Synthesis and In Vitro Anticancer Evaluation of

Synthesis and In Vitro Anticancer Evaluation of Symmetrically Bridged 1,3-thiazine Derivatives

Month 2018

Tamer El Malah,a*

Esam A. Ishak,b Hany F. Nour,a Raafat M. Shaker,c Mamdouh M. Ali,d Abeer E. Mahmoud,d and Salwa M. Solimane

a

Photochemistry Department, Chemical Industries Research Division, National Research Centre, 33 El Buhouth Street, Dokki, Cairo, P.O. Box:12622, Egypt b Chemistry Department, Faculty of Science, Al-Azhar University, Assiut, Egypt c Chemistry Department, Faculty of Science, Minia University, El-Minia 61519, Egypt d Biochemistry Department, Genetic Engineering and Biotechnology Division, 33 El Buhouth Street, 12622 Dokki, Egypt e Nawah Scientific, Pharmaceutical Research and Drug Design Department, Cairo, Egypt *E-mail: [email protected] Received November 7, 2017 DOI 10.1002/jhet.3212 Published online 00 Month 2018 in Wiley Online Library (wileyonlinelibrary.com). O O O O

O n

NC O

+

1a-c

R

NH2 TsOH/EtOH

N H

O O

O

O

Reflux

SH

H N

O

O n

H N

NC

NH

R

2a-e

O

S

S

CN

HN

3a-o

R

PhCH2Cl

CH3CH2I K2CO 3/DMF

KOH O O O NC Ph

H N N S

O

O

Ph

H N Ph

5a-c

O O

O n N

S

O CN Ph

O

H N N

NC S

O n

Ph

H N Ph

4a-c

N

O CN

S

A series of bis-1,3-thiazine derivatives 3a–o were synthesized from the condensation reactions of symmetric dialdehydes 1a–c possessing aliphatic ether spacer units with 3-substituted-amino-2-cyano-3mercaptoacrylamides 2a–e. The chemical structures of the products were fully characterized by using different spectroscopic techniques, such as 1H NMR, 13C NMR, IR, electron impact mass spectrometry, and elemental analysis. Compounds 3a, 3f, and 3k underwent ring opening followed by recyclization and alkylation in basic medium to afford bis-pyrimidinones 4a–c and 5a–c. The anticancer potential of the new bis-1,3-thiazines was assessed in vitro against six different human cell lines, including lung A549, colon HCT116, breast MCF-7, prostate PC3, liver HepG2, and normal melanocyte HFB4. The results revealed a potent activity of compounds 3e and 3k against breast and liver cancer cell lines in comparison with the reference drug doxorubicin with no noticeable toxicity on normal cells. J. Heterocyclic Chem., 00, 00 (2018).

INTRODUCTION Over decades, extensive efforts have been devoted toward developing new treatments for cancer as it counts as one of the most leading causes of mortality worldwide [1–3]. Although great advances in cancer research have already been achieved, the continued attempts of discovering new therapeutics for anticancer are still critically required. Combination treatments, including radio and chemotherapy, showed satisfactory victory over the disease, however, these were associated in many cases with serious side effects on the patient. On the other hand, the discovery of drugs that can target only cancer cells without affecting the normal ones is still believed to be a real challenge. A recent report by the World Health Organization recorded 8.8 million deaths with cancer-related diseases, only in 2015 [4]. Cases with

cancers are estimated to increase at an alarming rate over the next few decades [5]. Therefore, there is an urgent demand to develop and discover novel anticancer agents with a broader spectrum of cytotoxicity that can be effective toward different types of cancer cell lines. Heterocyclic architectures, which contain sulfur and nitrogen elements, were reported to exhibit wide range of pharmacological activities [6–11]. In particular, the 1,3thiazines have been given considerable interest in pharmaceutical research as anti-inflammatory [12], antitumor [13], antibacterial [14], and antiviral agents [15]. However, only a few studies exploring their anticancer potential have been reported [13,16–22]. Moreover, in recent years, attention has been increasingly paid to the synthesis of bis-heterocyclic compounds, which exhibit various biological activities [23–25]. In view of the promising biological profile of the 1,3-

© 2018 Wiley Periodicals, Inc.

T. El Malah, E. A. Ishak, H. F. Nour, R. M. Shaker, M. M. Ali, A. E. Mahmoud, and S. M. Soliman Vol 000 thiazines, a series of symmetrically bridged bis-thiazine derivatives were synthesized and their anticancer activities were evaluated against various cancer cells.

RESULTS AND DISCUSSION Chemistry. As part of our growing interest in synthesizing bis-heterocyclic compounds of interesting biological activities [26–29], we report here the versatile and hitherto unreported synthesis of a series of symmetrically functionalized bis-thiazines, in which the two substituted heterocycles were linked together through a flexible bridge of alkyl chains with variable number of methylene moieties and their anticancer activities were evaluated against various cancer cells. The bis-aldehydes 1a–c were chosen as ideal candidates for the synthesis of the target bis-thiazine scaffolds 3a–c because they can undergo nucleophilic attack upon reaction with the bisnucleophile 2a–c followed by subsequent facial cyclization. More importantly, the distance between the two thiazine rings can be tailored though varying the number of methylene groups of the spacer units of the dialdehydes 1a–c. The bridged dialdehydes 1a–c, which provide the complementary carbons of the six 1,3thiazine rings, were prepared from the alkylation reactions of the 4-hydroxy-3-methoxybenzaldehyde with dihaloalkanes according to the literature [30]. Compounds 2a–e, which possess two nucleophilic terminals of amide and thiol functionalities, were obtained from the addition reactions of the cyanoacetamide with the corresponding aryl or cyclohexyl isothiocyanates according to the literature [31]. The bis-1,3-thiazine derivatives 3a–o were obtained

in good to excellent yields (79–91%) from the condensation reactions of dialdehydes 1a–c with two equivalents of the 3-substituted-amino-2-cyano-3mercaptoacrylamides 2a–e in ethanol in the presence of tosyl chloride as shown in Scheme 1. It is noteworthy that symmetrical bis-1,3-thiazines with rigid phenyl linkages have been reported from the condensation reactions of terephthalaldehyde with 3arylamino-2-cyano-3-mercapto-acrylamide [28]. To the best of our knowledge, bis-1,3-thiazines connected through flexible alkyl bridges were not frequently discussed in the literature with only one exceptional case reported by Wang and co-workers [22]. The chemical structures of the new bis-thiazine derivatives 3a–o were fully characterized by using different spectroscopic techniques and elemental analysis. The 1H NMR spectrum of compound 3a as an example showed a signal at δ 3.88 ppm with an integration of six, corresponding to the two methoxy groups. It also showed a signal at δ 4.48 ppm for the two methylene moieties. The methine signal appeared at δ 5.70 ppm. The two NH groups of the 1,3-thiazine rings appeared at δ 8.43 ppm, while the exocyclic NH moieties appeared at δ 10.21 ppm. On the other hand, the IR spectrum of 3a showed the expected absorption bands of the NH groups at νmax 3215 cm1 and 3182 cm1. It also showed the characteristic sharp absorption band of the cyano group at νmax 2199 cm1 and a strong absorption band, corresponding to the carbonyl functionality of the 1,3-thiazine ring at νmax 1675 cm1. The mass spectrum of 3a showed the anticipated molecular ion peak at m/z 732. The full characterizations of compounds 3b–o are shown in detail in the Experimental. In order to gain more insight into the through space orientation of two

Scheme 1. Acid-catalyzed condensation of bis-aldehyde 1a–c with compounds 2a–e in ethanol.

