Scientific paper
Synthesis and Reactions of Some Novel Nicotinonitrile
Derivatives for Anticancer and Antimicrobial Evaluation
Eman R. Kotb,1 M. A. El-Hashash,2 Mowafea A. Salama,1* Hemat S. Kalf1 and Naiera A. M. Abdel Wahed3
1 Photochemistry Department, National Research Centre, Dokki, Cairo, Egypt. 2 Chemistry Department, Faculty of Science, Ain Shams University, Abassia, Cairo, Egypt. 3 Natural and Microbial Products Department, National Research Centre, Dokki, Cairo, Egypt * Corresponding author: mowafsalam@yahoo.com
Received: 13-03-2009
Abstract
New diaryl-3-cyano-1ff-pyridinone derivatives 2a-c were synthesized. They were reacted with phosphorous oxychlori-de to give the chloro derivatives 3b,c. On the other hand, the pyridine derivative 2a was used for the preparation of thie-nopyridine derivative 4a. Further 2b,c were glycosidated with 2,3,4,6-tetra-O-acetyl-a-glucopyranosyl bromide (a-ABG) to afford the corresponding nucleosides 5b,c. Also 2a,b reacted with ethyl chloroacetate to afford the O-ethyl glycolate derivatives 6a,b. Compounds 6a,b upon treatment with hydrazine hydrate, gave the hydrazide derivatives 7a,b which condensed with the appropriate aldehyde to afford the arylmethylene hydrazone derivatives 8a-d. The latter compounds were cyclised with thioglycolic acid or acetic anhydride to afford 9c,d and 10c, respectively. The hydrazone derivatives 11a-c were prepared by reaction of hydrazide derivative 7b with some monosaccharides. The behaviour of compounds 7a,b towards phenyl isothiocyanate has been investigated and gave 12a,b, the latter compounds were condensed with chloroacetic acid to produce 13a,b. Also compound 14b was prepared by the reaction with acetylacetone. Additionally, compounds 7a,b were reacted with aliphatic acids namely, formic and acetic acid to afford compounds 15a-d. Some of the newly prepared products showed potent anticancer and antimicrobial activity.
Keywords: Pyridinones, nicotinonitriles, cyclic and acyclic nucleosides, anticancer and antimicrobial activity.
1. Introduction
Numerous thieno[2,3-b]pyridines have been investigated in relation with their biological and pharmacological activities. Some of them proved to possess antibacte-rial,1,2 antiviral,3 antihypertensive and immunostimulating activities.4'5 Others are useful as gonadotropin releasing hormone antigonists6-12 and as lipoxygenases inhibitors.13 Recently, certain thieno[2,3-b]pyridine derivatives were prepared as antiinflammatory agents, particularly for treating arthritis and as bone-resorption agents.14 Pyridothie-nopyrimidine derivatives have found applications as analgesics15 and antipyretics.16 In view of all these facts and as the continuation of our work on the synthesis of new heterocyclic derivatives,1718 we undertook the synthesis and investigated reactions of some new pyridine derivatives,
which might have good biological and medicinal applications.
2. Results and Discussion
Enones are excellent starting materials for the synthesis of pyridine derivatives via the reaction of enones 1 with ethyl cyanoacetate in the presence of ammonium acetate.1920 This procedure is time consuming and the reported yield does not exceed 30% (scheme 1). Here, a method for one step synthesis of the new naphthyl-2(1^)-pyridinone 2 is carried out;21 the method is easy to perform and uses almost all of the available starting materials to give product in high yield. Thus, heating an equimolar mixture of 2-acetylnaphthalene as ketone part and aroma-
tic aldehyde (4-chlorobenzaldehyde, 2-thiophenecarbo-xaldehyde or 3-methoxybenzaldehyde) with ethyl cyanoa-cetate in the presence of ammonium acetate afforded ni-cotinonitrile derivatives 2a,20 b,c, respectively. The formation of compounds 2a-c by this method takes place in high yields (around 80%). The elemental analyses and spectroscopic data are consistent with the assigned structures. IR spectrum of 2c exhibited absorption bands at 3200, 2217 and 1651 cm-1 due to NH (keto-enol),22 CN and C=O groups, respectively. 1H NMR spectrum of compound 2b showed signals at 5 7.12 (s, 1H, pyridine 5-H), 7.34 (d, J = 5.4 Hz, 1H, thiophene H), 7.63-7.66, 8.00-8.12 (2 X m, 8H, Ar-H), 8.55 (s, 1H Ar-H) and 12.85 (s, 1H, NH, D20 exchangeable). The mass spectrum of compound 2b showed molecular ion peak, as the base peak, at m/z 328 (100%).
The formation of compounds 2 possibly takes place as shown in Scheme 1, where the aldehyde condenses with the more reactive methylene group in ethyl cyanoa-cetate, rather than with the less reactive methyl group in 2-acetylnaphthalene. The Michael addition of the 2-acetyl-naphthalene on the produced cyanoacrylate takes place, followed by the replacement of enolic OH by NH2, then cyclization takes place with elimination of ethanol and finally dehydrogenation to produce compounds 2a-c.
Compounds 2 have been utilized as key starting materials in the synthesis of many novel interesting hete-
rocyclic compounds (Scheme 2), thus compounds 2b,c reacted with phosphorous oxychloride to afford the corresponding chloro substituted nicotinonitriles 3b,c. The structures of compounds 3b,c were confirmed with mi-croanalytical and spectroscopic data. The IR spectrum of the chloro derivative 3b revealed the disappearance of the carbonyl band at 1651 cm-1 and the presence of absorption bands at 2223 (CN) and 1067 cm-1 (strong, C-Cl aryl). The mass spectrum of compound 3b showed molecular ion peak (C20H11ClN2S) at m/z 346 as the base peak.
When compound 2a was treated with phosphorous pentasulfide, it afforded the corresponding pyridinethione derivative 4a. The structure of the latter compound was in agreement with spectral data (elemental analysis), besides showing absorption bands in the IR spectrum at 3428 (NH), 2213 (CN) and 1405 cm-1 (C=S). Mass spectrum showed molecular ion peak (C22H13ClN2S) at m/z 372.5 as the base peak.
It is well know that cyclic and acyclic nucleosides often enhance the biological activity of heterocyclic deri-vatives.23,24 Thus, when compounds 2 were glycosidated with 2,3,4,6-tetra-O-acetyl-a-glucopyranosyl bromide (a-ABG) in the presence of dimethylformamide, ß-glu-copyranosyl derivatives 5b,5c were isolated as shown by TLC analysis (Scheme 2). The structures of the new products were established according to their microanalytical and spectroscopic data. The IR spectrum of compound 5b
revealed the disappearance of NH group as well as the existence of C=O groups at 1747, 1658 and CN at 2222 cm-1. 1H NMR spectrum of compound 5c showed signals at 2.08, 2.09, 2.10, 2.10 (4 x s, 12H, 4 x CH3CO), 3.50 (s, 3H, 0CH3), 5.23 (m, 2H, 6'-CH2), 4.25 (m, 2H, 5'-H, 4'-H), 5.28-5.48 (m, 2H, 3'-H, 2'-H), 6.03 (d, J1,2, = 9.0 Hz, 1H, 1'-H), 7.24 (s, 1H, pyridine 5-H), ,^.39-7.65, 8.01-8.40 (2 x m, 10H, Ar-H) and 8.54 (s, 1H Ar-H). The mass spectrum of compound 5c showed molecular ion peak (C37H34N2011) at m/z 352 (M+ - C14H19O9, sugar molecule, 100%) and 331 (M+ - C23H16N202, ^0%%).
