,4,9,9a-Tetrahydro-2-(phenyl methyl)-4,9-[10,20]-

benzeno-1H-benz-[f]-isoindole-1,3-(2H)-dione (4e)

(Schumann et al., 1964)

Yield—96 %, mp: 236–238 �C, IR (KBr) tmax: 1781 &

1632 (–CO–N–CO–), 1458 (Ar) cm-1. 1H NMR

(500 MHz, DMSO-d6) d: 2.848 (s, 2H, –CH2–), 3.666

(s, 2H, CH ? CH), 4.882 (s, 2H, CH ? CH), 7.180–7.213

(m, 4H, Ar), 7.254–7.279 (t, 3H, Ar), 7.301–7.354 (m, 6H,

Ar), 13C NMR (125 MHz, DMSO-d6) d: 44.30, 47.90,

119.03, 124.47, 124.84, 125.10, 126.52, 127.06, 128.52,

139.06, 141.08 and 171.57 GC–MS: m/z 365 (M?, 20 %),

178 (. +

, 100 %). Anal. Calcd for C25H19NO2: C 82.17,

H 5.24, N 3.83; Found C 82.15, H 5.20, N 3.79.

General procedure for the synthesis of bis imide

derivatives 6a–e

9,10 Ethanoanthracene-11,12-dicarboximide,N,N0-ethylene bis [9,10-dihydro] (6a)

9, 10-Dihydroanthracene-9,10-a,b-succinic anhydride (2;

552 mg, 2 mmol) and ethylene diamine (5a; 0.065 mL,

1 mmol) were grinded together in a mortar with a pestle

for 15 min. TLC of the reaction mixture on silica gel

using methanol/ethyl acetate (1:19) as mobile phase

exhibited that the reaction is complete. Crude product so

obtained was washed with diethyl ether and then recrys-

tallized from methanol to get pure condensation product

9,10 ethanoanthracene-11,12-dicarboximide, N,N0-ethyl-

ene bis [9,10-dihydro] (6a) (Kitahonoki and Kido, 1970).

Yield—515 mg 90 %, mp: [300 �C, IR (KBr) tmax: 1768

and 1711 (–CO–N–CO–), 1462 (Ar) cm-1. 1H NMR

(500 MHz, DMSO-d6) d: 2.678 (s, 4H, CH2 ? CH2),

3.148–3.179 (t, 4H, 4 9CH), 4.721–4.750 (d, 4H, 4

9CH), 7.045–7.141 (m, 8H, Ar), 7.182–7.231 (m, 4H,

Ar), 7.418–7.449 (m, 4H, Ar), 13C NMR (125 MHz,

DMSO-d6) d: 44.55, 45.00, 46.26, 124.19, 124.61, 126.24,

126.47, 139.23, 141.80, and 176.73 GC–MS: m/z 576

(M?, 1.29 %), 302 ( N

O

O

CH2CH2

+ , 2 %), 288. ( N

O

O

CH2

+ , 3 %),

96 ( N

O

O

+, 25 %). Anal. Calcd for C38H28N2O4: C 79.16, H

4.86, N 4.86; Found C 79.14, H 4.85, N 4.88.

9,10-Ethanoanthracene-11,12-dicarboximide, N,N0-trimethylene bis [9,10-dihydro] (6b) (Kitahonoki and Kido,

1970)

Yield—94 %, mp:[300 �C, IR (KBr) tmax: 1768 and 1696

(–CO–N–CO–), 1458 (Ar) cm-1. 1H NMR (500 MHz,

DMSO-d6) d: 0.424–0.451 (t, 2H, –CH2–), 2.512–2.551 (t,

4H, CH2 ? CH2), 3.175–3.185 (d, 4H, 4 9CH), 4.694 (s,

4H, 4 9CH), 7.052–7.068 (t, 4H, Ar), 7.149–7.212 (m, 8H,

Ar), 7.448–7.465 (t, 4H, Ar), 13C NMR (125 MHz, DMSO-

d6) d: 35.27, 44.38, 46.14, 48.15, 124.21, 124.57, 126.22,

126.49, 139.04, 141.83, and 176.32 GC–MS: m/z 590 (M?,

0.4 %), 178, (. +

, 55 %), 96 ( N

O

O

+, 20 %). Anal. Calcd

for C39H30N2O4: C 79.32, H 5.08, N 4.74; Found C 79.30,

H 5.10, N 4.76.

