In view of the facts, and based on in silico studies carried out on thiazolidine-2,4-diones as HIV-1- RT inhibitors, a novel series of 2,4-thiazolidinedione analogs have been designed and synthesized.Structural modifications of thiazolidinediones at 3et al. Antimicrobial activity was determined by two fold serial dilution method. Docking study was performed for the highest active compounds by using Glide 5.0.Title compounds were prepared by the reported method. Conformations of the structures were assigned on the basis of results of different spectral data. The assay of HIV-1 RT was done as reported by Silprasit 24 showed significant HIV-1 RT inhibitory activity with 73% of inhibition with an IC50 value of 1.31 μM. Compound 10 showed highest activity against all the bacterial strains.The newly synthesized compounds were evaluated for their HIV-1 RT inhibitory activity. Among the synthesized compounds, compound 24, 10 and 4 with the non nucleoside inhibitory binding pocket(NNIBP) of RT, active site of GlcN-6-P synthase and cytochrome P450 14-α-sterol demethylase from Candida albicans (Candida P450DM) as the target receptors respectively using the Extra Precision (XP) mode of Glide software.A molecular modeling study was carried out in order to investigate the possible interactions of the highest active compounds N-(phenyl)propanamides (4–31) have been synthesized and evaluated for their HIV-1 RT inhibitory activity, antibacterial and antifungal activities. Some of the compounds have shown significant activity. Molecular docking studies showed very good interaction.A series of novel substituted 2-- The thiazolidinedione scaffold has been identified to play an essential role in medicinal chemistry ,2. CompoHIV is the causative organism for AIDS and is continuously evolving and rapidly spreading throughout the world as a global infection. The HIV infection targets the monocytes expressing surface CD4 receptors and produces profound defects in cell-mediated immunity . OvertimReverse transcription of the single-stranded (+) RNA genome into double-stranded DNA is an essential step in the HIV-1 replication life-cycle and requires the concerted function of both the DNA polymerase and ribonuclease H (RNase H) active sites of HIV-1 reverse transcriptase (RT). Due to its essential role in HIV-1 replication, RT is a major target for anti-HIV drug development and two classes of inhibitors, (1) the nucleoside and nucleotide RT inhibitors and (2) the nonnucleoside RT inhibitors (NNRTIs) have been approved by the United States Food and Drug Administration (FDA) for the treatment of HIV-1 infection . Though With the advent of AIDS and as a result of promiscuous use of drug therapy, antibacterial cytotoxins, steroids, or due to underlying disease or medical manipulation the normal defenses conferred by the microbial flora breaks down resulting in the prevalence of opportunistic bacterial and fungal infections. Bacterial diseases such as tuberculosis, typhus, plague, diphtheria, typhoid fever, cholera, dysentery and pneumonia have taken a high toll on humanity . Along wOpportunistic fungal infections have emerged as important causes of morbidity and mortality in immunocompromised patients and such infections include candidiasis, aspergillosis and mucormycosis . A dramaGlucosamine-6-phosphate synthase , is a new target for antibacterials  and antiRecent modeling studies report that azoles may be acting as antimicrobials by inhibition of GlcN-6-P synthase .The fungal cell wall, a structure essential to fungi and lacking in mammalian cells, is an obvious target for antifungal agents. Its major macromolecular components are chitin, ß-glucan, and mannoproteins . In fungrd and 5th position have exhibited significant biological activities [in silico studies carried out on thiazolidine 2,4-diones as HIV-1- RT inhibitors [2-CH2-CO-NH-) constituting the “body (hydrophilic)” flanked by aryl rings (hydrophobic) linked to the 3rd and 5th position of the thiazolidinedione ring and to that of substituted aromatic amines as the “wings” to enhance the hydrophobicity of the molecules  as eluents. Iodine chamber and Shimadzu (UV-254) spectrometer were used for visualization of TLC spots. Ashless Whatmann No.1 filter paper was used for vacuum filtration. Melting points were determined on an SRS Opti-melting point automatic apparatus and were uncorrected. Elemental data of C, H and N were within ±0.4% of the theortical value as determined by Perkin Elmer Model 240 analyzer. IR spectra  were recorded on SHIAMADZU FT/IR 8400 and were reported in cm.−11 H-NMR and 13C NMR spectra were respectively recorded at 400 and 100 MHz with BRUKER Advance Digital Spectrophotometer. Chemical shifts are expressed in δ-values (ppm) relative to TMS as an internal standard, using DMSO-d6. Chemical shifts are expressed in δ-values (ppm) relative to TMS as an internal standard, using DMSO-d6 and Mass spectra were recorded with a AZILANT Q-TOF Micromass LC-MS by using (ESI+).All reagents were purchased from commercial suppliers like Sigma Aldrich, Merck India Ltd., Himedia and Rankem chemicals. All reagents were GR or AR grade and were used without purification. The purity and homogeneity of the compounds were assessed by the TLC performed on Merck silica gel 60 F4–31 were synthesized as per the reported procedure [(2a-l) (0.01 mol) and the corresponding 3-chloro-N-phenylpropanamides (3a-l) (0.01 mol) were dissolved in 20 ml of acetonitrile. 0.02 mol of triethylamine was added dropwise to this solution with stirring. The reaction mixture was refluxed for 12 h, evaporated in rotary evaporator, cooled and poured into crushed ice and then basified with solid potassium carbonate. The resulting precipitate was filtered, washed with water (3 × 100 ml) and further washed with n-hexane (3 × 20 ml). The solid residue obtained was recrystallized from methanol to yield the desired compounds.Thiazolidine-2,4-dione (1)−1: 3132 (NH stretching), 1741, 1681, 1586 (C = O), 1H-NMR : 12.50 , 4.39 .IR (KBr) cm5-(benzylidene) thiazolidine-2,4-dione (2)−1: 3146 (NH stretching), 3039 (Ar-CH stretching), 2789 (C-CH stretching), 1741, 1693 (C = O stretching).IR (KBr) cm1H-NMR : 9.94 , 8.11 , 8.09-6.91 .2-chloro-N-phenylpropionamide (3)−1: 3138 (NH stretching), 1689 (C = O stretching), 1303 (C-CN stretching), 1H-NMR : 8.60 , 8.12-7.24 , 4.82 ,1.58 .IR (KBr) cm3--N-(3-hydroxyphenyl)propanamide (4)−1: 3353 (OH stretching), 3153 (NH stretching), 1741,1676, 1648 (C = O stretching), 1329 , 1H-NMR : 10.39 , 9.83, 7.80 , 7.51-6.85, 3.93 , 2.66 , 13C NMR(δ)DMSO-d6: 167.0, 166.0, 164.5 (C = O), 149.1 , 140.0 , 136.0 (=CH-), 135.1 , 130.0 , 129.2 , 128.5 , 128.1, 124.3 , 121.5 (C-5 TZD), 119.8 , 117.1 , 41.5 (CH2-CH2), 31.2 (CH2-CH2), MS (ESI+) m/z 369.0 (M+).IR (KBr) cm3-(5-(3-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-phenylpropanamide (5)−1: 3363 (OH stretching), 3153 (NH stretching), 3045, 2962 (Ar-CH stretching), 1676, 1648, 1640 (C = O stretching), 1H-NMR : 10.02 , 9.84 , 7.80 , 7.51-6.85 , 3.93 , 2.63 , MS (ESI+) m/z 369.0 (M+).IR (KBr) cm3-(5-(2-fluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-phenylpropanamide (6)−1: 3313 (NH stretching), 3051, 2929 (Ar-CH stretching), 1753, 1687, 1661 (C = O stretching), 1136 (C-F stretching), 1H-NMR : 10.03 , 7.88 , 7.58-6.98 , 3.93 , 2.66 , MS (ESI+) m/z 371.0 (M+).IR (KBr) cm3-(5-(3-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(3-hydroxyphenyl)propanamide (7)−1: 3525, 3416 (OH stretching), 3232 (NH stretching), 3053, 2947 (Ar-CH stretching),1722, 1664, 1652 (C = O stretching), 1H-NMR : 10.00 , 9.79 , 9.78 , 7.80 , 7.51-6.85 , 3.90 , 2.63 , MS (ESI+) m/z 385.1 (M+).IR(KBr) cm3--N-(2-chlorophenyl)propanamide (8)−1: 3306 (NH stretching), 1743, 1683, 1649 (C = O stretching), 821 (C-Cl stretching), 1H-NMR : 10.02 , 8.01, 7.91 -7.32 , 3.93, 2.63 , MS (ESI+) m/z 386.0 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-p-tolylpropanamide (9)−1: 3386 (NH stretching), 2935, 2852 (C-CH3 stretching), 1747, 1703, 1685 (C = O stretching), 1H-NMR : 10.05 , 8.06 , 8.02-7.20 , 3.33 , 2.66 , 2.30 , 2.24 , MS (ESI+) m/z 395.2 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-hydroxyphenyl)propanamide (10)−1: 3389 (OH stretching), 3198 (NH stretching), 2995, 2885 (C-CH3 stretching) 1668, 1646, 1640 (C = O stretching),1H-NMR : 10.20 , 10.11 , 7.90 , 7.61-7.29 , 3.93 , 2.70 , 2.45 , 2.24 , 13C NMR(δ)DMSO-d6: 166.7, 166.4, 164.5 (C = O), 148.1 , 135.8, 134.7, 133.0 , 132.8 (=CH-), 130.2, 129.0, 127.2, 125.4 , 122.2 , 121.5 (C-5 TZD), 120.2, 119.2 , 116.0 , 121.0, 119.4 (C-CH3), 41.5 (CH2-CH2), 31.2 (CH2-CH2), MS (ESI+) m/z 397.2 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-hydroxyphenyl)propanamide (11)−1: 3404 (OH stretching), 3136 (NH stretching), 2742 (C-CH3 stretching), 1734, 1681, 1641 (C = O stretching), 665 (C-S stretching), 1H-NMR : 10.15 , 10.03 , 8.06 , 8.03 -7.20 , 3.92 , 2.66 , 2.29 , 2.23 , MS (ESI+) m/z 397.2 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-hydroxyphenyl)propanamide (12)−1: 3304 (OH stretching), 3203 (NH stretching), 2980 (C-CH3 stretching), 1658, 1650, 1642 (C = O stretching), 690 (C-S stretching), 1H-NMR : 10.20 , 10.11 , 7.90 , 7.61-7.27 , 3.90 , 2.63 , 2.24-2.27 , MS (ESI+) m/z 397.0 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-hydroxyphenyl)propanamide (13)−1: 3657, 3566, 3412 (OH stretching), 3390 (NH stretching), 3087, 3032 (Ar-CH stretching), 1687, 1669, 1656 (C = O stretching), 1H-NMR : 12.44 , 10.70 , 10.01 , 9.98 , 8.01 , 8.03-7.24 , 3.98 , 2.23 , MS (ESI+) m/z 401.2 (M+).IR (KBr) cm3--N-(2-chloro-4-methylphenyl)propanamide (14)−1: 3281 (NH stretching), 1664, 1650 (C = O stretching), 1329 (C-N aromatic stretching), 783 (C-Cl stretching), 1H-NMR : 10.17 , 7.93 , 7.64 -6.44 , 2.66-3.96 , 2.31 , 2.26 , MS (ESI+) m/z 401.5 (M+).IR (KBr) cm3--N-(4-chloro-3-methylphenyl)propanamide (15)−1: 3283 (NH stretching), 1684, 1654, 1609 (C = O stretching), 1305 (C-N aromatic stretching), 761 (C-Cl stretching), 1H-NMR : 10.19 , 7.93 , 7.93-6.23 , 2.67-3.95 , 2.30 , 2.27 , MS (ESI+) m/z 401.3 (M+).IR (KBr) cm3-(5-(2-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-p-tolylpropanamide (16)−1: 3215 (NH stretching) 1666, 1655, 1645 (C = O stretching), 717 (C-Cl stretching),1H-NMR : 10.02 , 7.88 , 7.58 -7.23 , 3.94 , 2.67 , 2.30 , MS (ESI+) m/z 401.3 (M+).IR (KBr) cm3-2,4-dioxothiazolidin-3-yl)-N-p-tolylpropanamide (17)−1: 3649, 3629, 3595 (OH-stretching), 3312 (NH-stretching), 2869 (C-CH3 stretching), 1745, 1683,1656 (C = O stretching), 1H-NMR : 14.09 , 10.09 , 9.50 , 9.31 , 8.06, 8.04 -7.20 , 3.84 , 2.39 , 2.33 , MS (ESI+) m/z 415.3 (M+).IR(KBr)cm3-(5-(3-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-chloro-5-methylphenyl)- propanamide (18)−1: 3308 (OH stretching), 3223 (NH stretching), 3057, 2960 (Ar-CH stretching), 2924, 2854 (C-CH3 stretching), 1691,1656, 1646 (C = O stretching), 752 (C-Cl stretching), 1H-NMR : 10.19 , 10.11 , 7.93 , 7.64-6.23 , 3.96-2.66 , 2.30 , 2.27 , MS (ESI+) m/z 417.2 (M+).IR (KBr) cm3-(5-(2-fluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-chloro-4-methylphenyl)- propanamide (19)−1: 3306 (NH stretching), 2926, 2856 (C-CH3 stretching), 1743, 1693, 1654 (C = O stretching), 1282 (C-F stretching), 754 (C-Cl stretching), 1H-NMR : 10.02 , 7.88 , 7.58-7.23 , 3.99 , 2.60 , 2.28 , MS (ESI+) m/z 419.2 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-chloro-4-methylphenyl)- propanamide (20)−1: 3267 (NH stretching), 3039 (Ar-CH stretching), 2916, 2858 (C-CH3 stretching), 1681, 1651,1644 (C = O stretching), 742 (C-Cl stretching) 1H-NMR : 10.07 , 8.04 , 7.84-7.20 , 3.92 , 2.66 , 2.30 , 2.24 .IR (KBr) cm3-(5-(2-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-nitrophenyl)propanamide (21)−1: 3145 (NH stretching), 1685, 1657, 1644 (C = O stretching), 1311 (C-NO2 stretching), 776 (C-Cl stretching),1H-NMR : 10.39 , 8.01, 7.91-7.32 , 3.91 , 2.72 , MS (ESI+) m/z 432.8 (M+).IR (KBr) cm3-(5-(2-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-nitrophenyl)propanamide (22)−1: 3306 (NH stretching), 1743, 1693, 1612 (C = O stretching), 1342 (C-NO2 stretching), 754 (C-Cl stretching), 1H-NMR : 10.00 , 7.80 , 7.51-6.85 , 3.90 , 2.63 .IR (KBr) cm3-(5-(4-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-nitrophenyl)propanamide (23)−1: 3267 (NH stretching), 1702, 1664, 1650 (C = O stretching), 1373 (C-NO2 stretching), 752 (C-Cl stretching), 1H-NMR : 10.00 , 7.80 , 7.51 -6.87 , 3.90 , 2.63 .IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(2-mercaptophenyl) propanamide (24)−1: 3649, 3629, 3587(OH stretching), 3312(NH stretching) 2975, 2931 (Ar-CH stretching), 2896 (C-CH3 stretching), 2546 (SH-stretching), 1688, 1647, 1638 (C = O stretching), 1H-NMR : 13.96 , 10.09 , 9.50 , 9.31 , 8.01 , 7.90-7.18 , 3.84 , 3.49 , 2.30 , 13CNMR(δ)DMSO-d6: 167.2, 165.5, 164.5 (C = O), 148.2, 145.2, 142.1, 136.0 , 136.1 (=CH-), 130.2, 129.0, 125.8 , 124.3 , 123.6, 123.0 , 121.5 (C-5 TZD), 110.1, , 109.3 , 41.5 (CH2-CH2), 31.2 (CH2-CH2), MS (ESI+) m/z 432.8 (M+).IR (KBr) cm3--2,4-dioxothiazolidin-3-yl)-N-(3-chloro-2-methylphenyl)propanamide (25)−1: 3630, 3444 (OH stretching), 3273 (NH stretching), 3053(Ar-CH stretching), 2793 (C-CH3 stretching), 1730, 1674, 1648 (C = O stretching), 792 (C-Cl stretching), 1H-NMR : 10.51 , 10.19 , 9.69 , 8.09 , 7.41-7.16 , 3.95 , 2.68 , 2.19 , MS (ESI+) m/z 432.9 (M+).IR (KBr) cm3-(5-(4-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-chloro-4-methylphenyl)propanamide (26)−1: 3198 (NH stretching), 2918, 2848 (C-CH3 stretching), 1750, 1672, 1658 (C = O stretching), 1307 (C-N aromatic stretching), 748 (C-Cl stretching), 1H-NMR : 9.65 , 7.94 , 7.90 -6.53 , 3.94-3.91 , 2.70-2.76 , 2.32 , 13C NMR(δ)DMSO-d6: 167.6, 165.8, 164.5 (C = O), 135.7 (=CH-), 135.6, 133.0, 131.9, 132.0, 131.0 , 130.0  127.8, , 124.2 , 121.5 (C-5 TZD), 113.1, , 112.2, 109.5, 103.7 , 41.5 (CH2-CH2), 31.2 (CH2-CH2), 20.7 (CH3), MS (ESI+) m/z 436.2 (M+).IR (KBr) cm3-(5-(4-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-chloro-5-methylphenyl)propanamide (27)−1: 3273 (NH stretching), 1739, 1670, 1652 (C = O stretching), 808 (C-Cl stretching), 1H-NMR : 9.64 , 7.95 , 7.62 -6.20 , 3.94-3.20 , 2.76-2.32 , 2.23 , MS (ESI+) m/z 435.9 (M+).IR (KBr) cm3-(5-(2-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(2-hydroxyphenyl)propanamide (28)−1: 3411 (OH stretching), 3372 (NH stretching), 1688, 1669, 1646 (C = O stretching), 668 (C-Br stretching), 1H-NMR : 9.97 , 9.84 , 7.80 , 7.51-6.85 , 3.91 , 2.64 , 13CNMR(δ)DMSO-d6: 167.2, 165.2, 164.5 (C = O), 148.9, 138.1, 135.3 (=CH-), 135.1 , 132.3, 130.1, 128.1, 127.1, 127.0 , 121.5 (C-5 TZD), 113.0, , 112.9, 109.5, 104.8 , 41.5 (CH2-CH2), 31.2 (CH2-CH2), MS (ESI+) m/z 448.0.IR (KBr) cm3-(5-(2-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(3-hydroxyphenyl)propanamide (29)−1: 3444 (OH stretching), 3315 (NH stretching), 1752, 1680, 1650 (C = O stretching), 680 (C-Br stretching), 1H-NMR : 9.84 , 9.61 , 7.81 , 7.52-6.86 , 3.91 , 2.64 , MS (ESI+) m/z 448.2 (M+).IR (KBr) cm3-(5-(2-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-nitrophenyl)propanamide (30)−1: 3207 (NH stretching), 1710, 1685, 1658 (C = O stretching), 1319 (C-NO2 stretching), 750 (C-Br stretching), 1H-NMR : 9.73 , 8.12 , 8.09 -7.53 , 3.83 , 2.20 .IR (KBr) cm3-(5-(3-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-chloro-3-methylphenyl) propanamide (31)−1: 3284 (NH stretching), 3014, 2922 (C-CH3 stretching) 1702, 1676, 1652 (C = O stretching), 779 (C-Br Stretching), 675 (C-Br stretching), 1H-NMR : 10.11 , 7.90 , 7.61 -7.29 , 3.90 , 2.64 , 2.45 , MS (ESI+) m/z 480.2 (M+).IR (KBr) cmCompounds rocedure . SubstitStaphylococcus aureus (NCIM 2122), Bacillus subtilis (MTCC 121), Gram-negative bacteria: Escherichia coli (MTCC118), Pseudomonas aeruginosa (MTCC 647), Salmonella typhi (NCIM 2501), Klebsiella pneumonia (MTCC 3384) and fungus Candida albicans (MTCC 227), Aspergillus niger (NCIM 1056). Test compounds were dissolved in 10% DMSO, to produce a 2000 μg/ml stock solution. These test tubes were serially diluted to give a concentration of 100, 50, 25, 12.5, 6.25, 3.125, 1.56, and 0.78 μg/mL. MHB (Mueller-Hinton Broth) was used for bacteria and SDB (Sabouraud Dextrose Broth) was used for fungus. The cell density of each inoculum was adjusted in sterile water of a 0.5 McFarland standard. A final concentration of ~107 CFU/mL and ~106 CFU/mL was obtained for bacteria and fungus, respectively. Microbial