When compounds 2a,b reacted with ethyl chloroa-cetate in dry acetone, in the presence of anhydrous potassium carbonate, they produced ethyl ester derivatives 6a,b (Scheme 2), that gave correct elemental analysis besides displaying the expected carbonyl absorption bands in the IR spectrum. Compound 6b showed absorption bands at
2215 (CN), 1739 (C=0 of ester) and 1585 cm-1 (C=N). Its 1H NMR spectrum showed signals at 1.18-1.23 (t, J = 7.5 Hz, 3H, CH3), 4.14-4.21 (q, J = 7.5 Hz, 2H, 0CH2CH3), 5.13 (s, 2H, 0CH2C00), 7.24 (s, 1H, pyridine 5-H), 7.36 (d, J = 5.4 Hz, 1H, thiophene H), 7.55-8.12, 8.21-8.36 (2 x m, 8H, Ar-H) and 8.48 (s, 1H Ar-H). Mass spectrum of compound 6b showed molecular ion peak (C24H18N203S) at m/z 414, as the base peak.
Compounds 6a,b reacted with hydrazine hydrate in ethanol to produce the hydrazide derivatives 7a,b (Scheme 2). IR spectrum of compound 7a displayed absorption bands near 3250, 3160 (NH2, NH), 2220 (CN) and 1675 cm-1 (C=0). 1H NMR spectrum of compound 7b showed signals at 4.22 (s, 2H, NH2, D20 exchangeable), 5.12 (s, 2H, 0CH2), 7.33 (s, 1H, pyridine 5-H), 7.62 (d, J = 5.4 Hz, 1H, tliiophene H), 7.90-8.25 (m, 7H, Ar-H), 8.35 (m, 1H, thiophene H), 8.91 (s, 1H Ar-H) and 9.63 (s, 1H, NH,
D20 exchangeable). Its mass spectrum showed molecular ion peak at m/z 400 (25%) as well as the base peak at m/z 328 (M+-CH-C0NHNH2).
Condensation of compounds 7a,b with 4-chloroben-zaldehyde or 4-hydroxybenzaldehyde took place by heating under reflux in ethanol in the presence of piperidine, where substituted hydrazides 8a-d were produced, which in turn underwent cyclocondensation with thioglycolic acid in dry benzene, according to the reported method, to obtain thiazolidine derivatives 9c,d (Scheme 3).25 Compound 8c was refluxed in excess acetic anhydride for 3 h to give oxadiazole 10c (Scheme 3).
The IR spectrum of compound 8d displayed absorption bands near 3186 (OH), 3054 (NH), 2215 (CN) and 1669 cm-1 (C=O). Compounds 9c,d displayed absorption bands near 3225 (NH), 2220 (CN) and 1730, 1685 cm-1 (2C=O). 1H NMR spectrum of compound 8b showed signals at 5.12 (s, 2H, OCH2), 7.75-8.55, 8.60-8.89 (2 x m, 17H, Ar-H, pyridine 5-H, benzylidine H), 10.12 and 10.40 (2 x s, 2H, NH, OH, D2O exchangeable). The mass spectrum of 8d showed molecular ion peak (C29H20N4O3S) at m/z 504 (32%) and the base peak at m/z 3^7 (100%) corresponding to the loss of C9H8N2O2. That of compound 9d showed molecular ion peak (C31H22N4O4S2) at m/z 578.
Ar'
CN
Ar'

O
RCOOH
NH.NH,
Schema 4
7a, b
The hydrazone derivatives 11a-c were prepared by the reaction of hydrazide derivative 7b with some monosaccharides (D-glucose, D-galactose and arabinose), in et-hanol and glacial acetic acid (Scheme 3). The products revealed absorption frequencies due to OH, NH and C=N in IR spectra, and their 1H NMR spectra showed the presence of the sugar protons, NH and azo-methine (CH=N). The structures of nucleoside derivatives 11a-c were elucidated on the basis of their elemental and spectral data. The IR spectrum of compound 11a as displayed absorption bands at 3442-3320 (broad OH, NH), 2215 (CN) and 1676 cm-1 (C=O). The 1H NMR spectrum of compound 11a showed signals at 5.00 (s, 2H, OCH2), 3.00-3.80 (m, 4H), 3.50-3.90 (m, 2H, CH2OH), 4.30-4.50, 5.30-5.80 (2 X m, 5H, 5 X OH, D2O exchangeable), 7.30 (s, 1H, pyridine 5-H), 7.64 (d, J = 5.4 Hz, 1H, thiophene H), 7.93-8.55 (m, 8H, Ar-H, HC=N), 8.88 (m, 1H, thiophene H), 9.10 (s, 1H, Ar-H) and 10.00 (s, 1H, NH, D2O exchangeable). The IR spectrum of compound 11b displayed absorption bands at 3400-3300 (broad OH, NH), 2215 (CN) and 1680 cm-1 (C=O). The 1H NMR spectrum of compound 11b showed signals at 3.30-3.80 (m, 4H), 4.20-4.60 (m, 5H, 5 X OH, D2O exchangeable), 5.10 (s, 2H, CH2), 5.60-5.80 (m, 2H, 6'-H, 6"-H), 7.30 (s, 1H, pyridine 5-H), 7.65 (d, J = 5.4 Hz, 1H, thiophene H), 7.92-8.55 (m, 9H, Ar-H, HC=N), 9.00 (s, 1H, Ar-H) and 11.40 (s, 1H, NH, D2O exchangeable). The IR spectrum of compound 11c sho2wed absorption bands at 3400-3300 (broad OH, NH), 2215 (CN) and 1676 cm-1 (C=O). The 1H NMR spectrum of compound 11c showed signals at 3.35-3.80 (m, 3H of alditol), 4.40-4.80 (m, 4H, 4 X OH, D2O exchangeable), 5.20 (s, 2H, OCH2), 5.41 (m, 2H, 5'-H-, 5''-H), 7.38 (s, 1H, pyridine 5-H), 7.65 (d, J = 5.4 Hz, 1H, thiophene H), 8.00-8.40 (m, 9H, Ar-H, HC=N), 8.82 (s, 1H, Ar-H) and 11.60 (s, 1H, NH, D2O exchangeable).
When compounds 7a,b reacted with phenyl isot-hiocyanate in dry dioxane hydrazinecarbothioamide derivatives 12a,b (Scheme 3) were obtained,25-27 which showed besides correct values in elemental analysis, IR absorption bands near 3428, 3270 (NH), 2211 (CN), 1652 (C=O) and 1428 cm-1 (C=S). 1H NMR spectrum of compound 12b showed signals at 4.19 (s, 1H, NH, D2O exchangeable), 5.76 (s, 2H, CH2), 7.10 (s, 1H, pyridine 5-H), 7.23 (d, J = 5.4 Hz, 1H, Ar-H), 7.35 (d, J = 5.4 Hz, 1H, Ar-H), 7.58-8.25 (m, 12H, Ar-H), 8.84 (s, 1H, Ar-H) and 12.83 (s, 1H, NH, D-O exchangeable). The mass spectrum of compound 12b showed molecular ion peak (C-9H-0N5O-S-) at m/z 534.
	Ar" 1		
	A	.CN	H
		HN r	-N
Ar			
		15a-d	
	Ar	Ar'	R
	xr		H
^■■ccr		/y'	CH,
-c	xy		H
1:C	rr	-43	

Compounds 12a,b reacted with chloroacetic acid in ethanol to produce thiazolidine derivatives 13a,b (Scheme 3). The IR spectrum of compounds 13a,b displayed absorption bands near 3300, 3200 (NH), 2220 (CN) and 1720, 1685 cm-1 (2C=O). 1H NMR spectrum of compound 13b showed signals at 2.63 (s, 2H, methylene H), 5.35 (s, 2H, OCH-), 7.20 (s, 1H, pyridine 5-H), 7.25-8.36 (m, 14H, Ar-H), 8.80 (s, 1H, Ar-H) and 10.81 (s, 1H, NH, D2O exchangeable), while its mass spectrum showed molecular ion peak (C31H-1N5O3S-) at m/z 575.
