Indo American Journal of Pharmaceutical Research, 2014

ISSN NO: 2231-6876

SYNTHESIS AND STUDY OF BIOLOGICAL ACTIVITY OF NEW TETRALONE ACIDS

ANALOGUES OF PODOPHYLLOTOXIN
Santhekasalagere Basavaiah Shivakumar, Basavaiah Umesha, Mudeenahally Hucchegowda Krishna and
Yeriyur Basavaiah Basavaraju*.
Department of Studies in Chemistry, University of Mysore, Manasagangotri, Mysore - 570 006, Karnataka, India.
ARTICLE INFO
Article history
Received 22/11/2014
Available online
19/12/2014

Keywords
Ketones,
Stobbe Condensation,
Soponification,
Catalytic Hydrogenation,
Tetralone Acids,
Antimitotic Activity.

ABSTRACT
New tetralone acid analogues of podophyllotoxin 7a-c i.e substituted 4-oxo-7-phenyl-4,5,6,7tetrahydrobenzo[b]thiophene-6-carboxylic acid were synthesized by Genslers method with
some changes in reagent and experimental procedure. Substituted 2-(phenyl(thiophen-2yl)methylene)succinic acids 4a-c were synthesized by stobbe condensation of substituted
phenyl(thiophen-2-yl)methanone 2a-c in presence of sodium hydride, diethyl succinate ,
ethanol in dry benzene to form compounds 3a-c which were subjected into soponification in
presence of sodium hydroxide in water and methanol mixture. Compound 4a-c on catalytic
hydrogenation in presence palladium over carbon furnished substituted 2-(phenyl(thiophen-2yl)methyl)succinic acids 5a-c. The compounds 5a-c were refluxed with acetyl chloride to
afford benhydril succinic anhydride 6a-c, which on intramolecular cyclisation with Lewis
acid like aluminum chloride gave target compounds 7a-c. The structure and products were
confirmed by IR, 1H NMR, 13C NMR, mass spectra and elemental analysis data. The
synthesized tetralone acids were screened for their antimitotic activity by onion root tip
method. The antimitotic activity was compared with control, compounds 7a, 7b, and 7c
showed considerable activity. Among the synthesized compounds, 7a exhibited more
inhibition compared with remaining synthesized compounds 7b and 7c, at the same time
compound 7b showed moderate inhibitions than compound 7c exhibited less inhibitions.

Copy right 2014 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Please cite this article in press as Santhekasalagere B. Shivakumar et al. Synthesis and Study of Biological Activity of New
Tetralone Acids Analogues of Podophyllotoxin. Indo American Journal of Pharm Research.2014:4(12).

5733

Corresponding author
Prof. Y. B. Basavaraju
DOS in Chemistry, University of Mysore,
Manasagangotri,Mysore-570 006,
Karnataka, India.
+919480441804
+918212421263
basavaraju_yb@yahoo.co.in, sbshiv999@gmail.com

Vol 4, Issue 12, 2014.

Santhekasalagere B. Shivakumar et al.

ISSN NO: 2231-6876

INTRODUCTION
Cancer is the most dangerous disease to the mankind all over the world with its many forms [1]. Many organic compounds
have been used as anticancer agents including podophyllotoxin (I) and its analogous [2] in the last four decades. Podophyllotoxin
(Figure 1) and its several analogues are being used as cytotoxic spindle poisons and antitumor agents at clinical levels [3]. Several
analogues of the podophyllotoxin have been reported in literature. Podophyllotoxin has been extracted from two important medicinal
plants named podophyllum emodi an Indian species and podophyllum peltatum, a North American species [4]. It was also been
isolated from many other plants of podophyllum species. It belongs to the family of natural products called lignans. Podophyllotoxin
showed other biological activities such as cathartic, antitropical skin disease, antimalarial, anti-HIV (AIDS) etc [5].

