Microsoft Word - Hrasna_et_al 2012.doc Nova Biotechnologica et Chimica 11-1 (2012) 73 DOI 10.2478/v10296-012-0008-x ©University of SS. Cyril and Methodius in Trnava SYNTHESIS, COMPLEX COMPOUNDS AND ANTIMICROBIAL ACTIVITY OF SOME DERIVATIVES OF FURO[3,2-C]PYRIDINE AND THEIR STARTING COMPOUNDS MARTIN HRAŠNA1, EVA ÜRGEOVÁ2, ALŽBETA KRUTOŠÍKOVÁ1 1Department of Chemistry, University of SS. Cyril and Methodius, J. Herdu 2, Trnava, SK-917 01, Slovak Republic (alzbeta.krutosikova@ucm.sk) 2Department of Biotechnology, University of SS. Cyril and Methodius, J. Herdu 2, Trnava, SK-917 01, Slovak Republic Abstract: Some [3,2-c]pyridine derivatives were synthesized. 3-(Furan-2-yl)propenoic acid (1a) was prepared from furan-2-carbaldehyde under the Perkin’s conditions. Obtained acid was converted to the corresponding azide 3, which in turn was cyclized to give furo[3,2-c]pyridin-4(5H)-one (4a). The reaction of pyridone 4a with phosphorus oxychloride rendered the chloroderivative 7a, which was treated in the condition of Suzuki coupling reaction with boronic acid to give 4-phenylfuro[3,2-c]pyridine (8e) and an unexpected product 10. Some title compounds have shown moderate to good antimicrobial activity against tested bacteria Xanthomonas sp., Erwinia amylovora, and filamentous fungi Pyrenophora avenae, Fusarium graminearum. Keywords: furo[3,2-c]pyridines, nucleophilic substitution, coupling reaction, 1H and 13C NMR spectra, biological activity 1. Introduction Compounds with 2-trifluoromethyl group in molecule have interest biological activity. For example 2-(trifluoromethyl)benzimidazoles are known as an important class of compounds due to their wide range of biological activity acting as antiviral, antifungal, antibacterial and anticancer drugs (NAVARRETE-VÁZQUEZ et al., 2006). Studies about the antiparasitic activity of 2-(trifluoromethyl)benzimidazole derivatives have shown high potential as antiprotozoal agents (NAVARRETE- VÁZQUEZ et al., 2001). The main features of these compounds are the application of the 2-trifluoromethyl group in order to enhance solubility and absorption properties and therefore antiparasitic activity (NAVARRETE-VÁZQUEZ et al., 2003). Some 4- substituted furo[3,2-b]pyrrole-5-carbohydrazides bearing the 3- (trifluoromethyl)phenyl group at the C-2 position were prepared and their effect on the chlorophyll content in alga suspensions of Chlorella vulgaris and the inhibition of photosynthetic electron transport in spinach chloroplasts were studied (GAŠPAROVÁ et al., 2008). A variety of fused pyridines have been studied for a long time in the field of the chemistry of heterocyclic compounds (SHERMAN 1996; 2008). Furopyridines are very similar to such skeletons as quinoline and isoquinoline which are present in many Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC 74 Hrašna, M. et al. compounds possessing biological activity. It was reported that some pharmacophores with potential antipsychotic activity contain the thieno- and furo[3,2-c]pyridine ring systems (NEW et al., 1989). By studying (KENNIS et al., 2000) of biological activity of tetrahydrobenzofuropyridines and benzothienopyridines was determined that these two groups of compounds are a part of compounds which show a high affinity