Research Article Synthesis, Characterisation, and In...

Hindawi Publishing CorporationBioMed Research InternationalVolume 2013 Article ID 239354 14 pageshttpdxdoiorg1011552013239354

Research ArticleSynthesis Characterisation and In Vitro AnticancerActivity of Curcumin Analogues Bearing PyrazolePyrimidineRing Targeting EGFR Tyrosine Kinase

Mohamed Jawed Ahsan1 Habibullah Khalilullah2 Sabina Yasmin3

Surender Singh Jadav3 and Jeyabalan Govindasamy2

1 Department of Pharmaceutical Chemistry Maharishi Arvind College of Pharmacy Ambabari Jaipur Rajasthan 302 023 India2Department of Pharmaceutical Chemistry Alwar Pharmacy College Alwar Rajasthan 301 030 India3 Department of Pharmaceutical Chemistry Birla Institute of Technology Mesra Ranchi Jharkhand 835 215 India

Correspondence should be addressed to Mohamed Jawed Ahsan jawedpharmagmailcom

Received 24 April 2013 Revised 20 July 2013 Accepted 23 July 2013

Academic Editor Vickram Ramkumar

Copyright copy 2013 Mohamed Jawed Ahsan et alThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

In search of potential therapeutics for cancer we described herein the synthesis characterization and in vitro anticancer activityof a novel series of curcumin analogues The anticancer effects were evaluated on a panel of 60 cell lines according to the NationalCancer Institute (NCI) screening protocol There were 10 tested compounds among 14 synthesized compounds which showedpotent anticancer activity in both one-dose and 5-dose assays The most active compound of the series was 35-bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl(phenyl)methanone (10) which showed mean growth percent of minus2871 in one-dose assay and GI

50

values between 00079 and 186120583M in 5-dose assay

1 Introduction

Cancer is still continuing to be a major health problemworldwide The development of new anticancer therapeuticagents is one of the fundamental goals inmedicinal chemistryas cancer causes about 13 of all the death [1] Surpassingcardiovascular diseases it is taking the position numberone killer due to various factors [2] Also the treatment ofcancer is associated with various side effects which includebone marrow depression alopecia drug-induced cancerhepatotoxicity and many more Because of the need andvalue of anticancer drugs many laboratories are intensivelyinvestigating the chemistry and biology of novel anticanceragents Also the development of resistance against the exist-ing anticancer drugs and cytotoxicity and genotoxicity ofanticancer drugs to the normal cells are othermajor problemsin cancer therapy keeping research window open in searchfor newer anticancer molecules [3] But the window passagehas become narrower because it is rather hard to search

a molecule that can selectively inhibit the proliferation ofabnormal cells only with least or no affect on normal cells

In the last decade several pyrazole derivatives proved tohave anticancer activity [4ndash8] The other activities reportedfor pyrazole nucleus include antitubercular anticonvulsantanticancer antimicrobial anti-HIV antihepatotoxic anti-inflammatory and analgesic [9ndash16] Also curcumin a majoryellow pigment and active component of turmeric has beenshown to possess anti-inflammatory and anticancer activities[17] In some countries curcumin was consumed in thediet up to 4 g per adultday which appeared to lower theincidence rate of colorectal cancer In a study curcuminshowed autophagic and apoptotic death of K562 cell line(leukemia) [18] The cytotoxicity studies in different celllines indicated that the toxicity of curcumin was significantlyhigher in tumor cells if compared to the normal cells [19]Curcumin and its derivatives possess a wide variety ofpharmacological activities namely antibacterial anti-HIVanti-inflammatory antimalarial anticancer and many more

2 BioMed Research International

[20ndash24] Considering these recent discoveries curcumin canbe considered as an ideal lead compound for anticancer drugdevelopment Earlier we have reported the anticancer activityof pyrazoline and oxadiazole analogues [25 26] Receptortyrosine kinases (RTKs) have been shown to be key regulatorsof normal cellular processes and to additionally play a criticalrole in the development and progression of many types ofcancer by binding either with polypeptide growth factors orcytokines or hormones [27] Several transforming oncogeneslike src a gene obtained from Rous sarcoma virus abl a geneobtained from Abelson murine leukemia virus and so forthare known to possess tyrosine kinase activity Overexpressionof certain RTKs shows association with promotion andmaintenance of malignancies Thus inactivation of thespecific tyrosine kinase represents a potential approach fordesign of anticancer drugs Gefitinib and erlotinib usedin the treatment of certain types of cancer are epidermalgrowth factor receptor (EGFR) tyrosine kinase inhibitorsProtein tyrosine kinases occupy a central position in thecontrol of cellular proliferation and it is well recognizedthat the response of many cells to growth factors is initiatedby activation of tyrosine kinase [28] The involvement ofthe EGFR family of tyrosine kinases in cancer proliferationsuggests that an inhibitor which blocks the tyrosine kinaseactivity of the entire EGFR family could have significanttherapeutic potential [29] Hence enthused by all these factswe have synthesized a novel series of curcumin analoguesand evaluated their antitumor activity and selected EGFRfamily of tyrosine kinase as a biological target for carryingout the docking study of some of the active compounds

2 Materials and Methods

21 Chemistry All chemicals were supplied by E Merck(Germany) Konark Herbal (India) and S D Fine Chemicals(India) Melting points were determined by open tube capil-lary method and are uncorrected Purity of the compoundswas checked by elemental analysis and the progress ofreactions was monitored by TLC plates (silica gel G) usingmobile phase hexane ethylacetate (6 4) and the spots wereidentified by iodine vapours or UV light IR spectra wereobtained on a Shimadzu 8201 PC FT-IR spectrometer (KBrpellets) 1H NMR spectra were recorded on a Bruker AC300MHz spectrometer using TMS as internal standard inDMSO Mass spectra were recorded on a Bruker EsquireLCMS using ESI and elemental analyses were performed onPerkin-Elmer 2400 elemental analyzer

22 General Method for the Synthesis of 35-Bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carboxa-mide Analogues (1ndash9) 17-Bis(4-hydroxy-3-methoxyph-enyl)hepta-16-diene-35-dione (curcumin) (0005mol)and substituted phenyl semicarbazides (0005mol) wererefluxed in glacial acetic acid for 12 h The substitutedsemicarbazides were synthesized as per reported method[30] The excess of solvent was removed under reducedpressure and then the reaction mixture was poured intothe crushed ice The solid mass was filtered washed

dried and recrystallized with ethanol furnishing the title35-bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carboxamide analogues (1ndash9)

221 35-Bis(4-hydroxy-3-methylstyryl)-N-(2-chlorophenyl)-1H-pyrazole-1-carboxamide (1) IR (KBr) cmminus1 3381 (OH)3195 (NH) 1679 (C=O) 1569 (C=N) 1334 (CndashN) 745 (CndashCl)1H NMR (300MHz DMSO-d

6) 120575 383 (6H s OCH

3) 661

(1H s CH=C) 673 (2H d J = 158Hz CH and CH) 678(2H d J = 158Hz CH and CH) 631ndash762 (10H m ArH)972 (2H s OH) 983 (1H s CONH) mz = 517 (M+) 519(M + 2)+


6) 120575 385 (6H s OCH

3) 660

(1H s CH=C) 677 (2H d J = 158Hz CH and CH) 681(2H d J = 158Hz CH and CH) 691ndash772 (10H m ArH)974 (2H s OH) 993 (1H s CONH) 13C NMR (75MHzDMSO-d

6) ppm 16912 16903 14827 14707 13860 13849

13011 12904 12878 12820 12704 12098 12089 1205111597 10983 9974 5597mz = 517 (M+) 519 (M + 1)+

223 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-fluorophenyl)-1H-pyrazole-1-carboxamide (3) IR (KBr) cmminus1 3311 (OH)3181 (NH) 1667 (C=O) 1508 (C=N) 1370 (CndashN) 1032 (CndashF)1H NMR (300MHz DMSO-d

6) 120575 383 (6H s OCH

3) 661

(1H s CH=C) 668 (2H d J = 162Hz CH and CH) 678(2H d J = 162Hz CH and CH) 692ndash760 (10H m ArH)962 (2H s OH) 997 (1H s CONH) 1297 (OH) 13C NMR(75MHzDMSO-d

6) ppm 1901 1585 1513 1473 1473 1449

1334 1236 1315 1288 1232 1201 1168 1157 1121 1077562mz = 501 (M+) 503 (M + 2)+

224 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-bromophenyl)-1H-pyrazole-1-carboxamide (4) IR (KBr) cmminus1 3380 (OH)3202 (NH) 1689 (C=O) 1569 (C=N) 1330 (CndashN) 634 (CndashBr) 1H NMR (300MHz DMSO-d

6) 120575 383 (6H s OCH

3)

662 (1H s CH=C) 667 (2H d J = 166Hz CH andCH) 671(2H d J = 166Hz CH and CH) 692ndash767 (10H m ArH)975 (2H s OH) 997 (1H s CONH) mz = 562 (M+) 564(M + 2)+