9,10-Ethanoanthracene-11,12-dicarboximide, N,N0-tetramethylene bis [9,10-dihydro] (6c) (Kitahonoki

and Kido, 1970)

Yield—92 %, mp:[300 �C, IR (KBr) tmax: 1768 and 1698

(–CO–N–CO–), 1461 (Ar) cm-1. 1H NMR (500 MHz,

DMSO-d6) d: 0.737–0.765 (t, 4H, CH2 ? CH2),

2.957–2.980 (t, 4H, CH2 ? CH2), 3.210 (s, 4H, 4 9CH),

4.751 (s, 4H, 4 9CH), 7.096–7.162 (m, 8H, Ar),

7.228–7.245 (q, 4H, J = 3, 5 Hz, Ar), 7.444–7.461 (q, 4H,

J = 3, 5 Hz, Ar), 13C NMR (125 MHz, DMSO-d6) d:

24.20, 44.55, 46.19, 52.30, 124.20, 124.67, 126.23, 127.00,

139.23, 141.86 and 176.56 GC–MS:m/z 302 ( N

O

O

CH2CH2

+ ,

10 %), 96 ( N

O

O

+, 25 %). Anal. Calcd for C40H32N2O4:

C 79.47, H 5.29, N 4.63; Found C 79.50, H 5.27, N 4.65.

9,10-Ethanoanthracene-11,12-dicarboximide,N,N0-propylene bis [9,10-dihydro] (6d) (Kitahonoki and Kido,

1970)

Yield—92 %, mp:[300 �C, IR (KBr) tmax: 1780 and 1660

(–CO–N–CO–), 1571 and 1463 (Ar) cm-1. 1H NMR

(500 MHz, DMSO-d6 ? D2O) d: -0.126 to -0.138 (d, 3H,

Med Chem Res (2013) 22:4278–4285 4283

123

J = 6 Hz, CH3), 2.115–2.134 (d, 1H, J = 9.5 Hz, CH),

3.064–3.142 (m, 4H, 4 9CH), 3.172–3.195 (q, 2H, J = 3.5,

8.5 Hz, CH2), 4.658–4.681 (q, 4H, J = 3.5, 8.5 Hz, 4 9CH),

7.007–7.191 (m, 12H, Ar), 7.398–7.404 (d, 4H, J = 3 Hz,

Ar), 13C NMR (125 MHz, DMSO-d6) d: 21.13, 44.47, 46.27,

48.99, 50.32, 124.5, 124.6, 126.5, 126.6, 139.3, 141.9, and

176.8 GC–MS: m/z 575 (M? –CH3, 1.3 %), 302 ( N CHCH3

+

O

O

,

2 %), 96 ( N

O

O

+, 25 %). Anal. Calcd for C39H30N2O4: C 79.32,

H 5.08, N 4.74; Found C 79.30, H 5.10; N 4.75.

9,10-Ethanoanthracene-11,12-dicarboximide,N,N0-[1,4 di

(trimethylene)piperazine] bis [9,10-dihydro] (6e)

Yield—93 %, mp: 278–280 �C, IR (KBr) tmax: 1660–1631

(–CO–N–CO–), 1597–1514 (Ar) cm-1. 1H NMR

(500 MHz, DMSO-d6) d: 0.739–0.766 (t, 4H, 2 9CH2),

1.800–1.827 (t, 4H, 2 9CH2), 2.076–2.147 (bd, 8H, 4

9CH2), 2.969–2.998 (t, 4H, 2 9CH2), 3.222 (bs, 4H, 4

9CH), 4.770 (bs, 4H, 4 9CH), 7.103–7.120 (t, 4H, Ar),

7.154–7.170 (t, 4H, Ar), 7.241–7.257 (t, 4H, Ar),

7.459–7.475 (t, 4H, Ar), 13C NMR (125 MHz, DMSO-d6)

d: 23.80, 35.75, 44.52, 46.18, 52.29, 54.32, 124.20, 124.67,

126.23, 126.45, 139.22, 141.85 and 176.59 GC–MS: m/z

360 ( N

O

O

(CH2)3 N N N

O

O

(H2C)3

. +

, 0.65 %), 96 ( N

O

O

+, 65 %). Anal.

