Synhtesis, Characterization and Cytotoxic Activity of ...shodhganga.inflibnet.ac.in/bitstream/10603/21199/16/22 r paper -1... · Synhtesis, Characterization and Cytotoxic Activity

RGUHS J PharmSci |Vol 3 | Issue3 | Jul–Sep, 2013 65

50

50

Synhtesis, Characterization and CytotoxicActivity of Analogues of 3- MethylFlavone

Afroze Alam1, K. L. Dhar,1 B. S. Jayshree2

1School of Pharmaceutical Sciences, Shoolini University, Bajhol Solan-173229 Himachal Pradesh,India2Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Udupi, Karnataka, India

Research Article

ABSTRACTThe flavonoids are polyphenolic compounds and universally present as constituents of flowering plants, particularlyof food plants. The flavonoids are phenyl substituted chroman (benzopyran derivatives) consisting of a 15-carbonbasic skeleton (C -C -C ), composed of a chroman (C -C ) nucleus (the benzo ring A and the heterocyclicring 6 3 6 6 3C), also shared by the tocopherols, with a phenyl (the aromatic ring B), substitution usually at the 2-position.Different substitutions can typically occur in the rings, A and B. Several plants and spices containingflavonoid derivatives have found application as disease preventive and therapeutic agents in traditional medicinein Asia for thousands of years. Various analogues of 3-methylflavones were synthesised, purified andcharacterised by UV, IR,1HNMR and mass spectrometry. All the synthesised compounds were evaluated for theircytotoxic activityby MTT assay against HeLa cell line. Most of the compounds show moderateactivity,whereas thecompounds likeFA-04, FA-20, FA-10 and FA-13 have shown good activity having IC value 22.00, 25.49.26.24 and 26.35 µg/ml respectively, in comparison to standard molecule Cisplastin had IC value 6- 12.5µg/ml. Thus it appears thepresence of 3-methyl group in the flavone nucleus significantly influences the log ‘P’ value of all the twenty testcompounds. The prominent activity the of above compounds is probably because of 3-methyl and N, N-dimethylamino substitution on 4’-positionof flavonoid nucleus.

Keywords: Benzopyrone, Cytotoxic activity, Flavonoids, HeLa cell line, log ‘P’ value, MTTassay.

1. INTRODUCTIONSuccess in the drug discovery is dependenton the ability to identity novel, patentablecompounds collectively called as NewChemical Entities (NCEs) that have poten-tial to treat a disease in a safe and efficaciousmanner.1

Benzopyrone class of compounds occursin nature as flavones, isoflavones, neofla-vones, coumarins, etc. Depending upontype of lactone ring, benzopyrone areavailable as α-benzopyrone, b-benzopy-rone and γ-benzopyrone. Among themα-benzopyrone (coumarines) and γ- benzo-pyrone (flavones, isoflavones, and neofla-vones) occurs widely in nature.Compoundshaving Chromones (γ-benzopyrone) moietyare associated with interesting physiological

activities such as antibacterial antiviral,anticancer, antioxidant, antifungal, anti-cholestermic, anti-diabetics, anti-allergicdiuretics etc.2 Flavonoids, thederivatives of chromones arepolyhydroxylated compounds, and theyare capable of selectively reacting with freeradicals or system related to the inductionof inflammatory process. Quercetin (3,3’, 4’, 5’, and 7-pentahy- droxy flavones)and related flavonoids are known to inhibitthe growth of tumor cell and potentiatesthe cytotoxicity of DNA demanding anti-cancer drugs such as cisplastin.3 Very fewreports are available on the influence oflipophilic substitution on the antioxidantsor anti-inflammatory activities of this classof natural products.4 Flavones inhibitCYPIA-mediated 7-ethoxyresurufin

Received Date : 14-09-2013Revised Date : 10-10-2013Accepted Date : 14-10-2013

DOI: 10.5530/rjps.2013.3.9

Address forcorrespondenceAFROZE ALAMSchool of Pharmacy, ShooliniUniversity,BajholSolan India.Mobil No: +918091974886Fax No: 01792308000Email:[email protected]

www.rjps.in


50

50




Research Article







DOI: 10.5530/rjps.2013.3.9


www.rjps.in


50

50




Research Article







DOI: 10.5530/rjps.2013.3.9


www.rjps.in

mailto:[email protected]

www.rjps.in


www.rjps.in



Afroze Alam, et al.: Synhtesis, Characterization and Cytotoxic Activity of Analogues of 3- Methyl Flavone

5 5

5o-dethylase (EROD)activity in rat and human liver5

microsomes. Certain bromoflavones are found to sig- 5 2+

nificantly induce quinine reductase activity, which is animportant mechanism of chemoprevention.6 On study &+of an inhibitor from the class of xanthone, chromoneand flavone reports are available in the literature to beararomatase7 inhibition properties. Cancer is the causeof more than six million deathseach year in the world.In 2001, about 1,268,000 newcancer cases and 553,400deaths were reported in theUnited States.8 For a longtime, plants are being used in the treatment of cancer.9

