synthesis of new fused thienopyrimidines …agents [20–24], and antimalarial [25,26]. some...

Month 2017 Synthesis of New Fused Thienopyrimidines Derivatives asAnti-inflammatory Agents

Mahmoud S. Tolba,a Mostafa Ahmed,a* Adel M. Kamal El-Dean,b Reda Hassanien,a and Mahmoud Farouka

aChemistry Department, New Valley Faculty of Science, Assiut University, Assiut 72511, EgyptbChemistry Department, Faculty of Science, Assiut University, Assiut 71516, Egypt

*E-mail: [email protected] Supporting Information may be found in the online version of this article.

Received July 15, 2017DOI 10.1002/jhet.3056

Published online 00 Month 2017 in Wiley Online Library (wileyonlinelibrary.com).

5-Amino-2-(p-tolylamino)-4-phenylthieno[2,3-d]pyrimidine-6-carbonitrile 9, which was synthesized byan innovative method, was used as a versatile precursor for synthesizing pyrimido-thienopyrimidine,triazolopyrimidothienopyrimidine, and pyrimidothienotriazine compounds. Thus, reaction ofaminothienopyrimidinecarbonitrile 9 with chloroacetylchloride in dioxane afforded thechloroacetylaminocarbonitrile derivative 10, which underwent nucleophilic substitution reactions with vari-ous primary and secondary amines gave the corresponding N-alkyl-(aryl)amino acetamides 11a,b. On theother hand, the reaction of aminocarbonitrile 9 with triethyl orthoformate followed by cyclization withhydrazine yielded an aminoiminopyrimidine derivative 13. The latter was used as versatile precursor forsynthesis of new heterocyclic compounds. The structures of all the new compounds have been establishedon the basis of their analytical and spectral data (IR, 1H NMR, 13C NMR, and MS). Some of the synthesizedcompounds were evaluated in vitro for their anti-inflammatory activity. All the tested compounds exhibitedremarkable anti-inflammatory activity.

J. Heterocyclic Chem., 00, 00 (2017).

INTRODUCTION

In the past few years, thienopyrimidines and fusedthienopyrimidine derivatives have attracted great attentionowing to their inspiring array of pharmacological activities

[1]. Thienopyrimidines are interesting heterocycliccompounds, and a lot of derivatives of these compoundsdisplay therapeutic activity as antimicrobial [2–6],antiviral [7–9], anti-inflammatory [10–12], antidiabetic[13], antioxidant [14–16], antitumor [17–19], anticancer

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agents [20–24], and antimalarial [25,26]. Somethienopyrimidine used as analgesic [27,28]. Theantibacterial activity of this class of compounds has beenunderexplored.Moreover, thieno[2,3-d]pyrimidines have fascinated

importance in medicinal chemistry, exhibitingpharmacological and therapeutic properties such asantidepressant [29,30], antiplatelet [31], antihypertensive [32],herbicidal [33], and plant growth regulatory properties [34]. Incontinuation of our program for the synthesis of heterocycliccompounds containing thienopyrimidine moiety [35–39], inthe present work, we report on the efficient synthesis ofthienopyrimidine derivatives. This study focused on thesynthesis and biochemical evaluation of the newlysynthesized heterocyclic compounds that were subjectedthrough anti-inflammatory evaluations.

RESULTS AND DISCUSSION

5-Cyano-4-oxo-6-phenyl-2-thioxo-1,2,3,4-tetrahydropyrimidine (1) has been synthesized viamulticomponents reaction by one pot condensationreaction according to the reported procedure [40], wasreacted with ethyl iodide and sodium acetate in refluxingethanol gave 2-ethylmercapto-4-oxo-6-phenyl-pyrimidine-5-carbonitriles (2) according to the literature procedure[41]. The ethylmercaptopyrim-idinecarbonitriles derivative(2) was reacted with p-toluidine in ethanol under reflux togive 2-(p-tolylamino)-6-oxo-4-phenyl-1,6-dihydropyrimidine-5-carbonitrile (3). Chlorination of the latter compound,using phosphorusoxychloride, afforded 4-chloro-2-(p-tolylamino)-6-phenyl-5-pyrimidine carbonitrile (4).Compound (4) was converted into the corresponding 4-mercapto-2-(p-tolylamino)-6-phenyl-5-pyrimidine carbonitrile(7) by refluxing with thiourea to give thiurenium salt (5) thatrefluxed in sodium hydroxide solution (10%) to give sodiumsalt (6) which upon acidification with HCl afforded the

mercaptopyrimidine compound (7). The structure ofcompounds 3, 4, and 7 were established by their elementaland spectral analyses. IR of compound 3 showed absorptionband at 3450 cm�1 characteristic of 2NH groups. 1H NMRspectrum of compound 3 showed two singlet signals at δ10.20 and 11.69 characteristic of 2NH groups and the signalscharacteristic of ethyl group in compound 2, disappeared. Onthe other hand, IR spectrum of compound 7 revealed thedisappearance of absorption band characteristic of NH groupin the starting material. While 1H NMR spectrum showed theappearance of singlet signal at δ 11.36 characteristic of SHgroup (Scheme 1).

When compound (7) allowed to react withchloroacetonitrile in ethanol in the presence of sodiumacetate, alkylation of mercapto group was occurredaffording compound (8). Compound (8) undergoingThorpe–Ziegler cyclization reaction in refluxed ethanol inthe presence of sodium ethoxide giving 5-amino-4-phenyl-2-p-tolylaminothieno[2,3-d]pyrimidine-6-carbonitrile (9).The IR spectrum of compound 8 revealed two absorptionbands for (2CN) groups at 2187 and 2226 cm�1, whichone of them disappeared upon Thorpe–Ziegler cyclizationto afford compound 9 and appearing of new bandscharacteristic of (NH2) group at 3477 and 3387 cm�1. 1HNMR spectrum of compound 8 showed signal at δ 4.63corresponding to CH2. On the other hand, the 1H NMRspectrum of compound 9 revealed the disappearance ofsignal characteristic for methylene group in compound 8and appearance of new signals characteristic of NH2

group. Mass spectrum gave a molecular ion peak at375.03 as a base peak. 13C NMR spectrum of compound 9revealed signal at δ 117.67 characteristic of CN and at δ160.78 for (C–NH2) (Scheme 2).