Cpd

n

R

Cpd

n

R

Cpd

n

R

3a 3b 3c 3d 3e

2 2 2 2 2

C6H5 p-ClC6H5 C6H11 CH2C6H5 (CH2)2C6H5

3f 3g 3h 3i 3j

3 3 3 3 3


3k 3l 3m 3n 3o

4 4 4 4 4


Journal of Heterocyclic Chemistry

DOI 10.1002/jhet

Month 2018

Synthesis and In Vitro Anticancer Evaluation of Symmetrically Bridged 1,3-Thiazine Derivatives 5a showed an absence of the exocyclic NH group and its C NMR spectrum showed a considerable downfield shift of the pyrimidinone CH group at δ 69.80 ppm. In Scheme 3, we postulate a mechanism for the formation of compounds 5a–c. This reaction may occur via deprotonation of the nitrogen thiazine ring with potassium hydroxide followed by subsequent ring opening of the 1,3-thiazine ring to form intermediate A. Thereafter, recyclization of intermediate A took place through the nucleophilic attack of the NH group on the imine moiety to afford the more stable intermediate bispyrimidinones B. The alkylation of intermediate B with benzyl chloride occurred in situ to yield the stable bispyrimidinones 5a–c [21,35]. Anticancer activity. The anticancer activities were expressed by median growth inhibitory concentration (IC50), as shown in Table 1. The results revealed that all the evaluated compounds did not exert any activity against lung A549, colon HCT116, and prostate PC3 cancer cells, while compounds 3d–f and 3o had no anticancer effect against breast cancer cells. Compounds 3l, 3m, and 3n revealed moderate activity compared with the reference drug doxorubicin with IC50 values 11.35 ± 1.50, 9.90 ± 1.05, and 10.80 ± 2.60 μg/mL, respectively, versus 3.11 ± 0.38 μg/mL for doxorubicin. Otherwise, compounds 3e and 3k were found to be potent anticancer agents with IC50 values 3.60 ± 0.47 and 3.80 ± 0.50 μg/mL, respectively, and their IC50 values were found to be close to the doxorubicin drug (IC50: 3.11 ± 0.38 μg/mL). Moreover, evaluation of the anticancer effect of the tested compounds against human liver cancer cell line revealed that although compounds 3d–f, 3n, and 3o exhibited no effect on the cancer cells, compounds 3l and 3m revealed moderate activity in comparison with the standard reference drug doxorubicin

1,3-thiazines along the dialdehyde linkers, we performed molecular modeling simulations in vacuo at the MM+ level using HyperChem software (Release 8.0.8) as shown in Figure 1 [32,33]. The molecular structures of compounds 3a, 3f, and 3k were constructed and drawn in HyperChem based on the X-ray structure reported by Liu et al. [34]. The structures were energy minimized in vacuo at the MM+ molecular mechanics level using the Polak-Ribiera algorithm and a root-mean-square of 0.01 kcal.mol1. It can be inferred from the modeling calculations that the flexibility as well as the dimensions of the molecules increases by introducing the spacer units holding the two heterocycles. Also, the two 1,3-thiazine rings adopt an anti-orientation relative to each other along the linker axis. This may enable them interacting ideally with the biological target with the aid of the remarkable flexibility of the aliphatic ether linkages. The 1,3-thiazines were reported to undergo facile ring opening under basic reaction conditions and recyclization to the corresponding pyrimidine derivatives [31,35]. As similar to the 1,3-thiazines, pyrimidinones were reported to exhibit interesting biological properties including antiinflammatory [36], antitumor [37], antibacterial [37], and antiviral activities [38]. The basic treatment of compounds 3a, 3f, and 3k with ethyl iodide and benzyl chloride afforded the corresponding bis-pyrimidinones 4a–c and 5a–c, respectively (Scheme 2). The structures of the alkylated products 4a–c and 5a–c were elucidated by using various spectroscopic methods. The 1H NMR of compound 4a, for example, did not show the exocyclic NH group and in addition, its 13C NMR spectrum showed a significant downfield shift of the pyrimidinone CH group at δ 69.24 ppm. Furthermore, the quartet-triplet signals of the ethyl moieties appeared in the 1H NMR of 4a at δ 1.12 and 2.85 ppm. As similar, the 1H NMR of

13

(b)

(a)

(c)

Figure 1. MM+ energy minimized structures of bis-thiazines (a) 3a, (b) 3f, and (c) 3k using the Polak-Ribiera algorithm conjugate gradient and root1 mean-square gradient of 0.01 kcal.mol . [Color figure can be viewed at wileyonlinelibrary.com]


DOI 10.1002/jhet

T. El Malah, E. A. Ishak, H. F. Nour, R. M. Shaker, M. M. Ali, A. E. Mahmoud, and S. M. Soliman Vol 000 Scheme 2. Basic alkylation of the bis-1,3-thiazines 3a, 3f, and 3k to afford bis-pyrimidinones 4a–c and 5a–c.

Scheme 3. Proposed mechanism of formation of bis-pyrimidinones 5a–c.

with IC50 values 9.13 ± 1.27 and 11.28 ± 1.70 μg/mL, respectively, versus 3.75 ± 0.48 μg/mL for doxorubicin. In addition, compounds 3e and 3k were found to be potent anticancer agents against liver cancer cell line with IC50 values 4.20 ± 0.49 and 4.50 ± 0.60 μg/mL, respectively, near to the doxorubicin (IC50: 3.75 ± 0.48 μg/mL). The effect of the synthesized compounds on the toxicity of normal HFB4 cells revealed that compounds 3e and 3k showed no noticeable activity against the HFB4 cells. Structure–activity relationship. The structures show a little variation in the biological activities that highlight some vital details of the structure–activity relationship.

Firstly, the dimensions of the compounds seem to be significantly important for the anticancer activity. Potent anticancer activity is attained by either the ethyl spacer between the two phenyl groups holding the aromatic or cyclohexyl imino-substituted 1,3-thiazine rings of compound 3e, or by the butyl linker in the middle of compound 3k. Another example is compound 3f that has a propyl linker in the middle and show anticancer activity better than 3a (with ethyl linker) but still less than 3k. Secondly, aromaticity at the two poles of the compounds might be crucial for the anticancer activity. This was observed from the low anticancer activity exhibited by compounds 3h and 3m bearing cyclohexyl substituents.