2

(W2+

2+&

.2+

5

According to an estimate, 50% of breast cancer and37% of prostate cancer patients use herbal products.10

In current study, 20 derivatives of 3-methyl flavoneshave beensynthesized with varioussubstitution on ben-zopyrone nucleus. The starting material was taken as

5 2+

2

5&+

5

$& $&

few substituted propiophenone and substituted benz- 5 2+ 2&+ 5 &O &+ 2&+

aldehyde. All the synthesized compounds were purified, 5 2+ 2&+ &+ &O ) 5 2+ &O ) 2&+ &+characterized and evaluated for cytotoxic activity on theHeLacell line.

2. MATERIALS AND METHODS

2.1 ReagentsAll the chemicals and solvents used were of AR-gradeand LR-grade and obtained from Sigma-Aldrich, SiscoResearch Laboratories, Qualigens, Rankem, S.D. Fine,Hi-Media and Merck and wereused without furtherpurification.2.1.1 Synthesis of Substituted 3-methyl Chalcones.11

(CA1–CA20)To a solution of 0.01 mole of orthohydroxy Propiophe-nones in 10 ml of 40% KOH and 20 ml of ethyl alco-hol, 0.01 mole of substituted benzaldehyde was addedand mixture was stirred for 48–72 hours. The colouredsolution was poured into crushed ice and acidified with1N HCl. The precipitate so obtained was washed withcold water, filtered, dried and recrystallized with abso-lute alcohol. Percentage yield and other physical data arepresent in Table No; 1.2.1.2.Experimental &TLC analysisMelting points were determined on a melting pointapparatus (Shital Scientific Industries, Mumbai) and areuncorrected. The reactions were monitored by Thin-

(HPLC grade). The FTIR studies were done on a Shi-madzu FTIR8310 spectrophotometer as KBr pellets.1HNMR spectra were taken on a NMR (AMX400) inDMSO-d6 (Merck) andthe mass spectra were recordedon a GC-MS(Shimadzu GCMSQP5050 corporationJapan.)2.2.1 Synthesis of Substituted 3-Methyl Flavones12

(FA1–FA20)To a solution of 0.01 mole of chacone in a 50 ml ofdimethyl sulphoxide (DMSO) taken in a 100 ml ofround bottom flask ,fitted with reflux condenser wasadded of 15–20 granules of iodine. The reaction mix-ture was reflux for 3–4 hours and kept overnight. Theprecipitate was neutralized with sodium thiosulphate toremove unreacted I2 washed with water, filtered, driedand recrystallized with absolute alcohol. Percentageyield and other physical data are present in Table No; 3.2.2.2 General Scheme of Synthesis

5HDFWLRQ

5

layer chromatography (TLC) was performed on pre-coated aluminium plates (Silica gel 60 F254, Merck).Plates were visualized by UV light and iodine vapourand the Rf values were determined on pre-coated TLCusing the solvent system n-hexane; ethyl acetate (9: 1).max ‘and €max for the synthesized test compounds were

5

5 2+

2

5

5

&+

'062 ,

5()/8;

5 5

5 2

&+

2

obtained on a UV-visible spectrophotometer (ShimadzuUV, Vis spectrophotometer UV-1650 PC) in methanol

5 2+ 2&+ 5 &O &+ 2&+

5 2+ 2&+ &+ &O ) 5 2+ &O ) 2&+ &+

www.rjps.in



FA-05 H CH3 H H

FA-06

FA-07

FA-08

H

H

H

Cl

H

H

H

H

H

H

OCH3

OH

3

3 2

3 2

3

2

2.1.3 Physical data of Synthesized 3-methyl Chalcones (CA1–CA20) Table-1Compound Yield (%) M.P. (0C) max € max Rf* logP

AC-01 73 75-80 266.7 1.048x104 0.72 1.03AC-02 62 100-105 265.9 2.39x104 0.68 1.69

AC -03 72 65-70 301.4 4.28x103 0.61 1.00

AC -04 73 80-85 280.0 1.35x104 0.64 1.21

AC -05 80 120-125 292.7 3.28x104 0.54 1.05

AC -06 68 110-112 256.8 1.76x104 0.70 1.24

AC -07 72 135-140 310.0 1.33x104 0.64 1.06

AC -08 64 112-115 221.0 6.05x103 0.61 0.61

AC -09 60 130-135 268.0 3.19x104 0.40 1.89

AC -10 85 65-75 230.0 6.55x103 0.33 1.05

AC-11 80 116-118 280.0 1.05x104 0.54 1.20

AC-12 85 85-90 279.8 1.06x104 0.46 1.15

AC -13 80 70-75 284.0 7.507x103 0.48 1.30

AC -14 70 80-85 270.0 2.84x104 0.34 1.34

AC -15 55 107-110 274.0 6.39x103 0.53 1.37

AC -16 65 256-259 291.5 5.23x104 0.44 1.63

AC -17 80 1252-253 300.0 1.072x104 0.37 1.70

AC -18 80 267-270 382.5 1.60x104 0.59 1.75

AC -19 70 270-73 352.0 1.340x104 0.68 1.05

AC -20 60 250-253 310.0 2.620x104 0.55 1.04

Satisfactory CHN data.*Solvent system-n-hexane: ethyl acetate (9: 1)