Fig. 1. Podophyllotoxin.
In view of the above facts, it was decided to modify the structure of podophyllotoxin and synthesized tetralone acids as
analogues [6-9]. They were synthesized by replacing 3,4,5-trimethoxyphenyl ring with substituted phenyl in podophyllotoxin , 1,3methylene dioxy ring with thiophene group and lactone ring with carboxylic acid group. The new analogues of podophyllotoxin were
synthesized using Genslers method [10] with some changes in reagents and experimental procedure.
EXPERIMENTAL
Materials and methods
All the reagents and chemicals were purchased. They were used without further purification. Melting points were taken in open
capillary tubes and are uncorrected. Thin layer chromatography (TLC) is performed with E. Merck pre-coated silica gel plates
(60F254). Acme, India silica gel, 60-120 mesh is used for column chromatography. IR spectra in KBr were recorded on Perkin-Elmer
model 683 spectrometers. 1H NMR (400 MHz) and 13C NMR (100 MHz) spectra were recorded using tetramethyl silane (TMS) as an
internal reference on Bruker spectrometer. Elemental analyses were performed on a Perkin-Elmer 2400. Mass spectra were obtained
by Water-Q-TOF ultima spectrometer. Micro analytical data were obtained by Elemental-Vario EL-III.
Synthetic procedure
General procedure for the preparation of substituted phenyl(thiophen-2-yl)methanones 2a-c
A solution of thiophene-2-carbonyl chloride (22.3 g, 0.1526 mol) in dichloromethane (75 mL) was added drop wise to the
stirred suspension of substituted benzene (15 g, 0.138 mol) 1a-c and anhydrous aluminum chloride (23.9 g, 0.179 mol) in
dichloromethane (75mL) at 0-5 C under inert atmosphere. The reaction mixture was stirred for 10 h at 25-30 C. The completion of
the reaction was known by TLC. The reaction mass was poured into 1 N HCl. The mixture was stirred for 2 h at 25-30 C. The
product was extracted into dichloromethane. The combined organic layer was washed with aq. sodium bicarbonate solution and then
with water, dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum using rotary evaporator. The products
were recrystallized from diethyl ether.

Thiophen-2-yl(p-tolyl)methanone: 2c: Color: Yellow solid. Yield: 61 %. M.P.:98-1000 C. IR (KBr, v, Cm-1 ): 1698 (C=O), 1594 (Ar
C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.32 (s, 3H, CH3), 7.1-7.77 (m, 7H, Ar-H phenyl and thiophen ring). 13C NMR (100 MHz,
CDCl3, , ppm):25.3, 127.3, 130.3, 132.2, 137, 138.2, 143, 145, 182.1. MS (ESI, m/z): 203.3 (M+). Anal. Calcd. for C12H10OS:
C,71.56; H, 4.98. Found: C, 71.9; H, 4.88; S, 15.6%.

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Phenyl(thiophen-2-yl)methanone: 2b: Color: white crystalline solid. Yield: 76.5 %. M.P.:56-580 C. IR (KBr, v, Cm-1 ): 1700 (C=O),
1596 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 7.1-7.88 (m, 8H, Ar-H phenyl and thiophen ring). 13C NMR (100 MHz, CDCl3, ,
ppm):128.1, 129.1, 132.3, 137.8, 138.1,142.3, 192.1. MS (ESI, m/z): 189.3 (M +). Anal. Calcd. for C11H8OS: C,70.18; H, 4.28. Found:
C, 70.11; H, 4.27; S, 16.9%.

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(4-methoxyphenyl)(thiophen-2-yl)methanone: 2a: Color: white solid. Yield: 67 %. M.P.:71-740C.IR (KBr, v, Cm-1): 1679 (C=O),
1613 (Aromatic C=C); 1H NMR (CDCl3-400MHz) ppm: 3.7 (s, 3H, OCH3), 7.2-7.8 (m, 7H, Ar-H phenyl and thiophen ring);13C
NMR (CDCl3 -100 MHz) ppm: 184.1, 165.5, 145.2, 137, 135, 130.8, 128.8, 126.8, 117.4, 56.6; MS (ESI) m/z: 219.3 (M +). Anal.
Calcd. for C12H10O2S; C, 66.03; H, 4.62 . Found: C, 65.08; H, 4.59; S, 14.5 %.

