research article ultrasonic irradiation: synthesis,...

Research ArticleUltrasonic Irradiation Synthesis Characterization andPreliminary Antimicrobial Activity of NovelSeries of 46-Disubstituted-135-triazine ContainingHydrazone Derivatives

Hessa H Al-Rasheed1 Monirah Al Alshaikh1 Jamal M Khaled23

Naiyf S Alharbi2 and Ayman El-Faham14

1Department of Chemistry College of Science King Saud University PO Box 2455 Riyadh 11451 Saudi Arabia2Department of Botany and Microbiology College of Science King Saud University Riyadh Saudi Arabia3Department of Biotechnology and Food Technology Thamar University Dhamar Yemen4Chemistry Department Faculty of Science Alexandria University PO Box 426 Ibrahimia Alexandria 12321 Egypt

Correspondence should be addressed to Jamal M Khaled gkhaledksuedusa andAyman El-Faham aymanel fahamhotmailcom

Received 1 September 2016 Revised 19 October 2016 Accepted 31 October 2016

Academic Editor Andrea Penoni

Copyright copy 2016 Hessa H Al-Rasheed et al This 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

Novel series of 46-disubstituted-135-triazines containing hydrazone derivatives were synthesized employing ultrasonic irradiationand conventional heatingTheultrasonication gave the target products in higher yields and purity in shorter reaction time comparedwith the conventional method IR NMR (1H and 13C) elemental analysis and LC-MS confirmed the structures of the newproducts The antimicrobial and antifungal activities were evaluated for all the prepared compounds against some selected Gram-positive and Gram-negative bacterial strains The results showed that only two compounds 7i (pyridine derivative) and 7k (4-chlorobenzaldehyde derivative) displayed biological activity against some Gram-positive and Gram-negative bacteria while therest of the tested compounds did not display any antifungal activity

1 Introduction

Ultrasound has been employed more and more frequentlyin organic synthesis [1 2] because it improved the reactionrate and could adjust the selectivity performance of thereaction [3] Comparing with traditional methods ultrasonicirradiation is suitable and simply controlled and it consideredas a green powerful synthetic technique in chemical pro-cesses Ultrasonic irradiation has proven to be a particularlyimportant tool for meeting goals of the green chemistrywhich is minimization of waste and decreasing of the energyrequired for the reaction [4] Applications of ultrasonicirradiation in organic synthesis are playing important roleespecially in cases where traditional methods require drasticconditions or elongated reaction time [5ndash10]

Richards and Loomis first reported the chemical effectsof ultrasound in 1927 [11] The effect of ultrasound duringorganic reaction is due to cavitation which led to sep-aration of molecules of liquids and then the collapse ofthe bubbles offers strong impulsions that generate short-lived regions with high pressure and temperature Suchlocalized hot spots act as microreactors in which thesound energy converted into beneficial chemical form[12ndash14]

Recently Schiff base hydrazone derivatives have attractedgreat attention in many applications [15ndash17] Compoundsbearing hydrazone moiety exhibit a broad range of biologicalactivities including antifungal antibacterial antiviral anti-malarial antiproliferative anti-inflammatory and antipyreticproperties [18ndash22]

Hindawi Publishing CorporationJournal of ChemistryVolume 2016 Article ID 3464758 9 pageshttpdxdoiorg10115520163464758

2 Journal of Chemistry

N

N

N

Cl

Cl Cl

N

N

N

1

HNu1 HNu2 HNu3 = N O P S F nucleophiles

HNu1 le0∘CminusHCl

minusHCl

minusHCl

HNu2 rtHNu3 60

∘Cge

Nu1

Nu2 Nu3

Scheme 1 Nucleophilic substitution reaction of cyanuric chloride

On the other hand cyanuric chloride 1 is themost impor-tant reagent for s-triazine derivatives because of the selectivereactivity of its chlorine atoms toward nucleophiles Cyanuricchloride 1 is commercially available and a very cheap reagentwhich makes its applications even more attractive [23ndash29]The easy displacement of chlorine atoms in cyanuric chlorideby various nucleophiles was controlled by temperature to runin a stepwise manner as shown in Scheme 1

Several derivatives of s-triazine have exhibited antimicro-bial [30] antibacterial [31] antifungal [32] anti-HIV [33]and anticancer [34 35] properties and a wide array of otherbiological activities [36 37] Recently some of 135-triazinendashSchiff bases have reported a significant activity againstMycobacterium tuberculosis H37Rv [38] and moderate toexcellent antiproliferative activity and high selectivity againstthe human lung cancer cell line H460 [39]

As part of our continuous research on s-triazine deriva-tives and their biological activities [40 41] we report here thesynthesis and the preliminary antimicrobial activity of novelseries of 46-disubstituted-s-triazine containing hydrazonederivatives

2 Experimental Section

21 Chemistry

211 Materials All solvents were of analytical reagent gradeand used without further purification The 1H NMR and13C NMR spectra were recorded on a JEOL 400MHz andAVANCE III 400MHz (Bruker Germany) spectrometer atroom temperature (see Figures S1ndashS10 S12 and S14ndashS17 inSupplementary Material available online at httpdxdoiorg10115520163464758) in CDCl

3andor DMSO-d

6using

internal standard 120575 = 0 ppm Elemental analysis was per-formed on Perkin-Elmer 2400 elemental analyzer Meltingpoints were determined on a Mel-Temp apparatus and areuncorrected Fourier transform infrared spectroscopy (FTIR)spectra were recorded on Nicolet 6700 spectrometer fromKBr discs The reaction was follow-up and checks of thepurity using TLC on silica gel-protected aluminum sheets(Type 60 GF254 Merck) using a mixture of methanol-chloroform (1 9) as an eluent LC-MS (see Figures S11S13 and S18 in Supplementary Material) was performed onShimadzu 2020 UFLC-MS using an YMC Triart C

18(5 120583m

46times150mm) column and data processingwas carried out bythe LabSolution software Buffer A 01 formic acid in H

2O

and buffer B 01 formic acid in CH3CN in 30min at 120582max

254 nm were used High-resolution mass spectrometric data

were obtained using a Bruker micrOTOF-Q II instrumentoperating at room temperature and a sample concentrationof approximately 1 ppm Ultrasonic bath was purchased fromSelecta (Barcelona Spain) All compounds were named byusing ChemBioDraw Ultra version 140 Cambridge SoftCorporation (Cambridge MA USA)

(i) General Method for the Synthesis of 2-Hydrazino-46-di-morpholinio or Dimethoxy-135-triazine (3 and 5) Hydrazinehydrate (10mL 80) was added dropwise to a solution of2-chloro-46-dimorpholino or dimethoxy-135-triazine 2 or4 (20mmol) in 50mL ethanol at room temperature andthen the reaction mixture was sonicated for 60min at 60∘CEthanol and excess hydrazine were removed under vacuumand then excess diethyl ether was added to afford the productas a white solid in yield gt90 The spectral data of thetwo products 3 and 5 were in good agreement with thereported data [40 42] and were used directly without furtherpurification

(ii) General Method for the Synthesis of 135-Triazine-hydra-zone Derivatives

Method A Conventional Method 2-Hydrazino-46-disubsti-tuted-135-triazine 3 or 5 (10mmol) was added to a solu-tion of aldehyde or ketone 6 (10mmol) in ethanol (30mL)containing 2-3 drops of acetic acid and then the reactionmixturewas stirred under reflux for 3 hours After completionof the reaction the solvent was reduced under vacuumand the precipitated product was filtered off dried at roomtemperature and then recrystallized from ethyl acetate toafford the target product (Table 1)