2.2.4 Physical data of synthesised 3-methylflavones (FA1-FA20). Table No: 3

2.2.3 List of Synthesized Substituted Flavones; TableNo: 2

Compound Yield(%)

M.P. (0C) max € max Rf* LogP

Compound R R1 R2 R3

FA-01 H H H N(CH )

FA-02 H H H CH

FA-03 H H H H

FA-04 OH H H N (CH )

FA-1 72 120-125 338.0 1.026x104 0.531.57

FA-2 80 125-132 339.0 1.524x104 0.441.34

FA-3 55 110-112 298.4 6.621x103 0.331.01

FA-4 65 160-170 339.0 1.707x104 0.831.22

FA-5 72 117-120 253.2 1.663x104 0.451.33

FA-6 90 95-100 282.4 5.450x104 0.571.03

FA-7 85 90-95 312.0 8.764x104 0.381.36

FA-8 60 105-108 225.0 8.496x104 0.860.93

FA-9 45 160-110 273.4 3.155x104 0.531.66

FA-09 OCH H H Cl FA-10 85 65-75 235.0 7.785x194 0.331.20

FA-10 H H H NO

FA-11 H Br H H

FA-12 H H H Cl

FA-13 H H H F

FA-14 H H H Br

FA-11 72 82-85 282.4 1.344x104 0.530.96

FA-12 88 70-75 281.8 1.689x104 0.441.03

FA-13 90 90-95 285.0 9.587x103 0.331.10

FA-14 85 120-125 278.8 1.203x104 0.831.05



3

3

3

3

FA-15 72 85-90 275.0 2.048x104 0.451.16

FA-16 65 139-142 292.4 1.944x104 0.770.94

FA-15 OCH H H HFA-17 60 105-110 323.6 2.378x104 0.55

1.62FA-16 H F H H

FA-17 OH H H CH

FA-18 H OCH H H

FA-18 60 129-132 347.4 2.405x104 0.541.12

FA-19 65 121-125 341.9 1.340x104 0.561.61

FA-20 60 115-120 312.8 2.620x104 0.430.91

FA-19 H H OCH H

FA-20 H OH H H The compounds gave satisfactory CHN data.*Solvent system - n-hexane : ethyl acetate (9: 1)



2

2

3. CYTOTOXIC ACTIVITY13-15

MTT assay is a laboratory test and a standard colomet-ric assay (an assay which measures changes in colour) formeasuring cellular growth. It can also be used to deter-mine cytotoxicity of potential medicinal agents and othertoxic materials. Yellow MTT (3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide, a tetrazole) isreduced to purple formazan in the mitochondria of liv-ing cells. A solubilisation solution (usually either dimethylsulphoxide, an acidified ethanol solution, or a solution ofthe sodium dodecyl sulphate in dilute hydrochloric acid,0.04N acidified isopropyl alcohol is added to dissolve theinsoluble purple product into a coloured solution. Theabsorbance of this coloured solution can be quantified by

MTT.Plates are incubate (370C, 5% CO ) for next 4hr.Flick off MTT and insoluble formazan product isdis-solved in 100μl of DMSO.Leave at room temperaturefor a few minutes to ensure all crystals are dissolved.Read plate using a wavelength of 540 nm. Be sure toread plates within one hour of adding the DMSO.Reagents MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphe-nyl tetrazolium bromide (Sigma #M2128): dissolve at1 mg/mL in PBS, filter and sterilize. PBS = Phosphatebuffer saline pH 7.4.3.1.1 Growth inhibition of HeLa cells.Table No: 4All the synthesized test compounds (FA-01–FA-20)were screened for their cytotoxic activity. They werescreened by MTT assay method. The 50% cytotoxic

measuring at wavelength (540 nm) by ELISA reader. This concentration (CTC50(mcg/ml) ) of the all test com-reduction takes place only when mitochondrial reduc-tase enzymes are active, and therefore conversion can bedirectly related to the number of viable (living) cells.