DOI 10.1002/jhet

Month 2018

Synthesis and In Vitro Anticancer Evaluation of Symmetrically Bridged 1,3-Thiazine Derivatives Table 1 The anticancer activity of the synthesized compounds (3a–o) on different cell lines IC50 (μg/mL)

Compounds Doxorubicin 3a 3b 3c 3d 3e 3f 3g 3h 3i 3j 3k 3l 3m 3n 3o

A549 HCT116 MCF-7 PC3 HepG2 HFB4 4.36 ± 53.00 ± 46.80 ± 37.45 ± 48.00 ± 34.90 ± 33.76 ± 40.20 ± 39.13 ± 51.08 ± 61.22 ± 41.31 ± 39.27 ± 63.11 ± 34.75 ± 28.83 ±

0.52 6.70 6.16 4.91 6.05 4.86 4.30 5.65 5.12 5.83 6.72 4.83 4.32 7.23 4.50 3.91

4.90 ± 38.26 ± 31.85 ± 41.11 ± 36.73 ± 55.39 ± 35.78 ± 43.19 ± 39.10 ± 29.90 ± 41.47 ± 49.00 ± 44.00 ± 46.82 ± 60.070 ± 37.81 ±

0.56 4.85 4.34 5.01 4.88 6.86 3.72 5.17 4.90 3.50 6.00 5.86 6.00 5.00 7.90 4.20

3.11 ± 0.38 39.44 ± 4.88 33.70 ± 4.90 42.81 ± 5.90 60.22 ± 7.76 3.60 ± 0.47 26.90 ± 3.77 41.80 ± 6.10 60.00 ± 7.00 32.67 ± 4.85 66.00 ± 7.12 3.80 ± 0.50 11.35 ± 1.50 9.90 ± 1.05 10.80 ± 2.60 33.60 ± 4.80

5.11 ± 42.03 ± 76.21 ± 47.82 ± 39.76 ± 48.61 ± 68.00 ± 72.11 ± 43.90 ± 45.25 ± 67.34 ± 49.90 ± 63.88 ± 46.81 ± 50.75 ± 66.40 ±

0.60 4.80 8.12 6.00 4.85 6.02 7.35 8.00 5.26 5.00 7.72 6.13 7.87 5.71 6.00 7.11

3.75 ± 42.20 ± 36.80 ± 29.90 ± 44.13 ± 4.20 ± 31.26 ± 25.76 ± 52.10 ± 28.31 ± 32.80 ± 4.50 ± 9.13 ± 11.28 ± 21.00 ± 45.20 ±

0.48 5.33 4.11 4.03 5.82 0.49 3.90 2.90 6.75 3.95 4.11 0.60 1.27 1.70 3.40 3.87

85.00 ± 10.11 11.79 ± 2.12 23.00 ± 3.11 20.06 ± 2.80 23.25 ± 4.22 81.11 ± 9.32 18.20 ± 1.90 12.33 ± 1.50 15.45 ± 1.77 19.60 ± 2.08 17.90 ± 2.00 79.65 ± 8.90 67.25 ± 7.50 61.85 ± 8.00 38.70 ± 1.90 21.40 ± 3.10

Data were expressed as mean ± standard error of three experiments.

Finally, chlorine substitution may not be favorable for the anticancer activity of this series of molecules. The anticancer activity of compound 3l dramatically decreased by introducing a chloride atom at the paraposition of the phenyl group in comparison with activity of the potent anticancer analogous 3k.

CONCLUSION In summary, we had synthesized a novel series of bis1,3-thiazine derivatives. The anticancer activity of the new compounds was evaluated in vitro against lung A549, colon HCT116, breast MCF-7, prostate PC3, liver HepG2, and normal melanocyte HFB4. Three compounds 3l, 3m, and 3n showed moderate activity, while two 3e and 3k displayed potent anticancer activity in comparison with the reference doxorubicin drug. The number of methylene groups of the spacer units connecting the 1,3thiazine rings and the substituents on the 1,3-thiazine groups contributed in controlling the dimensions of the compounds and consequently the anticancer activity.

EXPERIMENTAL Chemistry. All melting points are uncorrected and were measured using an electrothermal IA 9100 apparatus, Shimadzu (Japan). Microanalytical data were performed by using a Vario El-Mentar apparatus, Organic Microanalysis Section, Microanalytical Center, Cairo University, Giza, Egypt. The results of the microanalysis were found to be in agreement with the calculated values (±0.3).

The IR spectra (KBr) were recorded on a Perkin-Elmer 1650 spectrophotometer, Microanalytical Center, Cairo University, Giza, Egypt. 1H and 13C NMR spectra were determined on a JEOL 300 MHz in DMSO-d6, Microanalytical Center, Cairo University, Giza, Egypt. The chemical shifts were expressed in ppm relative to TMS as an internal reference. Mass spectra were recorded on 70 eV EI Ms-QP 1000 EX (Shimadzu, Japan), Microanalytical Center, Cairo University, Giza, Egypt. General procedure for the synthesis of the bis-1,3-thiazines 3a–o. A mixture of bridged dialdehydes 1a–c (0.01 mol),

2a–e (0.02 mol), and p-toluenesulfonic acid (0.076 g, 0.01 mol) in ethanol (20 mL) was refluxed. A pale yellow precipitate was formed after 30 min, and stirring was continued for extra 2 h. The precipitate was filtered off, washed with ethanol, dried, and recrystallized from DMF/EtOH to yield the corresponding products 3a–o. General procedure bis-pyrimidinones 4a–c.

for

General procedure bis-pyrimidinones 5a–c.

for

the

synthesis

of

the

Ethyl iodide (40 mmol) was added to a mixture of compounds 3a, 3f, or 3k (10 mmol) and anhydrous potassium carbonate (4 mmol) in DMF (5 mL). The reaction mixture was stirred for 18– 20 h at room temperature and then the mixture was poured into cold water. After stirring for additional 15 min, the resulting precipitate was collected by filtration, washed with water, dried, and recrystallized from ethanol to give compounds 4a–c. the

synthesis

of

the

To a stirred 0.75 N aqueous KOH solution (20 mL), compounds 3a, 3f, or 3k (10 mmol) and benzyl chloride (40 mmol) were added successively. The resulting precipitate was filtered off, washed with water, dried, and recrystallized from DMF/EtOH to afford compounds 5a–c.