inocula were added to the twofold diluted samples. The test tubes were incubated 18–24 h at 37° C ±1°C for bacteria and 2–5 days at 25°C ±1°C for fungus. Ciprofloxacin and fluconazole were used as standard drugs. The highest dilution of the test compound that completely inhibited the growth of test organism was considered as the MIC value of the test compound and was expressed in μg/ml.The standard strains were procured from Institute of Microbial Technology, Chandigarh and National Chemical Laboratory, Pune. Antimicrobial activity was determined by two fold serial dilution method  in dupliet al. [16 primer in a nuclease-free microcentrifuge tube were incubated at room temperature (25°C) for 1 hour to allow the primer/template annealing. The primer/template was prepared by 200-fold dilution in polymerization buffer. The primer/template was aliquoted and kept at −20°C until used. Five microliter of 8 μM stock purified HIV-1 reverse transcriptase was aliquoted and kept at −80°C until used. The working enzyme was diluted to 400 nM with 50 mM Tris–HCl, 20% glycerol, 2 mM DTT, pH 7.5.The assay of HIV-1 RT was done as reported by Silprasit et al. . All reaThe assays were performed in a total volume 15 μL of the polymerization reaction. The reaction containing 3 μl of 400 nM recombinant HIV-1 RT , 2 μL of TE buffer (10 mM Tris–HCl at pH 7.5) were added and gently mixed on ice prior. The polymerization reaction was initiated by the addition of 10 μL of the primer/template and incubated at room temperature for 30 minutes. After the reactions reached the desired incubation time, 5 μL of 0.2 M EDTA was added to stop the polymerization reaction (RTControl). The blank reaction was prepared by mixing 5 μL of 0.2 M EDTA with enzyme before adding primer/template (RTBlank). After termination of the reactions, the plate was gently shaken and incubated at room temperature for 3 min to allow the formation of a stable heteroduplex DNA/RNA complex, followed by addition of 180 μL of PicoGreen reagent diluted 700-fold with TE buffer for each well, making the final volume 200 μL and incubation at room temperature in the dark for 3 min, during which PicoGreen binds to double-stranded DNA and RNA-DNA hybrids, was followed by measurement of fluorescence with a fluorometer .To test the inhibition efficiency of the compounds, all the compounds were dissolved in dimethyl sulfoxide (DMSO) to make a 20 mM stock solution. Ten micromolar working solution of each inhibitor was further diluted by 10 mM Tris–HCl, pH 7.5 containing 50% DMSO. Two microliter of each inhibitor and 3 μL of 400 nM recombinant HIV-1 RT were added and gently mixed on ice prior. The reaction was initiated by the addition of 10 μL of the primer/template and incubated at room temperature for 30 minutes. After the reactions reached the desired incubation time, 5 μL of 0.2 M EDTA was added to stop the polymerization reaction. The relative inhibitory effect of HIV-1 RT activity was compared by using percent inhibition, which was calculated via the following eq   installeThe X-ray crystallographic structure of  was obtained from Brookhaven Protein Data Bank (RCSB) . All watMycobacterium tuberculosis (Mycobacterium P450DM) and fluconazole was present in the Protein Data Bank with the ID 1EA1 [Candida P450DM according to reported method [Crystallographic structure of the complex between cytochrome P450 14-R-sterol demethylase from  ID 1EA1 . The hig ID 1EA1  suggested method . Only 12All the compounds used in the docking study with Glide were built within maestro by using build module of Schrodinger Suite 2008. These structures were geometry optimized by using the Optimized Potentials for Liquid Simulations-2005 (OPLS_2005) force field with the steepest descent protocol followed by truncated Newton conjugate gradient protocol. Partial atomic charges were computed using the OPLS_2005 force field.All docking calculations were then performed using the “Extra Precision” (XP) mode of Glide Program 5.0. A grid was prepared with the center defined by the co-crystallized ligand. During the docking process, initially Glide performed a complete systematic search of the conformational, orientational and positional space of the docked ligand and eliminated unwanted conformations using scoring followed by energy optimization. Finally the conformations were further refined via Monte Carlo sampling of pose conformation. Predicting the binding affinity and rank-ordering ligands in database screens was implemented by modified and expanded version of the Glide scoring function. The most suitable method of evaluating the accuracy of a docking procedure is to determine how closely the lowest energy pose predicted by the scoring function resembles an experimental binding mode as determined by X-ray crystallography. Docking validation was performed with an obtained RMSD value of 0.370 Å for 1RT2, 1.674 Å for 2VF5 and 2.094 Å for 1EA1 ensuring precision and reproducibility of the docking process.rd and 5th position of the thiazolidinedione structure according to Scheme 1). Knoevenagel condensation of 2,4-thiazolidinedione and appropriate aryl aldehydes was carried out in ethanol under reflux conditions containing catalytic amount of piperidine, as base, to form the corresponding substituted 5-benzylidene-2,4-thiazolidinediones (2) [N-chloro-3-(phenylamino)propanamides (3) were prepared by the reported method [N-chloro-3-(phenylamino)propanamides (3) were thus prepared by reacting appropriate aryl amines with 3-chloropropionyl chloride in the presence of glacial acetic acid in cold condition. The substituted 5-benzylidene-2,4-thiazolidinediones (2) were condensed with substituted N-chloro-3-(phenylamino)propanamides (3) in the presence of triethylamine using acetonitrile as the solvent to get substituted 3--N-phenylpropanamides 4–31.An attempt has been made to incorporate aryl groups in the 3ones (2) . N-chlord method . Substit1H-NMR and mass spectral data for all the synthesized compounds are given in the above experimental protocols. The IR spectrum of all the final compounds exhibited very similar features and showed the expected bands for the characteristic groups present in the compounds such as N-H stretching in the range of 3390–3136 cm−1 and C = O stretching in the range of 1750–1645 cm−1 to confirm the presence of thiazolidinedione ring system. 1H NMR showed a characteristic singlet peak assigned to a δ value in the range of 10.39-9.31 thus confirming the presence of NH proton of thiazolidinedione scaffold in compounds 4–31. The methylenic (C = CH) protons of compounds 4–31 were seen as a singlet between 8.12-7.80 δppm while aromatic protons appeared as multiple peaks within the range 8.04-6.20 δ ppm. Characteristic triplet of 2 protons was assigned at δ ranging 3.99-2.66 to the methylenic protons of N-CH2-CH2-CO. Similarly; a triplet of 2 protons was assigned to the methylenic protons of N-CH2-CH2-CO observed at δ ranging 2.76-2.20 for all the compounds 4–31. The 13C NMR depicted the peaks of thiazolidine-2,4-dione (TZD) nucleus within the range δ 166.2-164.3 . 