Calcd for C46H44N4O4: C 77.09, H 6.14, N 7.82; Found

C 77.10, H 6.12, N 7.80.

Pharmacology

Anti-inflammatory activity (Winter et al., 1962)

Paw oedema inhibition test was used on albino rats of

Charles Foster by adopting the method of Winter et al.,

(1962). Groups of five animals of both sexes (body weight

120–160 g), excluding pregnant females, were given dose

of the compound to be tested. Thirty min later, 0.20 mL of

1 % freshly prepared carrageenan suspension in 0.9 %

NaCl solution was injected subcutaneously into the planter

aponeurosis of the hind paw and the volume was measured

by a water plethysmometer apparatus and then measured

again 1–3 h later. The mean increase of paw volume at

each interval was compared with that of control group (five

rats treated with carrageenan but not with tests compound)

at the same intervals and percent inhibition value calcu-

lated by the formula given below.

% anti-inflammatory activity ¼ 1� Dt=Dc½ � � 100

Dt and Dc are paw volumes of oedema in tested and control

groups, respectively.

In vitro cytotoxicity against human cancer cell lines

(Monks et al., 1991; Skehan et al., 1990)

The human cancer cell lines procured from National

Cancer Institute, Frederick, USA were used in this study.

Cells were grown in tissue culture flasks in complete

growth medium (RPMI-1640 medium with 2 mM gluta-

mine, pH 7.4 supplemented with 10 % fetal bovine serum,

100 lg/mL streptomycin and 100 units/mL penicillin) in a

carbon dioxide incubator (37 �C, 5 % CO2, 90 % RH). The

cells at subconfluent stage were harvested from the flask by

treatment with trypsin (0.05 % in PBS (pH 7.4) containing

0.02 % EDTA). Cells with viability of more than 98 %, as

determined by trypan blue exclusion, were used for

determination of cytotoxicity. The cell suspension of

1 9 105 cells/mL was prepared in complete growth med-

ium. Stock 4 9 10-2 M compound solutions were pre-

pared in DMSO. The stock solutions were serially diluted

with complete growth medium containing 50 lg/mL of

gentamycin to obtain working test solution of required

concentrations.

In vitro cytotoxicity against various human cancer cell

lines was determined (Monks et al., 1991) using 96-well

tissue culture plates. Then 100 lL of cell suspension was

added to each well of the 96-well tissue culture plates.

The cells were allowed to grow in CO2 incubator (37 �C,

5 % CO2, 90 % RH) for 24 h. The test materials in

complete growth medium (100 lL) were added after 24 h

incubation to the wells containing cell suspension. The

plates were further incubated for 48 h (37 �C in an

atmosphere of 5 % CO2 and 90 % relative humidity) in a

carbon dioxide incubator after addition of test material

and then the cell growth was stopped by gently layering

trichloroacetic acid (50 % TCA, 50 lL) on top of the

medium in all the wells. The plates were incubated at

4 �C for 1 h to fix the cells attached to the bottom of the

wells. The liquid of all the wells was gently pipetted out

and discarded. The plates were washed five times with

distilled water to remove TCA, growth medium low

molecular weight metabolites, serum proteins, etc., and air

dried. Cell growth was measured by staining with sulfo-

rhodamine B dye (Skehan et al., 1990). The adsorbed dye

was dissolved in Tris–HCl buffer (100 lL, 0.01 M, pH

10.4) and plates were gently stirred for 10 min using a

mechanical stirrer. The optical density (OD) was recorded

on ELISA reader at 540 nm.

Acknowledgments We are thankful to technical staff of the

Chemistry Department, I. I. T. Roorkee, for spectroscopic studies and

elemental analysis. Thanks also due to Head I. I. C. for providing

NMR facility. We are thankful to Prof. G. Bhattacharjee and Prof.

Ravi Bhushan of the Chemistry Department, I. I. T. Roorkee for

helpful discussion. Ms. Surbhi Arya (SRF) and Reshma Rani are

thankful to CSIR, New Delhi, and Mr. Sandeep Kumar to MHRD,

New Delhi for financial assistance.

4284 Med Chem Res (2013) 22:4278–4285

123

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