DOI 10.1002/jhet

T. El Malah, E. A. Ishak, H. F. Nour, R. M. Shaker, M. M. Ali, A. E. Mahmoud, and S. M. Soliman Vol 000 Anticancer assessment. Materials and methods, Fetal bovine serum and Lchemicals, and cell culturing.

glutamine were obtained from Gibco Invitrogen Company (Scotland, UK). Dulbecco’s modified Eagle’s medium was provided from Cambrex (New Jersey, USA). Dimethyl sulfoxide (DMSO), doxorubicin, penicillin, streptomycin, and Sulfo-Rhod-amine-B stain (SRB) (3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) were obtained from Sigma chemical company (St. Louis, MO, USA). All other chemicals and reagents used in this study were of analytical grade and purchased from SigmaAldrich chemical Co. (St. Louis, MO, USA). The lung A549, colon HCT116, breast MCF-7, prostate PC3, and liver HepG2 cancer cell lines as well as the normal cell line (human normal melanocyte, HFB4) were obtained from the American Type Culture Collection (Rockville, MD, USA). Cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% heat inactivated fetal calf serum (GIBCO), penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37°C in a humidified atmosphere containing 5% CO2. Cells at a concentration of 0.50 × 106 were grown in a 25 cm2 flask in 5 mL of culture medium. Anticancer activity assay. The anticancer activity of the synthesized compounds 3f–o were measured in vitro using the SRB assay according to Skehan et al. [39]. Briefly, cells were inoculated in 96-well microtiter plate (104 cells/well) for 24 h before treatment with the tested compounds to allow attachment of cell to the wall of the plate. Test compounds were dissolved in DMSO at 1 mg/mL immediately before use and diluted to the appropriate volume just before addition to the cell culture. Different concentration of tested compounds and doxorubicin were added to the cells. Four wells were prepared for each individual dose. Monolayer cells were incubated with the compounds for 48 h at 37°C and in an atmosphere of 5% CO2. After 48 h, cells were fixed, washed, and stained for 30 min with 0.4% (w/v) SRB dissolved in 1% acetic acid. Unbound dye was removed by four washes with 1% acetic acid and attached stain was recovered with Tris-EDTA buffer. Color intensity was measured in an ELISA reader. The relation between surviving fraction and drug concentration is plotted to get the survival curve for each cell line after the specified time. The concentration required for 50% inhibition of cell viability (IC50) was calculated and the results are given in Table 1. The results were compared with the effect of the reference drug, doxorubicin [40,41]. 2,20 -(4,40 -(Ethane-1,2-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenylamino)-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3a). Yield 86%; m.p. 222–224°C;

IR (KBr): νmax/cm1 = 3215, 3182 (NH), 2199 (CN), 1675 (CO); 1H NMR (DMSO-d6, ppm): δH = 3.88 (s, 6H,

2OCH3), 4.48 (s, 4H, 2OCH2), 5.70 (s, 2H, 2NHCHS), 6.98–7.41 (m, 14H, CHAr), 7.52 (s, 2H, CHAr), 8.43 (d, 2H, J = 2.4 Hz, 2NH), 10.21 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 52.46 (NHCHS), 57.15 (OCH3), 68.36 (COCCN), 70.01 (OCH2), 112.07 (CAr), 113.64 (CAr), 115.25 (CN), 122.86 (CAr), 124.57 (CAr), 127.28 (CAr), 130.92 (CAr), 133.27 (CAr), 138.60 (CAr), 149.09 (OCCHAr), 151.86 (CAr), 166.12 (CO), 171.48 (SCNH); MS, m/z (%): 732 (M+, 85). Anal. Calcd. for C38H32N6O6S2 (732.83) required C, 62.28; H, 4.40; N, 11.47; S, 8.75; found: C, 62.36; H, 4.35; N, 11.55; S, 8.84. 2,20 -(4,40 -(Ethane-1,2-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(4-chlorophenylamino)-4-oxo-3,4-dihydro2H-1,3-thiazine-5-carbonitrile) (3b). Yield 84%; m.p. 226–

228°C; IR (KBr): νmax/cm1 = 3224, 3194 (NH), 2027 (CN), 1671 (CO); 1H NMR (DMSO-d6, ppm): δH = 3.82 (s, 6H, 2OCH3), 4.47 (s, 4H, 2OCH2), 6.08 (s, 2H, 2NHCHS), 6.99–7.09 (m, 6H, CHAr), 7.22–7.48 (m, 8H, CHAr), 8.85 (d, 2H, J = 2.4 Hz, 2NH), 10.20 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 52.42 (NHCHS), 57.20 (OCH3), 68.43 (COCCN), 70.24 (OCH2), 112.10 (CAr), 113.75 (CAr), 115.31 (CN), 122.79 (CAr), 123.84 (CAr), 128.05 (CAr), 130.87 (CAr), 133.57 (CAr), 135.34 (CAr), 150.11 (OCCHAr), 152.27 (CAr), 166.52 (CO), 171.59 (SCNH); MS, m/z (%): 801 (M+, 90). Anal. Calcd. for C38H30Cl2N6O6S2 (801.72) required C, 56.93; H, 3.77; N, 10.48; S, 8.00; found: C, 56.85; H, 3.86; N, 10.56; S, 8.10.

2,20 -(4,40 -(Ethane-1,2-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(cyclohexylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3c). Yield 82%; m.p. 252–254°C;

IR (KBr): νmax/cm1 = 3218, 3185 (NH), 2199 (CN), 1678 (CO); 1H NMR (DMSO-d6, ppm): δH = 0.95–1.85 (m, 20H, CH2 cyhex), 2.15 (s, 2H, CHcyhex), 3.83 (s, 6H, 2OCH3), 4.31 (s, 4H, 2OCH2), 6.06 (s, 2H, 2NHCHS), 7.06 (d, 2H, J = 3.6 Hz, CHAr), 7.15 (d, 2H, J = 3.7 Hz, CHAr), 7.21 (s, 2H, CHAr), 8.22 (s, 2H, 2NH), 9.86 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 26.56 (CH2 cyhex), 26.85 (CH2 cyhex), 33.74 (CH2 cyhex), 50.12 (CH cyhex), 52.84 (NHCHS), 57.17 (OCH3), 68.59 (COCCN), 70.61 (OCH2), 112.54 (CAr), 113.74 (CAr), 115.73 (CN), 122.62 (CAr), 135.68 (CAr), 150.27 (OCCHAr), 152.32 (CAr), 166.95 (CO), 174.46 (SCNH); MS, m/z (%): 744 (M+, 90). Anal. Calcd. for C38H44N6O6S2 (744.92) required C, 61.27; H, 5.95; N, 11.28; S, 8.61; found: C, 61.34; H, 5.81; N, 11.35; S, 8.52. 2,20 -(4,40 -(Ethane-1,2-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(benzylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3d). Yield 87%; m.p. 238–240°C;

IR (KBr): νmax/cm1 = 3221, 3189 (NH), 2030 (CN), 1678 (CO); 1H NMR (DMSO-d6, ppm): δH = 3.49 (s, 4H, NHCH2Ph), 3.83 (s, 6H, 2OCH3), 4.48 (s, 4H, 2OCH2), 6.07 (s, 2H, 2NHCHS), 6.82–7.58 (m, 16H, CHAr), 8.12 (s, 2H, 2NH), 9.85 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 47.30 (NHCH2Ph), 52.37 (NHCHS), 57.43


DOI 10.1002/jhet

Month 2018

Synthesis and In Vitro Anticancer Evaluation of Symmetrically Bridged 1,3-Thiazine Derivatives

(OCH3), 68.46 (COCCN), 70.21 (OCH2), 112.57 (CAr), 113.62 (CAr), 115.40 (CN), 122.61 (CAr), 125.97 (CAr), 127.12 (CAr), 129.54 (CAr), 134.75 (CAr), 136.80 (CAr), 150.28 (OCCHAr), 152.34 (CAr), 167.95 (CO), 173.32 (SCNH); MS, m/z (%): 760 (M+, 62). Anal. Calcd. for C40H36N6O6S2 (760.88) required C, 63.14; H, 4.77; N, 11.05; S, 8.43; found: C, 63.22; H, 4.68; N, 11.12; S, 8.52. 2,20 -(4,40 -(Ethane-1,2-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenethylamino)-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3e). Yield 81%; m.p. 240–242°C;