13C NMR spectrum showed signals for thiazolidine-2,4-dione-C-5 atom at δ 121.5.The physical data are given in Table in vitro antibacterial activity against Gram-positive bacteria: Staphylococcus aureus (NCIM 2122), Bacillus subtilis (MTCC 121), Gram-negative bacteria: Escherichia coli (MTCC118), Pseudomonas aeruginosa (MTCC 647), Salmonella typhi (NCIM 2501), Klebsiella pneumonia (MTCC 3384) and fungus Candida albicans (MTCC 227), Aspergillus niger (NCIM 1056), by using the twofold serial dilution technique and the results are summarized in Table All the synthesized compounds were also evaluated for eus NCIM 122, Baci10 showed highest activity against all the bacterial strains. Compound 4 and 10 showed highest activity against E. coli while compound 24 exhibited moderate activity against the same bacterial strain while exhibited weak activity against all the other bacterial strains. Compound 5 and 7 exhibited highest activity against B. subtilis and K. pneumoniae respectively while compound 17 exhibited moderate activity against all the three bacterial strains B. subtilis, E. coli and S. typhi. Compound 8 exhibited moderate activity against S. aureus. However, all these compounds exhibited activity less than that of standard drug ciprofloxacin. Rest of the compounds showed mild to moderate activity against all other bacterial strains.Compound 4 exhibited excellent activity against the fungal strains C. albicans and A. niger. In fact, the activity was higher than that of standard fluconazole. Compound 7, 8, 15, 19, 21, 24 and 27 also exhibited very high activity against both the fungal strains comparable to the standard drug. Compounds 5, 10, 11, 18 and 28 exhibited significant activity against C. albicans while showing moderate activity against A. niger. All other compounds showed moderate to weak antifungal activity against both the fungal strains.Compound 19 and 27. The introduction of two methyl groups in the phenyl ring directly substituted at the 5th position of the thiazolidine −2,4-dione nucleus results in slight decrease in the antifungal activity but increases antibacterial activity as is evident from the activity of compound 10.The introduction of phenyl groups substituted at different positions with electronegative groups such as chloro, hydroxyl and nitro enhances the antimicrobial activity, particularly evident in case of antifungal activity. However, the introduction of one methyl group does not have much influence in the increase or decrease of antifungal activity as in case of compounds 24 showed significant HIV-1 RT inhibitory activity with 73% of inhibition with an IC50 value of 1.31 μM. Compound 23 also showed 58% inhibition, however its IC50 value was negligent. Rest of compounds showed weak activity.The newly synthesized compounds were evaluated for their HIV-1 RT inhibitory activity. Percentage of inhibition has given in Table 24 with hydroxyl groups substituted at 2, 3 and 4 positions of the phenyl ring attached at the 5th position of the thiazolidinedione ring with a linker group CH2CH2CONH at the N-3 position linked to a second phenyl ring substituted with a 2-mercapto group positively influenced the activity.From the results of HIV-1-RT inhibitory activity, it is evident that compound 4–31 exhibited promising antimicrobial activities. In case of HIV-1-RT inhibitory activity, only compound 24 was found to be significantly active while the others exhibited weak non-nucleoside reverse transcriptase activity.According to the above obtained data, we found that compounds in vitro results for the compounds 4–31, a molecular modeling study was carried out in order to investigate the possible interactions of the highest active compounds 24, 10 and 4 with the non nucleoside inhibitory binding pocket(NNIBP) of RT, active site of GlcN-6-P synthase and cytochrome P450 14-α-sterol demethylase from Candida albicans (Candida P450DM) as the target receptors respectively using the Extra Precision (XP) mode of Glide software [With the aim of rationalizing the biological data obtained and considering the best obtained software .24 to analyze the binding mode and key protein ligand interactions and was compared with that of the experimentally determined binding mode and interactions of the bound ligand TNK-651 and the standard efavirenz. The key interactions were mainly hydrogen bonding interactions with Lys103 and Lys101 respectively. The carbonyl oxygen at position 4 in the thiazolidinedione moiety forms a strong H-bond interaction with the NH terminal group of Lys103.Another strong H-bond interaction of the hydroxyl group at the ortho position of the first phenyl ring with one of carbonyl oxygen atoms of Lys101 was observed. The phenyl ring in the 2, 3, 4-trihydroxybenzaldehyde moiety along with the thiazolidinedione moiety was oriented in the bigger hydrophobic pocket formed by Phe227, Pro225 Leu234 Tyr181, Tyr188, Leu100 and Val179 while the CH2CH2CONH linker showed favorable interaction with the amino acid residues Tyr318, Pro236 and Val106. Docking score of the compound 24 (Glide XP score-11.30) was lower than the bound ligand TNK-651(Glide XP score-13.29) but comparable to that of standard efavirenz (Glide XP score-11.33) .Possible interactions for the reference ligand TNK-651, efavirenz and compound 24 have been shown in Figure To validate the Glide software, firstly the interaction between TNK651 and HIV-1 RT was modeled. Superimposition of the experimental bound  conformation of TNK651  and that10 respectively. The binding mode of GlcN-6-P synthase (2VF5) with its bound inhibitor glucosamine-6-Phosphate  has been depicted in Figure 10. The hydroxyl group on the benzylidene moiety forms a hydrogen bond with the carbonyl oxygen of Thr302. The NH in the CH2CH2CONH linker forms a strong hydrogen bond with the carbonyl oxygen of Val399. Another strong H-bond is evident between the carbonyl oxygen of thiazolidinedione with the NH terminal group of Ala602. The 2-hydroxyphenyl group makes favorable interaction with the side chain of Ala 400. The thiazolidinedione ring shows favorably oriented towards the residues Leu601, Cys300, Val605 and Ala353. The linker (CH2CH2CONH) shows favorable interactions with Leu484.The active site of GlcN-6-P synthase PDB code 2VF5) consists of 16 amino acid residues as Glu488, Ser303, Ala602 Ser347, Ser349, Gln348, Thr302, Thr352, Val605, Ala400, Cys300, Val399, Leu601, Leu484, Ser401 and Lys603 as shown in Figure VF5 consiCandida P450DM is a membrane-bound enzyme, it is difficult to crystallize by X-ray analysis; therefore, no experimental data has been available for the structure of this enzyme. However, the crystallographic structure of the complex between cytochrome P450 14-α-sterol demethylase from Mycobacterium tuberculosis (Mycobacterium P450DM) and fluconazole is present in the Protein Data Bank with the ID 1EA1. A perusal of the literature showed that high homology exists between the two analogous enzymes, Candida P450DM and Mycobacterium P450DM [As the target enzyme m P450DM .Candida P450DM complexed with fluconazole was modeled following the procedure of Rosello et al. [A chimeric enzyme of o et al. . Flucona4 (Glide XP score −8.13) has been analysed as follows (Figure 2CH2CONH linker forms a strong coordinate bond with Hem460. The aromatic ring forms π–π stacking interactions with Phe83 residue. The thiazolidinedione ring is oriented in the hydrophobic pocket formed by Tyr181, Phe100, Met255, Val395 and Leu79. Both phenyl rings attached to the thiazolidinedione nucleus makes favorable interactions with the side chains of Leu96, Al173, Tyr76, Cys394, Phe387, Ile324, Ala389 and Leu321.To illustrate the binding mode of the newly synthesized compounds in the active site of chimeric 1EA1, the docked pose of the highest active compound s Figure g. The ke4 with the active site residues and may be responsible for the very high antifungal activity as shown by compound 4.These docking results demonstrate that hydrogen bond interactions, hydrophobic interactions and the coordinate bond with the Hem residue in 1EA1 are very important for binding of compound th position of the thiazolidinedione ring system plays a significant role while in case of antifungal activity the electronegative groups, particularly hydroxyl groups substituted in the various positions of both the phenyl groups are responsible for enhanced antifungal activity. The study encourages us to consider a new molecular skeleton of thiazolidinediones substituted at the 3rd and 5th position by aryl groups with adequate spacers may be identified as a potential lead compound for the development of ant-HIV agents with the ability to combat opportunistic bacterial and fungal infections.In the present study, a series of thiazolidinedione analogs have been synthesized and their structures have been characterized by IR, NMR and mass spectroscopy. All the newly synthesized compounds were tested for HIV-1- RT inhibitory activity by microplate assay method and for antimicrobial activity by two fold serial dilution method. From the modeling studies as well as from the SAR, electronegative groups substituted at various positions of the phenyl rings with a thiazolidinedione scaffold may be responsible for the very high HIV-1-RT inhibitory activity of compound 24. In case of antibacterial activity the methyl groups substituted in the phenyl group attached to the 5