Further condensation of the hydrazide derivative 7b with acetylacetone in the presence of ethanol gave pyrazole 14b (Scheme 3). The structure was confirmed by analytical and spectral data. The IR spectrum of compound 14b displayed absorption bands at 3431 (OH), 2213 (CN) and 1665 cm-1 (C=O). 1H NMR spectrum of compound 14b showed signals at 1.74, 2.09 (2 x s, 6H, 2 x CH3), 6.57 (s, 1H, OH, D-O exchangeable), 7.36 (s, 1H, pyridine 5-H), 7.55 (s, 1H, pyrazole H), 7.64-8.33 (m, 10H, Ar-H, C=CO) and 8.75 (s, 1H, Ar-H). From 1H NMR data it seems that compound 14b really exists in keto-enol dynamic equilibrium in ratio 68:32 (17:8 or 2:1). Its mass spectrum showed molecular ion peak (C-7H-QN4O-S) at m/z 464.
Heating compounds 7a,b under reflux with aliphatic acids (formic or acetic acid) resulted in the formation of 1,3,4-oxadiazolidine derivatives 15a-d (Scheme 4). The structures of these compounds were confirmed according to their microanalytical and spectroscopic data. The IR spectrum of compound 15c displayed absorption bands at 3441, 3177 (NH), 2215 cm-1 (CN), those of compound 15d displayed absorption bands at 3190 (NH), 2215 cm-1 (CN). The 1H NMR spectrum of compound 15c showed signals at 5.05 (s, 1H, CH), 5.24, 5.18 (2 x s, 4H, 2 x CH-), 7.24 (s, 1H, pyridine 5-H), 7.38-8.88 (m, 10H, Ar-H), 10.20 and 10.56 (2 x s, 2H, 2 x NH, D-O exchangeable). Its mass spectrum showed molecular ion peak (C-3H18N4O-S) at m/z 414. Those of compound 15d sho-
wed signals at 1.88 (s, 3H, CH3), 5.18 (s, 2H, CH2), 7.38-8.90 (m, 11H, Ar H), 9.95 and 10.32 (2 x s, 2H, 2 x NH, D20 exchangeable). The mass spectrum of compound 15d showed molecular ion peak (C24H20N4O2S) at m/z 429.
3. Anticancer Evaluation
Six selected new compounds 1b, 5b, 5c, 8c, 11c and 12a were tested for cytotoxic activity against the McF7 (Breast Carcinoma Cell Line) and HEPG^ (Liver Carcinoma Cell Line). All new compounds tested were dissolved in DMSO in different concentrations (0, 1, 2.5, 5 and 10 ^g/mL). Preliminary experiments were made using the human tumour cell line to identify the cytotoxicity of the selected compounds according to Skehan et al.28
Material and Method: (i) Tumor (Human tumor cell); (ii) HEPG2 (Liver Carcinoma Cell Line); (iii) McF7 (Breast Carcinoma Cell Line).
Measurement of Potential Cytotoxicity by (SRB) Assay:
-	Cells were plated in 96-multiwell plates (104 cells/well) for 24 h before treatment with the compounds to allow attachment of cells to the wall of the plate.
-	Different concentrations of the tested compound (0, 1, 2.5, 5 and 10 ^g/mL) were added to the cell monolayer; triplicate wells were prepared for each individual dose.
-	Monolayer cells were incubated with the compounds for 48 h at 37 °C and in atmosphere of 5% C02.
-	After 48 h, cells were fixed, washed and stained with Sulfo-Rhodamine-B stain.
-	Excess stain was washed with 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 shown in Tables 1 and 2.
Results.
-	IC50: dose of the compound which reduces survival to 50%.
-	All tested compounds were proven to have cytotoxic activity against the McF7 (Breast Carcinoma Cell Line) and HEPG2 (Liver Carcinoma Cell Line) at the chosen drug concentrations (Tables 1 and 2). Only compound 11c had no cytotoxic activity against the HEPG2 (Table 2).
Table 1: Cytotoxic activity against the McF7 of tested compounds Tested
Compo- Concentration (^g/mL)/ Surviving fraction
Table 2: Cytotoxic activity against the HEPG2 of tested compounds
Tested Compo-
Concentration (^g/mL)/ Surviving fraction
unds	0.0	1.0	2.5	5.0	10.0	IC50
1b	1.000	0.668	0.480	0.468	0.407	2.35
5b	1.000	0.960	0.831	0.412	0.413	4.50
5c	1.000	0.817	0.748	0.560	0.397	6.31
8c	1.000	0.682	0.530	0.273	0.270	3.02
11c	1.000	0.958	0.879	0.580	0.441	7.72
12a	1.000	0.903	0.806	0.662	0.321	7.32
unds	0.0	1.0	2.5	5.0	10.0	IC50
1b	1.000	0.636	0.468	0.352	0.351	2.01
5b	1.000	0.926	0.586	0.406	0.405	3.89
5c	1.000	0.789	0.700	0.476	0.365	4.77
8c	1.000	0.618	0.498	0.362	0.327	2.68
11c	1.000	0.958	0.584	0.541	0.636	-
12a	1.000	0.977	0.685	0.200	0.185	3.42
4. Antimicrobial Evaluation
The target compounds were tested for their antimicrobial activity against Escherichia coli NRRL B-210 (Gram negative bacteria), Bacillus subtilis NRRL B-543 (Gram positive bacteria), Aspergillus niger and Candida albicans NRRL Y-477 (fungi). These microorganisms were obtained from Northern Utilisation Research and Development Division, U. S. Department of Agriculture, Peo-ria, Illionis, USA. Chloramphenicol and Fluconazole were purchased from Egyptian market and used at the concentration of 25 mg/mL as references for antibacterial and antifungal activities. These compounds were assayed by the agar diffusion method.29 The assay medium flasks containing 50 mL of nutrient agar medium for bacteria and Czapek's-Dox agar media for fungi, respectively were allowed to reach 40-50 °C and then inoculated with 0.5 mL of the last organism cell suspension.
The flasks were mixed well and then the content of each flask was poured into a Petri dish (15 x 2 cm) and allowed to solidify. Thereafter, holes (1 cm diameter) were made in the agar plate by the aid of a sterile corkborer. In these holes, 100 pL of the 1.8 mg/mL DMSO of each compound was placed using an automatic micropipette. The Petri dish were left at 5 °C for 1 h to allow diffusion of the samples through the agar medium and retard the growth of the test organism, then incubated at 30 °C for 24 h for bacteria and 72 h of incubation at 28 °C for fungi. The diameter of the resulted inhibition zone was measured in cm.
4. 1. Antibacterial Activity:
DMSO showed no inhibition zones. Result of antibacterial activity test against Escherichia coli (Gram negative bacteria) and Bacillus subtilis (Gram positive bacteria) showed that compounds 5c, 7b, 8c, 10c and 15a of fifteen compounds have an antibacterial activity while the other tested compounds were generally inefficient.
4. 2. Antifungal Activity
The prepared compounds were evaluated in vitro against two strains of fungi, Candida albicans and Asper-gillus niger. Results of antimicrobial activity showed that
compounds 2b and 10c have an antifungal activity while the other tested compounds were generally inefficient.
Table 3: In vitro antimicrobial activity by agar diffusion method of tested compounds
Tested	Inhibition Zone (mm)
Compounds	Microorganism
and Standards	Bacteria	Fungi
Gram- Gramnegative positive Eschericha Bacillus Aspergillus Candid
	coli	subtilsi	niger	albicans
Chloramphenicol	+++	+++	+	+
Fluconazole	-	-	+++	+++
2a	-	-	-	-
2b	-	-	+	+
5b	-	-	-	-
5c	-	++	-	-
6b	-	-	-	-
7a	-	-	-	-
7b	-	++	-	-
8a	-	-	-	-
8b	-	-	-	-
8c	-	++	-	-
9c	-	++	-	-
10c	++	++	+	-
11a	-	++	-	-
11b	-	++	-	-
11c	-	++	-	-
12a	-	-	-	-
14a	-	-	-	-
14b	-	-	-	-
15a	++	++	-	-
15b	-	-	-	-
15d	-	-	-	-
+++ Highly sensitive (inhibition zone 21-25 mm). ++ Fairly sensitive (inhibition zone 16-20 mm). + Slightly sensitive (inhibition zone 10-15 mm). - Not sensitive.