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General procedure for the preparation of substituted 2-(phenyl(thiophen-2-yl)methylene)succinic acids 4a-c

Sodium hydride (2.198 g, 0.096 mol) was added in portions to the stirred suspension of substituted phenyl(thiophen-2yl)methanone (10 g, 0.0458 mol) 2a-c and diethyl succinate (23.9 g, 0.137 mol) in dry benzene (30 mL) under nitrogen gas
atmosphere. The reaction mixture was stirred for 10 min at 25-30 C (until the evolution of hydrogen gas ceased). Absolute ethanol
(10.5 g, 0.229 mol) was added drop wise during 1 h to the above solution. The reaction mass was stirred at 25-30 C for 20 h under
nitrogen gas condition. The completion of the reaction was confirmed by TLC. The reaction mixture was cooled and acidified by
addition of glacial acetic acid. The compounds were extracted into ether and then into saturated sodium carbonate solution. The
sodium carbonate extract was neutralized by dilute acetic acid. The precipitated gummy residue was extracted into dichloromethane.
The organic layer was washed with water and concentrated under vacuum using rotary evaporator to give compounds (3a-c) as yellow
gummy mass in good yields.
The above substituted succinic acid half esters (5 g, 0.0144 mol) 3a-e were added to the stirred solution of methanol (20 mL)
and sodium hydroxide solution (20 mL) (2.8 g, 0.0722 mol). The resultant reaction mixture was refluxed at 75-80 C for 8 h. The
completion of the reaction was known by TLC. The reaction mixture was concentrated to give yellow residue. The resultant residue
was dissolved in 50 mL water. The aqueous layer was neutralized with dilute hydrochloric acid to obtain gummy light yellowish mass
and extracted to dichloromethane. The separated organic layer was washed with water, dried over anhydrous sodium sulphate and
concentrated under reduced pressure to afford light yellow gummy mass. The residue was purified by column chromatography on
silica gel (EtoAc and Cyclohexane 6:4) to afford colorless gummy compound. It was dissolved in diethyl ether and kept overnight in
fridge to get crystals and were filtered.
2-((4-methoxyphenyl)(thiophen-2-yl)methylene)succinic acid: 4a: White solid. Yield: 77 %. M.P.:121-1240 C. IR (KBr, v, Cm-1 ):
3200-3600 (OH), 1712 (Acid C=O), 1600 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.9 (s, 2H, CH2), 3.83 (s, 3H, OCH3), 6.77.66 (m, 7H, Ar-H phenyl and thiophen ring), 12.08 (bs, 2H, COOH). 13C NMR (100 MHz, CDCl3, , ppm):37.9, 57.2, 114.5, 121.3,
127.4, 128.3, 130.5, 132.4, 137.8, 161.3, 170.2, 172.3. MS (ESI, m/z): 319.3 (M+). Anal. Calcd. for C16H14O5S: C, 60.37; H, 4.43.
Found: C, 60.35; H, 4.41; S, 9.9 %.
(2-(phenyl(thiophen-2-yl)methylene)succinic acid: 4b: colorless oil. Yield: 78 %. IR (KBr, v, Cm-1 ): 3200-3500 (OH), 1710 (Acid
C=O), 1610 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 3.1 (s, 2H, CH2), 7.21-7.66 (m, 8H, Ar-H phenyl and thiophen ring). 13C
NMR (100 MHz, CDCl3, , ppm):33.3, 114.9, 127.2, 130.3, 133.3, 137.8, 151.7, 171.3, 178.7. MS (ESI, m/z): 289.09 (M +). Anal.
Calcd. for C15H12O4S: C, 62.49; H, 4.2. Found: C, 62.44; H, 4.15; S, 11.1 %.
2-(thiophen-2-yl(p-tolyl)methylene)succinic acid: 4c: Slight yellow oil. Yield: 69 %. IR (KBr, v, Cm-1): 3200-3400 (OH), 1711 (Acid
C=O), 1600 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.25 (s, 3H, CH3), 3.2 (s, 2H, CH2), 7.01-7.69 (m, 7H, Ar-H phenyl and
thiophen ring), 12.5 (bs, 2H, COOH). 13C NMR (100 MHz, CDCl3, , ppm):32.8, 118.3, 127.4, 127.9, 129.2, 130.4, 132.3, 137.7,
155.1, 177.1. MS (ESI, m/z): 303.06 (M +). Anal. Calcd. for C16H14O4S: C, 63.56; H, 4.67. Found: C, 63.45; H, 4.37; S, 10.34 %.
General procedure for the preparation of substituted 2-(phenyl(thiophen-2-yl)methyl)succinic acid 5a-c
In an autoclave vessel, substituted substituted 2-(phenyl(thiophen-2-yl)methylene)succinic acid (3 g, 0.0094 mol) 4a-c in
methanol (30 mL) and 10% Pd/C (0.3 g) were taken under nitrogen gas atmosphere. The reaction mixture was hydrogenated at 3-4
kg/cm3 hydrogen pressure at 25-30 C for 3-4 h. The Pd/C was filtered through hyflobed under nitrogen gas atmosphere. The mixture
was concentrated under reduced pressure. The products were recrystallized from ethanol.
2-((4-methoxyphenyl)(thiophen-2-yl)methyl)succinic acid: 5a: white solid. Yield: 68 %. M.P.:131-1330 C. IR (KBr, v, Cm-1): 35903200 (OH), 1710 (Acid C=O), 1606 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.78 (d, 2H, CH2), 3.48 (q, 1H, CH), 3.82 (s, 3H,
OCH3), 4.39 (d, 1H, CH), 6.8-7.3 (m, 7H, Ar-H phenyl and thiophen ring), 11.9 (bs, 2H, COOH). 13C NMR (100 MHz, CDCl3, ,
ppm): 35.2, 39.8, 56.7, 114.4, 123.2, 125.3, 126.2, 129.3, 131.2, 152.3, 158.2, 177.7, 178.2. MS (ESI, m/z): 321.3 (M +). Anal. Calcd.
for C16H16O5S: C, 59.99; H, 5.03. Found: C, 59.93; H, 5.01; S, 10.01 %.