N-acyl protected 2´-deoxy-5´-O-ribonucleosides were phosphorylated with bis(benzotriazol-1-yl) 2-chlorophenyl phosphate. The “outdated” phosphotriester strategy, based on coupling of PV building blocks in conjunction with quantitative precipitation of the oligodeoxyribonucleotide with MeOH is applied. Subsequent release of the resulting phosphate and base-protected oligodeoxyribonucleotide trimer 3’-pTpdCBzpdGibu-5’ as its 3’-(2-chlorophenyl phosphate) was achieved by reductive cleavage of the disulfide bond.The preparation of a disulfide-tethered precipitative soluble support and its use for solution-phase synthesis of trimeric oligodeoxyribonucleotide 3´-(2-chlorophenylphosphate) building blocks is described. To obtain the building blocks,  O-[4-methylphenyl]pentaerythritol support that precipitates quantitatively from MeOH -1H-1,2,3-triazol-1-ylmethylphenyl}pentaerythritol support (3) is outlined in S-propargyl thioacetate was first conjugated to the tetrakis-O-[4-(azidomethylphenyl)pentaerythritol support (1) [O-({4-[4-[(acetylthiomethyl)]-1H-1,2,3-triazol-1-ylmethyl]phenyl}pentaerythritol (2) as a thick oil. Careful aminolysis of the thioacetates with butylamine in degassed methanol under inert atmosphere followed by immediate reaction of the exposed mercapto groups with 2-(pyridine-2-yldisulfanyl)ethanol gave the desired tetrapodal soluble support 3.The synthesis of the disulfide tethered tetrakis-port (1)  by Cu(I)port (1) –25, yielN4-benzoyl-2’-deoxycytidine and N2-isobutyryl-2’-deoxyguanosine into their 3’-(benzotriazol-1-yl 2-chlorophenyl phosphates)  was prepared as described in our previous report -1H-1,2,3-triazol-1-ylmethyl]phenyl}pentaerythritol (3) has been synthesized and used as a tetrapodal soluble support for the synthesis of a fully protected trimeric oligodeoxyribonucleotide as a 3´-(2-chlorophenyl phosphate) by the 1-hydroxybenzotriazole promoted phosphotriester chemistry. The support enabled efficient purification of the support-bound intermediates and product by virtually quantitative precipitation from MeOH after each coupling and 5´-O-deprotection. However, on assembling the trimeric 3´-(2-chlorophenyl phosphate) building blocks, the 5´-terminal nucleoside should be introduced in a 5´-O-(4-methoxytrityl) protected form to prevent detritylation during the reductive cleavage of the trimer from the support.Tetrakis-−1 Et3NHOAc to 70% MeCN in 0.1 mol L−1 Et3NHOAc in 25 min, then continued with 70% MeCN in 0.1 mol L−1 Et3NHOAc to 100% MeCN in 35 min; (B) gradient elution from 25% MeCN in 0.1 mol L−1 Et3NHOAc to 100% MeCN in 25 min, then continued with MeCN; An analytical C-18 RP column  was used. Reactions were monitored by TLC , using short wavelength UV or charring with 10% aq H2SO4 for detection . Mass spectra were recorded with a Bruker Daltonics MicrOTOFQ spectrometer using ESI mode.General: NMR spectra were recorded on a Bruker Avance spectrometer (500 or 400 MHz) at 25 °C. Chemical shifts are given in ppm and referenced relative to the residual solvent signals. RP HPLC conditions: (A) gradient elution from 25% MeCN in 0.1 mol LTetrakis-O-(4-azidomethylphenyl)pentaerythritol (1) was prepared as described previously phenyl}pentaerythritol (2): S-Propargyl thioacetate  was added to the solution of compound 1 , sodium ascorbate  and CuI  in dry DMAc (5.0 mL) in a Pyrex tube. The tube was degassed via three freeze–pump–thaw cycles, and the mixture was stirred at 50 °C for 12 h. Water (10 mL) was added to the reaction mixture and extracted with ethyl acetate (20 mL × 3). The combined organic layer was washed with saturated NaHCO3, dried with Na2SO4, and the solvents were evaporated to dryness. The residue was purified by silica gel chromatography  to give 2  as a white foam. 1H NMR  δ 7.35 , 7.18 , 6.89 , 5.38 , 4.32 , 4.13 , 2.31 ; 13C NMR  δ 195.2, 158.9, 144.6, 129.7, 127.2, 122.1, 115.1, 66.4, 53.6, 44.7, 30.4, 23.9 ppm; ESIMS m/z: [M + H]+ calcd for C53H57N12O8S4, 1117.33, found, 1117.36; m/z: [M + Na]+ calcd for C53H56N12NaO8S4, 1139.31; found, 1139.32.Tetrakis-O-{4-phenyl}pentaerythritol (3). Degassed butylamine in MeOH  was added to the stirred solution of compound 2  in degassed MeOH. The mixture was stirred under N2 for 12 h. Degassed MeCN/DCM  was added and the mixture was neutralized with dry acidic ion exchange resin and filtered off. The solvent was removed by evaporation and the crude product was used in the next reaction without purification. 2-(Pyridine-2-yldisulfanyl)ethanol  was added to the crude compound in degassed MeCN/DCM/MeOH  under N2. The mixture was stirred for 30 min and the progress was monitored by TLC. Solvents were removed by evaporation and the product was purified by silica gel chromatography  to give 3  as a thick oil. 1H NMR  δ 7.40 , 7.18 , 6.88 , 5.42 , 4.30 , 3.97 , 3.79 , 2.73  ppm; 13C NMR  δ 158.9, 144.2, 129.6, 127.2, 121.9, 115.2, 66.4, 59.9, 53.7, 44.8, 42.5, 32.8 ppm; ESIMS m/z: [M + H]+ calcd for C53H65N12O8S8, 1253.28; found, 1253.27; m/z: [M + Na]+ calcd for C53H64N12NaO8S8, 1275.26; found, 1275.26.Bis(benzotriazol-1-yl) 2-chlorophenyl phosphate: The couplings were carried out essentially as described previously + calcd for C58H59N10O22P3, 1445.21; found, 1445.20; m/z: [M + Na]+ calcd for C58H58N10NaO22P3, 1467.19; found, 1467.18.File 11H NMR, 13C NMR and DEPT spectra for 2, and 3, 31P NMR spectra of 10, mass spectra for 2, 3, 7a–10.


Further these synthesized derivatives were subjected to antibacterial activity against all the selected microbial strains in comparison with amoxicillin and cefixime. The antibacterial activity of synthesized derivatives was correlated with their physicochemical and structural properties by QSAR analysis using computer assisted multiple regression analysis and four sound predictive models were generated with good R2, Radj2, and Fischer statistic. The derivatives with potent antibacterial activity were subjected to molecular docking studies to investigate the interactions between the active derivatives and amino acid residues existing in the active site of peptide deformylase to assess their antibacterial potential as peptide deformylase inhibitor. We report the synthesis and biological assessment of 1,3,4-oxadiazole substituted 24 derivatives as novel, potential antibacterial agents. The structures of the newly synthesized derivatives were established by the combined practice of UV, IR,  Oxadiazoles are the heterocyclic compounds containing one oxygen and two nitrogen atoms in a five membered ring , 2 posseResearchers have already reported that gram positive bacteria are much more susceptible to antimicrobial agents as compared to gram negative bacteria . These d2+ mediated catalysis which leads to inhibition of protein synthesis in bacteria . Th. Th 12a −1): 3030.17 (CH), 1627.92 (C=O), 1600.92 (C=N), 1519.91 (C=C), 1219.09 (C–O–Casymm), 1043.49 (C–O–Csymm), 920.05, 779.24, 738.74 (CH); 1H NMR : 8.3 , 7.9 , 7.7 , 7.6 , 7.5 , 7.3 ; 13C NMR (CDCl3): 136.2 (C-6), 128.6 (C-7), 138.3 (C8), 130.1 (C-9), 128.7 (C-10), 130.7 (C-11), 136.5 (C-12), 127.0 , 129.0 , 128.5 (C-15), 187.0 (C-18), 137.8 (C-20), 130.1 , 128.2 , 132.2 (C-23); MS: m/z 326.14, 326.11 (M + 1), 327.11 (M + 2).Yield 72.45%, white crystals, mp (°C): 291.00; IR : 3034.03, 2918.30 (CH), 1683.86 (C=O), 1583.56 (C=N), 1521.84 (C=C), 1257.59 (C–O–Casymm), 1176.58 (CH), 1058.92 (C–O–Csymm), 925.83, 811.78, 756.10 (CH), 545.85 (C–Cl); 1H NMR : 8.2  (J = 7.8 Hz), 7.8 , 7.6 , 7.4 , 7.2 ; 13C NMR (CDCl3): 136.4 (C-6), 128.5 (C-7), 138.7 (C-8), 130.5 (C-9), 128.2 (C-10), 130.9 (C-11), 134.6 (C-12), 128.4 , 129.4 , 133.8 (C-15), 187.2 (C-18), 137.4 (C-21), 130.6 , 128.8 , 131.9 (C-24); MS: m/z 360.07, 362.06 (M + 1), 363.10 (M + 2).Yield 76.60%, yellow powder, mp (°C): 283–285; IR : 3115.04, 2920.23 (CH), 1651.07 (C=O), 1620.21 (C=N), 1571.99 (C=C), 1496.76 (N=Oasymm), 1365.60 (N=Osymm), 1209.37 (C–O–Casymm), 1120.64 (CH), 1047.35 (N=Osymm), 798.53 (CH); 1H NMR : 8.6 , 8.3 , 7.9 , 7.7 , 7.4 ; 13C NMR (CDCl3): 136.0 (C-6), 128.2 (C-7), 138.3 (C-8), 130.9 (C-9), 128.0 (C-10), 130.4 (C-11), 