MethodBUltrasoundAssistedMethod Amixture of aldehydeor ketone 6 (10mmol) in ethanol (30mL) 2-hydrazino-46-disubstituted-135-triazine 3 or 5 (10mmol) and 2-3 dropsof acetic acid in a flask was heated into a sonicator at 40∘Cfor 30ndash60min After completion of reaction (TLCmethanol-chloroform 1 9) UV lamp was used for spot visualization at120582max 254 the solvent was removed under vacuum and theprecipitated product was recrystallized from ethyl acetate toafford the target product (Table 1)

212 (E)-2-(2-Benzylidenehydrazinyl)-46-dimethoxy-135-triazine (7a Supporting Information Figure S1 in Supple-mentary Material) The product was obtained as a yellowsolid in yield 74 (A) 93 (B) mp 235ndash237∘C IR (KBrcmminus1) 3219 (NH) 1561 (C=N) 1467 1386 (C=C) 1H NMR(DMSO-d

6) 120575 = 389 (s 6H 2 OCH

3) 741ndash743 (m 3H

Ar) 766 (d 2H J = 64Hz Ar) 817 (s 1H CH) 1168 (s1H NH) 13C NMR (DMSO-d

6) 120575 = 545 1268 1288 1298

1344 1449 1663 ppm Anal Calc for C12H13N5O2(25927)

C 5559 H 505 N 2701 Found C 5589 H 516 N 2723

213 (E)-2-(2-(4-Chlorobenzylidene)hydrazinyl)-46-dime-thoxy-135-triazine (7b Supporting Information Figure S2in Supplementary Material) The product was obtained asa white solid in yield 71 (A) 92 (B) mp 215ndash217∘C IR(KBr cmminus1) 3228 (NH) 1614 (C=N) 1577 1473 (C=C) 1H

Journal of Chemistry 3

Table 1 Yield reaction time and melting point of 7andasho deriva-tivesusing conventional method and ultrasonic irradiation

Compdnumber

Method AConventional

method

Method BUltrasonicirradiation Mp (∘C)

Yield () Time (h) Yield () Time(min)

7a 74 4-5 93 30 235ndash2377b 71 4-5 92 30 215ndash2177c 70 4-5 93 30 204ndash2067d 68 4-5 93 30 188ndash1907e 75 4-5 95 30 220ndash2227f 75 4-5 93 60 225ndash2277g 75 4-5 93 60 227ndash2297h 71 4-5 92 60 230ndash2327i 71 4-5 92 30 212ndash2147j 75 4-5 95 30 225ndash2277k 70 4-5 94 30 233ndash2357l 72 4-5 94 60 236ndash2387m 74 4-5 96 60 213ndash2157n 70 4-5 92 60 228ndash2307o 71 4-5 90 60 232ndash234

NMR (DMSO-d6) 120575 = 389 (s 6H 2 OCH

3) 747 (d 2H 119869 =

88Hz Ar) 769 (d 2H J = 88Hz Ar) 815 (s 1H CH) 1173(s 1H NH) 13C NMR (DMSO-d

6) 120575 = 545 1284 12891334 1341 1436 1663 ppm Anal Calc for C12H12ClN5O2(29307) C 4907 H 412 N 2384 Found C 4892 H 422N 2403

214 (E)-24-Dimethoxy-6-(2-(4-methylbenzylidene)hydrazi-nyl)-135-triazine (7c Supporting Information Figure S3 inSupplementaryMaterial) Theproductwas obtained as a lightyellow solid in yield 70 (A) 93 (B) mp 204ndash206∘C IR(KBr cmminus1) 3219 (NH) 1564 (C=N) 1465 1367 (C=C) 1HNMR (DMSO-d

6) 120575 = 233 (s 3H CH

3) 390 (s 6H 2

OCH3) 724 (d 2H J = 80Hz Ar) 758 (d 2H J = 808Hz

Ar) 815 (s 1H CH) 1162 (s 1HNH) 13CNMR (DMSO-d6)

120575 = 210 545 1268 1294 1317 1396 1451 1662 ppm AnalCalc for C

13H15N5O2(27312) C 5713 H 553 N 2563

Found C 5733 H 541 N 2580

215 (E)-24-Dimethoxy-6-(2-(4-methoxybenzylidene)hy-drazinyl)-135-triazine (7d Supporting Information Figure S4in Supplementary Material) The product was obtained as alight yellow solid in yield 68 (A) 93 (B) mp 188ndash190∘CIR (KBr cmminus1) 3244 (NH) 1570 (C=N) 1479 1364 (C=C)1H NMR (DMSO-d6) 120575 = 380 (s 3H OCH3) 391 (s6H 2OCH

3) 704 (d 2H J = 80Hz Ar) 762 (d 2H J =

808Hz Ar) 812 (s 1H CH) 1156 (s 1H NH) 13C NMR(DMSO-d

6) 120575 = 545 553 1143 1270 1285 1300 1449

1606 1661 ppm Anal Calc for C13H15N5O3(28930) C

5397 H 523 N 2421 Found C 5373 H 507 N 2400

216 (E)-2-(2-(4-Bromobenzylidene)hydrazinyl)-46-dime-thoxy-135-triazine (7e Supporting Information Figure S5in Supplementary Material) The product was obtained asa white solid in yield 75 (A) 95 (B) mp 220ndash222∘C IR(KBr cmminus1) 3217 (NH) 1578 (C=N) 1460 1368 (C=C) 1HNMR (DMSO-d

6) 120575 = 389 (s 6H 2 OCH3) 764 (s 4H Ar)815 (s 1H CH) 1174 (s 1H NH) 13C NMR (DMSO-d

6)

120575 = 545 1229 1287 13187 1337 1438 1663 ppm AnalCalc for C

12H12BrN5O2(33817) C 4262 H 358 N 2071

Found C 4288 H 364 N 2098

217 (E)-4-((2-(46-Dimethoxy-135-triazin-2-yl)hydrazon-o)methyl) phenol (7f Supporting Information Figure S6 inSupplementary Material) The product was obtained as awhite solid in yield 75 (A) 93 (B) mp 225ndash227∘C IR(KBr cmminus1) 3229 (OH) 3135 (NH) 1609 (C=N) 1567 1493(C=C) 1H NMR (DMSO-d

6) 120575 = 388 (s 3H OCH

3) 390

(s 3H OCH3) 681 (d 2H J = 80Hz Ar) 752 (d 2H J =

80Hz Ar) 807 (s 1H CH) 993 (s 1H OH) 1147 (s 1HNH) 13C NMR (DMSO-d

6) 120575 = 545 1158 1255 1288

1455 1592 1661 1718 ppm Anal Calc for C12H13N5O3

(27527) C 5236 H 476 N 2544 Found C 5221 H 489N 2563

218 (E)-4-((2-(46-Dimethoxy-135-triazin-2-yl)hydrazon-o)methyl)-NN-dimethylaniline (7g Supporting InformationFigure S7 in Supplementary Material) The product wasobtained as a yellow solid in yield 75 (A) 93 (B) mp227ndash229∘C IR (KBr cmminus1) 3211 (NH) 1567 (C=N) 14591362 (C=C) 1HNMR (DMSO-d

6) 120575= 295 (s 6HN(CH

3)2)

386 (s 3H OCH3) 389 (s 3H OCH

3) 671 (d 2H J =

72Hz Ar) 752 (d 2H J = 88Hz Ar) 803 (s 1H CH)1138 (s 1H NH) 13C NMR (DMSO-d

6) 120575 = 404 550

1125 1224 1288 1466 1519 1665 ppm Anal Calc forC14H18N6O2 (30234) C 5562 H 600 N 2780 Found C5588 H 620 N 2804