5. Experimental
All melting points are uncorrected and measured using Electro-thermal lA 9100 apparatus (Shimadzu, Tokyo, Japan). IR spectra were recorded as KBr pellets on a Per-kin-Elmer 1650 spectrophotometer (Perkin-Elmer, Nor-walk, CT, USA). 1H NMR was determined on a Jeol-Ex-300 NMR spectrometer (JE0L, Tokyo, Japan) and chemical shifts are expressed as ppm (5 values) against TMS as internal reference. Mass spectra were recorded on VG 2AM-3F mass spectrometer (Thermo electron corporation, USA). Microanalyses were obtained by Mario El Mentar apparatus, 0rganic Microanalysis Unit, and the results were within the accepted range (± 0.20) of the calculated values. Reaction progress and purity checks of the compounds were made by TLC on silica gel-precoated
aluminum sheets (Type 60 F254, Merck, Darmstadt, Germany).
General procedures for the synthesis of compounds 1a-c
A mixture of 2-acetylnaphthalene (0.01 mol) and aromatic aldehydes (4-chlorobenzaldehyde, 2-thiophene-carboxaldehyde or 3-methoxybenzaldehyde (0.01 mol)) in ethanol (30 mL) and 10% Na0H (15 mL) was added drop-wise within 15 min. The reaction mixture was stirred for 3 h and left overnight at room temperature. The mixture was poured onto ice cold water and the obtained precipitate was filtered off and recrystallized from ethanol to give the corresponding derivatives 1a-c, respectively. Compounds 1a,c were identified via their melting points.30,31
1-(2-Naphthyl)-3-(2-thienyl)-2-propen-1-one (1b)
Yield 80%; mp 100-101°C (Et0H). IR (KBr) v 1640 (C=0), 1606 (C=C) cm1. 1H NMR (DMS0-d6) 5 7.21 (d, J = 5.4 Hz, 1H, thiophene H), 7.55-8.30 (m, 10H, Ar-H), 8.88 (s, 1H, Ar-H). MS m/z 264 (M+, 100%). Anal. Calcd for C17H110S: C, 77.24; H, 4.58; S, 12.13. Found: C, 77.18; H, 4.^1; S, 12.16.
General procedures for the synthesis of compounds 2a-c
Method A:
A mixture of 2-acetylnaphthalene (0.01 mol), aromatic aldehydes (4-chlorobenzaldehyde, 2-thiophene-car-boxaldehyde or 3-methoxybenzaldehyde (0.01 mol)), ethyl cyanoacetate (0.01 mol) and ammonium acetate (0.08 mol) in ethanol (40 mL) was refluxed for 8 h. The obtained precipitate was filtered off, washed successively with water and recrystallized from DMF to give the corresponding derivatives 2a-c, respectively.
Method B:
A mixture of enones 1 (0.01 mol), ethyl cyanoaceta-te (0.01 mol) and ammonium acetate (0.08 mol) in ethanol (40 mL) was refluxed for 10 h. The obtained precipitate was filtered off and recrystallized from DMF to give products identical in all aspects with compounds 2a-c obtained before.
4-(4-Chlorophenyl)-6-(2-naphthyl)-2-oxo-1,2-dihy-dropyridine-3-carbonitrile (2a)10
Yield 60% (method A), 40% (method B); mp 340-341 °C (Et0H). IR (KBr) v 3300 (NH), 2210 (CN), 1650 (C=0) cm-1. 1H NMR (DMS0-d6) 5 7.16 (s, 1H, pyridine 5-H), 7.53-7.67 (m, 4H, Ar-H), 7.87-8.54 (m, 4H, Ar-H), 8.68 (d, J = 7.5 Hz, 2H, Ar-H), 8.89 (s, 1H, Ar-H), 12.91 (s, 1H, NH, D10 exchangeable). MS m/z 356 (M+, 100%). Anal. Calcd f(lr C11H13ClN10: C, 74.06; H, 3.67; N, 7.85; Cl, 9.94. Found: C, 773.99; H, 3.66; N, 7.87; Cl, 9.97.
6-(2-Naphthyl)-2-oxo-4-(2-thienyl)-1,2-dihydropyridi-ne-3-carbonitrile (2b)
Yield 60% (method A), 30% (method B); mp 328-329 °C (Et0H). IR (KBr) v 3350 (NH), 2220 (CN),
1660 (C=O) cm-1. 1H NMR (DMSO-d6) 5 7.12 (s, 1H, pyridine 5-H), 7.34 (d, J = 5.4 Hz, 1H, thiophene H), 7.63-7.66 (m, 2H, Ar-H), 8.00-8.12 (m, 6H, Ar-H), 8.55 (s, 1H Ar-H), 12.85 (s, 1H, NH, D2O exchangeable). MS m/z 328 (M+, 100%). Anal. Calcd for C20H12N2OS: C, 73.15; H, 3.68; N, 8.53; S, 9.76. Found: C, 773.00; H, 3.60; N, 8.49; S, 10.03.
4-(3-Methoxyphenyl)-6-(2-naphthyl)-2-oxo-1,2-dihy-dropyridine-3-carbonitrile (2c)
Yield 70% (method A), 35% (method B); mp 280-281 °C (EtOH). IR (KBr) v 3320 (NH), 2218 (CN), 1651 (C=O) cm-1. 1H NMR (DMSO-d6) 5 3.69 (s, 3H, OCH3), 7.11 (s, 1H, pyridine 5-H), 7.3 5-7.63 (m, 4H, Ar-H), 7.92-8.45 (m, 6H, Ar-H), 8.88 (s, 1H, Ar-H), 12.90 (s, 1H, NH, D2O exchangeable). MS m/z 351 (M+-H). Anal. Calcd for C23H1gN2O2: C, 78.39; H, 4.58; N, 7.95. Found: C, 78.47; H, 4.66; N, 7.91.
General procedure for the synthesis of compounds 3b,c.
A suspension of compound 2b or 2c (0.01 mol) and POCl3 (5 mL) was heated on a water bath for 3 h. The reaction mixture was poured gradually into ice-cold water and neutralized by diluted ammonia solution. The separated solid was filtered off and recrystallized from etha-nol/DMF to afford the corresponding derivatives 3b,c.
2-Chloro-6-(2-naphthyl)-4-(2-thienyl)nicotinonitrile (3b)
Yield 61%; mp 170-171 °C (EtOH/DMF). IR (KBr) v 2223 (CN), 1066 (C-Cl) cm-1. 1H NMR (DMSO-d6) 5 7.75-8.12 (m, 4H, Ar-H), 8.21-8.56 (m, 6H, Ar-H), 8.69 (s, 1H, Ar-H). MS m/z 348 (M+, Cl37, 33), 346 (M+, Cl35, 100), 312 (70). Anal. Calcd for C20H11ClN2S: C, 69.26; H, 3.20; Cl, 10.22; N, 8.08; S, 9.2^4. I^ound: C, 69.19; H, 3.22; Cl, 10.24; N, 8.10; S, 9.25.
2-Chloro-4-(3-methoxyphenyl)-6-(2-naphthyl)nicoti-nonitrile (3c)
Yield 64%; mp 169-170 °C (EtOH/DMF). IR (KBr) v 2220 (CN), 1066 (C-Cl) cm-1. 1H NMR (DMSO-d6) 5 3.32 (s, 3H, OCH3), 7.78-8.13 (m, 5H, Ar-H), 8.23-8.70 (m, 6H, Ar-H), 8.^2 (s, 1H, Ar-H). MS m/z 372 (M+, Cl,37 34), 370 (M+, Cl,35100). Anal. Calcd for C23H15ClN2O: C, 74.49; H, 4.08; Cl, 9.56; N, 7.55. Found: C, 74.^8; H, 4.05; Cl, 9.54; N, 7.52.