142.6 (C-12), 127.9 , 124.1 , 148.4 (C-15), 186.9 (C-18), 137.9 (C-21), 128.2 , 127.2 , 129.9 (C-24); MS: m/z 371.09, 372.06 (M + 1), 373.10 (M + 2).Yield 75.50%, brown crystals, mp (°C): 261–263; IR : 3000 (CH), 1655.96 (C=O), 1563.337 (C=N), 1521.90 (C=C), 1236.42 (C–O–Casymm), 1111.05 (CH), 1041.61 (C–O–Csymm), 858.36, 820.75 (CH); 1H NMR : 8.5 , 8.0 , 7.8 , 7.6 , 7.5 , 7.1 , 2.4 ; 13C NMR (CDCl3): 136.9 (C-6), 128.2 (C-7), 138.7 (C-8), 130.5 (C-9), 128.2 (C-10), 130.9 (C-11), 133.5 (C-12), 126.9 , 129.7 , 137.7 (C-15), 187.2 (C-18), 20.9 (C-20), 137.4 (C-21), 130.6 , 128.8 , 131.9 (C-24); MS: m/z 340.10, 341.12 (M + 1), 342.11 (M + 2).Yield 75.50%, yellow powder, mp (°C): 257–259; IR : 3605.11 (OH), 3073.70, 3032.23 (CH), 1662.71 (C=O), 1558.55 (C=N), 1510.33 (C=C), 1310.69 (OH), 1254.75 (C–O–C), 1117.80 (CH), 983.74, 835.21 (CH); 1H NMR : 8.4 , 7.9 , 7.75 , 7.4 , 7.2 , 6.9 , 5.5 , 3.7 ; 13C NMR (CDCl3): 130.1 (C-6), 114.0 (C-7), 149.7 (C-8), 142.9 (C-9), 117.2 (C-10), 120.7 (C-11), 136.8 (C-12), 127.9 (C-13), 138.0 (C-14), 129.4 (C-15), 128.4 (C-16), 130.1 (C-17), 56.3 (C-18), 188.1 (C-20), 137.8 (C-21), 129.9 , 128.5 , 131.9 (C-25); MS: m/z 372.11, 373.13 (M + 1), 374.12 (M + 2).Yield 78.80%, yellow powder, mp (°C): 268–271; IR : 3624.25 (OH), 3024.38 (CH), 1667.14 (C=O), 1589.34 (C=N), 1554.63 (C=C), 1402.25 (OH), 1247.94 (C–O–C), 1126.43 (CH), 933.70, 862.18, 769.60 (CH); 1H NMR : 8.3 , 7.9 , 7.6 , 7.39 , 7.3 , 6.9 , 5.09 ; 13C NMR (CDCl3): 136.9 (C-6), 128.2 (C-7), 138.7 (C-8), 130.5 (C-9), 128.2 (C-10), 130.9 (C-11), 129.1 (C-12), 128.4 , 116.2 , 157.3 (C-15), 187.2 (C-18), 20.9 (C-20) 137.4 (C-21), 130.6 , 128.8 , 131.9 (C-24); MS: m/z 342.10, 343.18 (M + 1), 344.11 (M + 2).Yield 80.0%, light yellow powder, mp (°C): 294-295; IR : 2920.35 (CH), 1668.50 (C=O), 1522.87 (C=N), 1518.04 (C=C), 1511.29 (N=Oasymm), 1371.45 (N=Osymm), 1265.36 (C–O–C), 1160.23, 1108.15, 1098.51 (CH), 758.06 (CH); 1H NMR : 8.3 , 8.0 , 7.7 , 7.5 , 7.4 ; 13C NMR (CDCl3): 131.6 (C-6), 146.9 (C-7), 124.1 (C-8), 129.4 (C-9), 135.1 (C-10), 127.9 (C-11), 135.9 (C-12), 127.9 (C-13), 138.0 (C-14), 129.4 (C-15), 128.4 (C-16), 130.1 (C-17), 187.0 (C-19), 137.4 (C-21), 130.6 , 128.8 , 131.9 (C-24); MS: m/z 371.10, 372.09 (M + 1), 373.10 (M + 2).Yield 75.85%, yellow crystals, mp (°C): 237–239; IR : 3381.21 (OH), 2987.74 (CH), 1602.85 (C=O), 1591.27 (C=N), 1352.10 (OH), 1070.49 (C–O–C), 742.59 (CH); 1H NMR : 8.5 , 8.39 , 7.81 , 7.61 , 7.3 , 6.8 , 6.7 , 6.5 ; 13C NMR (CDCl3): 123.7 (C-6), 155.8 (C-7), 116.2 (C-8), 129.9 (C-9), 121.6 (C-10), 128.4 (C-11), 135.9 (C-12), 127.9 (C-13), 138.0 (C-14), 129.4 (C-15), 128.4 (C-16), 130.1 (C-17), 187.0 (C-19), 137.4 (C-21), 130.6 , 128.8 , 131.9 (C-24); MS: m/z 342.10, 343.12 (M + 1), 344.11 (M + 2).Yield 78.0%, yellow crystals, mp (°C): 254-255; IR . Solutions containing 10, 12.5, 25, 50, 100, 200, 400, 800, and 1600 μg/mL of the test derivatives in dimethyl formamide (DMF) were added to each cup. Amoxicillin and cefixime were taken as positive control and DMF was taken as blank. The plates were incubated at 37°C for 24 hours and the results were recorded. The zones of inhibition of the microbial growth produced by different concentration of test compounds were measured in millimeters [The antibacterial evaluation was carried out by using agar cup-plate method. The microorganismsμg/mL). The drug solutions were allowed to diffuse for some time into the medium. The plates were incubated at 30–35°C for 24–48 hours. The incubation chamber was kept sufficiently humid. MIC values were determined at the end of the incubation period. The MIC values for synthesized compounds are summarized in Nutrient agar was prepared, sterilized, and cooled to 45°C with gentle shaking to bring about uniform cooling. It was inoculated with 0.5-0.6 mL of culture and mixed well by gentle shaking before pouring into the sterilized petri dishes. The poured materials were allowed to set and thereafter the cups were made by punching into the agar surface with sterile cork borer and scooping out the punched part of the agar. 0.1 mL of each test compounds was added into the cups with the help of sterile syringe. Twofold diluted solutions of the compounds and reference drugs were used  is stated as a mean of at least three experiments for 24 compounds [Biological activity data was converted to the logarithmic value. The biological activities used in the present studies were expressed as pMICompounds , 27.P, SAS, MR, ovality, MSA, and MW were selected in QSAR studies which are having significant effect on biological activity.The physicochemical parameters were computed  using ChDocking studies reported here IN were performed using software Molegro Virtual Docker 5.0.0. E. coli PDF.Ni enzyme by molecular docking [The newly synthesized 1,3,4-oxadiazoles derivatives were computationally designed and optimized to investigate the interactions between the target compounds and the amino acid residues of the docking . Target 1H-NMR, 13C NMR, and mass) methods. Both analytical and spectral data of all the synthesized derivatives were in full agreement with the proposed structures. The characteristic C=N band (1680–1520 cm−1) of medium intensity and a medium-strong band at 1300–1050 cm−1 were identified in each IR spectra; the latter could be attributed to the C–O–C vibration or heteroatom ring deformation of the oxadiazole ring. 1H NMR showed characteristic doublets at (7.8–8.2) and 7.0-7.1) due to aromatic protons of p-methoxy phenyl for derivatives (6a–h). The presence of two methylene groups was confirmed by presence of doublets at (3.0–3.2) and (2.8–3.0). All the other aliphatic and aromatic protons were observed within the expected regions. 13C data also supported the structures of the synthesized derivatives. In the mass spectra, molecular ions of medium intensity and the base peak usually belonged to the corresponding acylium ions and nitrile radical ions, which are formed by the cleavage of heteroatom ring. Both of these have been reported as a characteristic for this family. This part concluded the synthesis of 1,3,4-oxadiazole derivatives.The structures of all the newly synthesized derivatives were confirmed by chromatographic and spectroscopic  were recorded in comparison with amoxicillin and cefixime. The zone of inhibition of 6e , 8h , 13b , 13c , and 13e  was found to be good which is comparable with zone of inhibition of amoxicillin  and cefixime  indicated that these derivatives have potent antibacterial activity. No inhibitory effect was observed for DMF. All the derivatives were found to be active against Pseudomonas aeruginosa and Staphylococcus aureus. The zone of inhibition and MIC and zone of inhibition of all the synthesized derivatives is summarized in The antibacterial activity of the synthesized derivatives was carried out by agar cup plate method and the average radius of zone of inhibition (mm) and MIC  which affect drug absorption, distribution, bioavailability, drug-receptor interactions, metabolism, and toxicity profile.At one side of 1,3,4-oxadiazole nucleus substitution with phenyl ring or bulkier aromatic groups may also offer significant increase in the biological activity.  