219 (E)-4-(1-(2-(46-Dimethoxy-135-triazin-2-yl)hydrazon-o)ethyl)phenol (7h Supporting Information Figure S8 in Sup-plementary Material) The product was obtained as a yellowsolid in yield 71 (A) 92 (B) mp 230ndash232∘C IR (KBrcmminus1) 3466 (OH) 3364 (NH) 1576 (C=N) 1475 1374 (C=C)1H NMR (DMSO-d6) 120575 = 226 (s 3H CH3) 391 (s 6H2OCH3) 678 (d 2H J = 88Hz Ar) 768 (d 2H J = 84HzAr) 978 (s 1H OH) 1047 (s 1H NH) 13C NMR (DMSO-d6) 120575 = 139 544 1152 1279 1292 1513 1586 1669 ppm

Anal Calc for C13H15N5O3 (28930) C 5397 H 523 N242 Found C 5377 H 533 N 2443

2110 (E)-24-Dimethoxy-6-(2-(pyridin-2-ylmethylene)hy-drazinyl)-135-triazine (7i Supporting Information Figure S9in Supplementary Material) The product was obtained as apale yellow solid in yield 69 (A) 91 (B) mp 212ndash214∘CIR (KBr cmminus1) 3236 (NH) 1572 (C=N) 1467 1366 (C=C)1H NMR (DMSO-d

6) 120575 = 392 (s 6H 2OCH

3) 739(t 1H J

= 56 Ar) 788 (t 1H J = 72Hz Ar) 796 (d 1H J = 80HzAr) 822 (s 1H CH) 859 (d 1H J = 52Hz Ar) 1138 (s 1H


NH) 13CNMR (DMSO-d6) 120575 = 541 1197 1239 1369 1444


(26026) C 5077 H 465 N 3229 Found C 5096 H 473N 3240

2111 (E)-441015840-(6-(2-Benzylidenehydrazinyl)-135-triazine-24-diyl)dimorpholine (7j Supporting Information Figure S10in Supplementary Material) The product was obtained asa white solid in yield 75 (A) 95 (B) mp 215ndash217∘C IR(KBr cmminus1) 3267 (NH) 1546 (C=N) 1479 1450 (C=C) 1HNMR (CDCl

3) 120575 = 370ndash374 (m 8H 4 OCH

2-) 375ndash384

(m 8H 4 NCH2-) 735ndash741 (m 3H Ar) 770ndash773 (m 2H

Ar) 782 (s 1H CH) 831 (s 1H NH) 13CNMR (CDCl3) 120575 =

436 437 668 669 1271 1286 1296 1342 1425 1654 ppmAnal Calc for C

18H23N7O2(36943) C 5852 H 628 N

2654 Found C 5874 H 643 N 2679 (mz) Calcd36943 LC-MS (M+H) Found 370 at 119877119905 1411min (supportinginformation Figure S11 in Supplementary Material)

2112 (E)-441015840-(6-(2-(4-Chlorobenzylidene)hydrazinyl)-135-triazine-24-diyl)dimorpholine (7k Supporting InformationFigure S12 in Supplementary Material) The product wasobtained as a white solid in yield 70 (A) 94 (B) mp233ndash235∘C IR (KBr cmminus1) 3270 (NH) 1516 (C=N) 14801442 (C=C) 1H NMR (CDCl

3) 120575 = 371ndash374 (m 8H 4

OCH2-) 375ndash384 (m 8H 4 NCH2-) 736 (d 2H J =

40Hz Ar) 772 (d 2H J = 40Hz Ar) 782 (s 1H CH)831 (brs 1H NH) 13C NMR (CDCl

3) 120575 = 436 437 668669 1282 1289 1328 1354 1411 1654 ppm Anal Calc forC18H22ClN7O2 (40387) C 5353H 549N 2428 Found C5378 H 560 N 2451 (mz) Calcd 40387 LC-MS (M+H)Found 404 at 119877119905 158min (supporting information Figure S13in Supplementary Material)

2113 (E)-441015840-(6-(2-(4-Bromobenzylidene)hydrazinyl)-135-triazine-24-diyl) dimorpholine (7l Supporting InformationFigure S14 in Supplementary Material) The product wasobtained as a white solid in yield 72 (A) 94 (B) mp 236ndash238∘C IR (KBr cmminus1) 3273 (NH) 1515 (C=N) 1479 1441(C=C) 1H NMR (DMSO-d6) 120575 = 362 (m 8H 4 OCH2-)370 (m 8H 4 NCH2) 759 (s 4H Ar) 803 (s 1H CH)1090 (s 1H NH) 13C NMR (CDCl

3) 120575 = 438 666 12261288 1323 1349 1410 1647 1653 ppm Anal Calc forC18H22BrN7O2(44833) C 4822 H 495 N 2187 Found

C 4844 H 508 N 2203

2114 (E)-4-((2-(46-Dimorpholino-135-triazin-2-yl)hydra-zono)methyl)phenol (7m Supporting Information Figure S15in Supplementary Material) The product was obtained as awhite solid in yield 74 (A) 96 (B) mp 213ndash215∘C IR (KBrcmminus1) 3448 (OH) 3250 (NH) 1543 (C=N) 1493 1443 (C=C)1HNMR (CDCl3) 120575 = 362ndash374 (m 16H 4 N-CH2-CH2-O)674 (d 2H J = 856Hz Ar) 751 (d 2H J = 860Hz Ar) 768(s 1H CH) 802 (brs 1H NH) 13C NMR (CDCl

3) 120575 = 436

437 668 669 1156 1268 1289 1574 1654 ppm Anal Calcfor C18H23N7O3(38543) C 5609 H 602 N 2544 Found

C 5623 H 621 N 2569

2115 (E)-4-((2-(46-Dimorpholino-135-triazin-2-yl)hydra-zono)methyl)-NN-dimethylaniline (7n Supporting Informa-tion Figure S16 in Supplementary Material) The product wasobtained as a yellow solid in yield 70 (A) 92 (B) mp228ndash230∘C IR (KBr cmminus1) 3277 (NH) 1529 (C=N) 14901441 (C=C) 1H NMR (DMSO-d6) 120575 = 362ndash369 (m 16H 4N-CH2-CH2-O) 672 (d 2H J = 804Hz Ar) 744 (d 2H J= 88Hz Ar) 795 (s 1H CH) 1052 (s 1H NH) 13C NMR(DMSO-d

6) 120575 438 666 1124 1232 1282 1433 1514 1645

1653 ppm Anal Calc for C20H28N8O2(41250) C 5824 H

684 N 2717 Found C 5851 H 699 N 2736

2116 (E)-4-(1-(2-(46-Dimorpholino-135-triazin-2-yl)hy-drazono)ethyl)phenol (7o Supporting Information Figure S17in Supplementary Material) The product was obtained asa white solid in yield 71 (A) 90 (B) mp232ndash234∘C IR(KBr cmminus1) 3452 (OH) 3350 (NH) 1563 (C=N) 1524 1487(C=C) 1H NMR (CDCl

3) 120575 = 224 (s 3H CH

3) 370ndash383

(m 16H 4 N-CH2-CH2-O) 680 (d 2H J = 88Hz Ar) 769

(d 2H J = 846Hz Ar) 791 (s 1H CH) 801 (brs 1H NH)13C NMR (CDCl

3) 120575 = 127 437 669 1153 1278 1282

1309 1469 1568 1647 1652 1654 ppm Anal Calc forC19H25N7O3(39946) C 5713 H 631 N 2455 Found C

5735 H 644 N 2480 (mz) Calcd 39946 LC-MS (M+H)Found 400 at 119877

119905134min (supporting information Figure S18

in Supplementary Material)

22 Biology

221 Antimicrobial Activity The antimicrobial activities ofall compounds 7andasho were evaluated against some selectedpathogenic Gram-positive Gram-negative and filamentousfungus strains by using the disc diffusion method [43]Staphylococcus aureus (ATCC 29213) Neisseria meningi-tides (ATCC 1302) Streptococcus mutans (ATCC 35668)Escherichia coli (ATCC 25922) Pseudomonas aeruginosa(ATCC 27584) Salmonella typhimurium (ATCC 14028)Brevibacillus laterosporus Wild strain Candida parapsilo-sis (ATCC 22019) Cryptococcus neoformans Wild strainCandida albicans (ATCC 60193) Penicillium chrysogenum(AUMC9476) Aspergillus niger (AUMC8777) andFusariumsp were used in the evaluation test