in face of subtypes of receptors α1 and α2. This fact incites chemists about more complete explication of the structure and functions of these systems. Biological activity of the copper(II) and cobalt(II) 3-methylsufanylnicotinate complexes with furopyridines against various strains of bacteria and filamentous fungi has been investigated (SEGĽA et al., 2008; 2009). For a long time we have been interested in studying of the synthesis and reactivity of various furo[3,2-c]pyridines (BOBOŠÍK et al., 1995; KRUTOŠÍKOVÁ and SLEZIAK 1996; MOJUMDAR et al., 2005; 2009; GAJDOŠ et al., 2006; BÚDOVÁ et al., 2006; BRADIAKOVÁ et al., 2008; 2009; TARABOVÁ et al., 2010). This type of the fused heterocycles can be readily coordinated to metal centers through N-donor atom. A few from these compounds were used as ligands in the preparation of coordination compounds with transition metals Cu(II), Co(II) and Ni(II). The spectral, magnetic, thermal properties, coordination chemistry and X-ray analysis of these compounds have already been outlined (KRUTOŠÍKOVÁ et al., 2001; MIKLOVIČ et al., 2004; BARAN et al., 2005; TITIŠ et al., 2007; VRÁBEL et al., 2007a; 2007b; 2007c; BOČA and TITIŠ 2008). In the past were published synthesis 2-methyl and 2-aryl substituted furo[3,2- c]pyridine-4(5H)-thiones by reaction of corresponding 2-substituted furo[3,2- c]pyridine-4(5H)-ones with phosphorus pentasulfide (BOBOŠÍK et al., 1995; KRUTOŠÍKOVÁ and SLEZIAK 1996). Methylation of the 2-methylfuro[3,2- c]pyridine-4(5H)-thione and 1-benzofuro[3,2-c]pyridine-1(2H)-thione with methyl iodide afforded 4-methylsufanylfuro[3,2-c]pyridine or 1-methylsulfanyl-1- benzofuro[3,2-c]pyridine but methylation corresponding pyridones gave N-methyl compounds (BOBOŠÍK et al., 1995; TARABOVÁ et al., 2010). The 4-substituted furo[3,2-c]pyridines were prepared by various methods. One method used as the starting compounds of the 4-chlorofuro[3,2-c]pyridines in which the chlorine atom was substituted with several nucleophiles (BOBOŠÍK et al., 1995; BÚDOVÁ et al., 2006; BRADIAKOVÁ et al., 2009). The reaction with sodium alkoxides in appropriate alcohol afforded the corresponding 4-alkoxyfuro[3,2- c]pyridines (BÚDOVÁ et al., 2006). Similarly, utilization of sodium methyltiolate gave methylsufanyl derivatives (BOBOŠÍK et al., 1995). Chloro derivatives were converted to 4-amino-substituted furo[3,2-c]pyridines by nucleophilic substitution chlorine atom with some heterocyclic secondary amines as piperidine, morpholine or pyrrolidine (BÚDOVÁ et al., 2006). The nucleophilic reactions of 1- chloro[1]benzofuro[3,2-c]pyridine with secondary heterocyclic amines proceeded analogously (MOJUMDAR et al., 2009; TARABOVÁ et al., 2010). Suzuki coupling reaction was realized with 1-chloro[1]benzofuro[3,2-c]pyridine and phenylboronic acid or pyridin-3-ylboronic acid in the presence of Pd(PPh3)4 catalyst in dichloromethane 1-phenyl[1]benzofuro[3,2-c]pyridine and 1-(pyridin-3- yl)[1]benzofuro[3,2-c]pyridine were formed (TARABOVÁ et al., 2010). Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC Nova Biotechnologica et Chimica 11-1 (2012) 75 In the paper (BENCKOVÁ and KRUTOŠÍKOVÁ 1999, MOJUMDAR et al., 2009) are described the 4-cyanofuro[3,2-c]pyridine and 1-cyano[1]benzofuro[3,2- c]pyridine derivatives via N-oxides by Reissert-Henze reaction. The alkaline hydrolysis of the cyano derivatives raised the corresponding furo[3,2- c]pyridinecarboxylic acids and their amides were prepared by hydrolysis in acid conditions. The aim of this work was to study the reaction of 4-chlorofuro[3,2-c]pyridine with boronic acid in the condition of Suzuki coupling reaction. The main target of this work were syntheses of the earlier known derivatives of furo[3,2-c]pyridine and their starting compounds for testing their antimicrobial activities. 2. Material and methods 2.1 Chemical, physical methods and instruments Melting points were determined using Kofler hot plate. All solvents were distilled and dried before use. All reagents were commercially available and were used without purification. Elemental analyses were determined using an EAGER 300 at Institute of Inorganic Chemistry, Technology and Materials, STU in Bratislava. IR spectra were taken on a FTIR Nicolet NEXUS 470 spectrophotometer using KBr pellets (0.5 mg in 300 mg KBr) in region 4000 – 400 cm-1 at Institute of Physical Chemistry and Chemical Physics, STU in Bratislava. For interpretation of IR spectra following abbreviations are used s = strong band (a value of transmittance: 0-35%), m = medium band (a value of transmittance: 36-50%) w = weak (a value of transmittance: over 50%). 1H NMR spectra were measured in DMSO-d6 using Varian INOVA 600 (for 1H 599.782 MHz and for 13C 150.830 MHz) spectrometer, at 25 °C, at Institute of Analytical Chemistry, Department of NMR and MS Spectroscopy, STU in Bratislava. Chemical shifts (δ-scale) are quoted in parts per million and following abbreviations are used: s = singlet; d = doublet; coupling constants (J) are given in Hz. 2.2 Phytopathogene microorganism In our work phytopathogene bacteria Xanthomonas sp. CCM 2888 from Czech collection of microorganism of Masaryk University Brno and Erwinia amylovora CPPB A203 obtained from the Collection of Phytopathogenic Bacteria and Referential Antidotes (CPPB) at the Crop Research Institute in Prague-Ruzyně, Czech Republic were used. Bacteria were kept as a stock culture in the refrigerator at the temperature of 8 °C. Isolates of the filamentous fungi Pyrenophora avenae, Fusarium graminearum were provided by the Institute of the plants production from Piešťany as pure cultures. Obtained isolates were reinoculated, and stored at temperature 6 ± 2 °C. 2.3 Testing the biocide effect of synthesized compounds The antimicrobial activity of synthesized compounds was determined in vitro against a variety of phytopathogenic microorganism. Antimicrobial activities were Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC 76 Hrašna, M. et al. tested by the standard plate diffusion method (PIDDOCK, 1989) and zones of inhibition were measured in mm. The biocide effect was compared with the effect of 1.2 % w/v solution of TMTD (tetramethylthiuram disulfide), active substance of commercial pesticides. The solution of compounds (15 μL) in DMSO (respectively methanol) with concentration 200, 100, 50 and 25 mg.L-1 was placed to metal cylinders on the surface of a media inoculated with the tested microorganism. The plates were incubated at 25 °C, and clear zones developed