3.1 ExperimentalAdd 100 μl of cells to a 96 well plate at a cell density of5-10x 10 4 cells per well. Incubate (37 0C, 5% CO ) over-night to allow the cells to attach to the wells. Flick offold media. Add 100 μl of (drug+media) solution in eachwell according to dilution.Same volume also keeps ofblank, positive & negative control. Incubate (370C, 5%CO2) for next 48–72 hr. to allow the cells to attach to thewells.Flick off old media and add 100μl (1 mg/ml) of

pounds are enlisted in table no; 5.

4. RESULTS ANDDISCUSSIONPreparation of various substituted 3-mety chalconebased on Claisen- Schmidt condensation20. The start-ing materials for the synthesis of various substituted3-methyl chalcones were 2-hydroxyl propiophenoneand substituted benzaldehyde. Substituted 3-methyl fla-vones were synthesized from respective chalcones in thepresence of DMSO/I2. The purified compound s afterrecrystallization were determined for their purity, bymelting point, thin layer chromatography, and structure

Compound % Growth inhibition

6.25µg/ml 12.50 µg/ml 25 µg/ml 50 µg/mlFA-1 4.52 9.08 41.45 91.62

FA-2 3.67 12.38 4 2.25 88.18

FA-3 0.9 7.46 38.17 85.64

FA-4 6.56 18.85 73.00 93.85

FA-5 1.41 4.77 14.29 58.19

FA-6 2.96 13.88 22.56 39.48

FA-7 2.78 10.28 40.58 87.49

FA-8 1.30 5.50 21.43 39.82

FA-9 0.88 2.66 16.08 29.76

FA-10 3.70 17.54 40.03 86.64

FA-11 3.80 19.29 40.35 86.68

FA-12 4.30 7.00 37.72 91.80

FA-13 8.77 31.75 47.70 91.90

FA-14 5.85 33.90 58.00 82.10

FA-15 2.90 28.50 60.00 92.20

FA-16 0.90 07.00 31.90 75.20

FA-17 0.52 04.25 27.80 72.90

FA-18 0.67 03.19 25.70 89.50

FA-19 2.10 8.80 39.90 84.50

FA-20 7.25 16.90 40.50 90.75



Compounds CTC (50 cg/ml)FA-01 46.89

FA-02 45.45

FA-03 43.41

FA-04 22.00

FA-05 44.32

FA-06 46.82

FA-07 34.56

FA-08 32.89

FA-09 33.56

FA-10 26.24

FA-11 30.41

FA-12 30.50

FA-13 26.35

FA-14 44.38

FA-15 49.92

FA-16 53.48

FA-17 55.40

FA-18 40.66

FA-19 43.94

FA-20 25.49

Table No: 5 4.1 Major Molecular Mechanism of ActionStudies in vitro and in vivo have shown that someflavonoids modulate the metabolism and dispositionofcarcinogens and can contribute to cancer prevention.21–25

One important mechanism by which flavonoids mayexert their effects is through their interactionwith phaseI metabolizing enzymes (e.g., cytochrome P450), whichmetabolically activate a largenumber of procarcinogensto reactive intermediates that can interact with cellularnucleophiles andultimately trigger carcinogenesis. Fla-vonoids are demonstrated to inhibit the activities of cer-tain P450 isozymes such as CYP1A1 and CYP1A2, 26-27

Thus, they are likely to have a protective role againsttheinduction of cellular damage by the activation of carcin-ogens. Another mechanism of action is the inductionof phase II metabolizing enzymes such as glutathione-S-transferase, Quinone reductase, and UDP-glucuronyltransferase,28-29 by whichcarcinogens are detoxified andthus more readily eliminated from the body. This wouldalso help to explain the chemo preventive effects offlavonoids against carcinogenesis.Moreover, some fla-vonoids have been reported as potent aromatase inhibi-tors.30-33 Substantialevidence supports the concept thatoestrogens be involved inmammary carcinomas. Estra-

were established by IR, 1H-NMR, and Mass spectralstudies. The yields of final synthesized compounds werein the range of 60-90%. max and €max were found werefound to be in the range of 225–347 nm and 6.621x103-8.49x104

respectively.

diol, the mostpotent endogenous oestrogen, is bio-synthesized from androgens by the cytochrome P450enzymecomplex called aromatase. Inhibition of aroma-tase is an important approach for reducing growthstim-ulatory effects of oestrogens in hormone-dependent