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2-(thiophen-2-yl(p-tolyl)methyl)succinic acid: 5c: yellow gummy solid. Yield: 78 %. M.P.:152-1540 C. IR (KBr, v, Cm-1 ): 35603200 (OH), 1720 (Acid C=O), 1605 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.2 (s, 3H, CH3 ), 2.87(d, 2H, CH2), 3,8 (q, 1H,
CH), 4.3(d, 1H, CH), 6.92-7.1 (m, 7H, Ar-H phenyl and thiophen ring), 11.5 (bs, 2H, OH). 13C NMR (100 MHz, CDCl3, , ppm):22.4,
34.5, 47.2, 123.4, 124.4, 126.9, 128.3, 129.2, 133.3, 152.3, 177.1, 178.9. MS (ESI, m/z): 304.9 (M+). Anal. Calcd. for C16H16O4S: C,
63.14; H, 5.3. Found: C, 63.08; H, 5.2; S, 10.54%.

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2-(phenyl(thiophen-2-yl)methyl)succinic acid: 5b: white solid. Yield: 82 %. M.P.:111-1140 C. IR (KBr, v, Cm-1 ): 3550-3200 (OH),
1700 (Acid C=O), 1690 (, , unsaturated C=O), 1610 (Ar C=C). 1H NMR (400MHz, CDCl3, , ppm): 2.79 (d, 2H, CH2 ), 3.72(q, 1H,
CH), 4.5 (d, 1H, CH), 6.73-7.24 (m, 8H, Ar-H phenyl and thiophen ring). 13C NMR (100 MHz, CDCl3, , ppm):38.2, 116.1, 127.3,
128.3, 130.3, 132.4, 137.4, 153.2, 172.1. MS (ESI, m/z): 291.5 (M +). Anal. Calcd. for C15H14O4S: C, 62.05; H, 4.86. Found: C, 62.02;
H, 5; S, 11 %.