around cylinders indicated the inhibition of microbial growth. The size of the zone of inhibition caused by diffusion of agent into agar is directly related to the degree of susceptibility of an organism. If the tested sample shows microbicidal activity, inhibition zone will appear on agar plate. Tetramethyl tiuram disulfide (TMTD) (1.2% w/w methanol solution) was used as a standard of antimicrobial activity. Tested plates were inoculated with 1 mL of microbial suspension (106–107 CFU.mL-1). Sterile cylinders were placed onto the plates and filled by 15 μL of the solution (DMSO, methanol respectively) of the tested compounds. Plates were incubated at 25 °C for 24 h and 4 d, respectively. The zone inhibition diameter (in millimeters) was recorded. Two parallel tests of the compounds inhibition activity were made. 3. Results and discussion 3.1 Chemistry 3-(Furan-2-yl)propenoic acid (1a) was prepared from furan-2-carbaldehyde under the Perkin’s conditions. The acid 1a was converted to the corresponding azide 3a, which was cyclized by heating in diphenyl ether to furo[3,2-c]pyridine-4(5H)-one (4a). The compound 4a was aromatized with phosphorus oxychloride to chloroderivative 7a which was treated in the condition of Suzuki coupling reaction with boronic acid to give 4-phenylfuro[3,2-c]pyridine (8e) and an unexpected product 4-(furo[3,2- c]pyridine-4-yl)furo[3,2-c]pyridine (10) (Fig. 1). The acids 1b-1d were synthesized by condensation of the appropriate carbaldehyde with malonic acid under Knoevenagel conditions. The compounds 4b-4f were prepared analogously by cyclisation of appropriated azides (SLEZIAK and KRUTOŠÍKOVÁ, 1996; GAJDOŠ et al., 2006; MOJUMDAR et al., 2009) Reaction of 4b and 4d with phosphorus pentasulfide led to corresponding thiones 5a, 5b, which were methylated in PTC conditions giving 6a and 6b (BRADIAKOVÁ et al., 2009; TARABOVÁ et al., 2010). 2- Methyl[1]benzofuro[3,2-c]pyridine-1-one (4e) and 5-metyl-2-[3-(trifluoromethyl)- phenyl]furo[3,2-c]pyridine-4(5H)-one (4f) were obtained by reaction of 4b, 4d with NaH and then methylated with methyl iodide. The compound 4a was aromatized with phosphorus oxychloride to chloroderivative 7a. Analogously were synthesized chloroderivatives 7b-7d and they are described in ref. (SLEZIAK and KRUTOŠÍKOVÁ, 1996; BRADIAKOVÁ et al., 2008; GAJDOŠ et al., 2006) Refluxing of appropriate chloroderivatives 7b and 7d with secondary heterocyclic amines gave 8a-8d (BRADIAKOVÁ et al., 2009; MOJUMDAR et al., 2009). The compounds 8f-8j were prepared according (TARABOVÁ et al., 2010; BRADIAKOVÁ et al., 2009; BENCKOVÁ and KRUTOŠÍKOVÁ 1999). N-Oxides 9a, 9b were prepared as it is described in (BRADIAKOVÁ et al., 2009). Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC Nova Biotechnologica et Chimica 11-1 (2012) 77 Fig. 1. Synthesis and reaction of 4-chlorofuro[3,2-c]pyridine. 4-Phenylfuro[3,2-c]pyridine (8e). The mixture of 4-chlorofuro[3,2-c]pyridine (7a) (0.767 g; 5 mmol), phenyl boronic acid (0.945 g; 8 mmol), water solution of sodium carbonate (2 M, 7.5 mL) and Pd(PPh3)4 (0.345 g, 0.3 mmol) in 1.2- dimethoxyethane (9 mL) was heated at 80 °C for 6 h. Then the reaction mixture was poured into mixture dichloromethane and ice water (1:1). The separate organic layer was washed with water and brine, dried with magnesium sulfate, the solvent was evaporated in vacuum. Then from the residue the product 8e after purification by column chromatography was isolated (silica gel, eluted with CHCl3). Yield 0.2 g, 20.5%, white crystals, m.p. 99-100 °C (methanol); RF = 0.2 (CHCl3). Anal. Calcd. for C13H9NO: C, 79.98; H, 4.65; N, 7.17. Found: C, 80.31; H, 4.55; N, 7.23%. 