5-Amino-2-(p-tolylamino)-4-phenylthieno[2,3-d]pyrimi-dine-6-carbonitrile (9) was treated with chloroacetylchloride in dioxane followed by treatment with sodiumcarbonate solution to afford the chloroacetamidederivative (10). The IR spectrum of compound 10 showed

Scheme 1. Synthesis of 4-mercaptopyrimidine carbonitrile 7. Reagents and conditions: (a) Etl, AcONa, EtOH, Reflux 5 h; (b) p-toluidine, EtOH, Reflux6h; (c) POCl3, Reflux 4h; (d)NH2CSNH2, EtOH, Reflux 5h; (e) NaOH (10%); (f) dil HCI

M. S. Tolba, M. Ahmed, A. M. Kamal El-Dean, R. Hassanien, and M. Farouk Vol 000

Journal of Heterocyclic Chemistry DOI 10.1002/jhet

disappearance of absorption bands characteristic of aminogroup and appearance of absorption bands characteristicof 2NH at 3350 cm�1, and band characteristic of C=Oamidic at 1678 cm�1. 13C NMR spectrum of compound10 revealed signal at δ 179.79 characteristic for CO. Thechloroacetyl derivative 10 underwent nucleophilicsubstitution reactions with various primary and secondaryamines in refluxing ethanol for 2 h to afford thecorresponding N-alkyl-(aryl)aminoacetamides (11a,b).While the reaction of 5-amino-2-(p-tolylamino)-4-phenylthieno[2,3-d]pyrimidine-6-carbonitrile (9) withexcess chloroacetyl chloride on water bath under neatcondition for 5 h followed by treatment with sodiumcarbonate solution gave 2-chloromethyl-3H-7-p-tolylamino-4-oxo-9-phenylpyrimido[40,50:4,5]thieno[2,3-d]pyrimidine (12). IR spectrum of compound 12 revealedappearance of absorption bands for 2NH and CO at 3278,3156, and 1668 cm�1

, respectively and disappearance ofabsorption band corresponding to cyano group incompound 9. Mass spectrum of compound 12 gave apeak at m/z = 432.65 corresponding to the molecular ionpeak and the base peak [M+�1, 65.23%].which inagreement with the postulated structure. Chloromethylderivatives (12) underwent nucleophilic substitutionreactions with morpholine in refluxing dioxane toafford 3H–p-tolylamino-2-morpholinomethyl-4-oxo-9-phenylpyrimido[40,50:4,5]thieno[2,3-d] pyrimidine (13).The formation of compound 13 was confirmed byelemental and spectral data. Mass spectrum showed peakat 484.02 as a molecular ion peak .1H NMR of compound13 showed multiplet signals at δ 3.3, 3.6, 3.7, 3.9 for fourmethylene groups of morpholine ring singlet signal at δ12.4 for (NH) group. 13C NMR spectrum of compound(13) revealed signal at δ 53.76 characteristic of methylenegroup and two signals at δ 52.76 and 67.98 for (4CH2) ofmorpholine ring (Scheme 3).The reaction of o-aminocyano derivative (9) with

triethyl orthoformate in acetic acid affordedethoxymethyleneamino derivative (14). The 1H NMRspectrum of 14 revealed the appearance of a triplet signalat δ 1.34 ppm for methyl group, quartet signal at3.66 ppm for methylene group, and a singlet signal at δ8.25 ppm for an azomethine proton. Reaction ofethoxymethyleneamino derivative (14) with an equivalent

amount of hydrazine hydrate in dioxane at roomtemperature by stirring afforded the correspondingaminoiminopyrimidine derivative (15). IR spectrum ofcompound 15 showed absorption bands at ν 3487, 3390,3297 cm�1 for NH, NH2 accompanied with disap-pearance of absorption band characteristic of cyano groupin the starting material. 1H NMR spectrum showedsinglet signal at δ 5.95 ppm for amino group in additionto the singlet signal at δ 9.16 ppm for NH group. Massspectrum of 15 showed a molecular ion peak at 399.25 asa base peak. The aminoiminopyrimidine compound (15)was used as starting material to synthesize differentheterocyclic compounds. Thus, the boiling ofpyrimidinaminoimino compound (15) with triethylorthoformate and diethyl malonate afforded thecorresponding triazolopyrimidine derivatives (16) and(17), respectively. Refluxing of aminoimino compound(15) with benzaldehyde in ethanol in the presence of fewdrops of piperidine gave the correspondingtriazolopyrimidopyrimidine (18). Reaction of (15) withacetylacetone gave the methyl triazolo compound (19)rather than the triazepine compound (20). Methyltriazoloderivative (19) was also obtained from the reaction ofcompound (15) with ethyl acetoacetate. The structure ofcompound 19 was established by mp, TLC, IR, and massspectra. Its IR spectrum showed absorption bands at

Scheme 3. Synthesis and reactions of chloroacetyl derivative 10 andChloromethyl derivatives 12. Reagents and conditions: (a) ClCH2COCI,dioxan,Steam bath, Reflux 3h; (b) R1R2NH, EtOH, Reflux 2h; (c)CICH2COCI, Steam bath 70°C, Reflux 5h; (d) morpholin, dioxan,Reflux 3h.

Scheme 2. Synthesis of aminocarbonitrilethienopyrimidine 9. Reagents and conditions: (a) CICH2CN, AcONa, EtOH, Reflux 3h; (j) EtONa, EtOH,Reflux 0.5 h.



3465 cm�1 characteristic of NH group. 1H NMR spectrumof (19) showed a sharp signal at δ 2.36 for methyl group.The mass spectrum showed a molecular ion peak at423.22 as the base peak (Scheme 4).The condensations proceed via cyclization of the imine

group with a retro aldol type elimination of acetone, inthe case of acetylacetone and elimination of ethyl acetatein the case of ethyl acetoacetate as shown in [42](Scheme 5).Reaction of aminoimino compound (15) with phenacyl

bromide in hot ethanol containing a few dropsof triethyl amine afforded the corresponding3-phenyltriazinopyrimidothienopyrimidine derivative (21)rather than the 2-phenyltriazinopyrimidothienopyrimidinederivative (22). The structure of compound 21 wasestablished by TLC, 1H NMR, and 13C NMR spectra.The 1H NMR spectrum of 21 showed a sharp signal atδ 4.54 characteristic of methylene group of triazino

ring. While the 13C NMR showed signal at δ 67.98 formethylene group. Also, aminoimine compound (15) wascyclized to the corresponding triazolopyrimidothienopyrimidinethione (23) by heating with carbondisulfide in the presence of pyridine on steam bath.Compound (23) was easily alkylated using α-halogenatedcarbonyl compounds in refluxing ethanol containing fusedsodium acetate to yield ethyl 7-phenyl-11-p-tolylamino[1,3,5]triazolo[1″,2″:10,60] pyrimido[40,50:4,5]thieno[2,3-d]pyrimidine-2-ylsulfanyl]acetate (24). The IRspectrum of compound 24 showed absorption band at1712 cm�1 for ester group. While the 1H NMR spectrumshowed triplet and quartet signals at δ 1.39–1.41 and4.21–4.24, respectively for ethyl ester group in additionto singlet signal at δ 3.94 for –CH2CO-group(Scheme 6).

Aminocyanothienopyrimidine (9) was reacted with amixture of AcOH/HCl in sodium nitrite solution under

Scheme 5. Suggested mechanism for synthesis of compound 16.