225 35-Bis(4-hydroxy-3-methylstyryl)-N-(3-chloro-4-fluor-ophenyl)-1H-pyrazole-1-carboxamide (5) IR (KBr) cmminus13305 (OH) 3122 (NH) 1604 (C=O) 1555 (C=N) 1370 (CndashN) 737 (CndashF) 1HNMR (300MHz DMSO-d

6) 120575 384 (6H s

OCH3) 661 (1H s CH=C) 675 (2H d J = 162Hz CH and

CH) 678 (2H d J = 162Hz CH and CH) 690ndash793 (9Hm ArH) 976 (2H s OH) 1014 (1H s CONH) 13C NMR(75MHzDMSO-d

6) ppm 15307 15195 14181 13478 13353

13294 12544 12433 12242 12183 12078 11457 104486074mz = 535 (M+) 537 (M + 1)+

226 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-methylphenyl)-1H-pyrazole-1-carboxamide (6) IR (KBr) cmminus1 3384 (OH)3162 (NH) 1683 (C=O) 1571 (C=N) 1332 (CndashN) 1H NMR

BioMed Research International 3

(300MHz DMSO-d6) 120575 209 (3H s CH

3) 381 (6H s


CH) 668 (2H d J = 168Hz CH and CH) 682ndash741 (10Hm ArH) 927 (2H s OH) 1002 (1H s CONH) mz = 497(M+)

227 35-Bis(4-hydroxy-3-methylstyryl)-N-(2-methylphenyl)-1H-pyrazole-1-carboxamide (7) IR (KBr) cmminus1 3361 (OH)3151 (NH) 1686 (C=O) 1569 (C=N) 1337 (CndashN) 1H NMR(300MHz DMSO-d

6) 120575 212 (3H s CH

3) 384 (6H s



228 35-Bis(4-hydroxy-3-methylstyryl)-N-(24-dimethylphe-nyl)-1H-pyrazole-1-carboxamide (8) IR (KBr) cmminus1 3310(OH) 3104 (NH) 1681 (C=O) 1559 (C=N) 1374 (CndashN) 1HNMR (300MHz DMSO-d

6) 120575 119 (6H s CH

3) 383 (6H

s OCH3) 652 (1H s CH=C) 661 (2H d J = 168Hz

CH and CH) 665 (2H d J = 168Hz CH and CH) 678ndash743 (9H m ArH) 971 (2H s OH) 1018 (1H s CONH)13C NMR (75MHz DMSO-d

6) ppm 16860 14833 14722

13449 13435 1320 13120 13003 12877 12818 1268212554 12055 11604 10984 9978 5599mz = 511 (M+)


6) 120575 122 (6H s CH

3) 385 (6H s


CH) 676 (2H d J = 62Hz CH and CH) 689ndash792 (9H mArH) 997 (2H s OH) 1014 (1H s CONH)mz = 511 (M+)

23 General Method for the Synthesis of 35-Bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl (substituted phenyl)methanoneAnalogues (10-11) 17-Bis(4-hydroxy-3-methoxyphenyl) he-pta-16-diene-35-dione (curcumin) (0005mol) and substi-tuted phenyl hydrazides (0005mol) were refluxed in glacialacetic acid for 12 h The excess of solvent was removed underreduced pressure and then the reaction mixture was pouredinto the crushed ice The solid mass was filtered washeddried and recrystallized with ethanol furnishing the title 35-bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl (substitutedphenyl)methanone analogues (10-11)

231 35-Bis(4-hydroxy-3-methoxystyryl)-1H-pyrazole-1-yl(phenyl)methanone (10) IR (KBr) cmminus1 3342 (OH) 3168(NH) 1755 (C=O) 1578 (C=N) 1378 (CndashN) 1H NMR(300MHz DMSO-d

6) 120575 396 (6H s OCH

3) 621 (1H s

CH=C) 673 (2H d J = 152Hz C2H and C

6H) 675 (2H d J

= 152Hz C1H and C

7H) 687ndash757 (11H m ArH) 992 (2H

s OH) 13C NMR (75MHz DMSO-d6) ppm 15906 15632

15366 14790 14680 14574 13769 13742 13515 1342412970 12831 11952 11560 11175 11028 5558 MS mz(M+) 468

232 35-Bis(4-hydroxy-3-methoxystyryl)-1H-pyrazole-1-yl(2-bromophenyl)methanone (11) IR (KBr) cmminus1 3339 (OH)

3121 (NH) 1757 (C=O) 1514 (C=N) 1371 (CndashN) 596 (CndashBr)1H NMR (300MHz DMSO-d

6) 120575 398 (6H s OCH

3) 620

(1H s CH=C) 675 (2H d J = 158Hz C2H and C

6H) 678

(2H d J = 158Hz C1H and C

7H) 682ndash742 (10H m ArH)

947 (2H s OH) MSmz (M+) 547 (M + 2)+ 549

24 General Method for the Synthesis of DihydropyrimidineAnalogues (12ndash14) 17-Bis(4-hydroxy-3-methoxyphenyl)hepta-1 6-diene-35-dione (curcumin) (0005mol) and ureaguanidinethiourea (0005mol) were refluxed in glacialacetic acid for 12 h The excess of solvent was removedunder reduced pressure and then the reaction mixture waspoured into the crushed ice The solid mass was filteredwashed dried and recrystallized with ethanol furnishingthe title 35-bis(4-hydroxy-3-methylstyryl)-N-(substitutedphenyl)-1H-pyrazole-1-carboxamide analogues (12ndash14)

241 46-Bis(4-hydroxy-3-methoxystyryl)pyrimidine-2(1H)-one (12) IR (KBr) cmminus1 3402 (OH) 3192 (NH) 1633(C=O) 1573 (C=N) 1371 (CndashN) 1H NMR (300MHzDMSO-d

6) 120575 379 (6H s OCH

3) 492 (1H s NH) 51 (1H

s NH) 623 (1H s CH=C) 661 (2H d J = 142Hz C2H and

C6H) 665 (2H d J = 142Hz C

1H and C

7H) 669ndash722 (6H

m ArH) 996 (2H s OH) MSmz (M+) 391 (M + 1)+ 392

242 441015840-[(2-Imino-12-dihydropyrimidine-46-diyl)diethe-ne-21-diyl]bis(2-methoxyphenol) (13) IR (KBr) cmminus1 3423(OH) 3124 (NH) 1583 (C=N) 1375 (CndashN) 1H NMR(300MHz DMSO-d

6) 120575 373 (6H s OCH

3) 49 (1H s NH)

52 (1H s NH) 621 (1H s CH=C) 666 (2H d J = 142HzC2H and C

6H) 669 (2H d J = 142Hz C

1H and C

7H) 671ndash

714 (6H m ArH) 975 (2H s OH) MSmz (M+) 391 (M +1)+ 392

243 46-Bis[2-(4-hydroxy-3-methoxyphenyl)ethenyl]pyrim-idine-2(1H)-thione (14) IR (KBr) cmminus1 3419 (OH) 3114(NH) 1581 (C=N) 1365 (CndashN) 1273 (C=S) 1H NMR(300MHz DMSO-d

6) 120575 374 (6H s OCH

3) 51 (1H s NH)

661 (1H s CH=C) 665 (2H d J = 146Hz C2H and C

6H)

668 (2H d J = 146Hz C1H and C

7H) 678ndash712 (6H m

ArH) 972 (2H s OH) MSmz (M+) 408 (M + 1)+ 409

25 Anticancer Activity There were 10 compounds amongthe series selected and screened for their anticancer activityboth in one-dose and 5-dose assays by National CancerInstitute (NCI) on leukemia melanoma lung colon CNSovarian renal prostate and breast cancers cell lines nearly60 in number according to their screening protocol reportedelsewhere [31ndash34] All the curcumin analogues were synthe-sized and the structure of the compounds was submittedonline to the official site of NCI for anticancer screeningAmong 14 compounds only 10 compounds were selected foranticancer screening NCI has its own selection procedure ofthe compounds for anticancer screening based on the noveltyof heterocyclic ring system drug-like properties utilizing the


Table 1 Physical constants of the curcumin analogues (1ndash14)

HO

ONHN

OH

O

X

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

1ndash9 12ndash1410-11

Compound R X NSC code Yield () Mp (∘C)1 2-Chloro mdash 763372 76 922 4-Chloro mdash 757924 66 783 4-Fluoro mdash 757925 72 704 4-Bromo mdash mdash 74 1365 3-Chloro-4-fluoro mdash 757927 78 586 4-Methyl mdash 763373 82 607 2-Methyl mdash mdash 71 988 24-Dimethyl mdash 757926 88 689 26-Dimethyl mdash mdash 82 16610 H mdash 757929 84 11011 2-Bromo mdash 757928 76 28812 mdash O 763374 72 10413 mdash NH 763375 78 8214 mdash S mdash mdash mdash

concept of privileged scaffolds structure based on computer-aided drug design and so forth while the structures con-taining problematic linkage or functional groups (eg nitronitroso ndashNndashNndash ndashN=Nndash imine semicarbazone thioamidesand thioureas) for successful drug development are avoided[31]