Mueller-Hinton agar (Scharlau Microbiology Spain) wasused and prepared according to manufacturerrsquos guidelineswhere 1 L of medium was prepared containing 175 15 and17 gm of peptone meat infusion solid starch and agarrespectively For preparation of 1 L of medium 21 gm ofpowder was dissolved in 1 L of distilled water and sterilizedat 121∘C for 15min by an autoclave (HL-321 Taiwan) Aftersterilization and cooling to 50∘C the medium dispensed inPetri dishes and left to cool down to 25∘C Then they wereinoculated with the bacterial strains by streaking In the anti-fungal test a potato dextrose agar (PDA) (Scharlau Spain)was used and prepared according to the manufacturerrsquosdirections where 39 gm PDA dissolved in 1 L of distilledwater and then followed the previously described method

The microbial inoculation suspensions were preparedin sterile sodium chloride solution (089) from activated


N

N

N

Cl

Cl Cl

1

N

N

N

Cl

N N

N

N

N

HN

N N

N

N

N

Cl

MeO OMe

N

N

N

HN

MeO OMe

EthanolUS

MeOH

O O

2 3

O O

4 5

and rt 12

NaHCO3

NaHCO3

0 ∘

60 ∘C (4 h)

C (1 h)

NH2NH2

NH2

NH2

NH2NH2

60min60∘C

EthanolUS 60min60∘C

Acetone-H2O

h2 eq Morpholine0∘ C 2 h

Scheme 2 Synthesis of 2-hydrazino-46-disubstituted-135-triazine derivatives and US ultrasonic irradiation

microbial culturesThe optical densities of microbial suspen-sions were adjusted to 064 at 600 nm A sterile swab wasmoistened with the microbial suspensions and inoculatingthe dried surface of the medium in a Petri dish Afterinoculation all Petri dishes were kept at 25∘C for 5ndash10minutes before dispensing the standard antibiotic agentsand the prepared compounds discs on the surface of themedia

In the present work tobramycin (10 120583gdesk) chlo-ramphenicol (30 120583gdisk) fusidic acid (10 120583gdisk) aug-mentin (10120583gdisk) cycloheximide (30120583gdisk) canesten(10 120583gdisk) and caspofungin (10120583gdisk) were used as stan-dard antibacterial and antifungal agents To prepare disks(4mgdisk) from the tested compounds 200mg from eachcompound was dissolved in dimethylsulfoxide (DMSO) andthen 20120583L of the solution was added on the sterile filter disk(6mm) and then allowed to dry at room temperature insidethe safety biological cabinet After 18ndash24 h of incubation at37∘C for bacteria and 48ndash72 h at 25∘C for fungi theminimuminhibitory concentration (MIC mgmL) of the preparedcompounds was measured [44] The microdilution assaywas applied in 96-well plate (Corning Incorporated USA)using twofold serial dilution The original concentration was4mgmL and the total volume was 200 120583L (1 1 chemicalsuspension bacterial suspension) 4-iodonitrotetrazoliumviolet (SigmaUSA) reagent was added after the incubation tomeasure the bacterial growth through the emergence of violetcolor

3 Results and Discussion

31 Chemistry Compounds 2 and4were obtained using one-step reaction where cyanuric chloride 1 was reacted withmorpholine (2 equiv) in acetone-water media or methanol(as a solvent) in the presence of NaHCO

3as hydrogen

chloride removal at 0∘C for 2 h In the case of the synthesis of

dimorpholino derivative 2 the reaction temperature raisedgradually to room temperature and kept under stirring for12 h at the same temperature while the reported method [4042] was used for the preparation of the dimethoxy derivative4 (Scheme 2) The products 2 and 4 were obtained in goodyields and their spectral data agreed with the reported data[40 42]

The hydrazine derivatives 3 and 5 were obtained by treat-ment of 2 or 4 with hydrazine hydrate (80) in ethanol for60min at 60∘C employing ultrasonic irradiation (Scheme 2)to afford the products in excellent yields (gt90) and purity

The products 7andasho were obtained by condensation of thehydrazine derivative 3 or 5 with substituted benzaldehyde oracetophenone 6 in ethanol containing 2-3 drops of glacialacetic acid using ultrasonic irradiation for 30ndash60min at 40∘C(Scheme 3) to afford the target products in excellent yieldsand purity as observed from LC-MS (see SupplementaryFigures S11 S13 and S18) Ultrasonic irradiation gave thetarget products in high yields in shorter reaction timecompared with the conventional heating as shown in Table 1

The 1H NMR spectrum of 7k as a prototype for thebenzaldehyde derivatives showed a multiplet peak in therange at 120575 371ndash374 ppm related to 4 methylene groups (4OCH2-) another multiplet peak in the range at 120575 375ndash384 ppm related to 4 methylene groups (4 NCH2-) doubletat 120575 736 ppm for the two aromatic protons H-3 and H-5doublet at 120575 772 for the twoaromatic protons H-2 and H-6singlet at120575 782 related to theCH (CH=N- group) and a broadsinglet at 120575 831 for the NH The 13C NMR spectrum of 7kshowed absorption peak for themorpholine residue at 120575 436437 668 and 669 related to 2 CH

2-N-CH

2 and 2 CH

2-O-

CH2respectively absorption peaks at 120575 1282 1289 1328 and

1354 ppm related to the aromatic carbons and absorptionpeaks at 120575 1425 and 1654 ppm for C=N

The LC-MS of compound 7k using buffer A 01 formicacid in H2O and buffer B 01 formic acid in CH3CN in


N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z

X = morpholine 3 X = OMe 5

Y

Y

NCHO N

N

N

HN

MeO OMe

N

N

7iEthanol (2-3) drops AcOH

Ethanol (2-3) drops AcOH

US60 min40∘ C

US30ndash60 min40∘C

7andashh Jndasho

Z = H Y = H Cl Br CH3 OH OCH3 NMe2

Z = CH3 Y = OH

NH2

Scheme 3 Synthesis of 46-disubstituted-s-triazine-Schiff base derivatives and US ultrasonic irradiation

N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z

Scheme 4 E- and Z-isomer of compound 7k

30min showed one peak at 119877119905158min with the expected

mass [M+H] 404 (mz cacld 40387 Figure S13 in the Sup-plementary Material)

Compound 7k as an example could adopt two differentgeometrical isomers (E Z Scheme 4) Therefore 7k wasdemonstrated usingmolecular mechanicsMM2 calculationsIn addition quantum chemical calculations were carriedout with the GAUSSIAN 98 suite of programs Geom-etry optimization was carried out using the DFT level(B3LYP6-31Glowastlowast) of theory to assess the relative stabilityof the E-Z isomeric species Computed relative energiesof 7k indicated that the E-isomer (total energy content477837 kcalmol) is more stable than the Z ones (totalenergy content 417295 kcalmol) by 60542 kcalmol Thisobservation agreed with our reported data in which the

s-triazine hydrazone preferred the E-isomer rather than theZ- ones [41]

As a prototype for acetophenone derivatives the 1HNMRspectrum of 7o showed a singlet peak at 120575 224 ppm forthe methyl group of the acetophenone moiety two multipletpeaks in the range 120575 370ndash383 ppm related to the 8methylenegroups of the two morpholino rings (4 -N-CH