Scheme 4. Synthesis and reactions of aminoimino compound 15. Reagents and condiotions: (a) CH(OEt)3,AcOH, Reflux 2h; (b) NH2NH2/dioxan, r.t.;(c) CH(OEt)3,AcOH, Reflux 3h; (d) CH2(CO2Et)2, Reflux 4h; (e) PhCHO, EtOH, piperidine, Reflux 3h; (f) CH2(CH3CO)2, Reflux 2h;(j) CH3COCH2CO2Et, Reflux 4h.



cooling for 3 h to afford the correspondingpyrimidothienotriazine derivative (25). The IR spectrumof compound 25 showed disappearance of absorptionbands of NH2 and CN groups. Nucleophilic substitutionreaction of the chlorine atom with various primaryaromatic amines and sulfa drugs in thepyrimidothienotriazine derivative (25) was performed byreacting the chloro compound with different aminesunder neat condition for short time followed by refluxingin ethanol to afford compounds (26) and (27). Thestructures of compounds 26 and 27 were established byelemental and spectral data. The IR spectrum of 26represented absorption band at ν 3341 cm�1 for NHgroup. Also, the IR spectrum of compound 27 showedabsorption bands at ν 3469, 3297, 3256, 3157 cm�1 for4NH, NH2 groups and absorption band at ν 1393 cm�1

for SO2NH group. The 1H NMR spectrum of compound26 showed two singlet signals at δ 2.33 for 2CH3 groupand two singlet signals at δ 9.33 and 10.05 for 2NH. Inaddition, the 1H NMR spectrum of compound 27 showed

four singlet signals at δ 2.95, 8.69, 9.42, and 10.17 for(4NH) group and singlet signal at 6.29 for NH2. On theother hand, the 13C NMR spectrum of compound 27showed signal at 159.89 for NH–C–NH2 (Scheme 7).

Anti-inflammatory evaluation. The results of anti-inflammatory activity are summarized in (Table 1)Figures 1 and 2. The activity was evaluated by usingcarrageenan-induced paw edema assay, and indomethacinwas employed as standard for comparison of results [43].The results revealed that it has been observed that severalnewly prepared compounds (9, 10, 15, 17, 25, and 27)showed excellent anti-inflammatory properties(61.42–82.85% inhibition of edema) as compared withthat by indomethacin (80.96% inhibition). In addition,conversion of the amino thienopyrimidinecarbonitrilederivative 9 to its triazinothieno-pyrimidine derivative 25,increase the activity of compound compared with thestarting material, which is the most effective anti-inflammatory agent, revealing higher activity (82.85%inhibition) than that of indomethacin. However,

Scheme 6. Reactions of aminoimino compound 15. Reagents and conditions: (9)PhCOCH2Br, EtOH, ET3N, Reflux 2h; (b) CS2, pyridine,Steam bath,Reflux 4h; (c) CICH2COOEt,EtOH,AcONa, Reflux 3h.

Scheme 7. Synthesis of and reactions of pyrimidothienotriazine derivative 25. Reagents and conditions: (a) AcOH, HCI, NaNO2, stirring 3h, r.t.;(b) p-toluidine, EtOH, Reflux 3h; (c) sulphaguanidine, EtOH, Reflux 4h.



comparing the activity of 2-chloro-N-(6-cyano-4-phenyl-2-(p-tolylamino) thieno[2,3-d]pyrimidin-5-yl)acetamide 10,it was observed that substitution with an electron-withdrawing group, chloro acetamideamino, in the sidechain 10 74.29% inhibition seems more favorable forconstructing an anti-inflammatory active agent than thecase of substitution amino group, as exhibitedin 9 (61.42% inhibition). While, theaminoiminothienopyrimidine derivative 15 showed asignificant increase in the anti-inflammatory (75.71%inhibition). However, replacement of the hydrogen atom

of compound 16 in position 2 by ethylacetate groupderivative 17 possessed higher activity (77.14%inhibition) than the disubstituted derivative 15 (75.71%inhibition). Finally, 27 exhibited more anti-inflammatoryactivity (78.57% inhibition) comparable to that ofindomethacin (80.96% inhibition). Compound 9 exhibitedremarkable anti-inflammatory activity comparable to thatof indomethacin.

CONCLUSION

The objective of the present work was to synthesize,characterize, and investigate the anti-inflammatoryand antimicrobial activities of some newly thieno[2,3-d]pyrimidine derivatives. The starting compound 5-amino-4-phenyl-2-p-tolylaminothieno[2,3-d]pyrimidine-6-carbonitrile(9) that was used as a versatile precursor for synthesis to thetarget compounds. Compounds 10, 15, 17, 25, and 27 werethe most anti-inflammatory activity comparable toindomethacin.

EXPERIMENTAL

All melting points are uncorrected and measured on aFisher-Johns apparatus. Elemental analyses weredetermined on an Elemental Analyzer system GmbH-VarioEL V2.3 micro-analyzer in the central lab of AssiutUniversity. Their results were found to be in goodagreement (±0.2%) with the calculated values. FT-IRspectra were recorded on a Pye-Unicam Sp-100spectrophotometer using KBr wafer technique and valuesrepresented in cm�1. 1H NMR and 13C NMR werecarried out on Varian Gemini 300 MHz. The electronimpact mass spectra were recorded on JEOL JMS-600

Figure 1. A plot of edema inhibition (mm) × time.

Figure 2. A plot of paw edema inhibition (%) × time.

Table 1

Anti-inflammatory activity of tested compounds (9, 10, 15, 17, 25, and 27) using acute carrageenan-induced paw edema in rats (statistical analysis).

Compounda

Paw edema inhibition (Swel ± SEM)a,b,c (%)

30 min 1(h) 2(h) 3(h) 4(h) Potencyd

Edema induced by carrageenan (% edema inhibition relative to control)9 0.73 ± 0.02 0.72 ± 0.02 0.68 ± 0.02 0.62 ± 0.02 0.57 ± 0.02 61.4210 0.72 ± 0.02 0.68 ± 0.02 0.63 ± 0.02 0.52 ± 0.03 0.48 ± 0.02 74.2915 0.72 ± 0.02 0.68 ± 0.02 0.58 ± 0.03 0.52 ± 0.03 0.47 ± 0.00 75.7117 0.70 ± 0.00 0.68 ± 0.02 0.57 ± 0.03 0.51 ± 0.03 0.46 ± 0.03 77.1425 0.70 ± 0.00 0.64 ± 0.00 0.53 ± 0.00 0.51 ± 0.00 0.42 ± 0.00 82.8527 0.70 ± 0.00 0.65 ± 0.03 0.57 ± 0.04 0.48 ± 0.02 0.45 ± 0.03 78.57Indomethacin 0.67 ± 0.02 0.58 ± 0.02 0.55 ± 0.03 0.45 ± 0.03 0.43 ± 0.02 80.96Control 0.73 ± 0.02 0.70 ± 0.00 0.75 ± 0.00 0.77 ± 0.02 0.70 ± 0.05 –

Swel means difference in rat paw volume between right and left paw ± SE; % inhibition.aDose 20 μmol/kg.bn = 6.cStatistically significant from the indomethacin at p < 0.05.dPotency was expressed as % edema inhibition of the tested compounds relative to % edema.



spectrometer. Compounds 1 and 2 were prepared accordingto literature procedure [40,41] with mp 298–300°C and238–240°C, respectively.