The anticancer screening was carried out as per theNCI US protocol Using the seven absorbance measurements(time zero (119879

119894) control growth (119862) and test growth in

the presence of drug at the five concentration levels (119879119891))

the percentage growth was calculated at each of the drugconcentrations level as [(119879

119891minus 119879

119894)(119862 minus 119879

119894)] times 100 for

concentrations for which 119879119891ge 119879

119894and [(119879

119891minus 119879

119894)119879

119894] times 100

for concentrations for which 119879119891lt 119879

119894

Three-dose response parameters (GI50 TGI and LC

50)

were calculated for each of the experimental agents Growthinhibition of 50 (GI

50) was calculated from 100 times [(119879

119891minus

119879

119894)(119862 minus 119879

119894)] = 50 which was the drug concentration

resulting in a 50 reduction in the net protein increase (asmeasured by sulforhodamine B SRB staining) in controlcells during the drug incubation The total growth inhibi-tion (TGI) was calculated from 119879

119891= 119879

119894 which was the

drug concentration resulting in total growth inhibition andsignified the cytostatic effect The LC

50was calculated from

100 times [(119879

119891minus 119879

119894)119879

119894] = minus50 indicating a net loss of cells

following treatment which indicated the concentration ofdrug resulting in a 50 reduction in the measured proteinat the end of the drug treatment as compared to that at thebeginning Values were calculated for each of these threeparameters at the level of activity however if the effect didnot reach to the level of activity the value of parameter was

expressed as less than the minimum concentration testedor if the effect exceeded the level of activity the valueof parameter was expressed as greater than the maximumconcentration tested [31 35 36] LogGI

50 log TGI and

log LC50

are the logarithm molar concentrations producing50 growth inhibition (GI

50) a total growth inhibition

(TGI) and a 50 cellular death (LC50) respectively

26 Molecular Docking Studies

261 Protein Structure X-ray crystal structure EGFR kinase(PDB 2J5F) with resolution 300 A 119877-value 0194 (obs)was obtained from the protein data bank (Research Collab-oratory for Structural Bioinformatics (RCSB) (httpwwwrcsborgpdb))

262 Protein Preparation The protein (PDB 2J5F) wasprepared using the Protein PreparationWizard Preprocessedbond orders were assigned hydrogens were added metalswere treated and water molecules were deleted Heterostatefor cocrystallized ligand was generated using Epik proto-nation state and optimization of H-bonding of the proteinside chains were assigned using ProtAssign Energy wasminimized (Impref minimization) using RMSD 030 A andconverged by OPLS2005 force field utilities of Schrodingerrsquossuite 93

263 Receptor Grid Generation Receptor Grid has beengenerated with GLIDE module of Schrodinger with default


Table 2 Sixty human tumor cell lines anticancer screening data of curcumin analogues (1ndash14)

Panelcell line Growth percent in one-dose assay1 2 3 5 6 8 10 11 12 13

LeukemiaCCRF-CEM minus454 445 682 087 044 154 minus512 352 1672 168HL-60(TB) minus2428 minus2094 minus2136 minus2892 minus2817 minus3036 minus4220 minus2389 6133 367K-562 227 minus730 022 minus390 229 minus718 minus1552 362 4203 549MOLT-4 minus457 minus200 minus356 minus1767 011 minus2068 minus3987 minus653 3634 417RPMI-8226 minus2231 291 332 minus1132 minus1446 minus1673 minus3360 1425 6249 288SR minus1772 152 253 144 minus1465 minus207 minus1389 minus477 2724 minus794

Non-small-cell lung cancerA549ATCC 875 403 1202 590 471 minus398 840 4173 7168 1009EKVX 1768 1685 2687 minus1767 2029 minus1472 minus3879 3055 7160 2216HOP-62 839 1706 3022 minus778 1162 minus3566 minus6773 4353 7077 485HOP-92 minus1442 minus470 minus345 NT minus1538 minus2702 NT NT 3370 minus2124NCI-H226 minus372 573 1588 minus432 minus089 minus370 minus819 3848 7620 1170NCI-H23 minus2582 316 1074 minus1287 minus1041 minus2609 NT 1766 6285 minus279NCI-H322M NT 2213 2732 1263 NT 813 minus364 3948 NT NTNCI-H460 739 941 835 443 1014 187 minus007 915 6970 554NCI-H522 minus1866 minus494 minus604 1089 minus1934 631 minus3023 420 5459 minus4131

Colon cancerCOLO 205 minus7349 757 873 5943 minus2581 minus6555 minus8405 2387 5181 minus5038HCC-2998 283 455 692 975 219 428 582 4094 7099 093HCT-116 116 461 495 215 160 148 minus583 690 3475 033HCT-15 1505 786 1063 664 1522 384 minus338 1621 8179 2955HT29 minus3495 minus1107 minus4184 minus3357 minus2235 minus3854 minus5239 038 minus641 minus7624KM12 009 353 244 008 minus113 minus935 minus2796 498 2926 minus2925SW-620 699 956 1052 203 812 minus1094 minus2272 2086 4358 186

CNS cancerSF-268 minus638 1387 2012 657 minus1150 380 minus1252 2797 4819 356SF-295 283 minus2189 minus2161 minus5532 053 minus5102 minus7147 minus2968 9276 minus525SF-539 406 016 368 minus1658 595 minus1958 minus3979 402 6902 1195SNB-19 1549 925 2015 442 2027 245 118 3622 8260 2319SNB-75 minus471 1875 2693 minus1260 953 minus3053 minus5318 2728 4432 550U251 233 431 1338 minus159 619 minus1227 minus147 3506 5564 297

MelanomaLOX IMVI 122 304 400 minus2087 189 minus2979 minus3711 929 4904 minus1609MALME-3M 827 394 915 minus1680 1727 minus2170 minus3898 2343 5357 minus2885M14 026 404 269 minus2186 230 minus3814 minus5595 542 7546 minus2613MDA-MB-435 408 246 minus232 minus2594 minus164 minus4091 minus6734 minus3053 5689 minus812SK-MEL-2 minus1227 minus2352 minus2707 minus2500 minus1398 minus2722 minus4811 minus033 8115 minus2101SK-MEL-28 1210 1272 1670 787 1322 557 minus673 2864 5089 955SK-MEL-5 minus2376 minus2966 minus836 minus7386 116 minus8344 minus8652 minus191 7074 minus1840UACC-257 128 311 1366 minus1589 480 minus2142 1603 8713 6246 minus190UACC-62 104 406 201 minus3742 049 minus4847 minus8439 348 6331 minus5766

Ovarian cancerIGROV1 minus658 993 1741 457 476 minus175 minus2118 2495 9312 342OVCAR-3 minus2677 minus1097 389 minus2644 minus3415 minus3733 minus4692 1255 5934 minus757OVCAR-4 659 1325 3555 926 1608 787 449 4980 5398 1831OVCAR-5 1318 1094 3132 088 2053 minus424 minus774 8449 10167 583OVCAR-8 150 minus1983 047 minus4400 minus298 minus3466 minus1311 2773 5871 933NCIADR-RES 2083 1746 2791 1201 2050 764 318 2813 9066 3932SK-OV-3 minus3332 minus037 minus1866 minus3116 minus2909 minus6358 minus5308 1655 7163 minus1739


Table 2 Continued


Renal cancer786-0 minus1480 minus220 minus690 minus3292 minus118 minus5907 minus7667 991 8257 minus5893A498 minus1183 minus060 3149 minus1365 1202 minus1470 minus2517 5641 6276 minus5692ACHN 283 929 2022 573 1612 525 277 2512 8936 1346CAKI-1 729 013 minus781 minus3910 865 minus3663 minus6045 minus3477 8861 1924RXF 393 minus5032 minus5360 minus4384 minus5761 minus4746 minus6729 minus7003 minus383 2375 minus6392SN 12C 1420 900 1795 206 1830 047 minus311 1423 6226 2214TK-10 1615 2310 3812 2404 2855 1586 618 6502 10274 2138UO-31 minus3484 271 1405 minus1695 097 minus3874 minus4464 2386 5425 316

Prostate cancerPC-3 654 424 901 minus1426 808 minus1082 minus3314 1292 6101 1299DU-145 123 1166 677 minus1184 220 minus2664 minus4574 2887 8037 1304

Breast cancerMCF7 441 536 949 206 630 minus378 minus571 2022 5210 759MDA-MB-231ATCC 1771 1435 3361 1224 1994 1025 523 3969 5419 339HS 578T minus471 823 2004 minus1292 minus1597 minus1433 minus1298 359 5391 minus100BT-549 minus2640 minus1435 minus833 minus2417 minus2300 minus3381 minus4716 1449 5503 minus2441T-47D 455 1850 3115 229 565 minus2255 minus2393 4899 5914 157MDA-MB-468 minus2419 minus2640 minus1460 minus2756 minus2914 minus3034 minus3186 297 634 minus3504