30

In the present work, effect of 3- methyl substitution on the breast cancer. Therefore, flavonoids couldbe consid-

solubility of flavones was studied. Out of the twenty methylflavones subjected for ‘P’ value determination, almost allof them showed log P value16-19 in the range of 0.912-1.66,as shown in the table no.3. Thus, it appears the presence ofmethyl group in the flavones significantly influences on the‘P’ value of all the twenty test compounds.Cytotoxic studies was determined by MTT assay at awavelength 540 nm .All test compounds (FA1-FA20)were screened for cytotoxic evaluation .Most of the testcompounds showed moderate cytotoxic activity againstHeLa cell line. All twenty test compounds showedCTC50 value between 22.00- 55.40µg/ml. Enlisted intable no; 5.The standard Cisplastin had a value of 6.5–12µg/ml.The test compounds FA-04, FA-20, FA-10, and FA-13showed CTC50 value between 22.00–26.38 µg/ml. Thetest compounds FA-11, FA-12, FA-08, FA-09, FA-07and FA-18 showed CTC50 value between 30–40 µg/ml. The test compounds FA-01, FA-02, FA-04, FA-05,FA-06, FA-14, FA-15, and FA-19 CTC50value between40–50 µg/ml. The test compounds FA-17 and FA-16CTC50 value between 50-56µg/ml.

ered potential agents against breast cancer through theinhibition of aromatase activity.

5. CONCLUSIONCytotoxic activities of all test compounds wereevalu-ated. All test compounds showed activity at all theconcentrations (6.25, 12.50, 25.00 50 µg/ml), prob-ably because ofbenzopyrone nucleus/flavonoids ring.According to the literature review compounds isolatedfrom plant sources havingbenzopyrone/flavonoidsringshow promising cytotoxic34-36 activity. The distancebetween two phenyl ring A and B of our test com-pounds is very much comparable to that of flavonols,flavones, isoflavones and flavanones (Flavonoids fam-ily) obtained from the natural products like kaempferol,chrysin, daidzein and hesperitin respectively as all arepotent anticancer agents.34-36 The structural activityrelationship of our test compounds showed cytotoxicactivity based on above observation and their probablemolecular mechanism. Thusthere is much evidence thatflavonoids have important effect on inhibitory carcino-genesis. Now we conclude that flavonoids are generally



non-toxic, non-steroidal and manifest a diverse rangeofbeneficial biological activities.

6. FUTURE SCOPE3-methyl flavones may become the probable potentialclass of compounds for future study. The recent reportsavailable on analogues of 3-methyl flavones for theirglutathione transferase37 and farnesyltransferase38

inhibitors, will initiate our study to synthesize manymore of such substituted flavones and test them forsuch activity, there is a future scope for the syntheticflavones to be targeted for newer targets such as CYP-450 aromatase to exploit their anticancer activity.

7. ACKNOWLEDGEMENTAuthors thank Saif Central Instrumentation Labora-tory, Punjab University, Chandigarh for getting spectraldata, like NMR and Mass spectra and School of Phar-maceutical Sciences, Shoolini University for researchfacilities.

(2H, d, 2”,), 2.35(3H,s,H-6”), 2.05(1H,S,3-H), 7.64(1H,d, 5’-H), 7.03 (1H, t,6’- H), 7.34(1H,t, H-7’), 6.90(1H, d,H-8’); GCMS: m/z 253 (M+1).FA-06 IR (KBr) cm-1 ( C-H) 2867, (ArC=C) 1573, (ArC-H) 3072, (C=O) 1690, (C-Cl) 710. ,1HNMR (400 MHzCDCI3); δ 7.04 (1H,t,H-4”-H), 7.09 (2H,t H-3”-H),7.22 (1H, d, 5’’-H), 7.24 (1H,d,2”-H) 2.95 (1H,S,3-H),7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’),7.00 (1H, d, H-8’); GCMS: m/z 273.5 (M+1).FA-07 IR (KBr) cm-1 (C-H) 2917 ,(ArC=C) 1570, (ArC-H) 3080, (C=O) 1685, (C-O) 1209;, 1HNMR(400MHz CDCI3); δ 3.70 (3H,s,H-4”-O-( CH3),6.73(2H,d H-3”,5”-H), 7.22 (2H, d,2”,6”-H),1.95(1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H),7.36 (1H,t, H-7’), 7.00 (1H, d, H-8’); GCMS:m/z268 (M-2).FA-08 IR (KBr) cm-1 (C-H) 2907 ,(ArC=C) 1580, (ArC-H) 3060 ,(C=O) 1675, (O-H) 3310:, 1HNMR(400MHz CDCI3); δ 2.40 (1H,s,4”-OH), 6.63 (2H, dH-3”,5”-H), 7.13 (2H, d, 2”,6”-H), 1.95 (1H,S,3-H), 7.64(1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’), 6.92(1H, d, H-8’); Molecular ion peak at m/e = 253(M+)

-18. SPECTRAL DATA FA-09 IR (KBr) cm (C-H) 2860, (ArC=C) 1560,

FA-01 IR (KBr) cm-1(C-H) 2794, (ArC=C) 1599,(Ar C-H) 3019 ,(C-N )1290 cm-1

(C=O) 1660;, 1HNMR(400 MHz CDCI3); δ 2.80(6H,s,H-4”-N-( CH3)2 ), 6.53(2H,d H-3”,5”-H), 7.13

(Ar C-H) 3092, (C=O) 1680, (C-Cl) 700. (C-O) 1209,1HNMR (400 MHz CDCI3); δ 7.23 (2H,d H-3”,5”-H),7.23 (2H, d, 2”,6”-H), 2.95 (1H,S,3-H), 7.54 (1H, d,5’-H), 7.08 (1H, d,6’- H), 3.73(3H,s,OCH3-7’), 6.40 (1H,d, H-8’); GCMS: m/z 306(M+2).