IR (KBr): νmax/cm1 = 3212, 3181 (NH), 2104 (CN), 1679 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.79–2.91 (m, 8H, 2NHCH2CH2Ph), 3.83 (s, 6H, 2OCH3), 4.47 (s, 4H, 2OCH2), 6.01 (s, 2H, 2NHCHS), 6.92–7.31 (m, 8H, CHAr), 7.33–7.55 (m, 8H, CHAr), 8.79 (s, 2H, 2NH), 9.84 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 36.64 (NHCH2CH2Ph), 44.52 (NHCH2CH2Ph), 52.35 (NHCHS), 57.45 (OCH3), 69.28 (COCCN), 70.35 (OCH2), 112.51 (CAr), 113.70 (CAr), 114.97 (CN), 122.67 (CAr), 126.24 (CAr), 127.43 (CAr), 129.87 (CAr), 135.62 (CAr), 140.13 (CAr), 150.32 (OCCHAr), 152.37 (CAr), 168.64 (CO), 175.01 (SCNH); MS, m/z (%): 788 (M+, 79). Anal. Calcd. for C42H40N6O6S2 (788.93) required C, 63.94; H, 5.11; N, 10.65; S, 8.13; found: C, 64.05; H, 5.21; N, 10.71; S, 8.18. 2,20 -(4,40 -(Propane-1,3-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenylamino)-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3f). Yield 87%; m.p. 245–247°C; 1

IR (KBr): νmax/cm = 3201, 3188 (NH), 2192 (CN), 1670 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.49–2.51 (m, 2H, OCH2CH2), 3.82 (s, 6H, 2OCH3), 4.14–4.19 (m, 4H, 2OCH2), 6.04 (s, 2H, 2NHCHS), 6.98 (d, 4H, J = 3.9 Hz, CHAr), 7.06–7.39 (m, 12H, CHAr), 8.41 (d, 2H, J = 2.7 Hz, 2NH),10.18 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 29.88 (OCH2CH2), 52.51 (NHCHS), 56.87 (OCH3), 66.04 (OCH2), 69.41 (COCCN), 112.18 (CAr), 113.68 (CAr), 114.19 (CN), 122.75 (CAr), 124.47 (CAr), 127.32 (CAr), 131.12 (CAr), 135.82 (CAr), 138.68 (CAr), 149.53 (OCCHAr), 152.47 (CAr), 167.86 (CO), 170.55 (SCNH); MS, m/z (%): 746 (M+, 92). Anal. Calcd. for C39H34N6O6S2 (746.85) required C, 62.72; H, 4.59; N, 11.25; S, 8.59; found: C, 62.80; H, 4.45; N, 11.31; S, 8.65. 0

0

2,2 -(4,4 -(Propane-1,3-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(4-chlorophenylamino)-4-oxo-3,4-dihydro2H-1,3-thiazine-5-carbonitrile) (3g). Yield 80%; m.p. 248– 1

250°C; IR (KBr): νmax/cm = 3211, 3185 (NH), 2200 (CN), 1677 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.48– 2.50 (m, 2H, OCH2CH2), 3.82 (s, 6H, 2OCH3), 4.10–4.12 (t, 4H, J = 5.1 Hz, 2OCH2), 6.08 (s, 2H, 2NHCHS), 6.99– 7.51 (m, 14H, CHAr), 8.46 (s, 2H, 2NH), 10.19 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 29.91 (OCH2CH2), 52.50 (NHCHS), 57.14 (OCH3), 66.21 (OCH2), 68.74 (COCCN), 112.19 (CAr), 113.67 (CAr), 114.25 (CN), 122.80 (CAr), 124.48 (CAr), 127.43 (CAr),

128.91 (CAr), 131.15 (CAr), 135.88 (CAr), 138.75 (CAr), 149.60 (OCCHAr), 152.42 (CAr), 168.57 (CO), 170.69 (SCNH); MS, m/z (%): 814 (M+, 95). Anal. Calcd. for C39H32Cl2N6O6S2 (814.12) required C, 57.42; H, 3.95; N, 10.30; S, 7.86; found: C, 57.31; H, 4.04; N, 10.36; S, 7.93.

2,20 -(4,40 -(Propane-1,3-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(cyclohexylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3h). Yield 88%; m.p. 216–218°C;

IR (KBr): νmax/cm1 = 3228, 3191 (NH), 2199 (CN), 1675 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.08–1.94 (m, 20H, CH2 cyhex), 2.09–2.16 (m, 2H, OCH2CH2), 3.40– 3.45 (m, 2H, CH2 cyhex), 3.82 (s, 6H, 2OCH3), 4.29 (t, 4H, J = 3 Hz, 2OCH2), 6.05 (s, 2H, 2NHCHS), 7.02 (d, 2H, J = 3.7 Hz, CHAr), 7.13 (d, 2H, J = 3.5 Hz, CHAr), 7.21 (s, 2H, CHAr), 8.20 (s, 2H, 2NH), 9.84 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 26.43 (CH2 cyhex), 26.81 (CH2 cyhex), 30.05 (OCH2CH2), 33.78 (CH2 cyhex), 50.24 (CH cyhex), 52.48 (NHCHS), 57.35 (OCH3), 66.24 (OCH2), 68.80 (COCCN), 112.34 (CAr), 113.27 (CAr), 114.87 (CN), 122.75 (CAr), 135.57 (CAr), 149.71 (OCCHAr), 152.39 (CAr), 168.61 (CO), 175.18 (SCNH); MS, m/z (%): 758 (M+, 78). Anal. Calcd. for: C39H46N6O6S2 (758.95) required C, 61.72; H, 6.11; N, 11.07; S, 8.45; found: C, 61.83; H, 5.98; N, 11.12; S, 8.53.