2-(p-Tolylamino)-6-oxo-4-phenyl-1,6-dihydropyrimidine-5-carbonitrile (3). Compound 2 (30 g, 99 mmol) fused with(20 g) p-toluidine for 3 h. Then ethanol (50 mL) was addedand the reflux was continued for 3 h. After cooling, thecrystals is filtrated, washed with ethanol, and dried in air,recrystallized from dioxane-ethanol (1:1) to give 3 as palegreen crystals in 73% yield; mp: 294–296°C; ir: NH3450, CH aromatic 3093, CN 2195, CO 1646 cm�1;1HNMR: (CDCl3) δ 2.32 (s, 3H, CH3), 7.08–7.40 (m, 9H,ArH), 10.20 (s, 1H, NH), and 11.69 ppm (s, 1H, NH);ms: m\z 301.25 (M+�1) as molecular ion peak. Anal.Calcd for C18H14N4O (302.34): C, 71.51%; H, 4.67%; N,18.53%. Found: C, 71.47%; H, 4.71%; N, 18.45%.

4-Chloro-2-(p-tolylamino)-6-phenyl-5-pyrimidinecarbonitrile (4). Compound 3 (20 g, 66 mmol) in POCl3(40 mL) was refluxed for 4 h. After cooling, the reactionmixture was poured into water/ice mixture (600 g), thenneutralized using sodium carbonate solution. The reactionmixture was stirred for 30 min; the formed precipitatewas collected by filtration, washed several times withwater, dried in air, and recrystallized from ethanol-water(1:2) to afford white crystals of 4 in 95% yield; mp:168–170°C; ir: NH 3450, CH aromatic 3069, CHaliphatic 2970, CN 2214 cm�1; 1H NMR (DMSO-d6) δ:2.31 (s, 3H, CH3), 6.95–7.67 (m, 9H, 2ArH), and 9.73(s, 1H, NH); ms: m\z 320.15 (M+) and base peak. Anal.Calcd for C18H13ClN4 (320.78): C, 67.42%; H, 4.06%;Cl, 11.05%; N, 17.47%. Found: C, 67.37%; H, 4.04%;Cl, 11.09%; N, 17.42%.

4-Mercapto-2-(p-tolylamino)-6-phenyl-5-pyrimidinecarbonitrile (7). A mixture of 4 (3.20 g, 10 mmol) andthiourea (20 mmol) in ethanol (100 mL) was heatedunder reflux for 5 h and then allowed to cool. The yellowprecipitate formed was filtered off and washed withethanol. The thiouronium salt was dissolved in NaOH(10%, 50 mL) and then acidified with dilute HCl to justneutral. The crude product was collected by filtration,washed several times with water, dried in air, andrecrystallized from ethanol-dioxane (1:2) to give 7 as ayellow crystals in 80% yield; mp: 317–319°C; ir: NH3177, CH aromatic 3051, CH aliphatic 2974, CN2212 cm�1; 1H NMR (DMSO-d6) δ: 2.31 (s, 3H, CH3),6.95–7.66 (m, 9H, 2ArH), 9.57 (s, 1H, NH), and 11.36(s, 1H, SH); ms: m\z 317.04 (M+�1). Anal. Calcd forC18H14N4S (318.40): C, 67.90%; H, 4.43%; N, 17.60%;S, 10.07%. Found: C, 67.86%; H, 4.48%; N, 17.64%; S,10.03%.

5-Cyano-6-phenyl-2-p-tolylamino-pyrimidine-4-sulfanylacetonitrile (8). A mixture of compound 7 (4 g,12 mmol), chloroacetonitrile (2 mmol), and sodiumacetate (0.75 g, 8.5 mmol) in ethanol (20 mL) was heated

under reflux for 3 h and then cooled. The solid productwas filtered off, washed with water, dried, andrecrystallized from ethanol-water (1:1) to afford 8 as a palegreen crystals in 55% yield; mp: 177–179°C; ir: NH 3387,CH aromatic 3047, CH aliphatic 2959, 2CN 2187,2216 cm�1; 1H NMR: δ (DMSO-d6) 2.31 (s, 3H, CH3),4.63 (s, 2H, CH2), 7.11–7.66 (m, 9H, ArH), and 11.87(s, 1H, NH); ms: m\z 374.13 (M+�1). Anal. Calcd forC20H15N5S (357.44): C, 67.25%; H, 4.23%; N, 19.55%; S,8.97%. Found: C, 67.28%; H, 4.25%; N, 19.53%; S, 8.94%.

5-Amino-4-phenyl-2-p-tolylamino-thieno[2,3-d]pyrimidine-6-carbonitrile (9). A solution of compound 8 (10 mmol)in absolute ethanol (20 mL) and few drops of sodiumethoxide prepared by 0.5 g of finely divided sodiummetal in absolute ethanol (20 mL) solution was refluxedfor 30 min. The precipitated that formed after coolingwas filtered off, dried, and recrystallized from ethanol-water (1:2) into 9 yellow crystals in 65% yield; mp:208–210°C; ir: NH2 3477, 3387, CH aromatic 3047, CHaliphatic 2959, CN 2187 cm�1; 1H NMR: (DMSO-d6) δ2.31 (s, 3H, CH3), 6.47 (s, 2H, NH2), 7.11–7.66 (m, 9H,ArH), and 11.87 (s, 1H, NH); 13C NMR: 21.13 (C21:CH3 p-toluidine), 83.56 (C6: C–CN), 110.98 (C4a),120.56 (C10, C14), 125.97 (C11, C13), 131.87 (C17,C19), 139.67 (C16, C20), 117.67 (C22: CN), 121.96(C12), 128.98 (C18), 142.55 (C9), 148.98 (C4),154.77 (C2), 160.78 (C5: C–NH2), 178.98 (C7a: N–C–S);ms:m\z 375.03 (M+) and base peak. Anal. Calcd forC20H15N5S (357.44): C, 67.25%; H, 4.23%; N, 19.55%; S,8.97%. Found: C, 67.29%; H, 4.26%; N, 19.51%; S, 8.93%.