Mean minus466 181 723 minus1225 minus066 minus1919 minus2871 1903 5994 minus546Range 9432 7670 8196 9790 7601 9930 10255 12190 10915 11556NT not tested

parameters and without any constraints Site has been speci-fied as centroid of the work space ligand (20 A) with van derWaals radius scaling factor 10 and partial charge cutoff 025

264 Molecular Docking Protocol The ligand docking wasperformed in GLIDE50 Ligands which show less than35 rotatable bonds and which are having less than 200atoms were selected The scaling factor will be 080 and thepotential charge cutoff is 015 All the conformers from theconfgen-ligprep output were docked in the EGFR TyrosineKinase active site All default parameters were used for extraprecision docking Glide extra precision mode was employedfor the current docking study Best poses were chosen forenergy minimization during docking a distance dependentdielectric constant of 20 and maximum number of min-imization step of 100 was used The docking simulations(Ligand receptor interactions) are scored using the Xtraprecision (XP) mode which is implemented in GLIDE50

27 Materials and Methods All computational analysis wascarried out on a Red Hat 50 Linux platform running on aDell Precision work station with Intel core 2 quad processorand 8GB of RAM

3 Results and Discussion

31 Chemistry The curcumin analogues (1ndash14) described inthe study are shown in Table 1 and the reaction sequencefor the synthesis is summarized in Scheme 1 In the first part

of the synthesis 17-bis(4-hydroxy-3-methoxyphenyl)hepta-16-diene-35-dione (curcumin) and substituted phenyl semi-carbazide were refluxed in glacial acetic acid to obtainthe 35-bis(4-hydroxy-3-methoxystyryl)-N-(substituted phe-nyl)-1H-pyrazole-1-carboxamide analogues (1ndash9) Substi-tuted phenyl semicarbazides (ArNHCONHNH

2) were syn-

thesized as per reported method [30] In the second partof the synthesis 17-bis(4-hydroxy-3-methoxyphenyl)hepta-16-diene-35-dione (curcumin) and substituted phenylhy-drazide (ArCONHNH

2) were refluxed in glacial acetic acid to

obtain the 35-bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl(substituted phenyl)methanone (10-11) In the third partof synthesis 17-bis(4-hydroxy-3-methoxyphenyl)hepta-16-diene-35-dione (curcumin) and ureaguanidinethioureawere refluxed in glacial acetic acid to obtain the pyrimidineanalogues (12ndash14) The plausible mechanism of reactions isgiven in Figure 1 The yields of the title compounds wereranging from 66 to 88 after recrystallization with absoluteethanol The completion of reaction was monitored by TLCusing mobile phase hexane ethylacetate (6 4) and purityof the compounds was checked by elemental analyses Boththe analytical and spectral data (IR NMR and MS) of thesynthesized compounds were in full accordance with theproposed structures In general IR spectra of the compoundsafforded C=N stretching at 1514ndash1583 cmminus1 and CndashN stretchat 1320ndash1335 cmminus1 and carbamoyl group NndashH stretching at3122ndash3210 cmminus1 andC=O stretching at 1680ndash1689 cmminus1 bandsfor carboxamide analogues (1ndash9) and at 1755ndash1757 cmminus1bands formethanone analogues (10-11)The 1HNMR spectra


Table 3 NCI in vitro testing results of compounds of the compounds 8 10 and 13 at five-dose level in 120583M

S no Panelcell line 8 (NSC 757926) 10 (NSC 757929) 13 (NSC 763375)GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50

(1)

LeukemiaCCRF-CEM 0232 gt100 gt100 0127 gt100 gt100 0543 NT gt100HL-60(TB) 0237 0806 gt100 00884 NT gt100 218 617 gt100MOLT-4 0364 NT gt100 0347 gt100 gt100 101 519 gt100RPMI-8226 0336 NT gt100 0275 NT gt100 129 418 gt100SR 0004 gt100 gt100 00353 gt100 gt100 072 358 gt100

(2)

Non-small cell lung cancerA549ATCC 0445 363 563 0403 191 gt100 222 604 764EKVX 0771 551 gt100 0702 238 NT 222 683 gt100HOP-62 0497 163 421 0441 162 423 196 428 NTHOP-92 0538 339 gt100 035 332 gt100 126 519 gt100NCI-H226 0389 21 129 0382 183 768 183 434 12NCI-H23 0275 125 gt100 0113 161 gt100 18 424 NTNCI-H322M 0571 28 858 0615 249 NT 222 539 gt100NCI-H460 0306 NT gt100 0274 NT gt100 205 520 gt100NCI-H522 013 0616 NT 0286 759 NT 124 279 631

(3)

Colon cancerCOLO 205 205 362 638 186 334 597 18 324 585HCC-2998 064 58 gt100 0591 271 gt100 175 319 579HCT-116 0323 NT NT 0269 114 NT 11 239 518HCT-15 0335 166 NT 0167 152 NT 282 103 gt100HT29 0696 28 994 0463 319 gt100 314 0948 421KM12 0298 125 NT 00092 119 NT 163 332 679SW-620 0441 NT gt100 00079 NT gt100 165 368 NT

(4)

CNS cancerSF-268 0571 NT NT 0256 NT NT 211 156 gt100SF-295 0546 NT gt100 0538 NT NT 193 454 gt100SF-539 0354 15 NT 00604 153 NT 205 573 gt100SNB-19 0634 201 NT 0572 212 NT 181 795 gt100SNB-75 0416 182 NT 00099 187 NT 126 334 884U251 0471 159 NT 0428 170 NT 179 569 663

(5)

MelanomaLOX IMVI 0284 149 NT 00626 156 NT 127 259 NTMALME-3M 0563 198 NT 0525 217 NT 271 676 gt100M14 0372 141 NT 0155 148 NT 165 344 NTMDA-MB-435 00448 0383 NT 00175 000815 gt100 171 NT gt100SK-MEL-2 0366 182 NT 0266 203 gt100 198 41 NTSK-MEL-28 0509 247 861 0176 272 905 185 453 26SK-MEL-5 0317 129 367 0171 130 369 149 289 559UACC-257 154 547 319 114 429 533 153 35 8UACC-62 0413 165 489 0371 164 NT 149 323 NT


Table 3 Continued


(6)

Ovarian cancerIGROV1 0454 262 gt100 0483 NT gt100 295 708 gt100OVCAR-3 0412 172 gt100 0242 116 gt100 265 862 635OVCAR-4 0481 332 gt100 0530 676 gt100 171 115 711OVCAR-5 151 685 gt100 127 451 gt100 203 433 922OVCAR-8 0563 235 971 0479 378 gt100 219 724 gt100NCIADR-RES 0338 401 gt100 00874 385 gt100 288 188 gt100SK-OV-3 0419 16 728 0358 135 388 26 823 gt100

(7)

Renal cancer786-0 0549 176 461 0688 203 499 17 4 NTA498 0575 198 463 0697 2 465 151 326 704ACHN 0412 211 gt100 0131 169 NT 254 788 gt100CAKI-1 0341 549 gt100 0282 324 gt100 223 646 gt100RXF 393 0227 0653 272 0263 0878 360 15 32 NTSN 12C 0339 188 NT 0313 220 gt100 274 gt100 gt100TK-10 0424 2 715 0446 176 519 339 954 534UO-31 0362 174 593 0258 165 473 209 757 gt100

(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)

Breast cancerMCF7 0317 137 486 0162 16 657 181 633 388MDA-MB-231ATCC 0473 471 gt100 0404 483 gt100 145 408 gt100HS 578T 0295 gt100 gt100 00779 gt100 gt100 0524 299 gt100BT-549 0693 168 gt100 0631 228 gt100 13 352 NTT-47D 079 294 NT 0793 293 979 283 955 gt100MDA-MB-468 0248 0825 631 0276 0969 gt100 NT NT NT

NT not tested

Table 4 The docking score and E model score of the reportedcompounds

S no Ligand Glide score E model score1 Reference minus8288 minus684912 1 minus7538 minus806383 2 minus5935 minus831814 3 minus6903 minus764375 5 minus7272 minus800806 6 minus6721 minus813427 8 minus6725 minus716928 10 minus6647 minus767109 11 minus6766 minus7859510 12 minus7677 minus5662611 13 minus7702 minus47220

showed singlet at 120575 119ndash128 ppm corresponding to CH3

a singlet at 120575 379ndash385 ppm corresponding to OCH3 a

singlet at 120575 652ndash662 ppm corresponding to CH=C proton(pyrazoledihydropyrimidine) a doublet at 120575 661ndash675 ppmcorresponding toCH=CHproton a doublet at 665ndash681 ppmcorresponding to CH=CH proton a multiplet at 120575 681ndash793 ppm corresponding to aromatic protons broad singlet at120575 993ndash1016 ppm corresponding to CONH