2-CH2-O)

and a broad singlet at 120575 801 ppm related to the NH besidetwo doublets forming an AB system at 120575 680 (J = 88Hz)and 7698 ppm (J = 846) for the aromatic protons H-3 H-5and H-2 H-6 respectively The 13C NMR spectrum showedabsorption peak at 120575 127 related to methyl group and twopeaks at 120575 437 and 669 ppm related to the methylene groups(-NCH2 and O-CH2 resp) the absorption peaks at 120575 15681647 1652 and 1654 ppm were assigned for carbons of


Table 2 Antimicrobial activity (zones of inhibition mm) and a minimum inhibitory concentration (MIC mgml) of 7i and 7k comparingwith several standard antimicrobial drugs

MicroorganismsAntifungal agent Antibacterial agents Compounds

mM mgmLCylowast Co Ca To Ch Fu 7i 7k 7i 7k

Bacteria

Salowastlowast NT NT NT 14 30 23 15 mdash 010 mdashSm NT NT NT mdash 35 20 10 mdash 020 mdashEc NT NT NT 20 40 mdash mdash lt8 mdash mdashPa NT NT NT 15 13 mdash mdash mdash mdash mdashSt NT NT NT 14 30 mdash mdash lt8 mdash mdashNm NT NT NT 21 34 20 15 17 010 010BL NT NT NT mdash 35 20 20 mdash 005 mdash

Fungi

Cp mdash mdash mdash NTlowastlowastlowast NT NT mdash mdash mdash mdashCn 32 25 mdash NT NT NT mdash mdash mdash mdashCa mdash 46 20 NT NT NT mdash mdash mdash mdashPc 23 30 mdash NT NT NT mdash mdash mdash mdashAn 22 23 20 NT NT NT mdash mdash mdash mdashFs 30 15 mdash NT NT NT mdash mdash mdash mdash

lowastCy cycloheximide Co canesten Ca caspofungin To tobramycin Ch chloramphenicol and Fu fusidic acid 119878119886lowastlowast Staphylococcus aureusATCC 29213 NmNeisseria meningitides ATCC 1302 Sm Streptococcus mutans ATCC 35668 EC Escherichia coli ATCC 25922 Pa Pseudomonas aeruginosa ATCC 27584 StSalmonella typhimurium ATCC 14028 Cp Candida parapsilosis ATCC 22019 Cn Cryptococcus neoformansWild strain C Candid albicans ATCC 60193 BLBrevibacillus laterosporusWild strain Pc Penicillium chrysogenum AUMC 9476 An Aspergillus niger AUMC 8777 and Fs Fusarium spNTlowastlowastlowast not tested

C=N and the remaining five peaks at 120575 1153 1278 12821309 and 1469 ppm related to the aromatic carbons of theacetophenone moiety

The LC-MS of compound 7o using buffer A 01 formicacid in H

2O and buffer B 01 formic acid in CH

3CN in


mass [M+H] 400 (mz calcd 39946 Figure S18 in theSupplementary Material)

32 Biology The antimicrobial activities of the preparedcompounds against several pathogenic tested organismsare represented in Table 2 The results revealed that onlytwo compounds 7i and 7k from all the tested compoundswere biologically active with different spectrum activity onthe other hand they did not show any antifungal activity(Table 2) The results in Table 2 showed that 7i had goodbiological activity against S aureus N meningitides andtobramycin resistant B laterosporus while it had poor bio-logical activity against S mutans (10mm) Compound 7kshowed a good biological activity against N meningitides(17mm) and a very weak activity (lt8) against E coli and Styphimurium The measured minimum inhibitory concen-trations (MIC) of 7i that inhibited S aureus S mutans Nmeningitides and B laterosporus were 010 020 010 and005mgmL respectively while the MIC of 7k that inhibitedN meningitides was 010mgmL

It is obvious from Table 2 that a small structural variationin s-triazine ring may induce an effect on antibacterialactivity By observing the antibacterial activities of all thederivatives it is interesting to note that the dimorpholino-4-chlorophenyl-s-triazine derivatives 7k show good antibacte-rial activity against N meningitides and weak activity against

E coli and S typhimurium The other derivatives in the samefamily did not show any biological activity this observationagreed with the reported data for substituted s-triazine [45]The presence of pyridine ring (compound 7o) in place ofthe benzaldehyde ring (compound 7a) changed the biologicalactivity significantly as observed from Table 2 on the otherhand all the tested compounds did not show any antifungalactivity this observation also agreed with the reported datafor s-triazine derivatives [45]

4 Conclusion

The target products (7andasho) were prepared using conventionalheating and ultrasonic irradiation The ultrasonic irradia-tion affords the products in higher yields and purity inshorter reaction timeThe biological activity for the preparedcompounds showed that only two compounds 7i (pyridinederivative) and 7k (4-chlorobenzaldehyde derivative) havebiological activities against some Gram-positive and Gram-negative bacteria while all tested compounds did not showany antifungal activity It is obvious that changing thesubstituent in the s-triazine ring as well as the benzylidenemoiety may affect the antimicrobial activity Further inves-tigations are run in our lab to get insights into the mode ofaction and the relation between the biological activity andsubstituent effect in the pattern in s-triazine

Competing Interests

The authors declare no conflict of interests


Acknowledgments

The authors thank the Deanship of Scientific Research atKing Saud University for funding this work through ProlificResearch Group Program (PRG-1437-33 Saudi Arabia)

References

[1] C Petrier J L Luche and J L Luche Synthetic OrganicSonochemistry Plenum Press New York NY USA 1998

[2] J-T LiW-Z Yang S-XWang S-H Li andT-S Li ldquoImprovedsynthesis of chalcones under ultrasound irradiationrdquo Ultrason-ics Sonochemistry vol 9 no 5 pp 237ndash239 2002

[3] K Shibata I Katsuyama M Matsui and H Muramatsu ldquoSyn-thesis of ferrocenyl-substituted 3-cyano-2-methylpyridinesrdquoBulletin of the Chemical Society of Japan vol 63 no 12 pp 3710ndash3712 1990

[4] P Cintas and J-L Luche ldquoGreen chemistry the sonochemicalapproachrdquo Green Chemistry vol 1 no 3 pp 115ndash125 1999

[5] T J Mason ldquoUltrasound in synthetic organic chemistryrdquoChemical Society Reviews vol 26 no 6 pp 443ndash451 1997

[6] R Cella and H A Stefani ldquoUltrasound in heterocycles chem-istryrdquo Tetrahedron vol 65 no 13 pp 2619ndash2641 2009

[7] M F Mady A A El-Kateb I F Zeid and K B JorgensenldquoComparative studies on conventional and ultrasound-assistedsynthesis of novel homoallylic alcohol derivatives linked tosulfonyl dibenzene moiety in aqueous mediardquo Journal of Chem-istry vol 2013 Article ID 364036 9 pages 2013

[8] J-T Li G-F Chen W-Z Yang and T-S Li ldquoUltrasoundpromoted synthesis of 2-aroyl-135-triaryl-4-carbethoxy-4-cyanocyclohexanolsrdquo Ultrasonics Sonochemistry vol 10 no 3pp 123ndash126 2003

[9] J-T Li G-F ChenW-Z Xu andT-S Li ldquoTheMichael reactioncatalyzed by KFbasic alumina under ultrasound irradiationrdquoUltrasonics Sonochemistry vol 10 no 2 pp 115ndash118 2003

[10] L Ding W Wang and A Zhang ldquoSynthesis of 15-dinitroaryl-14-pentadien-3-ones under ultrasound irradiationrdquoUltrasonicsSonochemistry vol 14 no 5 pp 563ndash567 2007

[11] W T Richards and A L Loomis ldquoThe chemical effects of highfrequency sound waves I A preliminary surveyrdquo Journal of theAmerican Chemical Society vol 49 no 12 pp 3086ndash3100 1927

[12] C Leonelli and T JMason ldquoMicrowave and ultrasonic process-ing now a realistic option for industryrdquo Chemical Engineeringand Processing Process Intensification vol 49 no 9 pp 885ndash900 2010