2-Chloro-N-(6-cyano-4-phenyl-2-(p-tolylamino)thieno[2,3-d]pyrimidin-5-yl)acetamide (10). A mixture compound 9(1.6 g, 4.2 mmol), and chloroacetyl chloride (0.8 mL,7 mmol) in dioxane (25 mL) was heated on steam bath for3 h. After cooling, the mixture was poured onto water andneutralized with diluted sodium carbonate solution. Thesolid thus formed was filtered, washed with water, dried,and recrystallized from ethanol-water (2:1) to afford 10 asa pale green crystals in 56% yield; mp: 176–178°C; ir:2NH 3350, CH aromatic 3058, CH aliphatic 2922, 2851,CN 2214, CO amidic 1678 cm�1; 1H NMR (DMSO-d6)δ 2.31 (s, 3H, CH3), 4.27 (s, 2H, CH2Cl), 6.94–7.62 (m,9H, 2ArH), 11.01 (s, 1H, NHCO), and 11.78 (s, 1H,NH); 13C NMR: 21.13 (C21: CH3 p-toluidine), 42.77(C24: CH2CO), 97.15 (C6: C–CN), 114.78 (C4a), 116.55(C10, C14), 125.89 (C11, C13), 126.89 (C17, C19),142.99 (C16, C20), 119.45 (C26: CN), 128.87 (C18),132.11 (C12), 138.46 (C15), 137.54 (C5), 143.34 (C9),152.69 (C4), 153.39 (C2), 137.99 (C23: CO),179.69 (C7a: N–C–S); ms: m\z 433.12 (M+) and basepeak. Anal. Calcd for C22H16ClN5OS (433.93): C,60.88%; H, 3.74%; Cl, 8.19%; N, 16.14%; S, 7.41%.Found: C, 60.93%; H, 3.70%; Cl, 8.16%; N, 16.19%;S, 7.44%.



N-(2-cyano-5-p-tolylamino-4-yl-thieno[2,3-d]pyrimidin-5-yl)-2-alkyl(aryl)-aminoacetamide (11a,b). A mixture of (0.5 g,1.15 mmol) of chloroacetamide derivative 10 and thecorresponding amine (2 mmol) was dissolved in 20 mLethanol and refluxed for 2 h. The solid precipitate thatwas formed, filtered off, dried, and recrystallized from theproper solvent.

N-(2-cyano-5-p-tolylamino-4-yl-thieno[2,3-d]pyrimidin-5-yl)-2-phenylaminoacetamide (11a). The solid thus formed wasfiltered, washed with water, dried, and recrystallized fromethanol-water (2:1) to give 11a as a yellow needles in77% yield; mp: 225–227°C; ir: 3NH 3436, 3261, CHaromatic 3053, CH aliphatic 2920, CN 2223, CO amidic1665 cm�1. 1H NMR (DMSO-d6) δ: 2.31 (s, 3H, CH3),3.97 (s, 2H, CH2NH), 5.92 (s, 1H, NHPh), 7.10–7.63(m, 14H, 3ArH), 11.33 (s, 1H, NHCO), and 11.69 (s, 1H,NH); ms: m\z 488.05 [M+�2]. Anal. Calcd forC28H22N6OS (490.59): C, 68.55%; H, 4.52%; N, 17.14%;S, 6.55%. Found: C, 68.58%; H, 4.56%; N, 17.18%;S, 6.51%.

N-(2-cyano-5-p-tolylamino-4-yl-thieno[2,3-d]pyrimidin-5-yl)-2-diethylaminoacetamide (11b). The solid thus formed wasfiltered, washed with water, dried, and recrystallized fromethanol-water (1:2) as a pale brown crystals of 11b in67% yield; mp: 118–120°C; ir: 2NH 3321, CN 2217,CO amidic 1685 cm�1; 1H NMR (DMSO-d6) δ 1.05(t, J = 7.5 Hz, 6H, 2CH3), 2.32 (s, 3H, CH3), 2.79(q, J = 6.4 Hz, 4H, 2CH2), 3.4 (s, 2H, COCH2N),6.96–7.63 (m, 9H, 2ArH), 9.65 (s, 1H, NHCO), and11.04 (s, 1H, NH); 13C NMR: 12.34 (C27, C29), 21.13(C21: CH3 p-toluidine), 47.78 (C26, C28), 57.66 (C24:CH2CO), 96.52 (C6: C–CN), 114.12 (C4a), 115.99(C10, C14), 125.55 (C11, C13), 126.23 (C17, C19),142.88 (C16, C20), 118.86 (C30: CN), 129.12 (C18),132.22 (C12), 138.12 (C15), 140.21 (C5), 143.21 (C9),150.98 (C4), 153.27 (C2), 161.88 (C23: CO), 175.66(C7a: N–C–S); ms: m\z 470.12 (M+). Anal. Calcd forC26H26N6OS (470.60): C, 66.33%; H, 5.55%; N,17.86%; S, 6.82%. Found: C, 66.29%; H, 5.51%; N,17.89%; S, 6.85%.

2-Chloromethyl-3H-7-p-tolylamino-4-oxo-9-phenylpyrimido[40,50:4,5]thieno[2,3-d]pyrimidine (12). Compound 9 (2 g,5.3 mmol) and chloroacetyl chloride (5 mL) was heated onsteam bath for 5 h. Then poured into cold water (100 mL),neutralized with sodium carbonate solution (10%) to justalkaline. The solid product was filtrated off, dried, andrecrystallized from dioxane-water (1:1) to give 12 as greenfine in 77.70% yield; mp: 330–332°C; IR: 2NH3278,3156, CH aromatic 3031, CH aliphatic 2934, 2867,CO 1668 cm�1; 1H NMR (DMSO-d6) δ: 3.6, 3.7, 3.9(s, 8H, 4CH2), 4.4 (s, 2H, CH2), 7.5 (m, 3H, ArH), 7.9 (d,2H, ArH), and 13.0 (s, 1H, NH); ms: m\z 432.65 (M+�1,65.23%). Anal. Calcd for C22H16ClN5OS (433.92): C,60.90%; H, 3.72%; Cl, 8.17%; N, 16.14%; S, 7.39%.

Found: C, 60.94%; H, 3.68%; Cl, 8.21%; N, 16.09%;S, 7.43%.

3H-p-Tolylamino-2-morpholinomethyl-4-oxo-9-phenylpyrimido[40,50:4,5]thieno[2,3-d]pyrimidine (13). A mixture ofcompound 12 (4.13 g, 9.5 mmol) and morpholin(20 mmol) was heated for 5 min, and then ethanol(20 mL) was added. The mixture was refluxed for 3 h.The solid product that is formed on hot was filtrated off,dried, and recrystallized from dioxane-ethanol (1:1) toafford 13 as a white crystals in 71.50% yield; mp:306–308°C; ir: 2NH 3276, 3178, CH aromatic 3053, CHaliphatic 2968, 2917, CO 1663 cm�1; 1H NMR(DMSO-d6) δ: 2.31 (s, 3H, CH3), 3.3, 3.6, 3.7, 3.9 (m,8H, 4CH2), 4.6 (s, 2H, CH2), 7.5 (m, 5H, ArH), 7.85 (d,4H, ArH), 10.2 (s, 1H, NH), and 12.4 (s, 1H, NH); 13CNMR: 21.13 (C23: CH3 p-toluidine), 52.76 (C26, C30),53.76 (C24: CH2–N), 67.98 (C27, C29), 113.77 (C9a),114.86 (C4a), 116.87 (C12, C16), 125.78 (C13, C15),130.65 (C19, C21), 142.65 (C18, C22), 128.65 (C20),132.43 (C14), 140.35 (C17), 142.87 (C4), 143.87 (C11),150.45 (C9), 155.99 (C7), 173.87 (C2), 181.09 (C5a:N-C-S). Anal. Calcd for C26H24N6O2S (484.58): C,64.45%; H, 5.00%; N, 17.32%; S, 6.61%. Found: C,64.41%; H, 5.04%; N, 17.36%; S, 6.57%.