2 The compounds

(1ndash14) in 1HNMRspectra exhibited two doublets with 119869 valuebetween 146 and 166Hz confirming the trans coupling Themass spectra of the compounds revealed in each case a peakcorresponding to their molecular ion peaks The elementalanalysis results were within plusmn04 of the theoretical values

32 Anticancer Activity Ten compounds were evaluated fortheir anticancer activity in both one-dose and 5-dose assaysThe observed anticancer screening data of the compoundsare given in Table 2 The 5-dose assay screening data ofthree compounds are given in Table 3 Compound 1 wasfound to be highly active on COLO 205 (colon cancer)with cell promotion of minus7349 followed by RXF 393


HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO

Scheme 1 Protocol for the synthesis of curcumin analogues

OO

HOO

OHO

OOH

HOO

OHO

Diketone (tautomer I) Keto-enol (tautomer II)

OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3

H3C CH3 H3C H3CCH3CH3

CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O

Figure 1 Plausible mechanism of reaction for the synthesis of pyrazole analogues (1ndash14)

(renal cancer) with cell promotion of minus5032 and HT29(colon cancer) with cell promotion of minus3495 while themaximum cell promotion was observed on NCIADR-RES(ovarian cancer) which showed 2083 growth promotion(7917 growth inhibition) at one-dose assay Compound2 was found to be highly active on RXF 393 (renal cancer)with cell promotion of minus5360 followed by SK-MEL-5(melanoma) with cell promotion of minus2966 and MDA-MB-468 (breast cancer) with cell promotion of minus2640 whilethe maximum cell promotion was observed on TK-10 (renalcancer) which showed 2310 growth promotion (7690growth inhibition) at one-dose assay The compound 3was found to be highly active on RXF 393 (renal cancer)

with cell promotion of minus4384 followed by HT29 (coloncancer) with cell promotion of minus4184 and SK-MEL-2(melanoma) with cell promotion of minus2707 while themaximum cell promotion was observed on TK-10 (renalcancer) which showed 3812 growth promotion (6188growth inhibition) at one-dose assay Compound 5 wasfound to be highly active on COLO 205 (colon cancer)with cell promotion of minus5943 followed by RXF 393 (renalcancer) with cell promotion of minus5761 and SF-295 (CNScancer) with cell promotion of minus5532 while the maximumcell promotion was observed on TK-10 (renal cancer)which showed 2404 growth promotion (7596 growthinhibition) at one-dose assay Compound 6 was found to


100

50

0

minus50

minus100

minus9 minus8 minus7 minus6 minus5 minus4

LeukemiaG

row

th (

)

log10 of sample concentration (molar)

CCRF-CEMMOLT-4HL-60(TB)

RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0

COLO 205HCT-15SW-620HCC-2998

HT29HCT-116KM12

Colon cancer

(c)

Figure 2 (a) Five-dose anticancer screening of compound 8 on leukemia cell line (b) Five-dose anticancer screening of compound 10 onleukemia cell line (c) Five-dose anticancer screening of compound 13 on colon cancer cell line

Figure 3 Green colour stick model indicates the reference ligand and atom macromodel stick indicates our ligands

be highly active on RXF 393 (renal) with cell promotionof minus4746 followed by MDA-MB-468 (breast cancer)with cell promotion of minus2914 and SK-OV-3 (ovariancancer) with cell promotion of minus2909 while the maximumcell promotion was observed on TK-10 (renal cancer)

which showed 2855 growth promotion (7145 growthinhibition) at one-dose assay Compound 8 was foundto be highly active on SK-MEL-5 (melanoma) with cellpromotion of minus8344 followed by RXF 393 (renal cancer)with cell promotion of minus6729 and SK-OV-3 (ovarian


(a) (b)

Figure 4 (a) Compound 13 is represented with macromodel stick modelThe hydrogen bonds are shown with yellow colour dotted lines andhydrophobic interactions with orange dotted lines (b) Compound 13 at binding site on surface

(a) (b)

Figure 5 (a) Compound 3 is represented with macromodel stick modelThe hydrophobic cavity residues are represented with magenta stickmodelThe hydrogen bonds are shownwith yellow colour dotted lines and hydrophobic interactions with orange dotted lines (b) Compound3 at binding site on surface

cancer) with cell promotion of minus6358 while the maximumcell promotion was observed on TK-10 (renal cancer)which showed 1584 growth promotion (8416 growthinhibition) at one-dose assay The compound 10 was foundto be highly active on SK-MEL-5 (melanoma) with cellpromotion of minus8652 followed by UACC-62 (melanoma)with cell promotion of minus8439 and COLO 205 (coloncancer) with cell promotion of minus8405 while the maximumcell promotion was observed on UACC-257 (melanoma)which showed 1603 growth promotion (8397 growthinhibition) at one-dose assay Compound 11 was found to behighly active on CAKI-1 (renal cancer) with cell promotionof minus3477 followed by MDA-MB-435 (melanoma) with cellpromotion of minus3053 and SF-295 (CNS cancer) with cellpromotion of minus2968 while the maximum cell promotionwas observed on UACC-257 (melanoma) which showed8713 growth promotion (1287 growth inhibition) at one-dose assay Compound 12 was found to be highly active onHT29 (colon cancer) with cell promotion of minus641 followedby MDA-MB-468 (breast cancer) with cell promotion ofminus634 and CCRF-CEM (leukemia) with cell promotion ofminus1672 while the maximum cell promotion was observedon TK-10 (renal cancer) which showed 10274 growthpromotion at one-dose assay Compound 13 was found to behighly active on HT29 (colon cancer) with cell promotion ofminus7624 followed by 786-0 and A498 (renal cancer) with cellpromotion of minus5893 and minus5692 respectively while themaximum cell promotion was observed on NCIADR-RES(ovarian cancer) which showed 3932 growth promotion(1287 growth inhibition) at one-dose assay Among the35-bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carboxamide analogues (1ndash9) Compound

8 was found to be the most active compound of theseries with mean growth percent of minus1919 followed bycompound 5 which showed mean growth percent of minus1225and compound 1 which showed mean growth percent ofminus466 Among the 35-bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl (substituted phenyl)methanone analogues(10-11) Compound 10 showed maximum activity with meangrowth percent of minus2871 while compound 13 was found tobe the most active compound among dihydropyrimidineanalogues (12ndash14) which showed mean growth percent ofminus546 Overall the most active compound of the serieswas compound 10 and most of the 35-bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carbox-amide analogues possessed potent anticancer activity Basedon our finding we can conclude that electronegative groupat position 2 (ie 2-chloro) on N-substituted phenyl ringamong 35-bis(4-hydroxy-3-methylstyryl)-N-(substitutedphenyl)-1H-pyrazole-1-carboxamide analogues (1ndash9) showedmore anticancer activity than at position 4 (ie 4-chloro and4-fluoro) Electron releasing group such as 24-dimethyl and4-methyl on N-substituted phenyl ring increased anticanceractivity and the activity was found to be more if thenumber of electron releasing groups was increased Amongthe 35-bis(4-hydroxy-3-methylstyryl)-1H-pyrazole-1-yl(substituted phenyl)methanone analogues (10 and 11) theactivity was found to bemore when there was no substitutionin the phenyl ring Among six member dihydropyrimidineanalogues (12ndash14) compound with X substitution as ldquoNHrdquoshowed more activity than as ldquoOrdquo substitution

The results of 5-dose assay of the most active compoundsamong their respective series are given in Table 3 Compound8 presented GI

50ranging between 0004 and 204 120583M The


best results were recorded on the leukemia cell lines withvalues ranging from 0004 to 0364 120583M (Figure 2(a)) Anumber of 3 tested cancer cell lines presented TGI value withgt100120583M the best result value being noted on the MDA-MB-435 (melanoma) with value 0383120583M Only in 23 cell linescompound 8 registered LC

50valuewithgt100120583MCompound

10 presented GI50ranging between 00079 and 186 120583M The

best results were recorded on the leukemia cell line withvalue ranging from 00353 to 0347 120583M (Figure 2(b)) Fourof the tested cancer cell lines presented TGI value withgt100120583M the best result value being noted on the MDA-MB-435 (melanoma) with value 000815120583M Only in 27 cell linescompound 10 registered LC

50value with gt100 120583M Com-

pound 13 presented GI50ranging between 0524 and 339 120583M

The best results were recorded on the colon cancer cell linewith values ranging from 0543 to 218 120583M(Figure 2(c)) Onlyone tested cancer cell line presented TGI value with gt100 120583Mthe best result value being noted on the HT29 (colon cancer)with value 0948 In 28 cell lines compound 13 registeredLC50value with gt100120583M

Some of the synthesized compounds were evaluated forEGFR tyrosine kinase inhibitory activity by recombinanttyrosine kinase assay using an ELISA-based assay withpoly(Glu Tyr 4 1) as a substrate [37] The results showedcompounds 8 10 and 13 shows inhibition of EGFR tyrosinekinase with IC