(2H, d, 2”,6”-H), 1.95 (1H,S,3-H), 7.66 (1H, d, 5’-H),7.0 (1H, dd,6’- H), 7.36 (1H,dd, H-7’), 6.90 (1H, d,H-8’); GCMS: m/z 281(M+1).FA-02 IR (KBr) cm-1:3178 (ArC-H), 2813 (C-H), 1680(C=O), 1570 (C=C); ,1HNMR(400 MHz CDCI3 ); δ2.40 (3H,s,H-4”-CH3), 7.03 (2H,d H-3”,5”-H), 7.23

FA-10 IR (KBr) cm-1 (C-H) 2910, (ArC=C) 1565, (ArC-H) 3069, (C=O) 1680, (C-N) 1295, 1HNMR (400MHz CDCI3); δ 8.03 (2H,d H-3”,5”-H), 7.53 (2H,d, 2”,6”-H), 1.95 (1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08(1H, t,6’- H), 7.36 (1H,t, H-7’), 6.930 (1H, d, H-8’);GCMS:m/z 284(M+1).

(2H, d, 2”,6”-H), 2.95 (1H,S,3-H), 7.64 (1H, d, 5’-H),7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’),7.00 (1H, d, H-8’);;GCMS: m/z 254(M+2)

FA-03 IR (KBr) cm-1:3072 (ArC-H), 2800 (C-H), 1690(C=O), 1580 (C=C); ,1HNMR(400 MHz CDCI3 );δ 7.14 (3H,s,H-4”-H), 7.21(2H,d H-3”,5”-H),7.30(2H, d, 2”,6”-H),2.0(1H,S,3-H), 7.64(1H, d, 5’-H),7.01 (1H, t,6’- H), 7.37(1H,t, H-7’), 6.90(1H, d, H-8’);GCMS :m/z 238 (M+1.).

FA-04 IR (KBr): 3390 (O-H), 3100 (ArC-H), 2800 (CH),1682 (C=O), 1561 (C=C), 1312 (C-N);, 1HNMR(400MHz CDCI3); δ 2.70 (3H,s,H-4”-N-(CH3)2), 6.93 (2H,d

FA-11 IR (KBr) cm-1(C-H) 2854 ,(Ar C=C) 1500,(ArC-H) 3090 ,(C=O) 1718, (C-Br) 604, (Ar=C-H) 801,1HNMR (400 MHz CDCI3); δ 7.40 (1H,t,H-4”), 7.33(2H,d H-3”,5”-H), 7.20 (2H, d, 2”-H),1.95 (1H,S,3-H),7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’),7.00 (1H, d, H-8’); GCMS: m/z 318(M+2).FA-12 IR (KBr) cm-1 (C-H) 2807, (ArC=C) 1553, (ArC-H) 3172 ,(C=O) 1700, (C-Cl) 700. 1HNMR (400MHz CDCI3); δ 7.23 (2H,d H-3”,5”-H), 7.24 (2H, d,2”,6”-H), 1.95 (1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H,t,6’- H), 7.36 (1H,t, H-7’), 6.92 (1H, d, H-8’GCMS: m/z274.5 (M+2).

H-3”,5”-H), 7.20 (2H, d, 2”,6”-H), 2.09 (1H,S,3-H),7.54 (1H, d, 5’-H), 6.88 (1H, t,6’- H), 5.0(1H,s, H-7’),6.390 (1H, d, H-8’);); GCMS: m/z 295 (M+1).FA-05IR (KBr) cm-1(C-H) 2794 ,(Ar C=C) 1589, (ArC-H) 3029 ,(C=O) 1660;, 1HNMR(400 MHz CDCI3);δ 7.04 (3H,s,H-4”-H ), 7.02(2H,d H-3”,5”-H), 7.28

FA-13 IR (KBr) cm-1 (C-H) 2790, (ArC=C) 1508 ,(ArC-H) 3142, (C=O) 1710, (C-F) 1198., 1HNMR (400MHz CDCI3); δ 6.73 (2H,d H-3”,5”-H), 7.27 (2H, d,2”,6”-H), 1.95 (1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H,t,6’- H), 7.3 (1H,t, H-7’), 6.91.00 (1H, d, H-8’); GCMS:m/z 257(M+1).