1-Ethoxymethyleneamino-5-p-tolylamino-7-phenylthieno[2,3-d]pyrimidine-2-carbonitrile (14). A mixture ofaminocarbonitrile compound 9 (3 g, 8 mmol) and triethylorthoformate (6 mL, 40 mmol) in acetic acid (10 mL) wasrefluxed for 2 h. The resultant solid product 14 wasfiltered off, washed with water, dried, and crystallizedfrom dioxane-water (1:1) to give 14 as a yellow crystalsin 83% yield; mp: 148–150°C; ir: NH 3302, CH aromatic3051, CH aliphatic 2959, 2918, 2869, CN 2207, C=N1644 cm�1; 1H NMR (DMSO-d6) δ: 1.34 (t, J = 8.2 Hz,3H, CH3), 3.66 (q, J = 7.2 Hz, 2H, CH2), 6.95–7.75 (m,9H, 2ArH), 8.25 (s, 1H, N=CH), and 11.86 (s, 1H, NH);13C NMR: 14.70 (C28: CH3 ester), 21.13 (C21: CH3 p-toluidine), 63.58 (C27: CH2 ester), 100.98 (C6), 114.78(C7a), 116.55 (C16, C20), 125.89 (C17, C19), 130.87(C9, C13), 145.76 (C10, C12), 119.45 (C22), 128.87(C18), 132.10 (C12), 143.65 (C9), 153.98 (C4), 155.18(C25), 167.71 (C5), 179.87 (C3a); ms: m/z 413.24 (M+,77.98%). Anal. Calcd for C23H19N5OS (413.50): C,66.81%; H, 4.64%; N, 16.94%; S, 7.75%. Found: C,66.77%; H, 4.68%; N, 16.91%; S, 7.79%.

3-Amino-4-imino-7-p-tolylamino-9-phenylpyrimido[40,50:4,5]thieno[2,3-d]pyrimidine (15). A solution of 14 (4 g, 1 mmol)in warm dioxane was stirred at room temperature, and0.8 mL, 1.6 mmol of hydrazine hydrate was added to thesolution. Stirring of the mixture continued for 2 h. The solidprecipitate that was formed filtered off, dried, andrecrystallized from ethanol-dioxane (2:1) as a white crystalsof 15 in 62% yield; mp: 228–230°C; ir: 2NH, NH2 3487,3390, 3297, 2960, CH aliphatic 2854 cm�1; 1H NMR



(DMSO-d6) δ: 2.31 (s, 3H, CH3), 4.15 (s, 1H, NH), 5.95 (s,2H, NH2), 6.94–7.70 (m, 9H, 2ArH), 9.16 (s, 1H, NH), and11.77 (s, 1H, NH); 13C NMR: 21.13 (C23: CH3 p-toluidine), 112.76 (C9a), 116.85 (C18, C22), 125.84 (C19,C21), 130.98 (C12, C14), 143.87 (C11, C15), 126.76 (C4a),128.78 (C13), 143.35 (C17), 132.10 (C20), 140.69 (C10),143.87 (C2), 148.77 (C9b), 151.79 (C5, C9),156.55 (C7),179.35 (C5a); ms: m/z 399.25 (M+, 94.98%). Anal. Calcdfor C21H17N7S: (399.48): C, 63.14%; H, 4.24%; N,24.54%; S, 8.08%. Found: C, 63.20%; H, 4.20%; N,24.53%; S, 8.04%.

9-Phenyl-7-p-tolylamino-[1,2,4]triazolo[1″,5″:10,60]pyrimido[40,50:4,5]thieno[2,3-d]pyrimidine (16). Compound 15(0.7 g, 1.75 mmol) and triethyl orthoformate (5 mL) wereheated under reflux for 3 h. The solid product obtained onhot was filtered off, dried, and recrystallized from ethanol togive 16 as a white crystals in 74% yield; mp: 278–280°C;ir: NH 3181, CH aromatic 3054, CH aliphatic 2966,2956 cm�1; 1H NMR (DMSO-d6) δ: 2.31 (s, 3H, CH3),6.96–7.69 (m, 9H, 2ArH), 9.32 (s, 1H, CH pyrimidine),9.85 (s, 1H, CH triazole), and 11.82 (s, 1H, NH); ms:m/z 409.12 (M+, 53.51%). Anal. Calcd for C22H15N7S(409.48): C, 64.53%; H, 3.69%; N, 23.94%; S, 7.83%.Found: C, 64.50%; H, 3.72%; N, 23.90%; S, 7.87%.

Ethyl-9-phenyl-7-p-tolylamino-[1,2,4]triazolo[1″,5″:10,60]pyrimido[40,50:4,5]thieno[2,3-d] pyrimidine-2yl acetate(17). A mixture of 15 (0.7 g, 1.75 mmol) and diethylmalonate (5 mL) was heated under reflux for 4 h. Aftercooling, the solid product was filtered, washed withwater, dried, and crystallized from ethanol: browncrystals; mp 146–148°C; yield 64%; IR: NH 3287, CHaliphatic 2934, 2982, CO ester 1753 cm�1; 1H NMR(DMSO-d6) δ: 1.37 (t, J = 8 Hz, 3H, CH3 ester), 2.31(s, 3H, CH3), 4.18 (s, 2H, CH2CO), 4.25(q, J = 6.7 Hz, 2H, CH2), 6.94–7.70 (m, 9H, 2ArH),9.19 (s, 1H, CH pyrimidine), 11.17 (s, 1H, NH); 13CNMR: 14.70 (C30: CH3 ester), 21.13 (C25: CH3 p-toluidine), 36.33 (C26: CH2CO), 61.18 (C29: CH2

ester), 113.98 (C6b), 115.78 (C14, C18), 126.43 (C15,C17), 131.43 (C21, C23), 143.87 (C20, C24), 129.34(C22), 132.21 (C11a), 132.66 (C16), 138.78 (C6a),141.02 (C19), 141.55 (C11b), 143.22 (C13), 147.21(C5), 152.66 (C7), 157.45 (C9), 165.12 (C2), 168.54(C27: CO), 181.99 (C10a); ms: m/z 495.12 (M+,99.90%). Anal. Calcd for C26H21N7O2S (495.57): C,65.23%; H, 4.05%; N, 23.15%; S, 7.57%. Found: C,65.19%; H, 4.09%; N, 23.19%; S, 7.53%.