2,20 -(4,40 -(Propane-1,3-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(benzylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3i). Yield 89%; m.p. 218–220°C;

IR (KBr): νmax/cm1 = 3223, 3185 (NH), 2199 (CN), 1681 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.19–2.25 (m, 2H, OCH2CH2), 3.67 (s, 4H, NHCH2Ph), 3.82 (s, 6H, 2OCH3), 4.26 (t, 4H, J = 6.3 Hz, 2OCH2), 6.06 (s, 2H, 2NHCHS), 7.21–7.55 (m, 16H, CHAr), 9.21 (d, 2H, J = 2.9 Hz, 2NH), 9.84 (s, 2H, 2NH). 13C NMR (DMSOd6, ppm): δC = 29.85 (OCH2CH2), 46.62 (NHCH2Ph), 52.46 (NHCHS), 56.81 (OCH3), 66.12 (OCH2), 69.34 (COCCN), 112.36 (CAr), 113.65 (CAr), 114.87 (CN), 122.65 (CAr), 126.97 (CAr), 127.36 (CAr), 131.24 (CAr), 135.87 (CAr), 138.81 (CAr), 149.58 (OCCHAr), 152.54 (CAr), 167.91 (CO), 174.05 (SCNH); MS, m/z (%): 774 (M+, 82). Anal. Calcd. for C41H38N6O6S2 (774.91) required C, 63.55; H, 4.94; N, 10.85; S, 8.28; found: C, 63.61; H, 5.02; N, 10.91; S, 8.19. 2,20 -(4,40 -(Propane-1,3-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenethylamino)-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3j). Yield 90%; m.p. 230–232°C;

IR (KBr): νmax/cm1 = 3211, 3173 (NH), 2200 (CN), 1661 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.11–2.26 (m, 2H, OCH2CH2), 2.85–2.93 (m, 8H, 2NHCH2CH2Ph), 3.83 (s, 6H, 2OCH3), 4.12–4.17 (m, 4H, 2OCH2), 5.99 (s, 2H, 2NHCHS), 7.06–6.93–7.58 (m, 16H, CHAr), 8.79 (s, 2H, 2NH), 9.84 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 29.78 (OCH2CH2), 36.65 (NHCH2CH2Ph), 44.54 (NHCH2CH2Ph), 52.60 (NHCHS), 56.86 (OCH3), 66.12 (OCH2), 69.42 (COCCN), 112.35 (CAr), 113.61 (CAr), 114.27 (CN), 122.73 (CAr), 126.18 (CAr), 127.54


DOI 10.1002/jhet

T. El Malah, E. A. Ishak, H. F. Nour, R. M. Shaker, M. M. Ali, A. E. Mahmoud, and S. M. Soliman Vol 000 (CAr), 130.81 (CAr), 135.84 (CAr), 140.75 (CAr), 149.87 (OCCHAr), 152.48 (CAr), 167.89 (CO), 174.24 (SCNH); MS, m/z (%): 802 (M+, 82). Anal. Calcd. for C43H42N6O6S2 (802.96) required C, 64.32; H, 5.27; N, 10.47; S, 7.99; found: C, 64.40; H, 5.32; N, 10.39; S, 8.04.

2,20 -(4,40 -(Butane-1,4-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenylamino)-3,4-dihydro-2H-1,3Yield 79%; m.p. 234– thiazine-5-carbonitrile) (3k).

2236°C; IR (KBr): νmax/cm1 = 3199, 3185 (NH), 2200 (CN), 1668 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.84–1.86 (m, 4H, OCH2CH2), 3.81 (s, 6H, 2OCH3), 4.01 (t, 4H, J = 6.01 Hz, 2OCH2), 6.05 (s, 2H, 2NHCHS), 6.96–6.94 (m, 4H, CHAr), 7.05–7.25 (m, 8H, CHAr), 7.34–7.40 (m, 4H, CHAr), 8.41 (d, 2H, J = 2.6 Hz, 2NH), 10.18 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 28.15 (OCH2CH2), 52.46 (NHCHS), 56.84 (OCH3), 66.16 (COCCN), 69.43 (OCH2), 112.21 (CAr), 113.67 (CAr), 114.21 (CN), 122.80 (CAr), 124.45 (CAr), 127.42 (CAr), 131.18 (CAr), 135.86 (CAr), 138.65 (CAr), 150.04 (OCCHAr), 152.63 (CAr), 167.92 (CO), 171.38 (SCNH); MS, m/z (%): 760 (M+, 89). Anal. Calcd. for C40H36N6O6S2 (760.88) required C, 63.14; H, 4.77; N, 11.05; S, 8.43; found: C, 63.21; H, 4.65; N, 10.94; S, 8.51. 2,20 -(4,40 -(Butane-1,4-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(4-chlorophenylamino)-4-oxo-3,4-dihydro2H-1,3-thiazine-5-carbonitrile) (3l). Yield 85%; m.p. 237–

239°C; IR (KBr): νmax/cm1 = 3194, 3178 (NH), 2195 (CN), 1665 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.89– 1.92 (m, 4H, OCH2CH2), 3.82 (s, 6H, 2OCH3), 4.09 (t, 4H, J = 6.05 Hz, 2OCH2), 6.08 (s, 2H, 2NHCHS), 6.97– 7.44 (m, 14H, CHAr), 8.43 (s, 2H, 2NH), 10.19 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 28.14 (OCH2CH2), 52.47 (NHCHS), 56.92 (OCH3), 66.27 (COCCN), 70.08 (OCH2), 112.35 (CAr), 113.75 (CAr), 114.28 (CN), 122.74 (CAr), 124.52 (CAr), 127.49 (CAr), 131.22 (CAr), 135.81 (CAr), 138.69 (CAr), 150.13 (OCCHAr), 152.78 (CAr), 168.05 (CO), 171.94 (SCNH); MS, m/z (%): 829 (M+, 96). Anal. Calcd. for C40H34Cl2N6O6S2 (829.77) required C, 57.90; H, 4.13; N, 10.13; S, 7.73; found: C, 58.02; H, 4.21; N, 10.20; S, 7.69. 2,20 -(4,40 -(Butane-1,4-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(cyclohexylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3m). Yield 81%; m.p. 226–228°C;

IR (KBr): νmax/cm1 = 3216, 3172 (NH), 2203 (CN), 1670 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.02–1.87 (m, 20H, CH2 cyhex), 2.05–2.14 (m, 4H, OCH2CH2), 3.41– 3.45 (m, 2H, CH2 cyhex), 3.83 (s, 6H, 2OCH3), 4.05 (t, 4H, J = 2.7 Hz, 2OCH2), 6.05 (s, 2H, 2NHCHS), 6.97– 7.03 (m, 2H, CHAr), 7.12 (d, 2H, J = 3.7 Hz, CHAr), 7.34 (s, 2H, CHAr), 8.20 (s, 2H, 2NH), 9.93 (s, 2H, 2NH). 13C NMR (DMSO-d6, ppm): δC = 26.44 (CH2 cyhex), 26.79 (CH2 cyhex), 29.12 (OCH2CH2), 33.74 (CH2 cyhex), 50.22 (CH cyhex), 52.50 (NHCHS), 57.37 (OCH3), 67.62 (COCCN), 69.24 (OCH2), 112.48 (CAr), 113.13 (CAr),

114.93 (CN), 122.67 (CAr), 135.62 (CAr), 149.60 (OCCHAr), 153.52 (CAr), 168.43 (CO), 175.27 (SCNH); MS, m/z (%): 772 (M+, 84). Anal. Calcd. for C40H48N6O6S2 (772.98) required C, 62.15; H, 6.26; N, 10.87; S, 8.30; found: C, 62.21; H, 6.17; N, 10.94; S, 8.22. 2,20 -(4,40 -(Butane-1,4-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(6-(benzylamino)-4-oxo-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3n). Yield 85%; m.p. 217–219°C;