FA-14 IR (KBr) cm-1 (C-H) 2890, (ArC=C) 1550, (ArC-H) 3092, (C=O) 1700, (C-Br ) 698. ( Ar=C-H), 785;,1HNMR (400 MHz CDCI3); δ7.31(2H,d H-3”,5”-H),7.19 (2H, d, 2”,6”-H), 1.95 (1H,S,3-H), 7.64 (1H, d,5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’), 7.00 (1H, d,H-8’); GCMS: m/z 318(M+2).FA-15 IR (KBr) cm-1 (C-H) 2920, (ArC=C) 1572, (ArC-H) 3082, (C=O) 1680, (C-O) 1200, 1HNMR (400MHz CDCI3); δ 7.14 (1H,t,H-4’’), 7.31 (2H,dd H-3”,5”-H), 7.33 (2H, d, 2”,6”-H), 1.94 (1H,S,3-H), 7.54 (1H, d,5’-H), 6.52 (1H, d,6’- H), 3.74 (3H,t, H-7’), 6.43 (1H, s,H-8’); GCMS: m/z 268 (M-2).FA-16 IR (KBr) cm-1 (C-H) 2810, (ArC=C) 1505, (ArC-H) 3140, (C=O) 1719, (C-F) 1200., 1HNMR (400MHz CDCI3); δ 7.12 (1H,t,H-4”), 6.98 (2H,d H-3”)6.92 (1H,d,5”-H), 7.28 (2H, d, 2”-H), 1.95 (1H,S,3-H),7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t, H-7’),7.00 (1H, d, H-8’); GCMS: m/z 257(M+1).FA-17 IR (KBr) cm-1:3175 (ArC-H), 2803 (C-H), 1690(C=O), 1570 (C=C); 3360 (ArO-H), 1HNMR (400 MHzCDCI3); δ 2.40 (3H,s,H-4”-CH3), 7.02 (2H,d H-3”,5”-H), 7.16 (2H, d, 2”,6”-H), 1.95 (1H,S,3-H), 7.47 (1H, d,5’-H), 6.48 (1H, d,6’- H), 5.(1H,s,OH-7’), 6.39 (1H, s,H-8’); GCMS: m/z 254(M+2)

FA-18 IR (KBr) cm-1 (C-H) 2915, (ArC=C) 1576, (ArC-H) 3109, (C=O) 1680, (C-O) 1206; 1HNMR (400MHz CDCI3); δ 7.03 (3H,t,H-4”), 7.03 (2H,d H-3”,5”-H), 7.23 (2H, d, 2”-H), 3.73 (3H,s,6’’-H) 1.95 (1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36 (1H,t,H-7’), 7.00 (1H, d, H-8’); GCMS: m/z 268(M-2).FA-19 IR (KBr) cm-1 (C-H) 2917, (ArC=C) 1570, (ArC-H) 3086, (C=O) 1683, (C-O) 1208; 1HNMR (400MHz CDCI3); δ 6.63 (3H,d,H-4”), 7.10 (2H,t H-3”)3.37 (3H,s,,5”-H), 6.83 (1H, d, 2”-H), 6.80 (1H,s,6’’-H)1.95 (1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H),7.36 (1H,t, H-7’), 7.00 (1H, d, H-8’); GCMS: m/z 268(M-2).FA-20 IR (KBr) cm-1 (C-H) 2900, (ArC=C) 1584, (ArC-H) 3064, (C=O) 1670, (O-H) 3300; 1HNMR (400MHz CDCI3); δ 6.93 (1H,t,H-4”), 6.77 (2H,t H-3”) 6.68(1H,d,5”-H), 7.13 (1H, d, 2”-H), 5.0 (1H,s,6’’-H) 1.95(1H,S,3-H), 7.64 (1H, d, 5’-H), 7.08 (1H, t,6’- H), 7.36(1H,t, H-7’), 6.92 (1H, d, H-8’); Molecular ion peak atm/e = 253(M+).

REFERENCES1. Foye WO. Principle of Medicinal chemistry. 3rd Ed. Verghese Publishing

house, Bombay; 1989.2. Karle BK, Hangargo RV ,Mane AS, Gill CH, Shingare MS. Ind.J.

Hetrocyclic chem. 2001; 11:81–82.3. Fritz B, Tobias S, Albrechts K, Charlotte B, Kent C, Anni A, Franz

J Joseph K. American Soc. Biochem and Mol. Bio. 1996 Jan;271(4):2262–2270.

4. Finar IL. Organic Chemistry volume 2. 5th ED. Pearson EducationAsia

Ltd.5. Ahluwalia VK,Parashar RK. Organic reaction, mechanism and

applications. 1st Ed .Narosa Publishing House.6. Lynda LS, Jerome KW, Sang K, Lee CG, Dial L, Richard CM, Robert

MM, John MP. Cancer Research. 1991 Feb;59.