4-(4-Chlorophenyl)-6-(2-naphthyl)-2-thioxo-1,2-dihy-dropyridine-3-carbonitrile (4a)
A mixture of compound 2a (0.01 mol) and P2S5 (2 g) in pyridine (10 mL) was refluxed for 6 h. The product was poured into ice-cold diluted HCl. The separated solid was filtered off and recrystallized from DMF to afford the corresponding thione derivative 4a. Yield 70%; mp 240241 °C (DMF). IR (KBr) v 3428 (NH), 2213 (CN), 1405 (C=S) cm-1. 1H NMR (DMSO-d6) 5 7.12 (s, 1H, pyridine
5-H),	7.55-7.69 (m, 4H, Ar-H), 76.88-8.64 (m, 4H, Ar-H),
8.67 (d, J = 7.5 Hz, 2H, Ar-H), 8.87 (s, 1H, Ar-H), 12.55 (s, 1H, NH, D2O exchangeable). MS m/z 372 (M+, 100%), 337 (20). Anal. Calcd for C22H13ClN2S: C, 70.87; H, 3.51; Cl, 9.51; N, 7.51; S, 8.60. F^oiind: CC, 70.80; H, 3.53; Cl, 9.55; N, 7.49; S, 8.63
General procedure for the synthesis of compounds 5b,c
To a solution of compound 2b or 2c (0.01 mol) in 1 mL triethylamine, a solution of 2,3,4,6-tetra-O-acetyl-a-glucopyranosyl bromide (0.02 mol) in 5 mL DMF was added, then the reaction mixture was stirred for 3 h at room temperature, evaporated under reduced pressure at 40 °C, the residue washed with distilled water, filtered off and dried to afford compounds 5b,c.
1-(2,3,4,6-Tetra-0-acetyl-ß-glucopyranosyl)-6-(2-naph-thyl)-2-oxo-4-(2-thienyl)pyridine-3-carbonitrile (5b)
Yield 64%; mp 170-171 °C (DMF). IR (KBr) v 2222 (CN), 1658 (C=O), 1747 (C=O) cm-1. 1H NMR (CDCl3) 5 1.87 (s, 3H, CH3CO), 2.08 (s, 3H, CH3CO), 2.10 (s, 3H, CH3CO), 2.10 (s, 3H, CH3CO), 4.11 (m, 2H, 6'-CH2), 4.22-43.30 (m, 2H, 5'-H, 4'-H), 5.23-5.48 (m, 2H, 3'-H, 2'-H), 6.03 (d, J1,2, = 9.0 Hz, 1H, 1'-H), 7.26 (s, 1H, pyridine 5-H), 7.36 (d, ' .1= 5.4 Hz, 1H, thiophene H), 7.59-8.01 (m, 6H, Ar-H), 8.12-8.22 (m, 2H, Ar-H), 8.54 (s, 1H Ar-H). MS m/z 328 (M+ - C14H19O9, 30%). Anal. Calcd for C34H30N2O10S (658): C, 62.00; H, 4.59; N, 4.25; S, 4.87. Found: C, 61.95; H, 4.60; N, 4.30; S, 4.84.
1-(2,3,4,6-Tetra-0-acetyl-ß-glucopyranosyl)-4-(3-met-hoxyphenyl)-6-(2-naphthyl)-2-oxopyridine-3-carboni-trile (5c)
Yield 67%; mp 170-171 °C (DMF). IR (KBr) v 2222 (CN), 1658 (C=O), 1747 (C=O) cm-1. 1H NMR (CDCl3) 5 2.08 (s, 3H, CH3CO), 2.09 (s, 3H, CH3CO), 2.10 (s, 3H, CH3CO), 2.10 (s, 3H, CH3CO), 3.50 (s, 3H, OCH3), 5.23 (m3 2H, 6'-CH2), 4.25 (m, 2H, 5'-H, 4'-H), 5.28-5.48 (m, 2H, 3'-H, 2'-H), 6.03 (d, J1,2, = 9.0 Hz, 1H, 1'-H), 7.24 (s, 1H, pyridine 5-H), 7.3^-7.65 (m, 4H, Ar-H), 8.01-8.40 (m, 6H, Ar-H), 8.54 (s, 1H Ar-H). MS m/z 352 (M+ - C14H19O5, 100%), 331 (M+ - C23H1g N2O2, 90%). Anal. Calcd for C37H34N2O11 (682): C, 65.10; H, 5.02; N, 4.10. Found: C, 65.15; H, 4.95; N, 4.07.
General procedure for the synthesis of compounds 6a,b
A mixture of compound 2a or 2b (0.01 mol), ethyl chloroacetate (0.01 mol), anhydrous potassium carbonate (0.04 mol) in dry acetone (30 mL) was refluxed for 30 h, on a water bath. The solvent was removed under reduced pressure, then water was added to the mixture and the solid formed was filtered off, recrystallized from appropriate solvent to afford the corresponding derivatives 6a,b.
Ethyl {[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}acetate (6a)
Yield 69%; mp 190-191°C (AcOH). IR (KBr) v
2214 (CN), 1743 (C=0) cm-1. 1H NMR (DMSO-d6) 5 1.17-1.21 (t, J = 7.5 Hz, 3H, CH3), 4.15-4.22 (q, J =7.5 Hz, 2H, 0CH2CH3), 5.12 (s, 2H, OCH2COO), 7.27 (s, 1H, pyridine 5-H), 7.55-7.68 (m, 4H, Ar-H), 7.88-8.15 (m, 6H, Ar-H), 8.51 (s, 1H, Ar-H). MS m/z 442 (M+, 100%), 369 (90), 339 (20). Anal. Calcd for C2gH19ClN2O3: C, 70.51; H, 4.32; Cl, 8.00; N, 6.32. Found: C, 70.44; H, 4.54; Cl, 8.08; N, 6.30.
Ethyl {[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyri-dinyl]oxy}acetate (6b).
Yield 70%; mp 160-161°C (EtOH). IR (KBr) v 2214 (CN), 1739 (C=0) cm-1. 1H NMR (DMSO-d6) 5 1.18-1.23 (t, J = 7.5 Hz, 3H, CH3), 4.14-4.21 (q, J == 7.5 Hz, 2H, 0CH2CH3), 5.13 (s, 2H, OCH2COO), 7.24 (s, 1H, pyridine 5-H), 7.3(5 (d, J = 5.4 Hz, 1H, thiophene H), 7.55-8.12 (m, 4H, Ar-H), 8.21-8.36 (m, 4H, Ar-H), 8.48 (s, 1H Ar-H). MS m/z 414 (M+, 100%), 341 (90), 369 (10), 311 (30). Anal. Calcd for C24H18N2O3S: C, 69.55; H, 4.38; N, 6.76; S, 7.74. Found: C, 69.^0; H, 4.30; N, 6.81; S, 7.75.
General procedure for the synthesis of compounds 7a,b
A solution of compound 6a or 6b (0.01 mol) in etha-nol (20 ml) and hydrazine hydrate (0.01 mol) was reflu-xed for 8 h. The separated solid after cooling was recry-stallized from dioxane to afford the corresponding hydra-zides 7a-b.