50values of 355 292 and 557120583Mrespectively

33 Molecular Docking The binding site of the referenceinhibitor is well defined by hydrophobic cavity The EGFRtyrosine kinase binding site contains the important residuesCys797 andThr790 re docking of reference ligand shown thatabove residues are important for hydrophobic interactionswith the same [38] The hydrophobic cavity with the residuesLeu788 Met766 Lys745 Glu762 Thr854 and Met793 makesthe binding site attractive for the design of new inhibitorsBased on our knowledge we designed some class of newpyrazole (1ndash11) dihydropyrimidine analogues (12ndash14) forEGFR tyrosine kinase inhibition But the binding sites ofour new molecules were found to be a little different withcomparison to reference inhibitor which is reported in thecrystal structure of EGFR tyrosine kinase (PDB 2J5F) Theseligandsrsquo interactions were shown in three different motifsThe first one is hydrophobic cavity in which the substitutedmethoxy hydroxy phenyl ring of the ligand is showinggood interactions as reference inhibitor The next bindingmotif contains the target residue Cys797 the N-substitutedpyrazole derivatives and six membered (dihydropyrimidine)are binding at this site Most of these interactions werehydrophobic The last binding motif contains the residuesLys875 and Asp831 which lie in side chain of the protein andshowed the side chain hydrogen bonding with the designedmolecules The docking score and Emodel score of some ofthe compounds selected for anticancer activity by NCI aregiven in Table 4 and binding modes are shown in Figure 3

Compound 13 showed the best hydrophobic interactionwith various residues such as Cys797 Met 793 Leu 844 Leu792 Ala 743 and Val 726 (Figures 4(a) and 4(b)) Negativelycharged Asp 837 interacted with methoxy hydroxy phenyl

ring (4-hydroxy-3-methoxyphenyl) Arg 841 showed 120587-120587cationic interaction The Met 793 present in the hydrophobiccavity showed H-bond backbone interaction and H-bond(side chain) interaction The positively charged Lys 845and Arg 841 destabilized interactions The interactions ofcompound 13 with the EGFR kinase receptor shown inribbon diagram the hydrophobic cavity is indicated withyellow colour loop motif-1 is indicated with green colourbeta sheet and the motif-2 is indicated with green colourloop The other ligands were also having good interactionswith the hydrophobic cavity and bindingmotifs the followingresidues leu718 Pro794 Met793 Ala743 Leu844 Val726and Cys797 were present around the ligand Thr790 Gln791having polar characteristics andMet 793 were responsible forbackbone H-bond interaction as well as H-bond (side chain)interaction Asp855 and Asp837 showed charged interactionslike destabilizing contacts Positively charged Lys875 andArg841 were found to have 120587-120587 cationic interaction and werepresent in bindingmotifmaking different binding patterns ofreported ligandsThe docking study for compound 3 is givenin Figures 5(a) and 5(b)

4 Conclusion

The novel series of curcumin analogues (pyrazoles) weresynthesized in satisfactory yields The anticancer activityshowed promising results The studies confirmed compound10 as potent lead compound for drug discovery and furtheroptimization The curcumin analogues discovered in thisstudy may provide valuable therapeutic intervention for thetreatment of cancer disease The pyrazole discovered in thisstudy may provide valuable therapeutic intervention for thetreatment of cancer

Acknowledgments

The authors are thankful to the staff members of the NationalCancer Institute (NCI) USA for in vitro anticancer screeningof the newly synthesized compounds They are especiallygrateful to Professor Dizon B Thelma Project ManagerNational Cancer Institute (NCI) USA

References

[1] WHO Cancer World Health Organization February 2006httpenwikipediaorgwikicancer

[2] M N Noolvi H M Patel V Bhardwaj and A ChauhanldquoSynthesis and in vitro antitumor activity of substituted quina-zoline and quinoxaline derivatives search for anticancer agentrdquoEuropean Journal of Medicinal Chemistry vol 46 no 6 pp2327ndash2346 2011

[3] N Aydemir and R Bilaloglu ldquoGenotoxicity of two anticancerdrugs gemcitabine and topotecan in mouse bone marrow invivordquoMutation Research vol 537 no 1 pp 43ndash51 2003

[4] I Bouabdallah L A MrsquoBarek A Zyad A Ramdani I Zidaneand A Melhaoui ldquoNew pyrazolic compounds as cytotoxicagentsrdquo Natural Product Research vol 21 no 4 pp 298ndash3022007


[5] D Havrylyuk B Zimenkovsky O Vasylenko L Zaprutko AGzella and R Lesyk ldquoSynthesis of novel thiazolone-based com-pounds containing pyrazoline moiety and evaluation of theiranticancer activityrdquo European Journal of Medicinal Chemistryvol 44 no 4 pp 1396ndash1404 2009

[6] M Shaharyar M M Abdullah M A Bakht and J MajeedldquoPyrazoline bearing benzimidazoles search for anticanceragentrdquo European Journal of Medicinal Chemistry vol 45 no 1pp 114ndash119 2010

[7] P-C Lv H-Q Li J Sun Y Zhou and H-L Zhu ldquoSynthesisand biological evaluation of pyrazole derivatives containingthiourea skeleton as anticancer agentsrdquo Bioorganic and Medici-nal Chemistry vol 18 no 13 pp 4606ndash4614 2010

[8] M S Christodoulou S Liekens K M Kasiotis and S AHaroutounian ldquoNovel pyrazole derivatives synthesis and eval-uation of anti-angiogenic activityrdquo Bioorganic and MedicinalChemistry vol 18 no 12 pp 4338ndash4350 2010

[9] M J Ahsan G J Samy H Khalillah R C Krit and SSoni ldquoMolecular properties prediction and synthesis of novelpyrazoline carboxamide analogs as antitubercular agentsrdquoAnti-Infective Agents vol 10 no 2 pp 117ndash123 2012

[10] M J Ahsan J G Samy H Khalilullah M A Bakht andM Z Hassan ldquoSynthesis and antimycobacterial evaluation of3a4-dihydro-3H-indeno [12-c] pyrazole-2-carboxamide ana-loguesrdquo European Journal of Medicinal Chemistry vol 46 no11 pp 5694ndash5697 2011

[11] M J Ahsan J G Samy K R Dutt et al ldquoDesign synthesis andantimycobacterial evaluation of novel 3-substituted-N-aryl-67-dimethoxy-3a4-dihydro-3H-indeno[12-c] pyrazole-2-carboxamide analoguesrdquo Bioorganic and Medicinal ChemistryLetters vol 21 no 15 pp 4451ndash4453 2011

[12] M J Ahsan J G Samy S Soni et al ldquoDiscovery of novelantitubercular 3a4-dihydro-3H-indeno[12-c]pyrazole-2-carndashboxamidecarbothioamide analoguesrdquo Bioorganic and Medici-nal Chemistry Letters vol 21 no 18 pp 5259ndash5261 2011

[13] M J Ahsan H Khalilullah J P Stables and J GovindasamyldquoSynthesis and anticonvulsant activity of 3a 4-dihydro-3H-indeno[1 2-c]pyrazole-2-carboxamide carbothioamide ana-loguesrdquo Journal of Enzyme Inhibition amp Medicinal Chemistryvol 28 no 3 pp 644ndash650 2013

[14] M J Ahsan J Govindasamy H Khalilullah et al ldquoPOMAanalyses as new efficient bioinformaticsrsquoplatform to predictand optimize bioactivity of synthesized 3a 4-dihydro-3H-indeno[1 2-c]pyrazole-2-carboxamidecarbothioamide anal-oguesrdquo Bioorganic amp Medicinal Chemistry vol 22 no 24 pp7029ndash7035 2012

[15] H Khalilullah S Khan M J Ahsan and B AhmedldquoSynthesis and antihepatotoxic activity of 5-(23-dihydro-14-benzodioxane-6-yl)-3-substituted-phenyl-45-dihydro-1H-pyrazole derivativesrdquo Bioorganic and Medicinal ChemistryLetters vol 21 no 24 pp 7251ndash7254 2011

[16] M J Alam M J Ahsan O Alam and S A KhanldquoSynthesis of 4-(5-chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)-6-(substituted phenyl)pyrimidin-2-ol analogues as 8 anti-inflammatory and analgesic agentsrdquo Letters in Drug Design ampDiscovery vol 10 no 8 pp 776ndash782 2013

[17] D P Chauhan ldquoChemotherapeutic potential of curcumin forcolorectal cancerrdquo Current Pharmaceutical Design vol 8 no 19pp 1695ndash1706 2002

[18] Y-L Jia J Li Z-H Qin and Z-Q Liang ldquoAutophagic andapoptotic mechanisms of curcumin-induced death in K562

cellsrdquo Journal of Asian Natural Products Research vol 11 no 11pp 918ndash928 2009

[19] A Kunwar A Barik BMishra K Rathinasamy R Pandey andK I Priyadarsini ldquoQuantitative cellular uptake localization andcytotoxicity of curcumin in normal and tumor cellsrdquoBiochimicaet Biophysica Acta vol 1780 no 4 pp 673ndash679 2008