and nitro group .The antimicrobial activity results indicated that the presence of electron-withdrawing halogen groups and nitro group at para and ortho position of the phenyl ring improved their biological activity. On the basis of biological data, improvement in biological activity was observed with an increase in electronegativity of the molecule due to the presence of halogen groupAn introspection of the active compounds revealed that different structural features of the compounds have more influence on biological activity. Three starting materials which have been selected for design and development of novel substituted 1,3,4-oxadiazole derivatives include methoxy benzene (anisole), N-phenyl anthranilic acid, andY  and independent variable X .Multilinear regression is one of the vital method of quantitative structure activity relationship which is used for displaying linear correlation among a dependent variable Consider the following:R2: coefficient of correlation, Radj2: coefficient of determination, F: Fischer statistics, N: numbers of synthesized derivatives, S: standard error of estimate, Press: predictive error sum of squares, Q2: cross validated R2, and BA: biological activity.Here and hereafter,  Escherichia coli are summarized in The observed and predicted antibacterial activity of synthesized 1,3,4-oxadiazole derivatives againstConsider the following: Pseudomonas aeruginosa are summarized in The observed and predicted antibacterial activities of synthesized 1,3,4-oxadiazole derivatives againstConsider the following: Staphylococcus aureus are summarized in The observed and predicted antibacterial activities of synthesized 1,3,4-oxadiazole derivatives againstConsider the following: Staphylococcus epidermidis are summarized in The observed and predicted antibacterial activities of synthesized 1,3,4-oxadiazole derivatives against50 values against observed pMIC50 for QSAR model of all bacterial strains is given in Plot of predicted pMICThe biological activity has shown dependence on LogP, SAS, MR, ovality, MSA, and MW. (6a–h), N-phenyl anthranilic acid (8a–h), and o-benzoyl benzoic acid (13a–h) in addition to 1,3,4-oxadiazole moiety which resulted in increased lipophilicity leads to better biological activity.The aromatic substituents like methoxy benzeneAlso the presence of electron withdrawing group at para and ortho positions in the compounds resulted in enhanced biological activities.The results suggested that the antimicrobial activity was highly dependent on LogP, SAS, MR, ovality, MSA, and MW.The derived models could be used in designing of more potent inhibitors against microbial infections.r2) and minimum standard error of estimate (S), developed against E. coli, P. aeruginosa, S. aureus, and S. epidermidis, could be used for the prediction of biological activities of unknown and unavailable compounds of this class.Six parameter correlation equations for antimicrobial activity, having good values of correlation coefficient , Lys 157 (13e), Arg 97 (13b and 13c), and Arg 153 (13e and 13g). The amino acids Lys 150 and Arg 153 were involved in interaction with the standard drugs (cefixime and amoxicillin) as well as the synthesized 1,3,4-oxadiazole derivatives. The docking score of derivatives 6f, 8e, 8f, 8h, and 13e was found to be higher than amoxicillin and 13e was found to have the highest dock scoring derivative than that of amoxicillin and cefixime.The results of docking study of synthesized 1,3,4-oxadiazole derivatives  Figures, and 9 wIn synthesized 1,3,4-oxadiazole derivatives replacement of free carboxylic group by oxadiazole nucleus enhanced the receptor interaction by formation of numerous hydrogen bonds and favourable steric interactions with peptide deformylase. These results could be used for the development of novel, potent, and effective antimicrobial agents.π-electron delocalization across the donor-acceptor links and affords significant electron transportation. Results also revealed that the hydrogen bond distance is important in docking studies. The distance more than 3.2 Å indicates feeble hydrogen bonding between ligand and receptor, 2.6 Å–3.2 Å represents virtuous hydrogen bonding, and less than 2.5 Å indicates robust bonding. Almost all the active derivatives showed good hydrogen bonding with enzymes.The carbonyl, nitro, and 1,3,4-oxadiazole functionalities (acting as acceptor), whereas –NH– and hydroxyl group (acting as donor) in the synthesized derivatives have played very imperative position in ligand-receptor interaction for the creation of numerous hydrogen bonds. Insertion of electron withdrawing 1,3,4-oxadiazole offers an upgraded 6e, 6f, 13b, 86, 8f, 8h 13c, and 13e were observed as the best antibacterial agents against all the selected microbial strains. While studying MIC against bacterial strains, compound 13b with p-chloro and 13e with m-methoxy and p-hydroxyl substitutions were found to be the most active among all the derivatives. In developed QSAR models, six physicochemical parameters  were instituted to be important for the antibacterial activity. All the developed models have good coefficient of correlation (0.796–0.885), coefficient of determination (0.616–0.815), and cross validated R2 (0.792–0.888) with good Fischer statistics (3.94–12.02). Among all the synthesized compounds 13e was found to be most potent peptide deformylase inhibitor with the highest dock score (−153.44) than that of amoxicillin and cefixime.Among all the synthesized derivatives


Musella lasiocarpa, led to the isolation of four rare bicyclic diarylheptanoids, musellarins B-E (2–5), two new phenylphenalenones, 2-methoxy-9--1Hphenalen-1-one (9), 2-methoxy-9-(3′-methoxy-4′-hydroxyphenyl)-1H-phenalen-1-one (10), a new acenaphtylene derivative, trans--3-(4′-methoxyphenyl)-acenaphthene-1,2-diol (13), and two new sucrose esters, 1,2′,3′,4′,6′-O-pentaacetyl-3-O-trans-pcoumaroylsucrose (16), 1,2′,3′,4′,6′-O-pentaacetyl-3-O-cis-p-coumaroylsucrose (17), together with nine known compounds. In addition, -1--7-(4″-hydroxyphenyl)-hepta-4,6-dien-3-one (15) was isolated for the first time from a natural source. The structures of new compounds were elucidated by analysis of their spectroscopic data. Compounds 2, 6, 8–10, 12, and 14 were cytotoxic toward several of the human tumor cell lines . Of these, the new compound 9 was the most potent one, with IC50 values of 5.8, 10.3, 6.3, 3.3, and 2.3 µM, respectively. Phytochemical investigation of the aerial parts of the monotypic plant, Supplementary material is available for this article at 10.1007/s13659-011-0007-7 and is accessible for authorized users. Supplementary material, approximately 1.23 MB.


Microcystis spp. collected in November, 2008, from a commercial fishpond near Kibbutz Kfar Blum, the Jordan Valley, Israel. The new natural products included the prenylated aeruginosin KB676 (1), microphycin KB921 (2), anabaenopeptins KB906 (3) and KB899 (4) and micropeptins KB928 (5), KB956 (6), KB970A (7), KB970B (8), KB984 (9), KB970C (10), KB1048 (11), KB992 (12) and KB1046 (13). Their structures were elucidated primarily by interpretation of their 1D and 2D nuclear magnetic resonance spectra and high-resolution mass spectrometry. Marfey’s and chiral-phase high performance liquid chromatography methods were used to determine the absolute configurations of their chiral centers. Aeruginosin KB676 (1) contains the rare -Choi and is the first prenylated aeruginosin derivative described in the literature. Compounds 1 and 5–11 inhibited trypsin with sub-μM IC50s, while Compounds 11–13 inhibited chymotrypsin with sub-μM IC50s. The structures and biological activities of the new natural products and our procedures of dereplication are described.Thirteen new and eighteen known natural products were isolated from a bloom material of an assembly of various  Cyanobacterial water blooms are initiated from dormant cells under appropriate environmental conditions, which develop into a stable population of different biomass intensities and toxin content, which eventually collapses, disintegrates and discharges toxins to the water environment . These bi.e., micropeptins, aeruginosins, anabaenopeptins, microginins and microviridins, have been shown to affect the normal life cycle of organisms that interact with cyanobacteria in the environment ,,Microcysazoles A , 915 D23 −14° ; UV (MeOH) λmax (log ε) 202 (4.29), 224 (3.79), 278 (3.12) nm; IR (ATR Diamond) νmax 2936, 1648, 1542, 1202, 1137 cm−1; 1H and 13C NMR . Retention times of amino acid (AA) Marfey’s derivatives: d-Phe 52.9 min (l-Phe 50.1 min), -Choi (l-6-epiChoi) 41.5 min, -Choi ) 38.3 min and -Choi (l-Choi)) 42.0 min. Retention time of d-hydroxyphenyllactic acid (Hpla) on the chiral-phase column 5.69 min (l-Hpla 5.41 min).Aeruginosin KB676 : Amorphous white solid; [α]D23 −36° ; UV (MeOH) λmax (log ε) 202 (4.25), 208 (4.15), 217 (3.94) nm; IR (ATR Diamond) νmax 2934, 1671, 1653, 1542, 1203, 1138 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Glu 36.8 min (d-Glu 37.7 min), l-Pro 41.2 min (d-Pro 49.0 min), 2 × l-Ala 45.4 min (d-Ala 49.0 min), 3 × l-Phe 50.0 min (d-Phe 52.9 min), l-Leu 50.0 min (d-Leu 53.9 min).Microphycin KB921 : Amorphous