2,7-Diphenyl-9-p-tolylamino-2,3dihydro[1,2,4]triazolo[1″,5″:10,60]pyrimido[40,50:4,5]thieno[2,3-d] pyrimidine (18). Compound15 (0.6 g, 1.5 mmol) and excess benzaldehyede (4 mL)and piperidine were fused for 15 min, and then absoluteethanol (10 mL) was added then reflux was continued foradditional 2 h. The solid product that was formed on hotfiltered off and recrystallized from dioxane-water (1:2) to

give 18 as pale yellow crystals in 75% yield; mp:271–273°C; ir: 2NH 3399, CH aromatic 3057, CHaliphatic 2918, C=N 1597 cm�1; 1H NMR (DMSO-d6) δ:2.31 (3H, s, CH3), 7.20–7.70 (14H, m, 3ArH), 8.48 (1H,s, CH pyrimidine), 9.11 (1H, s, NH), and 11.17 (1H, s,NH). Anal. Calcd for C28H21N7S (487.59): C, 68.97%;H, 4.34%; N, 20.11%; S, 6.58%. Found: C, 68.93%; H,4.39%; N, 20.15%; S, 6.54%.

2-Methyl-7-diphenyl-9-p-tolylamino[1,2,4]triazolo[1″,5″:10,60]pyrimido[40,50:4,5]thieno[2,3-d] pyrimidine (19). Method A. Amixture of compound 15 (0.7 g, 1.75 mmol) andacetylacetone (3 mL, 30 mmol) was heated under refluxfor 2 h. The product formed after cooling was filtered offand recrystallized from ethanol as yellow crystals of 19in 71% yield; mp: 254–256°C.

Method B. A mixture of compound 15 (0.7 g,1.75 mmol) and ethyl acetoacetate (4 mL, 30 mmol) inethanol (20 mL) in the presence of a few drops ofpiperidine was heated under reflux for 3 h. The resultantsolid product was filtered, washed with water, dried, andcrystallized from ethanol to afford 19 as yellow crystalsin 78% yield; mp: 254–256°C; ir: 2NH 3465, 3350, CHaliphatic 2921, 2850, C=N 1629 cm�1; 1H NMR(DMSO-d6) δ: 2.31 (s, 3H, CH3), 2.36 (s, 3H, CH3),6.94–7.69 (m, 9H, 2ArH), 9.18 (s, 1H, CH pyrimidine),11.40 (s, 1H, NH). Anal. Calcd for C23H17N7S (423.50):C, 65.23%; H, 4.05%; N, 23.15%; S, 7.57%. Found: C,65.27%; H, 4.02%; N, 23.11%; S, 7.61%.

3,8-Diphenyl-10-p-tolylamino-2H-[1,2,5]triazino[1″,6″:10,60]pyrimido[40,50:4,5]thieno[2,3-d] pyrimidine (21). A mixtureof compound 15 (0.6 g, 1.5 mmol) and phenacylbromide (0.4 g, 2.0 mmol) in ethanol (20 mL) in thepresence of a few drops of triethyl amine (0.3 mL) washeated under reflux for 2 h. The solid product thusobtained after cooling was filtrated off, dried, andrecrystallized from ethanol to give 21 as yellow crystalsin 63% yield; mp: 264–266°C; ir: NH 3345, CHaliphatic 2917, C=N 1647 cm�1; 1H NMR (DMSO-d6)δ: 2.30 (s, 3H, CH3), 4.54 (s, 2H, CH2 triazine), 7.27–7.67 (m, 14H, 3ArH), 9.77 (s, 1H, CH pyrimidine),11.51 (s, 1H, NH); 13C NMR δ: 21.76 (C33: CH3 p-toluidine), 67.98 (C2), 113.34 (C7b), 116.56 (C22, C26),125.64 (C23, C25), 143.22 (C28, C32), 130.66 (C29,C31), 129.26 (C16, C19), 129.46 (C15, C19), 126.87(C17), 129.07 (C12a), 128.89 (C30), 132.87 (C24),140.76 (C27), 141.22 (C12b), 143.34 (C21), 144.87(C7a), 146.89 (C6), 151.35 (C8), 155.45 (C10), 157.88(C3), 181.09 (C11a: N–C–S); ms: m/z 501.23 (M++2,78.89%). Anal. Calcd for C29H21N7S (499.71): C,69.72%; H, 4.24%; N, 19.63%; S, 6.42%. Found: C,69.68%; H, 4.27%; N, 19.56%; S, 6.38%.

7-Phenyl-11-p-tolylamino[1,3,5]triazolo[1″,2″:10,60]pyrimido[40,50:4,5]thieno[2,3-d]pyrimidine-2-(3H)-thion (23). Amixture of compound 15 (0.7 g, 1.75 mmol) and carbon



disulfide (1.5 mL) in pyridine (3 mL) was heated underreflux on a water bath for 4 h. The resultant solid productwas filtered, washed with water, dried, and crystallizedfrom ethanol to give 23 as orange crystals in 63% yield;mp: 274–276°C; ir: 2NH 3395, 3287, CH aliphatic 2917,2849, C=N 1655 cm�1; 1H NMR (DMSO-d6) δ: 2.30 (s,3H, CH3), 6.94–7.69 (m, 9H, 2ArH), 8.35 (s, 1H, CHpyrimidine), 11.60 (s, 1H, NH), and 12.77 (s, 1H, NH);13C NMR δ: 21.13 (C25: CH3 p-toluidine), 113.87 (C6b),116.87 (C14, C18), 125.76 (C15, C17), 130.76 (C21,C23), 143.66 (C20, C24), 128.98 (C22), 128.44 (C11a),132.11 (C16), 140.09 (C19), 143.23 (C13), 149.55 (C6a),146.56 (C11b), 149.55 (C5), 152.28 (C7), 155.32 (C9),177.87 (C2), 181.02 (C10a: N–C–S); ms: m/z 440.76(M+�1, 65.87%). Anal. Calcd for C22H15N7S2 (441.54):C, 59.85%; H, 3.42%; N, 22.21%; S, 14.52%. Found: C,59.88%; H, 3.46%; N, 22.17%; S, 14.56%.

Ethyl 7-phenyl-11-p-tolylamino[1,3,5]triazolo[1″,2″:10,60]pyrimido[40,50:4,5]thieno[2,3-d] pyrimidine-2-yl sulfanyl]acetate (24). A mixture of compound 23 (0.6 g,1.35 mmol) and ethyl chloroacetate (2 mmol) in ethanol(15 mL) in the presence of fused sodium acetate (0.85 g,0.01 mmol) was heated under reflux for 3 h. Aftercooling, the solid product was filtered, washed withwater, dried, and recrystallized from ethanol-water (1:1)to afford 24 as yellow crystals in 57% yield; mp:189–190°C; ir: 2NH 3324, CH aliphatic 2922, 2851, CO1712, C=N 1646 cm�1; 1H NMR (CDCl3) δ: 1.39–1.41(t, J = 7.5 Hz, 3H, CH3 ester), 2.31 (s, 3H, CH3), 3.94(s, 2H, CH2CO), 4.21–4.24 (q, J = 6.34 Hz, 2H, CH2

ester), 6.94–7.53 (m, 9H, 2Ar–H), 8.99 (s, 1H, CHpyrimidine), 10.66 (s, 1H, NH); 13C NMR δ: 14.70 (C31:CH3 ester), 21.13 (C25: CH3 p-toluidine), 36.86 (C27:CH2CO), 61.18 (C30: CH2 ester), 114.41 (C6b), 115.96(C14, C18), 126.02 (C15, C17), 131.66 (C21, C23),143.54 (C20, C24), 128.75 (C22), 131.93 (C11a), 132.33(C16), 140.00 (C19), 142.16 (C6a), 144.88 (C11b), 144.99(C5), 151.87 (C7), 156.02 (C9), 160.21 (C2), 168.73(C28: C = O ester), 181.57 (C10a: N–C–S); ms: m/z525.66 (M+�2, 98.89%). Anal. Calcd for C26H21N7O2S2(527.60): C, 59.19%; H 4.01%; N, 18.58%; S, 12.15%.Found: C, 59.15%; H, 4.05%; N, 18.54%; S, 12.11%.