1H NMR (DMSO-d6), δ: 8.56 (d, 1H, 3J(7,6) = 5.8 Hz, H-7), 8.19 (d, 1H, 3J(2,3) = 1.77 Hz, H-2), 8.01 (d, 2H, 3J(2´,3´) = 7.8 Hz, H-2´, H-6´), 7.67 (d 1H, H-6), 7.55 (dd, 2H, H-3´, H-5´), 7.49 (d 2H, 3J(4´,3´) = 7.2 Hz, H-4´), 7.32 (d, 1H, H-3). 13C NMR (DMSO-d6), δ: 159.75 (C-7a), 151.88 (C-4), 147.2 (C-2), 144.15 (C-6), 138.61 (C-1´), 128.98 (C-4´), 128.69 (C-3´, C-5´), 128.19 (C-2´, C-6´), 121.44 (C-4´), 106.12 (C-7), 105.58 (C-3). IR / cm-1: 3088s; 3038s; 1602s; 1495m; 1412s; 1260s; 1112m; 1056s; 992w; 818s; 702s; 686m. 4-(Furo[3,2-c]pyridin-4-yl)furo[3,2-c]pyridine (10). Yield 12.7%, m.p. 160-165 °C (methanol). Anal. Calcd. for C13H9NO: C, 71.18; H, 3.41; N, 11.86. Found: C, 71.13; H, 3.78; N, 11.48%. 1H NMR (DMSO-d6), δ: 8.68 (d, 2H, 3J(6,7) = 6.0 Hz, H-6, H-6´), 8.21 (d, 2H, 3J(2,3) = 2.4 Hz, H-2, H-2´), 7.86 (dd 2H, 3J(3,2) = 2.4 Hz, 5J(3,7) = 0.6 Hz, H-3, H-3´), 7.78 (dd 2H, 3J(7,6) = 6.0 Hz, 5J(7,3) = 0.6 Hz, H-7, H-7´). 13C NMR (DMSO-d6), δ: 160.0 (C-7a, C-7´a), 150.5 (C-4, C-4´), 147.2 (C-2, C-2´), 143.7 (C-6, C-6´), 122.8 (C-3a, C-3´a), 107.9 (C-3, C-3´), 107.4 (C-7, C-7´). IR/ (cm-1): 3472m; 3453m; 3173w; 3164w; 3140m; 1597m; 1567w; 1532w; 1443m; 1400m; 1317w; 1299w; 1263w; 1194w; 1160w; 1121w; 1111w; 1043w; 1013s; 903w; 879m; 816m; 794w; 783m; 749s; 625w; 607w; 590w; 466w; 425w. Suzuki coupling reaction was realized with 4-chlorofuro[3,2-c]pyridine (7a) and phenylboronic acid in the presence of Pd(PPh3)4 catalyst in dichloromethane, in which 4-phenylfuro[3,2-c]pyridine (8e) and 4-(furo[3,2-c]pyridine-4-yl)furo[3,2-c]pyridine (10) were formed (Fig. 1). The compounds 8e and 10 were purified on a silica gel column. Their structures were determined by 1H and 13C NMR spectra. The low yield of 8e (20.2%) comparing with 1-phenyl[1]benzofuro[3,2-c]pyridine (8f) (59%) O CH=CH-CO2HO CH=O CH3CO2Na ClCO2Et NaN3 O CH=CH-CON3 Δ O NH O POCl3 O N Cl PhB(OH)2 Pd(PPh3)4O N N O + Ph2O (CH3CO)2O 1a 3 4a 7a 8e10 O N 1 2 3 4 5 6 7 1´ 2´ 3´ 4´ 1 2 3 3a 3a4 6 7 7a7a 5 1´ 2´ 3´ 4´ 5´ 6´ 7´ 3´a 7´a Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC 78 Hrašna, M. et al. (TARABOVÁ et al., 2010) can be explained by the higher [1]benzofuro[3,2- c]pyridine system stability. The mechanism of the compound 10 formation is for us until now confused. 3.2 Antimicrobial activity The structures whose biological activity was studied are presented in Fig. 2. Fig. 2. Structures of compounds, in which biological activity was studied. Antimicrobial activity of tested compounds 1a-d (characterized by MIC) is summarized in Table 1. It was noticed that the tested compounds 1 inhibited growth of microorganism with the lowest concentration 25 mg.L-1, except the compound 1d, in O N O O NH S O R1 R R2 CO2H R1 R Compound R R1 1a H H 1b 3-CF3C6H4 H 1c CH3 H 1d CH=CH-CH=CH O CO2 - R1 R 2 Cu O CO2 - R1 R 2 Co Compound R R1 2a H H 2b CH=CH-CH=CH H 2c