IR (KBr): νmax/cm1 = 3215, 3192 (NH), 2205 (CN), 1676 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.81–1.96 (m, 4H, OCH2CH2), 3.82 (s, 6H, 2OCH3), 4.01 (s, 4H, NHCH2Ph), 4.18 (m, 4H, 2OCH2), 6.07 (s, 2H, 2NHCHS), 6.95–7.38 (m, 16H, CHAr), 8.25 (s, 2H, 2NH), 9.84 (s, 2H, 2NH). 13 C NMR (DMSO-d6, ppm): δC = 28.75 (OCH2CH2), 46.81 (NHCH2Ph), 52.67 (NHCHS), 56.87 (OCH3), 66.24 (COCCN), 68.72 (OCH2), 112.43 (CAr), 113.58 (CAr), 114.64 (CN), 121.78 (CAr), 125.92 (CAr), 127.60 (CAr), 131.23 (CAr), 135.65 (CAr), 138.50 (CAr), 150.43 (OCCHAr), 152.70 (CAr), 168.15 (CO), 175.76 (SCNH); MS, m/z (%): 788 (M+, 75). Anal. Calcd. for C42H40N6O6S2 (788.93) required C, 63.94; H, 5.11; N, 10.65; S, 8.13; found: C, 64.03; H, 5.16; N, 10.72; S, 8.05.

2,20 -(4,40 -(Butane-1,4-diylbis(oxy))bis(3-methoxy-4,1phenylene))bis(4-oxo-6-(phenethylamino)-3,4-dihydro-2H-1,3thiazine-5-carbonitrile) (3o). Yield 91%; m.p. 215–217°C;

IR (KBr): νmax/cm1 = 3211, 3185 (NH), 2202 (CN), 1679 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.91 (t, 4H, J = 6.6 Hz, OCH2CH2), 2.81–2.92 (m, 8H, 2NHCH2CH2Ph), 3.82 (s, 6H, 2OCH3), 4.17 (t, 4H, J = 4.5 Hz, 2OCH2), 5.97 (s, 2H, 2NHCHS), 6.97–7.55 (m, 16H, CHAr), 8.78 (s, 2H, 2NH), 9.83 (s, 2H, 2NH). 13 C NMR (DMSO-d6, ppm): δC = 28.62 (OCH2CH2), 36.62 (NHCH2CH2Ph), 43.75 (NHCH2CH2Ph), 52.54 (NHCHS), 56.41 (OCH3), 68.24 (OCH2), 70.58 (COCCN), 112.49 (CAr), 113.85 (CAr), 114.97 (CN), 122.68 (CAr), 126.07 (CAr), 128.65 (CAr), 130.78 (CAr), 136.34 (CAr), 140.79 (CAr), 150.76 (OCCHAr), 152.62 (CAr), 168.95 (CO), 175.25 (SCNH); MS, m/z (%): 816 (M+, 72). Anal. Calcd. for C44H44N6O6S2 (816.99) required C, 64.69; H, 5.43; N, 10.29; S, 7.85; found: C, 64.76; H, 5.32; N, 10.37; S, 7.94. 2-(4-(2-(4-(5-Cyano-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4tetrahydropyrimidin-2-yl)-2-methoxy-phenoxy)ethoxy)-3methoxyphenyl)-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4Yield 77%; m.p. tetrahydropyrimidine-5-carbonitrile (4a).

281–283°C; IR (KBr): νmax/cm1 = 3251, 3058 (NH), 2199 (CN), 1669 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.12 (t, 6H, 2CH3, J = 3.5 Hz), 2.85 (q, 4H, 2CH2, J = 3.2 Hz), 3.76 (s, 6H, 2OCH3), 4.41 (s, 4H, 2OCH2), 6.11 (s, 2H, 2NHCHN), 6.72–7.46 (m, 16H, CHAr), 8.71 (d, 2H, 2NH, J = 4.6 Hz). 13C NMR (DMSO-d6, ppm): δC = 15.42 (CH3), 26.54 (CH3CH2S), 58.37 (OCH3), 68.41 (COCCN), 69.24 (NHCHN), 70.85 (OCH2), 112.46 (CAr), 113.69 (CAr), 115.12 (CN), 121.48 (CAr), 123.05 (CAr), 123.25 (CAr), 129.74 (CAr), 137.69 (CAr),


DOI 10.1002/jhet

Month 2018

Synthesis and In Vitro Anticancer Evaluation of Symmetrically Bridged 1,3-Thiazine Derivatives

145.42 (NCAr), 150.16 (CAr), 151.76 (CAr), 168.81 (CO), 179.15 (SC─N–Ph); MS, m/z (%): 788 (M+, 87). Anal. Calcd. for C42H40N6O6S2 (788.93) required C, 63.94; H, 5.11; N, 10.65; S, 8.13; found: C, 63.84; H, 5.18; N, 10.57; S, 8.04. 2-(4-(3-(4-(5-Cyano-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4tetrahydropyrimidin-2-yl)-2-methoxy-phenoxy)propoxy)-3methoxyphenyl)-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4Yield 79%; m.p. tetrahydropyrimidine-5-carbonitrile (4b).

271–273°C; IR (KBr): νmax/cm1 = 3238, 3187 (NH), 2163 (CN), 1679 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.18 (t, 6H, J = 3.6 Hz, 2CH3,), 2.34–2.36 (m, 2H, OCH2CH2), 2.87 (q, 4H, J = 3.1 Hz, 2CH2), 3.75 (s, 6H, 2OCH3), 4.09–4.12 (m, 4H, 2OCH2), 6.05 (s, 2H, 2NHCHN), 6.69–7.48 (m, 16H, CHAr), 8.60 (d, 2H, J = 4.5 Hz, 2NH). 13C NMR (DMSO-d6, ppm): δC = 15.30 (CH3), 26.46 (CH3CH2S), 29.84 (OCH2CH2), 58.35 (OCH3), 68.43 (COCCN), 69.28 (NHCHN), 70.75 (OCH2), 112.48 (CAr), 113.75 (CAr), 114.83 (CN), 120.72 (CAr), 122.62 (CAr), 123.28 (CAr), 130.45 (CAr), 137.65 (CAr), 145.57 (NCAr), 150.22 (CAr), 151.75 (CAr), 168.94 (CO), 179.72 (SC–N–Ph); MS, m/z (%): 802 (M+, 92). Anal. Calcd. for C43H42N6O6S2 (802.96) required C, 64.32; H, 5.27; N, 10.47; S, 7.99; found: C, 64.25; H, 5.31; N, 10.54; S, 8.05. 2-(4-(4-(4-(5-Cyano-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4tetrahydropyrimidin-2-yl)-2-methoxyph-enoxy)butoxy)-3methoxyphenyl)-6-(ethylthio)-4-oxo-1-phenyl-1,2,3,4Yield 80%; m.p. tetrahydropyrimidine-5-carbonitrile (4c).