2-{[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}acetohydrazide (7a)
Yield 50%; mp 210-211°C (dioxane). IR (KBr) v 3250, 3160 (NH2, NH), 2220 (CN), 1675 (C=0) cm-1. 1H NMR (DMSO-dg) 5 4.43 (s, 2H, NH2, D20 exchangeable), 5.11 (s, 2H, OCH2), 7.13 (s, 1H, pyridine 5-H), 7.72 (d, J = 7.5 Hz, 2H, Ar-H), 7.83 (d, J = 7.5 Hz, 2H, Ar-H), 7.95-7.45 (m, 6H, Ar-H), 8.79 (s, 1H, Ar-H), 9.60 (s, 1H, NH, D20 exchangeable). MS m/z 428 (M+, 30%), 356 (100), 328 (40). Anal. Calcd for C24H17ClN4O2: C, 67.21; H, 4.00; Cl, 8.27; N, 13.06. Found: C:, 677.33; H, 3.92; Cl, 8.20; N, 13.05.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyri-dinyl]oxy}acetohydrazide (7b)
Yield 50%; mp 240-241 °C (dioxane). IR (KBr) v 3250, 3160 (NH2, NH), 2221 (CN), 1635 (C=0) cm-1. 1H NMR (DMSO-d6) 5 4.22 (s, 2H, NH2, D20 exchangeable), 5.12 (s, 2H, 0CH2), 7.33 (s, 1H, pyridine 5-H), 7.62 (d, J = 5.4 Hz, 1H, thiophene H), 7.90-8.25 (m, 7H, Ar-H), 8.35 (m, 1H, thiophene H), 8.91 (s, 1H Ar-H), 9.63 (s, 1H, NH, D20 exchangeable). MS m/z 400 (M+, 25%), 369 (80), 3^8 (100), 311 (25). Anal. Calcd for C22H15N4O2S: C, 65.99; H, 4.03; N, 13.99; S, 8.01. Found: C, 66.08; H, 3.99; N, 14.02; S, 7.99.
benzaldehyde or 4-hydroxybenzaldehyde) in ethanol (20 mL) was refluxed for 8 h. The solution was cooled, the precipitate formed was filtered off and recrystallized from the DMF to give compounds 8a-d.
2-{[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}-^'-[(4-chlorophenyl)methylidene]ace-tohydrazide (8a)
Yield 60%; mp 280-281 °C (DMF). IR (KBr) v 3200 (NH), 2217 (CN), 1672 (C=0) cm-1. 1H NMR (DMSO-d6) 5 5.12 (s, 2H, 0CH2), 7.75-8.55 (m, 13H, Ar-H), 8.60-8.89 (m, 4H, Ar-H, pyridine 5-H, benzylidine H), 10.12 (s, 1H, NH, D20 exchangeable). MS m/z 550 (M+, 30%). Anal. Calcd for C31H20Cl2N4O2: C, 67.52; H, 3.66; Cl, 12.86; N, 10.16. Found: C, 677.59; H, 3.61; Cl, 12.84; N, 10.10.
2-{[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}-^'-[(4-hydroxyphenyl)methylidene] acetohydrazide (8b)
Yield 62%; mp 280-281°C (DMF). IR (KBr) v 3200 (NH), 2217 (CN), 1672 (C=0) cm1; 1H NMR (DMSO-d6) 5 5.12 (s, 2H, 0CH2), 7.75-8.55 (m, 13H, Ar-H), 8.60-8.89 (m, 4H, Ar-H, pyridine 5-H, benzylidine H), 10.12 (s, 1H, NH, D20 exchangeable), 10.40 (s, 1H, OH, D20 exchangeable). MS m/z 532 (M+, 35%). Anal. Calcd for C31H21ClN4O3: C, 69.86; H, 3.97; Cl, 6.65; N, 10.51. Found: C, 69.91; H, 3.90; Cl, 6.68; N, 10.48.
^'-[(4-Chlorophenyl)methylidene]-2-{[3-cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl]oxy}acetohydrazi-
de (8c)
Yield 60%; mp 270-271°C (DMF). IR (KBr) v 3208 (NH), 2215 (CN), 1671 (C=0) cm1; 1H NMR (DMSO-d6) 5 5.05 (s, 2H, OCH2), 7.37-8.40 (m, 12H, Ar-H), 8.50-8.80 (m, 4H, Ar-H, pyridine 5-H, benzylidine H), 9.60 (s, 1H, NH, D20 exchangeable). MS m/z 522 (M+, 25%), 327 (100), ^68 (40). Anal. Calcd for C25H19Cl N402S: C, 66.60; H, 3.66; Cl, 6.78; N, 10.71; S, 6.13. Found: C, 66.76; H, 3.59; Cl, 6.75; N, 10.68; S, 6.10.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyri-dinyl]oxy}-^'-[(4-hydroxyphenyl)methylidene]ace-tohydrazide (8d)
Yield 60%; mp 300-301 °C (DMF). IR (KBr) v 3186 (OH), 3054 (NH), 2216 (CN), 1669 (C=0) cm1. 1H NMR (DMSO-d6) 5 5.17 (s, 2H, OCH2), 7.30-8.45 (m, 12H, Ar-H), 8.^0-8.87 (m, 4H, Ar-H, pyridine 5-H, benzylidine H), 9.96 (s, 1H, NH, D20 exchangeable), 10.33 (s, 1H, OH, D20 exchangeable). MS m/z 504 (M+, 32%), 368 (95), 327 (100), 340 (90). Anal. Calcd for C29H20N403S: C, 69.03; H, 4.00; N, 11.10; S, 6.35. Found: C2 68.99; H, 4.00; N, 11.08; S, 6.39.
General procedure for the synthesis of compounds 8a-d
A mixture of compound 7a or 7b (0.01 mol) and equimolar amount of the aromatic aldehydes (4-chloro-
General procedure for the synthesis of compounds 9c,d
A mixture of compound 8a or 8d (0.01 mol) and equimolar amount of the thioglycolic acid (0.01 mol) in
dry benzene (30 mL) was refluxed for 10 h. After evaporation of the solvent under reduced pressure, the obtained product was filtered off and recrystallized from the DMF to give compounds 9c,d.
^-[2-(4-Chlorophenyl)-4-oxo-1,3-thiazolidin-3-yl]-2-{[3-cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl]
oxy}acetamide (9c)
Yield 60%; mp 320-321 °C (DMF). IR (KBr) v 3208 (NH), 2215 (CN), 1665 (C=0), 1671 (C=0) cm1. 1H NMR (DMS0-d6) 5 4.30 (s, 1H, thiazolidine), 5.10 (d, 2H, CH2-thiazolidin6e), 5.60 (s, 2H, CH2), 7.20-7.43 (m, 3H, Ar-^, pyridine 5-H), 7.65-8.35 (m, 11H, Ar-H), 8.90 (s, 1H, Ar-H), 10.80 (s, 1H, NH, Dl0 exchangeable). MS m/z 596 (M+, 66%). Anal. Calcd for C31Hl1ClN403Sl: C, 62.36; H, 3.54; Cl, 5.94; N, 9.38; S, 10.^4. Found: C, 62.29; H, 3.59; Cl, 5.98; N, 9.33; S, 10.71.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl] oxy}-^-[2-(4-hydroxyphenyl)-4-oxo-1,3-thiazolidin-3-yl]acetamide (9d)
Yield 65%; mp 300-301°C (DMF). IR (KBr) v 3186 (0H), 3054 (NH), 2215 (CN), 1665 (C=0), 1671 (C=0) cm1. 1H NMR (DMS0-d6) 5 4.30 (s, 1H, thiazolidine), 5.10 (d, 2H, CHl-thiazolidine), 5.60 (s, 2H, CHl), 7.22-7.44 (m, 3H, Ar-H, pyridine 5-H), 7.65-8.35 (m, 11H, Ar-H), 8.91 (s, 1H, Ar-H), 10.80 (s, 1H, NH, Dl0 exchangeable), 10.33 (s, 1H, 0H, Dl0 exchangeable). M4S m/z 578 (M+, 66%). Anal. Calcd for C31HllN404Sl: C, 64.35; H, 3.83; N, 9.68; S, 11.06. Found: C, 64l.42; H, 3.88; N, 9.64; S, 11.00.
2-{[4-Acetyl-5-(4-chlorophenyl)-4,5-dihydro-1,3,4-oxadiazol-2-yl]methoxy}-6-(2-naphthyl)-4-(2-thienyl) nicotinonitrile (10c)
A mixture of (0.005 mol) of compound 8c and acetic anhydride (20 mL) was refluxed for 3 h. The excess acetic anhydride and acetic acid were removed in vacuo and the solid residue was filtered off, washed with water, dried and re-crystallized from dioxane to give compound 10c. Yield 40%; mp 201-202 °C (dioxane). IR (KBr) v 2215 (CN), 1654 (C=0) cm1. 1H NMR (DMS0-d6) 5 2.15 (s, 3 H, CH3), 5.12 (s, 2H, 0CHl), 5.08 (s, 1H, oxadiazole H), 7.75-8.53 (m, 15H, Ar-H). MS m/z 564 (M+, 60%). Anal. Calcd for C31Hl1ClN403S: C, 65.90; H, 3.75; Cl, 6.27; N, 9.92; S, 5.67. Found: C, (46.00; H, 3.70; Cl, 7.30; N, 9.89; S, 6.62.