CH3 H Compound R R1 2d H H 2e CH=CH-CH=CH H 2f CH3 H O CON3 3a Compound R R1 R2 4a H H H 4b 3-CF3C6H4 H H 4c 4-NO2C6H4 H H 4d CH=CH-CH=CH H 4e 3-CF3C6H4 H CH3 4f CH=CH-CH=CH CH3 R R1 Compound R R1 5a 3-CF3C6H4 H 5b CH=CH-CH=CH O N SCH3 R R1 Compound R R1 6a 3-CF3C6H4 H 6b CH=CH-CH=CH Compound R R1 7a H H 7b 3-CF3C6H4 H 7c 4-CH3OC6H4 H 7d CH=CH-CH=CH O N Cl R R1 O R N R1 R 2 Compound R R1 R2 8a 3-CF3C6H4 H piperidin-1-yl 8b 3-CF3C6H4 H morpholin-4-yl 8c CH=CH-CH=CH piperidin-1-yl 8d CH=CH-CH=CH morpholin-4-yl 8e H H phenyl 8f CH=CH-CH=CH phenyl 8g H H CO2H 8h H H CN 8i H H CONH2 8j 3-CF3C6H4 H CO2H O N O R Compound R 9a H 9b 3-CF3C6H4 Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC Nova Biotechnologica et Chimica 11-1 (2012) 79 which the antifungal effect was repressed. Substitution on a furan ring did not influence on antifungal and antibacterial effect. Table 1. Antimicrobial activity of tested compounds 1a-1d characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 1a 3.3 ± 1.3 25 1,6 ± 0.1 25 3.3 ± 0.0 25 1.5 ± 0.0 25 1b 3.3 ± 2.3 25 1,6 ± 0.4 25 1.0 ± 0.0 25 3.0 ± 1.3 25 1c 2.8 ± 0.3 25 2.0 ± 0.5 25 2.4 ± 0.1 25 2.0 ± 0.5 25 1d 2.8 ± 1.0 25 1.0 ± 0.0 25 0 N/A 3. 0 ± 0.0 25 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration, N/A compound does not show antimicrobial effect Antimicrobial activities of tested compounds 2a-2f are summarized in Table 2. In comparison, the effect of some of this compounds is lower (MIC 50-200 mg.L-1) than tested compounds of group 1a-1d. The complexes of copper(II) and cobalt(II) shown similar antibacterial effects but the antifungal effects are various. In comparison, copper(II) complexes 2a-2c are more effective on F. graminearum than cobalt(II) complexes. Cobalt(II) complexes 2d-2f are more effective on P. avenae. Sensitivity of microorganism on compounds 2a-2f decreased in the order: E. amylovora > Xanthomonas sp. > P. avenae > F. graminearum. Table 2. Antimicrobial activity of tested compounds 2a-2f characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas. sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 2a 1.5 ± 0.5 25 4.5 ± 1.5 25 1.0 ± 0.0 25 1.0 ± 0.0 50 2b 1.0 ± 0.0 25 1.0 ± 0.0 25 1.3 ± 0.0 50 1.5 ± 0.0 25 2c 1.3 ± 0.0 25 4.3 ± 0.0 25 4.3 ± 0.0 25 2.5 ± 0.0 100 2d 5.3 ± 0.0 200 2.0 ± 0.0 25 0 N/A 4.5 ± 0.0 25 2e 2.0 ± 1.0 25 1.3 ± 0.3 25 0 N/A 2.0 ± 0.0 25 2f 2.0 ± 1.0 25 2.8 ± 0.8 25 3.0 ± 0.0 25 1.5 ± 0.5 25 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration, N/A compound does not show antimicrobial effect The azide 3a inhibited the growth of bacteria Xanthomonas sp. and E. amylovora (MIC 25 mg.L-1). The same effect was registered on filamentous fungi F. graminearum and P. avenae with MIC 25 mg.L-1. In comparison, all of tested Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC 80 Hrašna, M. et al. microorganisms are more sensitive to azide 3a than to previous group of complexes 2a-2f. Table 3. Antimicrobial activity of 3a characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MICb Ø a MIC Ø a MIC Ø a MIC 3a 1.5 ± 0.3 25 2.3 ± 1.0 25 2.3 ± 0.4 25 1.0 ± 0.0 25 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration From the results we can assumed, that the derivatives of 3-(furan-2 yl)propenoic acid have