[13] G Cravotto and P Cintas ldquoPower ultrasound in organic synthe-sis moving cavitational chemistry from academia to innovativeand large-scale applicationsrdquo Chemical Society Reviews vol 35no 2 pp 180ndash189 2006

[14] T J Mason A J Cobley J E Graves and D MorganldquoNew evidence for the inverse dependence of mechanical andchemical effects on the frequency of ultrasoundrdquo UltrasonicsSonochemistry vol 18 no 1 pp 226ndash230 2011

[15] R S Yamgar Y R Nivid S Nalawade M Mandewale RG Atram and S S Sawant ldquoNovel zinc(II) complexes ofheterocyclic ligands as antimicrobial agents synthesis charac-terisation and antimicrobial studiesrdquo Bioinorganic Chemistryand Applications vol 2014 Article ID 276598 10 pages 2014

[16] M C Mandewale B R Thorat D Shelke R Patil and RYamgar ldquoSynthesis characterization and fluorescence study ofN-[(E)-(2-hydroxyquinolin-3-yl)methylidene]-1-benzofuran-

2-carbohydrazide and its metal complexesrdquoHeterocyclic Lettersvol 5 no 2 pp 251ndash259 2015

[17] MCMandewale B RThorat andR S Yamgar ldquoSynthesis andanti-mycobacterium study of some fluorine containing Schiffbases of quinoline and their metal complexesrdquo Der PharmaChemica vol 7 no 5 pp 207ndash215 2015

[18] D N Dhar and C L Taploo ldquoSchiff bases and their applica-tionsrdquo Journal of Scientific amp Industrial Research vol 41 no 8pp 501ndash506 1982

[19] P Przybylski A Huczynski K Pyta B Brzezinski and F BartlldquoBiological properties of schiff bases and azo derivatives ofphenolsrdquo Current Organic Chemistry vol 13 no 2 pp 124ndash1482009

[20] M Singh and N Raghav ldquoBiological activities of hydrazones areviewrdquo International Journal of Pharmacy and PharmaceuticalSciences vol 3 no 4 pp 26ndash32 2011

[21] B Kaya Y Ozkay H E Temel and Z A Kaplancıklı ldquoSyn-thesis and biological evaluation of novel piperazine containinghydrazone derivativesrdquo Journal of Chemistry vol 2016 ArticleID 5878410 7 pages 2016

[22] M C Mandewale B Thorat D Shelke and R Yamgar ldquoSyn-thesis and biological evaluation of new hydrazone derivativesof quinoline and their Cu(II) and Zn(II) complexes againstMycobacterium tuberculosisrdquo Bioinorganic Chemistry and Appli-cations vol 2015 Article ID 153015 14 pages 2015

[23] G Blotny ldquoRecent applications of 246-trichloro-135-triazineand its derivatives in organic synthesisrdquoTetrahedron vol 62 no41 pp 9507ndash9522 2006

[24] B Kolesinska and Z J Kaminski ldquoThe umpolung of substituenteffect in nucleophilic aromatic substitution A new approach tothe synthesis of NN-disubstituted melamines (triazine triske-lions) under mild reaction conditionsrdquo Tetrahedron vol 65 no18 pp 3573ndash3576 2009

[25] G R Gustafson C M Baldino M-M E OrsquoDonnell et alldquoIncorporation of carbohydrates and peptides into largetriazine-based screening libraries using automated parallelsynthesisrdquo Tetrahedron vol 54 no 16 pp 4051ndash4065 1998

[26] S S Machakanur B R Patil D S Badiger R P Bakale KB Gudasi and S W A Bligh ldquoSynthesis characterization andanticancer evaluation of novel tri-arm star shaped 135-triazinehydrazonesrdquo Journal of Molecular Structure vol 1011 pp 121ndash127 2012

[27] K Srinivas U Srinivas V J Rao K Bhanuprakash K HKishore andU SNMurty ldquoSynthesis and antibacterial activityof 246-tri substituted s-triazinesrdquo Bioorganic and MedicinalChemistry Letters vol 15 no 4 pp 1121ndash1123 2005

[28] G Nieddu and G Giacomelli ldquoA microwave assisted synthesisof benzoxazoles from carboxylic acidsrdquo Tetrahedron vol 69 no2 pp 791ndash795 2013

[29] C Courme N Gresh M Vidal et al ldquoSynthesis of aryl phos-phates based on pyrimidine and triazine scaffoldsrdquo EuropeanJournal of Medicinal Chemistry vol 45 no 1 pp 244ndash255 2010

[30] J P Raval A R Rai N H Patel H V Patel and P SPatel ldquoSynthesis and in vitro antimicrobial activity of Nrsquo-(4-(arylamino)-6- (pyridin-2-ylamino)-135-triazin-2-yl)ben-zohydraziderdquo International Journal of ChemTech Research vol1 no 3 pp 616ndash620 2009

[31] S Nishigaki F Yoneda H Matsumoto and K Morinaga ldquoSyn-thetic antibacterials I Nitrofurylvinyl-s-triazine derivativesrdquoJournal of Medicinal Chemistry vol 12 no 1 pp 39ndash42 1968


[32] L D S Yadav K N Shukla R Dwivedi and H SinghldquoSynthesis of new 1 3 4-oxadiazolo [3 2-a]-s-triazine-5 7-dithiones and the dithionone analogues as potential antifungalagentsrdquo Indian Journal of Pharmaceutical Sciences vol 54 no 1pp 33ndash37 1992

[33] D H Mahajan C Pannecouque E De Clercq and K HChikhalia ldquoSynthesis and studies of new 2-(coumarin-4-yloxy)-46-(substituted)-S-triazine derivatives as potential anti-HIV agentsrdquo Archiv der Pharmazie vol 342 no 5 pp 281ndash2902009

[34] Z Nie C Perretta P Erickson et al ldquoStructure-based designsynthesis and study of pyrazolo[15-a][135]triazine derivativesas potent inhibitors of protein kinase CK2rdquo Bioorganic andMedicinal Chemistry Letters vol 17 no 15 pp 4191ndash4195 2007

[35] Z Nie C Perretta P Erickson et al ldquoStructure-based designand synthesis of novel macrocyclic pyrazolo[15-a] [135]tri-azine compounds as potent inhibitors of protein kinase CK2and their anticancer activitiesrdquoBioorganic andMedicinal Chem-istry Letters vol 18 no 2 pp 619ndash623 2008

[36] V K Pandey S Tusi Z Tusi M Joshi and S Bajpai ldquoSynthesisand biological activity of substituted 246-s-triazinesrdquo ActaPharmaceutica vol 54 no 1 pp 1ndash12 2004

[37] A Agarwal K Srivastava S K Puri and P M S ChauhanldquoSynthesis of substituted indole derivatives as a new classof antimalarial agentsrdquo Bioorganic and Medicinal ChemistryLetters vol 15 no 12 pp 3133ndash3136 2005

[38] V R Avupati R P Yejella V R Parala et al ldquoSynthesis charac-terization and in vitro biological evaluation of some novel 135-triazinemdashSchiff base conjugates as potential antimycobacterialagentsrdquo Bioorganic and Medicinal Chemistry Letters vol 23 no21 pp 5968ndash5970 2013

[39] W Zhu Y Liu Y Zhao et al ldquoSynthesis and biological evalu-ation of novel 6-hydrazinyl-24-bismorpholino pyrimidine and135-triazine derivatives as potential antitumor agentsrdquo Archivder Pharmazie vol 345 no 10 pp 812ndash821 2012