7. Silvia G, Cavalli A. Cytochrome P450 aromatase inhibitors.J.Med.

Chem2006; 49:4777–4780.8. Izevbigie EB (2003). Discovery of Water-Soluble Anticancer

Agents (Edotides) from a Vegetable Found in Benin City, Nigeria. Exp.Biol. and Med. 228:293–298

9. Havsteen BH ( 2002). The biochemistry and medical significance of theflavonoids. Pharmacol. Therap.96. P. 67–202.

10. Richardson MA (2001). Biopharmacologic and herbal therapies forcancer: research update from NCCAM. J. Nut. 131:3037S–3040S.

11. Wolff ME .Burger ‘s Medicinal Chemistry and Drug Discovery 5th

Ed.John Wiley and Sons New York.

12. Finar IL. Organic Chemistry volume 2. 5th ED. Pearson EducationAsia

Ltd.13. Block, J. H.; Beale, J. M. Jr. Wilson and Gisvold’s Textbook of

Organic, Medicinal and Pharmaceutical Chemistry. 11th edition,Lippincott. Williams and Wilkins. 2004, 249, 310.

14. Jeffrey M Edmondson, Linda S. Armstrong and Andrew OMartinez.

A Rapid and Simple MTT-Based Spectrophotometric Assay ForDetermining Drug Sensitivity In Monolayer Culture, Tissue CultureMethods, 1988 11, 15–17,.

15. Rubinstein LV, Shoemaker RH, Paull KD, Sinon RM, Tosini S,Skehan P, Scudiero A Monks, Boyd MR., Comparison of in-vitroAnticancer Drug screening data generated with Tetrazolium AssayVersus a Protein assay Against a Diverse Panel of Human TumorCell Lines. Journal of National Cancer Institute, 1990. 82(13), 1113–1118,

16. ht t p : / /ecb. j rc . i t /DOCUMENTS/ Test i ng -Methods/A NNE XV/AO8web1992.pdf.17. Selassie, C. D. History of Quantitative Structure-Activity

Relationship.Burger’s Medicinal Chemistry and Drug Discovery. Vol I. 6th

edition. Wiley Intersciences 2003, 15–23.18. Taylor, P. J. Hydrophobic Properties of Drug. Comprehensive

MedicinalChemistry Vol IV. 1st Indian edition. Pergamon Press Elsevier,2005,270–273 .

19. Beckett, A. H.; Stenlake, J. B. Practical Pharmaceutical Chemistry.4th

edition part-II. CBS Publishers and Distributors. 2005, 275–278.20. Markandi JK , Shashi, Surender k. Ind.J. Chem. 2004 April

43B:895-896.

21. Wattenberg LW. Inhibition of carcinogenesis by minordietary constituents. Cancer Res 1992; 52: 2085s–2091s.

22. Calomme M, Pieters L, Vlietinck A, Vanden Berghe D. Inhibitionof

bacterial mutagenesis by Citrus flavonoids. Planta Med 1996; 62: 222–226.

23. Dashwood RH, Xu M, Hernaez JF, Hasaniya N, Youn K, RazzukA.

Cancer chemo preventiveechanisms of tea against heterocyclic aminemutagens from cooked meat. Proc Soc Exp Biol Med 1999;220:239–243

24. Williamson G, Faulkner K, PlumbGW. Glucosinolates and phenolicas antioxidants from plant foods.Eur J Cancer Prev 1998; 7:17–21.

25. Carroll KK, Guthrie N, So FV, Chambers AF. Anticancer properties offlavonoids, with emphasis on citrus flavonoids. In: Rice-Evans CA,Packer L, editors. Flavonoids in health and disease. New York:MarcelDekker Inc.; 1998. p 437–446.

26. Lahiri-Chatterjee M, Katiyar SK, Mohan RR, Agarwal R. A

http://ecb.jrc.it/DOCUMENTS/Testing-Methods/ANNEXV/



flavonoid antioxidant, Silymarin, afford exceptionally highprotection against tumor promotion in the SENCAR mouse skintumorigenesis model. Cancer Res 1999; 59:622–632.

27. Tsyrlov IB, Mikhailenko VM, Gelboin HV. Isozyme- and species-specific susceptibility of cDNA expressed CYP1A P-450s to differentflavonoids. Biochim Biophys Acta 1994; 1205: 325–335.

28. Bu-Abbas A, Clifford MN,Walker R, Ioannides C. Contributionof caffeine and flavanols in the induction of hepatic Phase II activitiesby green tea. Food Chem Toxicol 1998; 36:617–621.

29. Sun XY, Plouzek CA, Henry JP,Wang TT, Phang JM. IncreasedUDP-

glucuronyl transferase activity and decreased prostate specific antigen



Documents

Synhtesis, Characterization and Cytotoxic Activity of ...shodhganga.inflibnet.ac.in/bitstream/10603/21199/16/22 r paper -1... · Synhtesis, Characterization and Cytotoxic Activity