potential in the development of the new pesticides, effective on bacterial as well as fungal affections of the plants. Antimicrobial activities of compounds 4a-4f are summarized in Table 4. In comparison, these compounds are less sensitive to filamentous fungi than in previous groups of compounds 1-3. Inhibition of the growth of filamentous fungi P. avenae is registered only by pyridones 4d and 4e. The rest of microorganisms react sensitively on pyridones 4a-4f, except pyridone 4b (Xanthomonas sp., MIC 100 mg.L-1), 4c (E. amylovora, MIC 100 mg.L-1). Pyridone 4e has not antifungal effect on F. graminearum. Table 4. Antimicrobial activity of pyridones 4a-4f characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 4a 3. 0 ± 0.0 25 3.0 ± 0.0 25 1.0 ± 0.0 25 0 N/A 4b 2.0 ± 0.0 100 1.0 ± 0.0 25 2.5 ± 0.2 25 0 N/A 4c 1.0 ± 0.0 25 3.0 ± 0.0 100 4.5 ± 0.0 25 0 N/A 4d 4.2 ± 0.8 25 3.8 ± 0.0 25 4.0 ± 0.0 25 2.0 ± 0.0 50 4e 3.0 ± 0.0 25 4.0 ± 0.0 25 0 N/A 1.0 ± 0.0 25 4f 2.5 ± 0.5 25 1.0 ± 0.0 25 2.8 ± 0.8 25 0 N/A TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration, N/A compound does not show antimicrobial effect Antimicrobial activities of pyridine-thiones 5a a 5b are summarized in Table 5. In comparison, pyridine-thiones are more effective on all microorganisms than in groups 2 and 4. Both of these compounds have the same effect on microorganisms Xanthomonas sp., E. amylovora, and P. avenae. F. graminearum reacts sensitively on pyridine-thione 5b (MIC 25 mg.L-1). Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC Nova Biotechnologica et Chimica 11-1 (2012) 81 Table 5. Antimicrobial activity of 5a,b characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 5a 3.0 ± 0.0 25 1.0 ± 0.0 25 3.5 ± 0.0 50 3.5 ± 0.0 25 5b 2.5 ± 0.5 25 2.5 ± 0.0 25 2.3 ± 0.0 25 3.0 ± 0.0 25 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibtion diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration Antimicrobial effects of chloro derivatives 7a-7d are summarized in Table 6. Compounds of this group showed various antimicrobial effects. Filamentous fungi are the most sensitive on chloro derivatives 7a a 7b (MIC 25 mg.L-1). The biggest effect on bacteria Xanthomonas sp. and E. amylovora has chloro derivative 7c (MIC 25 mg.L-1). Table 6. Antimicrobial activity of compounds 7a-7d characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 7a 3.0 ± 0.0 50 2.3 ± 1.3 25 2.5 ± 0.5 25 3.8 ± 0.0 25 7b 2.7 ± 0.0 25 3.7 ± 0.0 50 3.3 ± 0.0 25 3.0 ± 0.0 25 7c 3.0 ± 0.5 25 2.8 ± 0.0 25 2.0 ± 0.0 100 1.5 ± 0.0 100 7d 2.5 ± 0.2 50 1.0 ± 0.0 100 4.0 ± 0.0 100 1.0 ± 0.0 50 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration Antimicrobial activities of 8a-8j are summarized in Table 7. In comparison, this group of compounds 8a-8j has lower antimicrobial effects than previous groups. Compounds 8a and 8f inhibited the growth of bacteria by higher concentrations (MIC 50 mg.L-1) and they are less effective. Compounds 8h and 8i with MIC 25 mg.L-1 have antifungal effect on F. graminearum, the effect of compounds 8c (MIC 100 mg.L-1), 8f (MIC 200 mg.L-1) and 8g (50 mg.L-1) is registered also. Sensitivity of microorganisms by these compounds decreased in the order: E. amylovora > Xanthomonas sp. > P. avenae > F. graminearum. The