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General procedure for the preparation of substituted 4-oxo-7-phenyl-4,5,6,7-tetrahydrobenzo[b]thiophene-6-carboxylic acid 7a-c

Substituted 2-(phenyl(thiophen-2-yl)methyl)succinic acid (2 g, 0.0062 mol) 5a-c and acetyl chloride (20 mL) mixture was
refluxed for 6 h. The completion of the reaction was known by TLC. The excess of the acetyl chloride was distilled off under
atmospheric pressure. The resultant mixture was dissolved in dichloromethane (25 mL) and washed with 5% cold sodium bicarbonate
solution. The separated organic layer was finally washed with water and dried over anhyd sodium sulphate. Distilled off solvent to
afford a gummy mass 6a-c in good yields.
Above compounds 6a-c (2 g, 0.0065 mol) were dissolved in dichloromethane (20 mL) and added drop wise to the stirred
suspensions of anhydride aluminum chloride (2.2 g, 0.0164 mol) in dichloromethane (20 mL) at 0-5 C under inert atmosphere. The
reaction mixture was stirred for 8h at 25-30 C. The completion of the reaction was known by TLC. The reaction mass was poured
into 1 N HCl (40 mL) at 0-50C. The product was extracted into dichloromethane and then into saturated Na 2CO3 solution. The sodium
carbonate extract was acidified by dil. HCl to give yellow solid. The crude solids were purified by column chromatography on silica
gel (EtoAc and cyclohexane 8:4). The residue was dissolved in ethanol and kept overnight in fridge to get crystal and were filtered to
afford pure compound.
7-(4-methoxyphenyl)-4-oxo-4,5,6,7-tetrahydrobenzo[b]thiophene-6-carboxylic acid: 7a: Color: slight brown solid. Yield: 50 %.
M.P.:144-1480 C.IR (KBr, v, Cm-1): 1710 (acid C=O),1679 (tetralone C=O), 1585 (Ar C=C); 1H NMR (CDCl3-400MHz) ppm: 2.53
(dd, 2H, CH2 ), 3.42 (q, 1H, CH), 3.79 (s, 3H, -OCH3), 4.4 (d, 1H, CH), 6.92-7.34 (m, 6H, Ar-H phenyl and thiophen ring), 12.14 (bs,
1H, OH). ;13C NMR (CDCl3 -100 MHz) ppm: 36.4, 35.3, 45.2, 56.2, 117.4, 121.3, 125.4, 129.3, 136.2, 153.4, 154.3, 158.0, 179.2,
193.2. MS (ESI) m/z: 303.3 (M+). Anal. Calcd. for C16H14O4S; C, 63.56; H, 4.67. Found: C, 63.49; H, 4.56; S, 10.45 %.
4-oxo-7-phenyl-4,5,6,7-tetrahydrobenzo[b]thiophene-6-carboxylic acid: 7b: Color: slight brown solid. Yield: 61 %. M.P.:163-1670
C.IR (KBr, v, Cm-1): 1714 (acid C=O),1687 (tetralone C=O), 1590 (Ar C=C); 1H NMR (CDCl3-400MHz) ppm: 2.82 (dd, 2H, CH2 ),
3.39 (q, 1H, CH), 4.52 (d, 1H, CH), 6.63-7.4 (m, 6H, Ar-H phenyl and thiophen ring), 12.6 (bs, 2H, -COOH). ; 13C NMR (CDCl3 -100