[20] J Lal S K Gupta DThavaselvam and D D Agarwal ldquoDesignsynthesis synergistic antimicrobial activity and cytotoxicityof 4-aryl substituted 34-dihydropyrimidinones of curcuminrdquoBioorganic and Medicinal Chemistry Letters vol 22 no 8 pp2872ndash2876 2012

[21] R K Singh D Rai D Yadav A Bhargava J Balzarini and EDe Clercq ldquoSynthesis antibacterial and antiviral properties ofcurcumin bioconjugates bearing dipeptide fatty acids and folicacidrdquo European Journal of Medicinal Chemistry vol 45 no 3pp 1078ndash1086 2010

[22] K Saja M S Babu D Karunagaran and P R SudhakaranldquoAnti-inflammatory effect of curcumin involves downregula-tion of MMP-9 in blood mononuclear cellsrdquo InternationalImmunopharmacology vol 7 no 13 pp 1659ndash1667 2007

[23] S Mishra K Karmodiya N Surolia and A Surolia ldquoSynthesisand exploration of novel curcumin analogues as anti-malarialagentsrdquo Bioorganic and Medicinal Chemistry vol 16 no 6 pp2894ndash2902 2008

[24] G Liang L Shao Y Wang et al ldquoExploration and synthesis ofcurcumin analogues with improved structural stability both invitro and in vivo as cytotoxic agentsrdquo Bioorganic and MedicinalChemistry vol 17 no 6 pp 2623ndash2631 2009

[25] M J Ahsan ldquoSynthesis and anticancer activity of 3a 4-dihydro-3H-indeno[1 2-c]pyrazole-2-carboxamide analoguesrdquo Lettersin Drug Design amp Discovery vol 9 no 9 pp 823ndash827 2012

[26] Salahuddin M Shaharyar A Majumdar and M J AhsanldquoSynthesis characterization and anticancer evaluation of 2-(naphthalen-1-ylmethylnaphthalen-2-yloxymethyl)-1-[5-(sub-stituted phenyl)-[134]oxadiazol-2-ylmethyl]-1H-benzimida-zolerdquo Arabian Journal of Chemistry 2013

[27] E Zwick J Bange andAUllrich ldquoReceptor tyrosine kinase sig-nalling as a target for cancer intervention strategiesrdquo Endocrine-Related Cancer vol 8 no 3 pp 161ndash173 2001

[28] H A Bhuva and S G Kini ldquoSynthesis anticancer activity anddocking of some substituted benzothiazoles as tyrosine kinaseinhibitorsrdquo Journal ofMolecular Graphics andModelling vol 29no 1 pp 32ndash37 2010

[29] J Mendelsohn and J Baselga ldquoThe EGF receptor family astargets for cancer therapyrdquo Oncogene vol 19 no 56 pp 6550ndash6565 2000

[30] M Amir M J Ahsan and I Ali ldquoSynthesis of N1-(3-chloro-4-flouropheny)-N4-substituted semicarbazones as novel anticon-vulsant agentsrdquo Indian Journal of Chemistry B vol 49 no 11 pp1509ndash1514 2010

[31] httpdtpncinihgov[32] A Monks D Scudiero P Skehan et al ldquoFeasibility of a high-

flux anticancer drug screen using a diverse panel of culturedhuman tumor cell linesrdquo Journal of theNational Cancer Institutevol 83 no 11 pp 757ndash766 1991

[33] M R Body and K D Paull ldquoSome practical considerations andapplications of the national cancer institute in vitro anticancerdrug discovery screenrdquoDrug Development Research vol 34 no2 pp 91ndash109 1995

[34] R H Shoemaker ldquoThe NCI60 human tumour cell line anti-cancer drug screenrdquo Nature Reviews Cancer vol 6 no 10 pp813ndash823 2006


[35] M C Alley D A Scudiero A Monks et al ldquoFeasibility ofdrug screening with panels of human tumor cell lines using amicroculture tetrazolium assayrdquo Cancer Research vol 48 no 3pp 589ndash601 1988

[36] M R Grever S A Schepartz and B A Chabner ldquoTheNationalCancer Institute cancer drug discovery and development pro-gramrdquo Seminars in Oncology vol 19 no 6 pp 622ndash638 1992

[37] P Dubreuil S Letard M Ciufolini et al ldquoMasitinib (AB1010)a potent and selective tyrosine kinase inhibitor targeting KITrdquoPLoS ONE vol 4 no 9 Article ID e7258 2009

[38] J A Blair D Rauh C Kung et al ldquoStructure-guided devel-opment of affinity probes for tyrosine kinases using chemicalgeneticsrdquo Nature Chemical Biology vol 3 no 4 pp 229ndash2382007

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

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AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


[20ndash24] Considering these recent discoveries curcumin canbe considered as an ideal lead compound for anticancer drugdevelopment Earlier we have reported the anticancer activityof pyrazoline and oxadiazole analogues [25 26] Receptortyrosine kinases (RTKs) have been shown to be key regulatorsof normal cellular processes and to additionally play a criticalrole in the development and progression of many types ofcancer by binding either with polypeptide growth factors orcytokines or hormones [27] Several transforming oncogeneslike src a gene obtained from Rous sarcoma virus abl a geneobtained from Abelson murine leukemia virus and so forthare known to possess tyrosine kinase activity Overexpressionof certain RTKs shows association with promotion andmaintenance of malignancies Thus inactivation of thespecific tyrosine kinase represents a potential approach fordesign of anticancer drugs Gefitinib and erlotinib usedin the treatment of certain types of cancer are epidermalgrowth factor receptor (EGFR) tyrosine kinase inhibitorsProtein tyrosine kinases occupy a central position in thecontrol of cellular proliferation and it is well recognizedthat the response of many cells to growth factors is initiatedby activation of tyrosine kinase [28] The involvement ofthe EGFR family of tyrosine kinases in cancer proliferationsuggests that an inhibitor which blocks the tyrosine kinaseactivity of the entire EGFR family could have significanttherapeutic potential [29] Hence enthused by all these factswe have synthesized a novel series of curcumin analoguesand evaluated their antitumor activity and selected EGFRfamily of tyrosine kinase as a biological target for carryingout the docking study of some of the active compounds

2 Materials and Methods

21 Chemistry All chemicals were supplied by E Merck(Germany) Konark Herbal (India) and S D Fine Chemicals(India) Melting points were determined by open tube capil-lary method and are uncorrected Purity of the compoundswas checked by elemental analysis and the progress ofreactions was monitored by TLC plates (silica gel G) usingmobile phase hexane ethylacetate (6 4) and the spots wereidentified by iodine vapours or UV light IR spectra wereobtained on a Shimadzu 8201 PC FT-IR spectrometer (KBrpellets) 1H NMR spectra were recorded on a Bruker AC300MHz spectrometer using TMS as internal standard inDMSO Mass spectra were recorded on a Bruker EsquireLCMS using ESI and elemental analyses were performed onPerkin-Elmer 2400 elemental analyzer

22 General Method for the Synthesis of 35-Bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carboxa-mide Analogues (1ndash9) 17-Bis(4-hydroxy-3-methoxyph-enyl)hepta-16-diene-35-dione (curcumin) (0005mol)and substituted phenyl semicarbazides (0005mol) wererefluxed in glacial acetic acid for 12 h The substitutedsemicarbazides were synthesized as per reported method[30] The excess of solvent was removed under reducedpressure and then the reaction mixture was poured intothe crushed ice The solid mass was filtered washed

dried and recrystallized with ethanol furnishing the title35-bis(4-hydroxy-3-methylstyryl)-N-(substituted phenyl)-1H-pyrazole-1-carboxamide analogues (1ndash9)


6) 120575 383 (6H s OCH

3) 661

(1H s CH=C) 673 (2H d J = 158Hz CH and CH) 678(2H d J = 158Hz CH and CH) 631ndash762 (10H m ArH)972 (2H s OH) 983 (1H s CONH) mz = 517 (M+) 519(M + 2)+


6) 120575 385 (6H s OCH

3) 660

(1H s CH=C) 677 (2H d J = 158Hz CH and CH) 681(2H d J = 158Hz CH and CH) 691ndash772 (10H m ArH)974 (2H s OH) 993 (1H s CONH) 13C NMR (75MHzDMSO-d

6) ppm 16912 16903 14827 14707 13860 13849

13011 12904 12878 12820 12704 12098 12089 1205111597 10983 9974 5597mz = 517 (M+) 519 (M + 1)+

223 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-fluorophenyl)-1H-pyrazole-1-carboxamide (3) IR (KBr) cmminus1 3311 (OH)3181 (NH) 1667 (C=O) 1508 (C=N) 1370 (CndashN) 1032 (CndashF)1H NMR (300MHz DMSO-d