265–267°C; IR (KBr): νmax/cm1 = 3226, 3129 (NH), 2213 (CN), 1670 (CO); 1H NMR (DMSO-d6, ppm): δH = 1.15 (t, 6H, J = 3.3 Hz, 2CH3,), 1.92–1.95 (t, 4H, J = 4.1 Hz OCH2CH2), 2.86 (q, 4H, 2CH2, J = 3.4 Hz), 3.80 (s, 6H, 2OCH3), 4.05–4.08 (m, 4H, 2OCH2), 6.07 (s, 2H, 2NHCHN), 6.62–7.44 (m, 16H, CHAr), 8.73 (d, 2H, 2NH, J = 4.1 Hz). 13C NMR (DMSO-d6, ppm): δC = 15.13 (CH3), 26.48 (CH3CH2S), 29.71 (OCH2CH2), 58.40 (OCH3), 68.27 (COCCN), 69.21 (NHCHN), 69.84 (OCH2), 112.51 (CAr), 113.74 (CAr), 114.94 (CN), 120.76 (CAr), 122.74 (CAr), 123.21 (CAr), 130.38 (CAr), 138.42 (CAr), 145.48 (NCAr), 150.31 (CAr), 151.70 (CAr), 168.74 (CO), 180.13 (SC-N-Ph); MS, m/z (%): 816 (M+, 87). Anal. Calcd. for C44H44N6O6S2 (816.99) required C, 64.69; H, 5.43; N, 10.29; S, 7.85; found: C, 64.53; H, 5.51; N, 10.17; S, 7.93. 6-(Benzylthio)-2-(4-(2-(4-(6-(benzylthio)-5-cyano-4-oxo-1phenyl-1,2,3,4-tetrahydropyrimidin-2-yl)-2-methoxyphenoxy) ethoxy)-3-methoxyphenyl)-4-oxo-1-phenyl-1,2,3,4Yield 75%; m.p. tetrahydropyrimidine-5-carbonitrile (5a).

274–276°C; IR (KBr): νmax/cm1 = 3237, 3094 (NH), 2199 (CN), 1674 (CO); 1H NMR (DMSO-d6, ppm): δH = 3.79 (s, 6H, 2OCH3), 3.99 (s, 4H, 2SCH2Ph), 4.59 (s, 4H, 2OCH2), 6.08 (s, 2H, 2NHCHN), 6.50–6.99 (m, 12H, CHAr), 7.15–7.53 (m, 14H, CHAr), 8.62 (d, 2H, 2NH, J = 5.1 Hz). 13C NMR (DMSO-d6, ppm):

δC = 39.85 (PhCH2S), 57.23 (OCH3), 68.54 (COCCN), 69.80 (NHCHN), 70.95 (OCH2), 112.32 (CAr), 113.51 (CAr), 115.09 (CN), 121.45 (CAr), 123.11 (CAr), 123.51 (CAr), 129.47 (CAr), 130.04 (CAr), 130.62 (CAr), 132.27 (CAr), 137.45 (CAr), 138.98 (CAr), 145.39 (NCAr), 150.37 (CAr), 151.69 (CAr), 170.24 (CO), 179.28 (SC-N-Ph); MS, m/z (%): 913 (M+, 75). Anal. Calcd. for C52H44N6O6S2 (913.07) required C, 68.40; H, 4.86; N, 9.20; S, 7.02; found: C, 68.32; H, 4.95; N, 9.25; S, 6.96. 6-(Benzylthio)-2-(4-(3-(4-(6-(benzylthio)-5-cyano-4-oxo-1phenyl-1,2,3,4-tetrahydropyrimidin-2-yl)-2-methoxyphenoxy) propoxy)-3-methoxyphenyl)-4-oxo-1-phenyl-1,2,3,4tetrahydropyrimidi-ne-5-carbonitrile (5b). Yield 76%; m.p.

278–280°C; IR (KBr): νmax/cm1 = 3245, 3137 (NH), 2118 (CN), 1668 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.18–2.22 (m, 2H, OCH2CH2), 3.77 (s, 6H, 2OCH3), 3.97 (s, 4H, 2SCH2Ph), 4.43 (s, 4H, 2OCH2), 5.87 (s, 2H, 2NHCHN), 6.49–7.02 (m, 12H, CHAr), 7.19–7.56 (m, 14H, CHAr), 8.84 (d, 2H, 2NH, J = 5.3 Hz). 13C NMR (DMSO-d6, ppm): δC = 31.04 (OCH2CH2), 40.21 (PhCH2S), 57.35 (OCH3), 68.47 (COCCN), 69.43 (NHCHN), 69.87 (OCH2), 112.74 (CAr), 112.98 (CAr), 115.26 (CN), 121.60 (CAr), 122.82 (CAr), 123.73 (CAr), 128.95 (CAr), 129.26 (CAr), 130.48 (CAr), 132.19 (CAr), 137.91 (CAr), 139.24 (CAr), 145.84 (NCAr), 150.64 (CAr), 152.47 (CAr), 171.35 (CO), 179.13 (SC-N-Ph); MS, m/z (%): 927 (M+, 94). Anal. Calcd. for C53H46N6O6S2 (927.10) required C, 68.66; H, 5.00; N, 9.06; S, 6.92; found: C, 68.59; H, 5.07; N, 8.91; S, 7.04.

6-(Benzylthio)-2-(4-(4-(4-(6-(benzylthio)-5-cyano-4-oxo-1phenyl-1,2,3,4-tetrahydropyrimidin-2-yl)-2-methoxyphenoxy) butoxy)-3-methoxyphenyl)-4-oxo-1-phenyl-1,2,3,4Yield 77%; m.p. tetrahydropyrimidine-5-carbonitrile (5c).

266–268°C; IR (KBr): νmax/cm1 = 3227, 3165 (NH), 2199 (CN), 1662 (CO); 1H NMR (DMSO-d6, ppm): δH = 2.03 (t, 4H, J = 4.5 Hz OCH2CH2), 3.79 (s, 6H, 2OCH3), 3.95 (s, 4H, 2SCH2Ph), 4.10–4.13 (m, 4H, 2OCH2), 5.90 (s, 2H, 2NHCHN), 6.51–7.09 (m, 12H, CHAr), 7.21–7.60 (m, 14H, CHAr), 8.92 (d, 2H, J = 5.6 Hz, 2NH). 13C NMR (DMSO-d6, ppm): δC = 29.72 (OCH2CH2), 40.26 (PhCH2S), 58.42 (OCH3), 68.42 (COCCN), 69.67 (NHCHN), 71.83 (OCH2), 112.58 (CAr), 112.92 (CAr), 115.43 (CN), 121.58 (CAr), 122.75 (CAr), 122.51 (CAr), 128.85 (CAr), 129.32 (CAr), 130.37 (CAr), 132.09 (CAr), 138.17 (CAr), 139.64 (CAr), 146.91 (NCAr), 150.76 (CAr), 152.45 (CAr), 172.43 (CO), 178.25 (SC–N–Ph); MS, m/z (%): 941 (M+, 81). Anal. Calcd. for C54H48N6O6S2 (941.13) required C, 68.92; H, 5.14; N, 8.93; S, 6.81; found: C, 69.01; H, 5.20; N, 8.85; S, 6.87.

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DOI 10.1002/jhet

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DOI 10.1002/jhet