MHz) ppm: 38.4, 44.7, 121.1, 125.8, 126.4, 129.3, 130.2, 140.1, 153.7, 155.3, 179.4, 195.3. MS (ESI) m/z: 273.3 (M+). Anal. Calcd.
for C15H12O3S; C, 66.16; H, 4.44. Found: C, 65.98; H, 4.42; S, 11.7%
4-oxo-7-p-tolyl-4,5,6,7-tetrahydrobenzo[b]thiophene-6-carboxylic acid: 7c: Color: slight brown solid. Yield: 64 %. M.P.:132-1340
C.IR (KBr, v, Cm-1): 1707 (acid C=O),1675 (tetralone C=O), 1610 (Ar C=C); 1H NMR (CDCl3-400MHz) ppm: 2.34 (s, 3H, CH3 ),
2.76 (dd, 2H, CH2 ), 3.47 (q, 1H, CH), 4.86 (d, 1H, CH), 6.92-7.24 (m, 6H, Ar -H phenyl and thiophen ring);13C NMR (CDCl3 -100
MHz) ppm: 23.9, 35.3, 37.4, 121.2, 127.4, 128.3, 129.9, 138.3, 154.8, 155.9, 179.2,194.3. MS (ESI) m/z: 286.35 (M+). Anal. Calcd.
for C16H14O3S; C, 67.11; H, 4.93. Found: C, 67.08; H, 4.83; S, 11.1 %.
Antimitotic study
The novel tetralone acid analogues of podophyllotoxin 7a-c were screened for antimitotic activity by onion root tip method
and the ID50 was determined. The materials required are acetoorcein solution, compound microscope, glass slides, cover slips,
hydrochloric acid (0.1 N), Carney's solution II, 70% ethanol and tested samples (100, 200 and 400 ppm). To study the effect of novel
tetralone acids analogues of podophyllotoxin on somatic cells, onion base was immersed to an extent of about half a centimeter in a
sample tube and control solution tube (73), after removing the old roots from them and immersion is continued for 24 h. for
germination. After different time intervals, the germinated root tips were removed and were fixed in Carney's solution II (alcohol and
acetic acid in 3:1 ratio respectively) for 24 h. After 24 h. Carney's solution II was decanted carefully and the root tips were washed
with preserving solvent (70% ethanol). The fixed root tips were preserved in 70% ethanol in refrigerator. The root tips were taken in
watch glass and stained with a drop of acetoorcein stain and a drop of 1 N HCl (7:1, v:v). The glasses were warmed and kept for 1 hr.
The roots were taken on a clean glass slide and squashed using 45% acetic acid following the method of Levan [11]. A microscope
cover glass was placed on the material and then pressure was applied on a cover glass to ensure uniform spreading. The cover glass
was sealed with molten paraffin wax and slide was observed under microscope. Mitotic Index (MI) was calculated by following
method of Fissceja [12]. The mitotic index was determined by examination of minimum of zone cells. Three replicates were made for
each calculation. The slides were observed under microscope and photographed.