General procedure for the synthesis of compounds 11a-c A mixture of compound 7a (0.01 mol) and mono-saccharides (D-glucose, D-galactose or arabinose (0.01 mol)) in ethanol (50 mL) and a catalytic amount of acetic acid was heated at 80 °C for 1 h. The precipitate formed was filtered off hot, washed with ethanol several times and dried to give compounds 11a-c.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl]
oxy}-^'-(glucosylidene)acetohydrazide (11a)
Yield 65%; mp 210-211 °C (Et0H). IR (KBr) v 3442-3320 (broad 0H, NH), 2215 (CN), 1676 (C=0) cm1. 1H NMR (DMS0-d6) 5 5.00 (s, 2H, 0CHl), 3.00-3.80 (m, 4H, protons of alditol congregated with the water signals), 3.50-3.90 (m, 2H, CHl0H), 4.30-4.50 (m, 2H, 2 x 0H, D20 exchangeable), 5.320-5.80 (m, 3H, 3 x 0H, Dl0 exchangeable), 7.30 (s, 1H, pyridine 5-H), 7.64 (d, J = 5.4 Hz, 1H, thiophene H), 7.93-8.55 (m, 8H, Ar-H, HC=N), 8.88 (m, 1H, thiophene H), 9.10 (s, 1H, Ar-H), 10.00 (s, 1H, NH, Dl0 exchangeable). MS m/z 562 (M+, 31%). Anal. Calcd for Cl8HlgN407S: C, 59.78; H, 4.66; N, 9.96; S, 5.70. Found: C, 59.70; H, 4.60; N, 10.02; S, 5.72.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl] oxy}-^'-(galactoylidene)acetohydrazide (11b)
Yield 65%; mp 210-211 °C (Et0H). IR (KBr) v 3400-3300 (broad 0H, NH), 2215 (CN), 1680 (C=0) cm-1. 1H NMR (DMS0-d6) 5 3.30-3.80 (m, 4H, protons of alditol congregated with the water signals), 4.20-4.60 (m, 5H, 5 x 0H, Dl0 exchangeable), 5.10 (s, 2H, CHl), 5.60-5.80 (m, 2H, 6'-H, 6"-H), 7.30 (s, 1H, pyridine 5-H), 7.65 (d, J = 5.4 Hz, 1H, thiophene H), 7.92-8.55 (m, 9H, Ar-H, HC=N), 9.00 (s, 1H, Ar-H), 11.40 (s, 1H, NH, D20 exchangeable). MS m/z 562 (M+, 45%). Anal. Calcd for Cl8HlgN407S: C, 59.78; H, 4.66; N, 9.96; S, 5.70. Found: C, 59.82; H, 4.58; N, 10.01; S, 5.60.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyridinyl] oxy}-^'-(arabinosylidene)acetohydrazide (11c)
Yield 65%; mp 220-221 °C (Et0H). IR (KBr) v 3400-3300 (broad 0H, NH), 2215 (CN), 1676 (C=0). 1H NMR (DMS0-d6): 5 3.35-3.80 (m, 3H of alditol), 4.40-4.80 (m, 4H6, 4 x 0H, D20 exchangeable), 5.20 (s, 2H, 0CHl), 5.41 (m, 2H, 5'-Hl, 5"-H), 7.38 (s, 1H, pyridine 5-H), 7.65 (d, J = 5.4 Hz, 1H, thiophene H), 8.00-8.40 (m, 9H, Ar-H, HC=N), 8.82 (s, 1H, Ar-H), 11.60 (s, 1H, NH, Dl0 exchangeable). MS m/z 532 (M+, 44%). Anal. Calcd for Cl7Hl4N40gS: C, 60.89; H, 4.54; N, 10.52; S, 6.02. Found: C, ^0.88; H, 4.55; N, 10.48; S, 6.08.
General procedure for the synthesis of compounds 12a,b
A mixture of compound 7a or 7b (0.01 mol), phenyl isothiocyanate (0.01 mol) and a catalytic amount of tri-ethyl amine in dry benzene (20 mL) was refluxed for 7 h. The reaction mixture was concentrated and the precipitate formed was filtered off and recrystallized from the DMF to give compounds 12a,b.
2-({[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}acetyl)-N-phenylhydrazinecarbothioa-mide (12a)
Yield 55%; mp 230-231 °C (DMF). IR (KBr) v 3428, 3270 (2NH), 2211 (CN), 1652 (C=0), 1428 (C=S) cm1. 1H NMR (DMS0-d6) 5 4.19 (s, 1H, NH, Dl0 exchangeable), 5.76 (s, 2H, CHl), 7.10 (s, 1H, pyridine 5-H),
7.22	(d, J = 5.4 Hz, 1H, Ar-H), 7.32 (d, J = 5.4 Hz, 1H, Ar-H), 7.55-8.22 (m, 13H, Ar-H), 8.80 (s, 1H, Ar-H), 12.83 (s, 2H, 2 x NH, D2O exchangeable). MS m/z 563 (M+, 56%). Anal. Calcd for C31H22ClN5O2S: C, 66.01; H, 3.93; Cl, 6.29; N, 12.42; S, 5.68. F'ound: (C, 66.15; H, 3.97; Cl, 6.33; N, 12.35; S, 5.60.
2-({[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyri-dinyl]oxy}acetyl)-^-phenylhydrazinecarbothioamide (12b)
Yield 66%; mp 340-341 °C (DMF). IR (KBr) v 3428, 3270 (2NH), 2211 (CN), 1652 (C=O), 1428 (C=S) cm-1. 1H NMR (DMSO-d6) 5 4.19 (s, 1H, NH, D2O exchangeable), 5.76 (s, 2H, CH2), 7.10 (s, 1H, pyridine 5-H),
7.23	(d, J = 5.4 Hz, 1H, Ar-H), 7.35 (d, J = 5.4 Hz, 1H, Ar-H), 7.58-8.25 (m, 12H, Ar-H), 8.84 (s, 1H, Ar-H), 12.83 (s, 1H, NH, D2O exchangeable). MS m/z 535 (M+, 66%), 328 (100), 339 (25). Anal. Calcd for C25H21N5O2S2: C, 65.03; H, 3.95; N, 13.07; S, 11.97. Found: C, 65.14; H, 3.88; N, 13.00; S, 11.99.
General procedure for the synthesis of compounds 13a,b
A mixture of compound 12a or 12b (0.01 mol) and chloroacetic acid (0.01 mol) in ethanol (30 mL) was reflu-xed for 7 h. The reaction mixture was concentrated and the precipitate formed was filtered off and recrystallized from the DMF to give compounds 13a,b.
2-{[4-(4-Chlorophenyl)-3-cyano-6-(2-naphthyl)-2-pyridinyl]oxy}-^-[4-oxo-2-(phenylimino)-1,3-thiazoli-din-3-yl]acetamide (13a)
Yield 66%; mp 250-251 °C (DMF). IR (KBr) v 3300, 3200 (NH), 2220 (CN), 1720, 1685 (2C=O) cm-1. 1H NMR (DMSO-d6) 5 2.57 (s, 2H, methylene H), 5.35 (s, 2H, OCH2), 7.10 (s, 1H, pyridine 5-H), 7.20-8.26 (m, 15H, Ar-H), 8.88 (s, 1H, Ar-H), 10.80 (s, 1H, NH, D2O exchangeable). MS m/z 603 (M+, 66%), 328 (100), 339 (25). Anal. Calcd for C33H22ClN5O3S: C, 65.61; H, 3.67; Cl, 5.87; N, 11.59; S, 5:^1. Found: C, 65.55; H, 3.60; Cl, 5.95; N, 11.62; S, 5.34.