[40] A El-Faham S M Soliman S M Osman et al ldquoOne potsynthesis molecular structure and spectroscopic studies (X-rayIR NMR UV-Vis) of novel 2-(46-dimethoxy-135-triazin-2-yl) amino acid ester derivativesrdquo Spectrochimica ActamdashPart AMolecular and Biomolecular Spectroscopy vol 159 pp 184ndash1982016

[41] A El-Faham S M Soliman H A Ghabbour et al ldquoUltrasonicpromoted synthesis of novel s-triazine-Schiff base derivativesmolecular structure spectroscopic studies and their prelimi-nary anti-proliferative activitiesrdquo Journal ofMolecular Structurevol 1125 pp 121ndash135 2016

[42] S T Asundaria S A Patel K M Mehta and K C PatelldquoSynthesis characterization and antimicrobial studies of somenovel 134-thiadiazolium-2-thiolate derivativesrdquo South AfricanJournal of Chemistry vol 63 pp 141ndash144 2015

[43] D G Gerbig Jr J E Ndong and H Aubihl ldquoA new twist to thekirby-bauer antibiotic susceptibility test activitymdashincreasingantibiotic sensitivity of pseudomonas fluorescens through ther-mal stressrdquo Journal of Microbiology and Biology Education vol14 no 2 pp 269ndash270 2013

[44] S J Cavalieri Manual of Antimicrobial Susceptibility TestingAmerican Society for Microbiology 2005

[45] K Srinivas U Srinivas K Bhanuprakash K Harakishore U SN Murthy and V Jayathirtha Rao ldquoSynthesis and antibacterialactivity of various substituted s-triazinesrdquo European Journal ofMedicinal Chemistry vol 41 no 11 pp 1240ndash1246 2006

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


N

N

N

Cl

Cl Cl

N

N

N

1

HNu1 HNu2 HNu3 = N O P S F nucleophiles

HNu1 le0∘CminusHCl

minusHCl

minusHCl

HNu2 rtHNu3 60

∘Cge

Nu1

Nu2 Nu3

Scheme 1 Nucleophilic substitution reaction of cyanuric chloride

On the other hand cyanuric chloride 1 is themost impor-tant reagent for s-triazine derivatives because of the selectivereactivity of its chlorine atoms toward nucleophiles Cyanuricchloride 1 is commercially available and a very cheap reagentwhich makes its applications even more attractive [23ndash29]The easy displacement of chlorine atoms in cyanuric chlorideby various nucleophiles was controlled by temperature to runin a stepwise manner as shown in Scheme 1

Several derivatives of s-triazine have exhibited antimicro-bial [30] antibacterial [31] antifungal [32] anti-HIV [33]and anticancer [34 35] properties and a wide array of otherbiological activities [36 37] Recently some of 135-triazinendashSchiff bases have reported a significant activity againstMycobacterium tuberculosis H37Rv [38] and moderate toexcellent antiproliferative activity and high selectivity againstthe human lung cancer cell line H460 [39]

As part of our continuous research on s-triazine deriva-tives and their biological activities [40 41] we report here thesynthesis and the preliminary antimicrobial activity of novelseries of 46-disubstituted-s-triazine containing hydrazonederivatives

2 Experimental Section

21 Chemistry

211 Materials All solvents were of analytical reagent gradeand used without further purification The 1H NMR and13C NMR spectra were recorded on a JEOL 400MHz andAVANCE III 400MHz (Bruker Germany) spectrometer atroom temperature (see Figures S1ndashS10 S12 and S14ndashS17 inSupplementary Material available online at httpdxdoiorg10115520163464758) in CDCl

3andor DMSO-d

6using

internal standard 120575 = 0 ppm Elemental analysis was per-formed on Perkin-Elmer 2400 elemental analyzer Meltingpoints were determined on a Mel-Temp apparatus and areuncorrected Fourier transform infrared spectroscopy (FTIR)spectra were recorded on Nicolet 6700 spectrometer fromKBr discs The reaction was follow-up and checks of thepurity using TLC on silica gel-protected aluminum sheets(Type 60 GF254 Merck) using a mixture of methanol-chloroform (1 9) as an eluent LC-MS (see Figures S11S13 and S18 in Supplementary Material) was performed onShimadzu 2020 UFLC-MS using an YMC Triart C

18(5 120583m

46times150mm) column and data processingwas carried out bythe LabSolution software Buffer A 01 formic acid in H

2O

and buffer B 01 formic acid in CH3CN in 30min at 120582max

254 nm were used High-resolution mass spectrometric data

were obtained using a Bruker micrOTOF-Q II instrumentoperating at room temperature and a sample concentrationof approximately 1 ppm Ultrasonic bath was purchased fromSelecta (Barcelona Spain) All compounds were named byusing ChemBioDraw Ultra version 140 Cambridge SoftCorporation (Cambridge MA USA)

(i) General Method for the Synthesis of 2-Hydrazino-46-di-morpholinio or Dimethoxy-135-triazine (3 and 5) Hydrazinehydrate (10mL 80) was added dropwise to a solution of2-chloro-46-dimorpholino or dimethoxy-135-triazine 2 or4 (20mmol) in 50mL ethanol at room temperature andthen the reaction mixture was sonicated for 60min at 60∘CEthanol and excess hydrazine were removed under vacuumand then excess diethyl ether was added to afford the productas a white solid in yield gt90 The spectral data of thetwo products 3 and 5 were in good agreement with thereported data [40 42] and were used directly without furtherpurification

(ii) General Method for the Synthesis of 135-Triazine-hydra-zone Derivatives

Method A Conventional Method 2-Hydrazino-46-disubsti-tuted-135-triazine 3 or 5 (10mmol) was added to a solu-tion of aldehyde or ketone 6 (10mmol) in ethanol (30mL)containing 2-3 drops of acetic acid and then the reactionmixturewas stirred under reflux for 3 hours After completionof the reaction the solvent was reduced under vacuumand the precipitated product was filtered off dried at roomtemperature and then recrystallized from ethyl acetate toafford the target product (Table 1)

MethodBUltrasoundAssistedMethod Amixture of aldehydeor ketone 6 (10mmol) in ethanol (30mL) 2-hydrazino-46-disubstituted-135-triazine 3 or 5 (10mmol) and 2-3 dropsof acetic acid in a flask was heated into a sonicator at 40∘Cfor 30ndash60min After completion of reaction (TLCmethanol-chloroform 1 9) UV lamp was used for spot visualization at120582max 254 the solvent was removed under vacuum and theprecipitated product was recrystallized from ethyl acetate toafford the target product (Table 1)

212 (E)-2-(2-Benzylidenehydrazinyl)-46-dimethoxy-135-triazine (7a Supporting Information Figure S1 in Supple-mentary Material) The product was obtained as a yellowsolid in yield 74 (A) 93 (B) mp 235ndash237∘C IR (KBrcmminus1) 3219 (NH) 1561 (C=N) 1467 1386 (C=C) 1H NMR(DMSO-d

6) 120575 = 389 (s 6H 2 OCH

3) 741ndash743 (m 3H

Ar) 766 (d 2H J = 64Hz Ar) 817 (s 1H CH) 1168 (s1H NH) 13C NMR (DMSO-d

6) 120575 = 545 1268 1288 1298

1344 1449 1663 ppm Anal Calc for C12H13N5O2(25927)

C 5559 H 505 N 2701 Found C 5589 H 516 N 2723

213 (E)-2-(2-(4-Chlorobenzylidene)hydrazinyl)-46-dime-thoxy-135-triazine (7b Supporting Information Figure S2in Supplementary Material) The product was obtained asa white solid in yield 71 (A) 92 (B) mp 215ndash217∘C IR(KBr cmminus1) 3228 (NH) 1614 (C=N) 1577 1473 (C=C) 1H