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The percentage of the number of dividing cells compared to the control and the percent inhibition of mitosis by antimitotic
agent at a different concentration such as 100, 200, and 400 ppm against a control were calculated. The concentration needed for 50%
inhibition (ID50) was extrapolated from the graph of the concentration verses percentage inhibition. ID 50 values for novel tetralone acid
analogues of podophyllotoxin for antimitotic activity were calculated individually following Hakala [13] method.

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RESULTS AND DISCUSSION

Chemistry
The synthesis of novel tetralone acid analogues of podophyllotoxin has been carried by using Genslers method with some
changes in experimental conditions and reagent Scheme. Herein, we reported five step synthesis of tetralone acid analogues of
podophyllotoxin (7a-c) starting from the readily available starting materials substituted benzene 1a-c and 2-thiophenic acid chloride.
The substituted benzenes 1a-c were taken up for Friedel Craft acylation with 2-thiophenic acid chloride in presence of lewis acid
aluminum chloride to obtain desired para-acylated product 2a-c exclusively in good yields [14]. The structure of the compounds 2a-c
was confirmed by IR and 1H NMR spectral studies. IR spectra of the compounds 2a-c showed the C=O stretching frequency in the
range 1679-1700 cm-1 and 1H NMR showed aromatic protons of phenyl and thiophene ring at 7.2-7.8 ppm. The condensation of the
ketonic compounds 2a-c was carried out using diethyl succinate, sodium hydride in dry benzene and absolute ethanol in presence of
nitrogen gas atmosphere [15] through stobbe condensation to obtain the compounds 3a-c which on subjected into soponification in
sodium hydroxide and water mixture to furnish the compound 4a-c. The structure of the compounds 4a-c was confirmed by IR and 1H
NMR spectral studies. IR spectra of the compounds 4a-c showed the carboxylic acid -OH group stretching frequency at 3200-3600
cm-1 and 1H NMR showed -CH2 and di carboxylic acid peak at the range 2.9 and 12.08ppm respectively. Reduction of the double
bond in compounds 4a-c was carried by hydrogenation with 10% palladium carbon under 3-4 kg hydrogen pressure in methanol using
autoclave at room temperature to furnish the compounds 5a-c [16, 17]. In its IR spectra appeared absorption band in the range 35903200 cm-1 and 1606 cm-1 corresponds respectively to carboxylic acid -OH and Aromatic C=C stretching frequencies and 1H NMR
showed -CH proton in the range 4.39ppm. The compounds 6a-c were easily prepared on refluxing compounds 5a-c with acetyl
chloride. Subsequently, compounds 6a-c subjected into intramolecular acylation using lewis acid aluminum chloride in dry
dichloromethane [18, 19]. For its full characterization, these compounds purified by using ethyl acetate and were recrystallized by
ethanol. The compounds 7a-c exhibited C=O (tetralone) stretching band at 1679cm-1and proton NMR showed broad singlet carboxylic
acid peak at 12.14ppm. Based on this, the synthesized compounds were conformed.

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Antimitotic activity
As regards the relationships between the structure of the podophyllotoxin scaffold and antimitotic properties, it showed
varied antimitotic activity (Table 1). The presence of different substituents on the ring C causes a certain changes in activity. The
compound 7a and 7c has methoxy and methyl, moiety on ring C, which is accounted for the enhanced antimitotic activity than when
compared to control solution. On the other hand, the remaining compound 7b have showed less activity compared to control. From the
obtained results, it is clear that the major role for antimitotic activity is played by substituents on ring C moiety. It is evident that novel
tetralone acid analogues of podophyllotoxin were showed good antimitotic activity.

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Scheme: Protocol for synthesis of new tetralone acid analogues of podophyllotoxin 7a-c.

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Table 1. Antimitotic activity of the compounds (7a-c) by onion root tip method.
Comp.
no.
7a

7b

7c

Control

Conc. in
ppm
100
200
400
100
200
400
100
200
400
-

% dividing
cells
16.40
14.78
10.35
22.74
18.19
19.74
17.81
14.78
12.28
34.00

% dividing cells
compared to control
48.23
43.47
30.44
66.88
53.53
58.05
52.38
43.47
35.99
100

% inhibition
compared to control
51.77
56.77
69.56
33.12
46.50
41.95
47.62
56.77
64.01
0.0

ID50 in ppm

95

235

155

CONCLUSION
The novel tetralone acid analogues of podophyllotoxin (7a-c) were synthesized using following Genslers method with some
changes in reagents and reaction conditions. The structures of synthesized compounds were confirmed and characterized by analytical
datas such as IR, 1H NMR, 13C NMR, Mass spectra and elemental analysis. The novel tetralone acid analogues of podophyllotoxin
were synthesized by replacing methylene dioxy group in podophyllotoxin with thiophene ring and the trimethoxyphenyl ring with
substituted phenyl ring and lactone ring with carboxylic acid. It is interesting and significant to note from the data in table-1 that the
analogues 7a and 7c have increased activity, i.e., the compounds 7a and 7c with ID 50 of 95 ppm and 155 ppm respectively. The
compound 7c is less active than 7a and 7b with ID 50 value of 235ppm. Finally the antimitotic activity of the compound 7a with
methoxy substituent attached to the ring C exhibited more activity compared with remaining synthesized compounds.