2-{[3-Cyano-6-(2-naphthyl)-4-(2-thienyl)-2-pyri-dinyl]oxy}-^-[4-oxo-2-(phenylimino)-1,3-thiazolidin-
3-yl]acetamide	(13b)
Yield 66%; mp 270-271 °C (DMF). IR (KBr) v 3300, 3200 (NH), 2220 (CN), 1720, 1685 (2C=O) cm-1. 1H NMR (DMSO-dg) 5 2.63 (s, 2H, methylene H), 5.35 (s, 2H, OCH2), 7.20 (s, 1H, pyridine 5-H), 7.25-8.36 (m, 14H, Ar-H), 8.80 (s, 1H, Ar-H), 10.81 (s, 1H, NH D2O exchangeable). MS m/z 575 (M+, 46%), 328 (100), 339 (25). Anal. Calcd for C31H21N5O3S2: C, 64.68; H, 3.68; N, 12.17; S, 11.14. Founc3; C, 64.74; H, 3.70; N, 12.12; S, 11.10.
A mixture of compound 7b (0.01 mol) and acetyl acetone (0.03 mol) in absolute ethanol (20 mL) was reflu-xed for 12 h. The reaction mixture was concentrated and the precipitate formed was filtered off and recrystallized from the DMF to give compound 14b. Yield 66%; mp 190-191 °C (DMF). IR (KBr) v 3431 (OH), 2213 (CN), 1665 (C=O) cm1. 1H NMR (DMSO-dg) 5 1.74 (s, 3H, CH3), 2.09 (s, 3H, CH3), 6.57 (s, 1H, OH, D2O exchangeable), 7.36 (s, 1H, py^ridine 5-H), 7.55 (s, 1H, pyrazole H), 7.64-8.33 (m, 10H, Ar-H, C=CH-O), 8.75 (s, 1H, Ar-H). MS m/z 464 (M+, 46%). Anal. Calcd for C27H20N4O2S: C, 69.81; H, 4.34; N, 12.06; S, 6.89. Found: C2 69.80; H, 4.21; N, 12.00; S, 6.83.
General procedure for the synthesis of compounds 15a-d A solution of compound 7a or 7b (0.01 mol) in acetic acid and/or formic acid (20 mL) was refluxed for 7 h. The precipitate formed was filtered off and recrystallized from the DMF to give compounds 15a-d.
4-(4-Chlorophenyl)-6-(2-naphthyl)-2-(1,3,4-oxadiazo-lidin-2-ylmethoxy)nicotinonitrile (15a)
Yield 66%; mp 260-261 °C (DMF). IR (KBr) v 3167 (NH), 2216 (CN) cm-1. 1H NMR (DMSO-d,) 5 5.06 (s, 1H, CH), 5.23 (s, 2H, CH2), 5.43 (s, 2H, CH2), 7.21 (s, 1H, pyridine 5-H), 7.57-8.88 (m, 11H, Ar-H), 10.20 (s, 1H, NH, D2O exchangeable), 10.57 (s, 1H, NH, D2O exchangeable). MS m/z 442 (M+, 85%). Anal. Calcd ^or C25H19ClN4O2: C, 67.80; H, 4.32; Cl, 8.00; N, 12.65. F(-und: C, ^7.74; H, 4.29; Cl, 8.05; N, 12.71.
4-(4-Chlorophenyl)-2-[(5-methyl-1,3,4-oxadiazolidin-2-yl)methoxy]-6-(2-naphthyl)nicotinonitrile (15b)
Yield 69%; mp 290-291 °C (DMF). IR (KBr) v 3181 (NH), 2217 (CN) cm-1. 1H NMR (DMSO-dg) 5 1.90 (s, 3H, CH3), 5.05 (s, 1H, CH), 5.15 (s, 1H, CH), 5.23 (s, 2H, CH2), 7.23 (s, 1H, pyridine 5-H), 7.37-8.89 (m, 11H, Ar-H), 9.97 (s, 1H, NH D2O exchangeable), 10.35 (s, 1H, NH, D2O exchangeable). MS m/z 456 (M+, 72%). Anal. Calcd ^or C2gH21ClN4O2: C, 68.34; H, 4.63; Cl, 7.76; N, 12.26. Found: C-; 68.;^2; H, 4.58; Cl, 7.80; N, 12.20.
6-(2-Naphthyl)-2-(1,3,4-oxadiazolidin-2-ylmethoxy)-4-(2-thienyl)nicotinonitrile (15c)
Yield 66%; mp 240-241°C (DMF). IR (KBr) v 3177 (NH), 2215 (CN) cm-1. 1H NMR (DMSO-dg) 5 5.05 (s, 1H, CH), 5.24 (s, 2H, CH2), 5.18 (s, 2H, CH2), 7.24 (s, 1H, pyridine 5-H), 7.38-8.88 (m, 10H, Ar-H), 10.20 (s, 1H, NH D2O exchangeable), 10.56 (s, 1H, NH, D2O exchangeable). MS m/z 414 (M+, 85%), 340 (55), 3^5 (50). Anal. Calcd for C23H18N4O2S: C, 66.65; H, 4.38; N, 13.52; S, 7.73. Found: C, 6(5.90; H, 4.16; N, 13.49; S, 7.68.
2-[2-(3,5-Dimethyl-1H-pyrazol-1-yl)-2-oxoethoxy]-6-(2-naphthyl)-4-(2-thienyl)nicotinonitrile (14b)
2-[(5-Methyl-1,3,4-oxadiazolidin-2-yl)methoxy]-6-(2-naphthyl)-4-(2-thienyl)nicotinonitrile (15d)
Yield 69%; mp 280-281 °C (DMF). IR (KBr) v 3190 (NH), 2215 (CN) cm-1. 1H NMR (DMSO-dg) 5 1.88 (s, 3H, CH3), 5.05 (s, 1H, CH), 5.15 (s, 1H, CH), 5.18 (s, 2H, CH-), 7.21 (s, 1H, pyridine 5-H), 7.38-8.90 (m, 10H, Ar-H), SI.95 (s, 1H, NH, D-O exchangeable), 10.32 (s, 1H, NH, D-O exchangeable). MS m/z 428 (M+, 62%). Anal. Calcd for C-4H-0N4O-S: C, 67.27; H, 4.70; N, 13.07; S, 7.47. Found: C, ^7.22; H, 4.73; N, 12.95; S, 7.39.
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Povzetek
Sintetizirali smo nove diaril-3-cia"0-1H-piridi"0"ske derivate 2a-c. Pri reakciji s fosforjevim oksikloridom so iz njih nastali klorovi derivati 3b,c. Piridinski derivat 2a smo uporabili za pripravo tienopiridinskega derivata 4a. Spojini 2b,c smo glikozidirali z -,3,4,6-tetra-O-acetil-a-glukopi^a"ozil bromidom (a-ABG) pri čemer so nastali ustrezni nukleozidi 5b,c. Iz spojin 2a,b sta z etil kloroacetatom nastala O-etil glikolatna derivata 6a,b. Spojini 6a,b sta ob obdelavi s hidra-zin hidratom dali hidrazinska derivata 7a,b ki sta z ustreznim aldehidom kondenzirala do arilmetilenskih hidrazonov 8a-d. Ti so s tioglikolno kislino v acetanhidridu ciklizirali v spojine 9c,d in 10c. Z reakcijo hidrazida 7b z nekaterimi monosaharidi so nastali hidrazonski derivati 11a-c. Raziskali smo tudi obnajanje spojin 7a,b ob prisotnosti fenil izotio-cianata ter tako pripravili spojini 12a,b, ki sta s kloroocetno kislino kondenzirali v spojini 13a,b. Spojino 14b smo pripravili z reakcijo z acetilacetonom. Spojini 7a,b sta reagirali z alifatskima kislinama (mravljična in ocetna kislina) do spojin 15a-d. Nekateri novi produkti so pokazali močno aktivnost proti rakastim celicam in mikrobom.