antimicrobial activities of tested N-oxides 9a, 9b are summarized in Table 8. Compounds 9a and 9b showed the lowest antimicrobial effect of all tested compounds. Inhibition of the growth of bacteria Xanthomonas sp. and E. amylovora is stronger for N-oxide 9b, which inhibits the growth of filamentous fungi P. avenae (MIC 100 mg.L-1) too. However, this compound has not antifungal effect on F. graminearum. Antifungal effect is not shown by N-oxides 9a. N-oxide 9b show bigger antimicrobial effect. Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC 82 Hrašna, M. et al. Table 7. Antimicrobial activity of compounds 8a-8j characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F. graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 8a 3.0 ± 0.0 50 4.0 ± 0.0 50 0 N/A 2.0 ± 0.0 100 8b 4.6 ± 0.2 50 2.5 ± 0.0 25 0 N/A 2.0 ± 0.0 25 8c 3.6 ± 0.0 25 3.5 ± 0,0 25 1.5 ± 0.5 100 4.3 ± 0.0 50 8d 2.2 ± 0.2 25 3.3 ± 0.3 25 0 N/A 3.5 ± 1.5 50 8e 0.3 ± 0.0 25 6.7 ± 0.0 25 0 N/A 4.9 ± 0.9 25 8f 2.7 ± 0.0 50 3.3 ± 0.0 50 10.3 ± 0.0 200 2.0 ± 0.0 25 8g 1.5 ± 0.3 25 2.3 ± 0.0 25 5.0 ± 1.0 50 1.7 ± 0.2 25 8h 1.7 ± 0.2 25 2.3 ± 0.3 25 1.0 ± 0.5 25 1.8 ± 0.3 25 8i 2.7 ± 0.0 25 2.4 ± 0.2 25 3.1 ± 0.4 25 1.8 ± 0.2 25 8j 3.3 ± 0.0 25 2.3 ± 0.0 25 0 N/A 2.0 ± 0.0 100 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration, N/A compound does not show antimicrobial effect Table 8. Antimicrobial activity of tested compounds 9a, 9b characterized by MIC [mg.L-1]. Bacteria Filamentous fungi Xanthomonas sp. E. amylovora F.graminearum P. avenae Compound Øa MIC Ø a MIC Ø a MIC Ø a MIC 9a 2.3 ± 0.8 50 1.0 ± 0.0 200 0 N/A 0 N/A 9b 2.3 ± 1.3 25 1.0 ± 0.0 50 0 N/A 3.0 ± 0.0 100 TMTD 0.8 ± 0.2 100 1.0 ± 0.0 200 4.6 ± 0.1 200 3.4 ± 0.9 200 a zone inhibition diameter expressed in mm over 24 h, 4 d respectively, ± SD MIC minimal inhibition concentration, N/A compound does not show antimicrobial effect. 4. Conclusions In summary, it was found that during study of the reaction of 4-chlorofuro[3,2- c]pyridine with boronic acid in the condition of Suzuki coupling reaction were formed two products. The main expecting product 4-phenylfuro[3,2-c]pyridine and the unexpected 4-(furo[3,2-c]pyridine-4-yl)furo[3,2-c]pyridine. The antimicrobial testing shown that earlier known synthesized derivatives of furo[3,2-c]pyridine and their starting compounds have various potential to inhibit the growth of phytopathogene microorganisms. In general it is shown, that the bacterial strains are more sensitive on these compounds than the fungal strains. Acknowledgements: This work was supported by the grants VEGA 1/233/12. NMR experimental part of this work was facilitated by support of Slovak National Research and Development Program No. 2003SP200280203. The authors are grateful to Prof. A. Gatial for IR spectra and Dr. N. Pronayová for NMR spectra measurements. Bereitgestellt von Slovenská poľnohospodárska knižnica | Heruntergeladen 16.01.20 12:17 UTC Nova Biotechnologica et Chimica 11-1 (2012) 83 References BARAN, P., BOČA, M., BOČA, R., KRUTOŠÍKOVÁ, A., MIKLOVIČ, J., PELIKÁN, J., TITIŠ, J.: Structural characterization, spectral and properties of isothiocyanate nickel(II) complexes with furopyridine derivatives. 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