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REFERENCES
[1]. Ahmedin Jemal D.V.M., Rebecca S., Elizabeth W., Yongping H., Jiaquan X., Taylor M., Michael J. T., Cancer Statistic. CA
Cancer J. Clin., 2008; 58: 7196.
[2]. Lee C.T., Lin V.C., Zhang S., Zhu X., VanVliet D., Hu H., Beers S.A., Wang Z., Mark Cosentino, L., Morris-Natschke
S.L., Lee K., Anti-AIDS agents. 291. Anti-HIV activity of modified podophyllotoxin derivatives, Bio. Organic. Medicinal.
Chemistry. Letter 1997; 7: 2897-2901.
[3]. Jordain I., Anticancer Agent Based on Natural Product Models. Cassady J. M., Douros J. D. Eds., New York: Academic Press
Inc; 1980.
[4]. Hartwell J.L., Schrecker A.W., The chemistry of podophyllum, Fortschritte der Chemie organischer Naturstoffe. 1958; 15: 83166.
[5]. Smissman E.E., Murray R.J., McChesney J.D., Houston L.L., Pazderni T.L., Synthesis and biological evaluation of certain
trans-2-aryl-trans-6-hydroxymethyl-3-cyclohexenecarboxylic acid .gamma.-lactones as antimitotic agents, Journal of Medicinal
Chemistry 1976; 19: 148-153.
[6]. Schreir E. A, 152nd National Meeting of the American Chemical Society. New York: 1966.
[7]. Anjanamurthy C., Lokanatha Rai K.M., Synthesis of podophyllotoxin and related analogs, synthesis of betaapopicropodophyllin analogs with modified lactone ring. Indian Journal of Chemistry Section B-Organic chemistry including
medicinal chemistry. 1987; 26(2): 131-135.
[8]. Ward R. S., The synthesis of lignans and neolignans. Chemical Society Reviews 1982; 11: 75-125.
[9]. Basavaraju Y.B., Anjanamurthy, C., Tetralone acids as intermediates for the synthesis of podophyllotoxin analogues. Indian
Journal of Chemistry 2003; 42B: 876-880.
[10]. Gensler W.J., Samour C.M., Wang S.Y., Johnson F., Compounds related to podophyllotoxin. X. Synthesis of picropodophyllin.
Journal of American Chemical Society 1960; 82: 1714-1727.
[11]. Levan A., The effect of colchicine on root mitosis in Allium, Hereditas 1938; 24: 471-86.
[12]. Fissceja G., The Allium test on a standard in environmental monitoring, Hereditas 1985; 102: 099-12.
[13]. Hakala TR, Lange PA, Fraley EF. Method in vitro in all mediated and tumor Immunity. In Bloom BR and David JR (Ed.). New
York: Academic Press; 1976.p.451.
[14]. Basavaraju Y.B., Hemakumar K.H., Sathish, A. D., Synthses and characterization of new diketone analogues of
podophyllotoxin. E-Journal Chemistry 2008 ; 5 : 114-119.
[15]. Daub G. H., Johnso, W. S., The Stobbe condensation with sodium hydride, Organic reactions, John Wiley & Sons, New York:
1950 pp.
[16]. Siegel S., Dunkel M., Smith G.V., Halpern W., Cozort, J., Stereochemistry and the mechanism of catalytic hydrogenation of
cycloalkenes. VII. Interaction mechanisms which control the ratio of stereoisomers, Journal of Organic Chemistry 1966; 31:
2802-2806.

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ACKNOWLEDGEMENT
The authors are thankful to University of Mysore for providing spectral data.

Vol 4, Issue 12, 2014.

Santhekasalagere B. Shivakumar et al.

ISSN NO: 2231-6876

[17]. Gault, F.G., Rooney, J.J., Kemball, C., The catalytic exchange with deuterium of polymethylcyclopentanes on metal films.
Evidence for -bonded intermediates. Journal Catalysis 1962; 1: 255-274.
[18]. Gensler W.J., Murthy C.D., Trammell M.H., Nonenolizable podophyllotoxin derivatives. Journal of. Medicinal Chemistry
1977; 20: 635-644.
[19]. Gensler, W.J., Gatsonis C.D., Synthesis of podophyllotoxin. Journal of Organic Chemistry 1966; 31: 4004-4008.

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