Compdnumber


method




NMR (DMSO-d6) 120575 = 389 (s 6H 2 OCH

3) 747 (d 2H 119869 =




6) 120575 = 233 (s 3H CH

3) 390 (s 6H 2




13H15N5O2(27312) C 5713 H 553 N 2563

Found C 5733 H 541 N 2580


3) 704 (d 2H J = 80Hz Ar) 762 (d 2H J =


6) 120575 = 545 553 1143 1270 1285 1300 1449


5397 H 523 N 2421 Found C 5373 H 507 N 2400



6)


12H12BrN5O2(33817) C 4262 H 358 N 2071

Found C 4288 H 364 N 2098


6) 120575 = 388 (s 3H OCH

3) 390



6) 120575 = 545 1158 1255 1288


(27527) C 5236 H 476 N 2544 Found C 5221 H 489N 2563


6) 120575= 295 (s 6HN(CH

3)2)

386 (s 3H OCH3) 389 (s 3H OCH

3) 671 (d 2H J =


6) 120575 = 404 550





6) 120575 = 392 (s 6H 2OCH

3) 739(t 1H J



NH) 13CNMR (DMSO-d6) 120575 = 541 1197 1239 1369 1444


(26026) C 5077 H 465 N 3229 Found C 5096 H 473N 3240


3) 120575 = 370ndash374 (m 8H 4 OCH

2-) 375ndash384




18H23N7O2(36943) C 5852 H 628 N



3) 120575 = 371ndash374 (m 8H 4






C 4844 H 508 N 2203


3) 120575 = 436


C 5623 H 621 N 2569


6) 120575 438 666 1124 1232 1282 1433 1514 1645


684 N 2717 Found C 5851 H 699 N 2736


3) 120575 = 224 (s 3H CH

3) 370ndash383



3) 120575 = 127 437 669 1153 1278 1282





22 Biology





N

N

N

Cl

Cl Cl

1

N

N

N

Cl

N N

N

N

N

HN

N N

N

N

N

Cl

MeO OMe

N

N

N

HN

MeO OMe

EthanolUS

MeOH

O O

2 3

O O

4 5

and rt 12

NaHCO3

NaHCO3

0 ∘

60 ∘C (4 h)

C (1 h)

NH2NH2

NH2

NH2

NH2NH2

60min60∘C


Acetone-H2O







3as hydrogen






2-N-CH

2 and 2 CH

2-O-





N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z


Y

Y

NCHO N

N

N

HN

MeO OMe

N

N



US60 min40∘ C


7andashh Jndasho


Z = CH3 Y = OH

NH2


N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z







2-CH2-O)






Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of



Compdnumber


method




NMR (DMSO-d6) 120575 = 389 (s 6H 2 OCH

3) 747 (d 2H 119869 =




6) 120575 = 233 (s 3H CH

3) 390 (s 6H 2




13H15N5O2(27312) C 5713 H 553 N 2563

Found C 5733 H 541 N 2580


3) 704 (d 2H J = 80Hz Ar) 762 (d 2H J =


6) 120575 = 545 553 1143 1270 1285 1300 1449


5397 H 523 N 2421 Found C 5373 H 507 N 2400



6)


12H12BrN5O2(33817) C 4262 H 358 N 2071

Found C 4288 H 364 N 2098


6) 120575 = 388 (s 3H OCH

3) 390



6) 120575 = 545 1158 1255 1288


(27527) C 5236 H 476 N 2544 Found C 5221 H 489N 2563


6) 120575= 295 (s 6HN(CH

3)2)

386 (s 3H OCH3) 389 (s 3H OCH

3) 671 (d 2H J =


6) 120575 = 404 550





6) 120575 = 392 (s 6H 2OCH

3) 739(t 1H J



NH) 13CNMR (DMSO-d6) 120575 = 541 1197 1239 1369 1444


(26026) C 5077 H 465 N 3229 Found C 5096 H 473N 3240


3) 120575 = 370ndash374 (m 8H 4 OCH

2-) 375ndash384




18H23N7O2(36943) C 5852 H 628 N



3) 120575 = 371ndash374 (m 8H 4






C 4844 H 508 N 2203


3) 120575 = 436


C 5623 H 621 N 2569


6) 120575 438 666 1124 1232 1282 1433 1514 1645


684 N 2717 Found C 5851 H 699 N 2736


3) 120575 = 224 (s 3H CH

3) 370ndash383



3) 120575 = 127 437 669 1153 1278 1282





22 Biology





N

N

N

Cl

Cl Cl

1

N

N

N

Cl

N N

N

N

N

HN

N N

N

N

N

Cl

MeO OMe

N

N

N

HN

MeO OMe

EthanolUS

MeOH

O O

2 3

O O

4 5

and rt 12

NaHCO3

NaHCO3

0 ∘

60 ∘C (4 h)

C (1 h)

NH2NH2

NH2

NH2

NH2NH2

60min60∘C


Acetone-H2O







3as hydrogen






2-N-CH

2 and 2 CH

2-O-





N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z


Y

Y

NCHO N

N

N

HN

MeO OMe

N

N



US60 min40∘ C


7andashh Jndasho


Z = CH3 Y = OH

NH2


N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z







2-CH2-O)






Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


NH) 13CNMR (DMSO-d6) 120575 = 541 1197 1239 1369 1444


(26026) C 5077 H 465 N 3229 Found C 5096 H 473N 3240


3) 120575 = 370ndash374 (m 8H 4 OCH

2-) 375ndash384




18H23N7O2(36943) C 5852 H 628 N



3) 120575 = 371ndash374 (m 8H 4






C 4844 H 508 N 2203


3) 120575 = 436


C 5623 H 621 N 2569


6) 120575 438 666 1124 1232 1282 1433 1514 1645


684 N 2717 Found C 5851 H 699 N 2736


3) 120575 = 224 (s 3H CH

3) 370ndash383



3) 120575 = 127 437 669 1153 1278 1282





22 Biology





N

N

N

Cl

Cl Cl

1

N

N

N

Cl

N N

N

N

N

HN

N N

N

N

N

Cl

MeO OMe

N

N

N

HN

MeO OMe

EthanolUS

MeOH

O O

2 3

O O

4 5

and rt 12

NaHCO3

NaHCO3

0 ∘

60 ∘C (4 h)

C (1 h)

NH2NH2

NH2

NH2

NH2NH2

60min60∘C


Acetone-H2O







3as hydrogen






2-N-CH

2 and 2 CH

2-O-





N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z


Y

Y

NCHO N

N

N

HN

MeO OMe

N

N



US60 min40∘ C


7andashh Jndasho


Z = CH3 Y = OH

NH2


N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z







2-CH2-O)






Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


N

N

N

Cl

Cl Cl

1

N

N

N

Cl

N N

N

N

N

HN

N N

N

N

N

Cl

MeO OMe

N

N

N

HN

MeO OMe

EthanolUS

MeOH

O O

2 3

O O

4 5

and rt 12

NaHCO3

NaHCO3

0 ∘

60 ∘C (4 h)

C (1 h)

NH2NH2

NH2

NH2

NH2NH2

60min60∘C


Acetone-H2O







3as hydrogen






2-N-CH

2 and 2 CH

2-O-





N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z


Y

Y

NCHO N

N

N

HN

MeO OMe

N

N



US60 min40∘ C


7andashh Jndasho


Z = CH3 Y = OH

NH2


N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z







2-CH2-O)






Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


N

N

N

HN

X X

O Z

6

+N

N

N

HN

X X

N

Z


Y

Y

NCHO N

N

N

HN

MeO OMe

N

N



US60 min40∘ C


7andashh Jndasho


Z = CH3 Y = OH

NH2


N

NN

NH

N

Cl

N

O

N

O

N

NN

NH

N

Cl

N

O

N

O

E Z







2-CH2-O)






Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of





Bacteria


Fungi






3CN in






4 Conclusion


Competing Interests



Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


Acknowledgments


References

























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

research article ultrasonic irradiation: synthesis,...

Documents