6) 120575 383 (6H s OCH

3) 661

(1H s CH=C) 668 (2H d J = 162Hz CH and CH) 678(2H d J = 162Hz CH and CH) 692ndash760 (10H m ArH)962 (2H s OH) 997 (1H s CONH) 1297 (OH) 13C NMR(75MHzDMSO-d

6) ppm 1901 1585 1513 1473 1473 1449

1334 1236 1315 1288 1232 1201 1168 1157 1121 1077562mz = 501 (M+) 503 (M + 2)+

224 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-bromophenyl)-1H-pyrazole-1-carboxamide (4) IR (KBr) cmminus1 3380 (OH)3202 (NH) 1689 (C=O) 1569 (C=N) 1330 (CndashN) 634 (CndashBr) 1H NMR (300MHz DMSO-d

6) 120575 383 (6H s OCH

3)

662 (1H s CH=C) 667 (2H d J = 166Hz CH andCH) 671(2H d J = 166Hz CH and CH) 692ndash767 (10H m ArH)975 (2H s OH) 997 (1H s CONH) mz = 562 (M+) 564(M + 2)+

225 35-Bis(4-hydroxy-3-methylstyryl)-N-(3-chloro-4-fluor-ophenyl)-1H-pyrazole-1-carboxamide (5) IR (KBr) cmminus13305 (OH) 3122 (NH) 1604 (C=O) 1555 (C=N) 1370 (CndashN) 737 (CndashF) 1HNMR (300MHz DMSO-d

6) 120575 384 (6H s


CH) 678 (2H d J = 162Hz CH and CH) 690ndash793 (9Hm ArH) 976 (2H s OH) 1014 (1H s CONH) 13C NMR(75MHzDMSO-d

6) ppm 15307 15195 14181 13478 13353

13294 12544 12433 12242 12183 12078 11457 104486074mz = 535 (M+) 537 (M + 1)+

226 35-Bis(4-hydroxy-3-methylstyryl)-N-(4-methylphenyl)-1H-pyrazole-1-carboxamide (6) IR (KBr) cmminus1 3384 (OH)3162 (NH) 1683 (C=O) 1571 (C=N) 1332 (CndashN) 1H NMR


(300MHz DMSO-d6) 120575 209 (3H s CH

3) 381 (6H s




6) 120575 212 (3H s CH

3) 384 (6H s




6) 120575 119 (6H s CH

3) 383 (6H

s OCH3) 652 (1H s CH=C) 661 (2H d J = 168Hz


6) ppm 16860 14833 14722

13449 13435 1320 13120 13003 12877 12818 1268212554 12055 11604 10984 9978 5599mz = 511 (M+)


6) 120575 122 (6H s CH

3) 385 (6H s





6) 120575 396 (6H s OCH

3) 621 (1H s


6H) 675 (2H d J

= 152Hz C1H and C

7H) 687ndash757 (11H m ArH) 992 (2H


15366 14790 14680 14574 13769 13742 13515 1342412970 12831 11952 11560 11175 11028 5558 MS mz(M+) 468



6) 120575 398 (6H s OCH

3) 620


6H) 678



947 (2H s OH) MSmz (M+) 547 (M + 2)+ 549



6) 120575 379 (6H s OCH

3) 492 (1H s NH) 51 (1H


C6H) 665 (2H d J = 142Hz C

1H and C

7H) 669ndash722 (6H

m ArH) 996 (2H s OH) MSmz (M+) 391 (M + 1)+ 392


6) 120575 373 (6H s OCH

3) 49 (1H s NH)


6H) 669 (2H d J = 142Hz C

1H and C

7H) 671ndash



6) 120575 374 (6H s OCH

3) 51 (1H s NH)


6H)

668 (2H d J = 146Hz C1H and C

7H) 678ndash712 (6H m

ArH) 972 (2H s OH) MSmz (M+) 408 (M + 1)+ 409




HO

ONHN

OH

O

X

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R








119891minus 119879

119894)(119862 minus 119879

119894)] times 100 for


119894and [(119879

119891minus 119879

119894)119879

119894] times 100


119894


50)



119891minus

119879

119894)(119862 minus 119879



119891= 119879




100 times [(119879

119891minus 119879

119894)119879




50 log TGI and

log LC50


















Table 2 Continued












2) were syn-







(1)


(2)


(3)


(4)


(5)



Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


(300MHz DMSO-d6) 120575 209 (3H s CH

3) 381 (6H s




6) 120575 212 (3H s CH

3) 384 (6H s




6) 120575 119 (6H s CH

3) 383 (6H

s OCH3) 652 (1H s CH=C) 661 (2H d J = 168Hz


6) ppm 16860 14833 14722

13449 13435 1320 13120 13003 12877 12818 1268212554 12055 11604 10984 9978 5599mz = 511 (M+)


6) 120575 122 (6H s CH

3) 385 (6H s





6) 120575 396 (6H s OCH

3) 621 (1H s


6H) 675 (2H d J

= 152Hz C1H and C

7H) 687ndash757 (11H m ArH) 992 (2H


15366 14790 14680 14574 13769 13742 13515 1342412970 12831 11952 11560 11175 11028 5558 MS mz(M+) 468



6) 120575 398 (6H s OCH

3) 620


6H) 678



947 (2H s OH) MSmz (M+) 547 (M + 2)+ 549



6) 120575 379 (6H s OCH

3) 492 (1H s NH) 51 (1H


C6H) 665 (2H d J = 142Hz C

1H and C

7H) 669ndash722 (6H

m ArH) 996 (2H s OH) MSmz (M+) 391 (M + 1)+ 392


6) 120575 373 (6H s OCH

3) 49 (1H s NH)


6H) 669 (2H d J = 142Hz C

1H and C

7H) 671ndash



6) 120575 374 (6H s OCH

3) 51 (1H s NH)


6H)

668 (2H d J = 146Hz C1H and C

7H) 678ndash712 (6H m

ArH) 972 (2H s OH) MSmz (M+) 408 (M + 1)+ 409




HO

ONHN

OH

O

X

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R








119891minus 119879

119894)(119862 minus 119879

119894)] times 100 for


119894and [(119879

119891minus 119879

119894)119879

119894] times 100


119894


50)



119891minus

119879

119894)(119862 minus 119879



119891= 119879




100 times [(119879

119891minus 119879

119894)119879




50 log TGI and

log LC50


















Table 2 Continued












2) were syn-







(1)


(2)


(3)


(4)


(5)



Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of



HO

ONHN

OH

O

X

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R








119891minus 119879

119894)(119862 minus 119879

119894)] times 100 for


119894and [(119879

119891minus 119879

119894)119879

119894] times 100


119894


50)



119891minus

119879

119894)(119862 minus 119879



119891= 119879




100 times [(119879

119891minus 119879

119894)119879




50 log TGI and

log LC50


















Table 2 Continued












2) were syn-







(1)


(2)


(3)


(4)


(5)



Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of











Table 2 Continued












2) were syn-







(1)


(2)


(3)


(4)


(5)



Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of




(1)


(2)


(3)


(4)


(5)



Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Table 3 Continued


(6)


(7)


(8)Prostate cancer

PC-3 0349 578 gt100 0331 127 gt100 148 557 gt100DU-145 0419 144 NT 0389 151 NT 24 631 88

(9)


NT not tested






2 The compounds




HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


HO

O

OH

O

HO

ONN

OH

ONH

OR

HO

ONN

OH

OO

R

HO

ONHN

OH

O

X

AcOHReflux

Curcumin

ArCONHNH2

ArNHCONHNH2

NH2CXNH2

1ndash9

12ndash14

10-11

OO


OO

HOO

OHO

OOH

HOO

OHO


OOH

HOO

OHO O

NH NH

N N

OHOO

HO

O

NH

R

OHOH

HOO

OHO

NHNH

ONH

OH

HOO

OHO

NNH

X

NH

R

R

R

Curcumin

OOH

HOO

OHO

R

CurcuminO

NH

OHOH

HOO

OHO

NHNH

O

R

R

OH

HOO

OHO

NNH

OR

OOH

HOO

OHO X

OHOH

HOO

OHO

NH

O

OH

HOO

OHO

N

X

HN N

X

OHOO

HO

Curcumin

N N

OHOO

HO

O

H3C H3C

H3C

H3C

H3C

CH3

CH3


CH3

CH3

CH3

CH3CH3CH3

H2N

H2N

H2N

H3CH3CH3CCH3H3CH2N

H2N

minusH2O

minusH2O

minusH2O

minusH2O minusH2O

minusH2O

NH2

1ndash9

12ndash1410-11

X = S O





100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


100

50

0

minus50

minus100


LeukemiaG

row

th (

)



RPMI-8226K-562SR

(a)

100

50

0

minus50

minus100


Leukemia

Gro

wth

()



RPMI-8226K-562SR

(b)

100

50

minus50

minus100


Gro

wth

()


0


HT29HCT-116KM12

Colon cancer

(c)






(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


(a) (b)


(a) (b)



















4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of














4 Conclusion


Acknowledgments


References














































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

Research Article Synthesis, Characterisation, and In...

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