4-Chloro-7-p-tolylamino-9-phenylpyrimido[50,40:4,5]thieno[3,2-d]triazine (25). Compound 9 (1.3 g, 3.4 mmol) in amixture of acetic acid (10 mL) and concentratedhydrochloric acid (7 mL), a sodium nitrite solution(10%), (4 mL, 6 mmol) was added with stirring during5 min. The stirring was continued at 5°C for 3 h. Theformed precipitate was collected and crystallized fromethanol-water (2:1) to afford 25 as white plates in 62.5%yield; mp: 186–188°C; ir: NH 3181, CH aromatic 3051,CH aliphatic 2848 cm�1; 1H NMR (CDCl3) δ: 2.36(s, 3H, CH3 p-toluidine), 6.90–7.54 (m, 9H, 2ArH), 10.22(s, 1H, NH); 13C NMR: 21.87(C23: CH3 p-toluidine),

116.09 (C12, C16), 125.87 (C13, C15), 127.87 (C19,C21), 141.87 (C19, C21), 130.44 (C20), 131.66 (C9a),132.76 (C14), 143.98 (C4a), 144.33 (C11), 148.36 (C17),150.90 (C7), 155.76 (C4), 164.76 (C9b), 168.88 (C5a:N–C–S); ms: m/z 405.12 (M++1, 42.43%) and the basepeak. Anal. Calcd for C20H13ClN6S (404.88): C, 59.33%;H, 3.24%; Cl, 8.76%; N, 20.76%; S, 7.92%. Found: C,59.29%; H, 3.21%; Cl, 8.79%; N, 20.79%; S, 7.85%.

4,7-Di p-tolylamino-9-phenylpyrimido[50,40:4,5]thieno[3,2-d]triazine (26). A mixture of compound 25 (0.25 g,6.17 mmol) and p-toluidine (2 mmol) was gently fusedfor 5 min, and then ethanol (10 mL) was added. Themixture was refluxed for 3 h. The solid precipitate thatformed during reflux was filtered, dried, andrecrystallized from ethanol to give 26 as buff fine in 92%yield; mp: 244–246°C; ir: 2NH 3461, 3339, CH aromatic3050, CH aliphatic 2918, 2865, C=N 1592 cm�1; 1HNMR (CDCl3) δ: 2.33 (s, 6H, 2CH3), 6.95–7.75 (m, 13H,3ArH), 9.33 (s, 1H, NH), and 10.05 (s, 1H, NH). Anal.Calcd for C27H21N7S (475.58): C, 68.19%; H, 4.45%; N,20.62%; S, 6.74%. Found: C, 68.16%; H, 4.42%; N,20.59%; S, 6.71%.

N-[7-p-tolylamino-9-phenylpyrimido[50,40:4,5]thieno[3,2-d]triazine]-sulfaguanidine (27). A mixture of compound 25(0.25 g, 6.17 mmol) and sulphaguanidine (2 mmol) wasgently fused for 5 min, and then ethanol (10 mL) wasadded. The mixture was refluxed for 3 h. The resultantsolid product was filtered, washed with water, dried, andrecrystallized from ethanol to afford 27 as white crystalsin 75% yield; mp: 266–268°C; ir: NH2, 4NH 3469, 3297,3256, 3157, CH aromatic 3050, CH aliphatic 2998, C=N1586, SO2NH 1393 cm�1; 1H NMR (DMSO-d6) δ: 2.32(s, 3H, CH3), 2.95 (s, 1H, NH), 6.29 (s, 2H, NH2),7.01–7.93 (m, 13H, ArH), 8.69 (s, 1H, NH), 9.42 (s, 1H,NH), 10.17 (s, 1H, NH); 13C NMR: 21.13 (C34: CH3 p-toluidine), 115.98 (C12, C16), 124.02 (C25, C29), 126.22(C13, C15), 128.55 (C19, C21), 129.66 (C26, C28),142.33 (C18, C22), 119.78 (C9a), 129.35(C4b), 131.33(C20), 134.33 (C27), 136.55 (C14), 144.42 (C11), 147.21(C17), 149.44 (C7), 152.32 (C5), 154.66 (C1), 155.44(C24), 159.89 (C4a), 174.66 (C8a: N-C-S); ms: m/z582.32 (M+, 65.64%). Anal. Calcd for C27H22N10O2S2(582.67): C, 55.66%; H, 3.81%; N, 23.04%; S, 11.01%.Found: 55.69%; H, 3.78%; N, 23.07%; S, 11.04%.

Anti-inflammatory Evaluation. Anti-inflammatoryactivity was measured using carrageenan-induced rat pawedema assay [43]. Edema was induced by subplantarinjection. Adult albino rats, weighing 150–200 g, wereused. The animals were allowed food and water adlibitum, except during the experiment. They were housedin a room at 23 ± 2°C with a 12 h light/dark cycle. Theanimals were randomly allocated into groups of sixanimals each at the beginning of the experiment and werefasted for 24 h before the experiment with free access to



water. All of the compounds and the reference drug weresuspended in a 0.5% carboxymethyl cellulose solution.The standard drug indomethacin was administered orallyat a dose of 20 μmol/kg�1. The tested compounds wereadministered orally at an equimolar oral dose relative to20 μmol/kg�1 of indomethacin. The control groupreceived a 0.5% carboxymethyl cellulose solution. Intothe subplantar region of the right hind paw of each rat,0.1 mL of 1% carrageenan solution in saline was injectedsubcutaneously, 1 h after the administration of the testcompounds and standard drug. The right paw volumewas measured using a digital plethysmometer(Model 7150, Ugo Basile, Varese, Italy), directly beforeand after 1, 2, 3, 4 h, intervals after administration of thetested compounds. The percent edema inhibition wascalculated from the mean effect in the control and treatedanimals according to the following equation.

Percent edema inhibition ¼ 1–Vt=Vcð Þ�100

where: Vt represents the mean increase in paw volume inrats treated with tested compounds and Vc represents themean increase in paw volume in the control group of rats.All the results are expressed as the mean ± standard errorof the mean. Statistical evaluation was performed usingone-way ANOVA followed by Newman–Keuls Test.

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synthesis of new fused thienopyrimidines …agents [20–24], and antimalarial [25,26]. some...

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