white solid; [α]D23 −62° ; UV (MeOH) λmax (log ε) 202 (4.18), 224 (3.67), 278 (2.91) nm; IR (ATR Diamond) νmax 3422, 1664, 1206 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 29.6 min (d-Arg 29.4 min), l-NMeHty 39.6 min, 2 × l-Ile 43.1 min (d-Ile 46.2 min), d-Lys 45.2 min (l-Lys 44.6 min), l-Hph 46.8 min (d-Hph 49.4 min).Anabaenopeptin KB906 : Amorphous white solid; [α]D23 −45° ; UV (MeOH) λmax (log ε) 202 (4.34), 225 (3.97), 278 (3.40) nm; IR (ATR Diamond) νmax 3310, 2934, 1638, 1542, 1204 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Val 44.4 min , l-NMeHty 46.4 min, l-Ile 48.1 min (d-Ile 50.7 min), d-Lys 49.0 min (l-Lys 50.46 min), l-Hph 52.0 min (d-Hph 55.2 min), l-Tyr 54.5 min (d-Tyr 57.1 min).Anabaenopeptin KB899 : Amorphous white solid; [α]D23 −82° ; UV (MeOH) λmax (log ε) 202 (4.37), 217 (4.00) nm; IR (ATR Diamond) νmax 3367, 2966, 1650, 1542, 1204, 1138 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 28.2 min (d-Arg 26.9 min), l-Asp 33.0 min (d-Asp 34.0 min), l-Thr 32.7 min (d-Thr 35.9 min), l-Glu 34.6 min (d-Glu 35.9 min), l-Val 43.5 min , l-Ile 47.3 min (d-Ile 52.1 min), l-NMePhe 48.2 min.Micropeptin KB928 : Amorphous white solid; [α]D23 −128° ; UV (MeOH) λmax (log ε) 202 (4.34), 217 (3.84) nm; IR (ATR Diamond) νmax 3367, 2962, 1639, 1527, 1200, 1177, 1133 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 27.8 min (d-Arg 26.6 min), l-Asp 28.6 min (d-Asp 29.5 min), l-Thr 27.8 min (d-Thr 31.2 min), l-Glu 30.1 min (d-Glu 31.2 min), l-Val 37.8 min , l-Ile 41.4 min (d-Ile 45.2 min), l-NMePhe 42.5 min.Micropeptin KB956 : Amorphous white solid; [α]D23 −54° ; UV (MeOH) λmax (log ε) 202 (4.21), 217 (3.57) nm; IR (ATR Diamond) νmax 3366, 2964, 1653, 1541, 1203, 1136 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 28.5 min (d-Arg 27.3 min), l-Asp 29.3 min (d-Asp 30.3 min), l-Thr 28.5 min (d-Thr 32.0 min), l-Glu 31.1 min (d-Glu 31.7 min), l-Val 38.3 min , l-Ile 41.8 min (d-Ile 45.2 min), l-NMePhe 42.8 min.Micropeptin KB970A : Amorphous white solid; [α]D23 −88° ; UV (MeOH) λmax (log ε) 202 (4.30), 217 (3.74) nm; IR (ATR Diamond) νmax 3366, 2933, 1652, 1557, 1202, 1135 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 28.5 min (d-Arg 27.3 min), l-Asp 29.2 min (d-Asp 30.2 min), l-Thr 28.5 min (d-Thr 31.9 min), l-Glu 31.0 min (d-Glu 32.0 min), l-Val 38.3 min , l-Ile 41.8 min (d-Ile 45.7 min), l-NMePhe 42.8 min.Micropeptin KB970B : Amorphous white solid; [α]D23 −62° ; UV (MeOH) λmax (log ε) 202 (4.32), 217 (3.74) nm; IR (ATR Diamond) νmax 3367, 2962, 1638, 1542, 1200, 1178 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 28.1 min (d-Arg 26.9 min), l-Asp 28.8 min (d-Asp 29.8 min), l-Thr 28.1 min (d-Thr 31.5 min), l-Glu 30.6 min (d-Glu 31.7 min), l-Val 37.8 min , l-Ile 41.4 min (d-Ile 45.2 min), l-NMePhe 42.4 min.Micropeptin KB984 : Amorphous white solid; [α]D23 −87° ; UV (MeOH) λmax (log ε) 202 (4.37), 217 (4.00) nm; IR (ATR Diamond) νmax 3432, 2922, 1645, 1208 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Arg 28.2 min (d-Arg 27.5 min), l-Asp 34.1 min (d-Asp 34.8 min), l-Thr 33.2 min (d-Thr 36.3 min), l-Glu 34.9 min (d-Glu 36.3 min), 2 × l-Val 43.8 min , l-NMePhe 48.4 min.Micropeptin KB970C : Amorphous white solid; [α]D23 −11° ; UV (MeOH) λmax (log ε) 202 (4.49), 230 (3.67), 282 (2.91) nm; IR (ATR Diamond) νmax 3366, 2962, 1652, 1541, 1201, 1136 cm−1; 1H and 13C NMR  [M + H]+, . Retention times of AA Marfey’s derivatives: l-Arg 28.7 min (d-Arg 27.5 min), l-Asp 29.4 min (d-Asp 30.4 min), l-Thr 28.7 min (d-Thr 32.7 min), l-Glu 31.3 min (d-Glu 32.3 min), 2 × l-Ile 42.0 min (d-Ile 46.6 min), l-NMeClTyr 49.9 min.Micropeptin KB1048 : Amorphous white solid; [α]D23 −47° ; UV (MeOH) λmax (log ε) 202 (4.29), 208 (4.14), 278 (2.92) nm; IR (ATR Diamond) νmax 3421, 2931, 1648, 1542 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Asp 33.9 min (d-Asp 34.6 min), l-Thr 33.3 min (d-Thr 36.3 min), l-Glu 35.5 min (d-Glu 36.5 min), l-Val 43.9 min , l-Ile 47.6 min (d-Ile 51.7 min), l-NMePhe 48.3 min, l-Leu 48.5 min (d-Leu 2.3 min). Retention time of d-hydroxyphenyllactic acid (Hpla) on the chiral-phase column 5.43 min (l-Hpla 5.27 min).Micropeptin KB992 : Amorphous white solid; [α]D23 −36° ; UV (MeOH) λmax (log ε) 202 (5.39), 225 (3.90), 276 (3.29) nm; IR (ATR Diamond) νmax 3367, 2933, 1638, 1533, 1202 cm−1; 1H and 13C NMR . Retention times of AA Marfey’s derivatives: l-Thr 28.8 min (d-Thr 32.2 min), 2 × l-Glu 31.4 min (d-Glu 32.2 min), l-Val 38.6 min , l-Ile 42.2 min (d-Ile 46.0 min), l-NMePhe 43.2 min. Retention time of HcAla derivatives with l-FDAA (46.9 min) and d-FDAA (50.2 min). Retention time of d-hydroxyphenyllactic acid (Hpla) on the chiral-phase column 5.36 min (l-Hpla 5.24 min).Micropeptin KB1046  were hydrolyzed in 6 N HCl (1 mL). The reaction mixture was maintained in a sealed glass bomb at 104 °C for 16 h. The acid was removed in vacuo, and the residue was re-suspended in 250 μL of H2O. FDAA solution -5-l-alanine amide) or -5-d-Alanine amide) in acetone  and NaHCO3  were added to each reaction vessel. The reaction mixture was stirred at 40 °C for 2 h. Then, HCl  was added to each reaction vessel, and the solution was evaporated in vacuo. The FDAA-amino acids derivatives from hydrolysate were dissolved in 1 mL CH3CN and compared with standard FDAA-amino acids by an HPLC analysis: Hibar LiChrospher 60, RP-select B , flow rate 1 mL/min, UV detection at 340 nm, linear gradient elution from 1:9 CH3CN:0.1% aq. TFA buffer (pH 3) to CH3CN, within 60 min. The absolute configuration of each amino acid was determined by spiking the derivatized hydrolysates with d,l-mixture of the standard derivatized amino acids. As with the advanced Marfey method [d- and l-derivatives were compared to determine the configuration of the constituent amino acid.Compounds y method , the ret8, 9 and 13 with ethyl ether separated Hpla from the amino acid salts. The ether was removed in vacuo, and the residue was dissolved in MeOH (1 mL). The MeOH solution was analyzed on an Astec, Chirobiotic T, LC stationary phase, 250 × 4.6 mm flow rate 1 mL/min, UV detection at 277 nm, linear elution with 19:1 MeOH:1% aq. triethylammonium acetate buffer (pH 4). The Hpla derivatives from 1, 12 and 13 were compared with an authentic standard of d,l-Hpla.The extraction of the acid hydrolysates of Compounds The procedures used to determine the inhibitory activity of the new compounds on trypsin and chymotrypsin were described in a previous paper . The ranMicrocystis blooms usually produce a large variety of short peptides, of which the micropeptins are dominant in quantity and diversity. In the current research, the 7:3 MeOH:H2O extract yielded 31 different peptides of six structural groups. As in the case of many other Microcystis blooms that we have investigated [1). These three groups of modified peptides are biosynthesized by non-ribosomal peptide synthetases that are flexible and capable of synthesizing series of analogues metabolites. The reason(s) for the biosynthesis of these metabolites and their high variability in cyanobacterial water blooms are intriguing, and our current research is aimed at revealing the purpose for the biosynthesis of these metabolites.stigated ,19,24,25


The products were easily transformed into the corresponding 3-(5-methoxycarbonyl-1H-imidazol-3-ium-3-yl)pyrrol-1-ides, which in turn can be hydrolyzed under basic conditions into the corresponding betaines. A carbene tautomeric form of the 4-methoxycarbonyl-substituted imidazolylpyrrolides was trapped by reaction with sulfur affording the corresponding imidazolethiones under very mild conditions.A simple approach was developed for the synthesis of methyl 4-imidazolylpyrrole-2-carboxylates from easily available compounds, 5-methoxyisoxazoles and phenacylimidazolium salts under hybrid Fe(II)/Et Furthermore, hydrolysis of 2 could provide an easy access to unknown carboxy-substituted ylides 4, and then could potentially be in the equilibrium with N-heterocyclic carbenes 5 and betaine 6 -1H-imidazoles based on the formation of a pyrrole ring via the reaction of 2H-azirines with 1-alkyl-3-phenacyl-1H-imidazolium bromides + calcd for C22Н19ClN3O2, 392.1160; found, 392.1168; IR  ν: 3389, 3042, 1706.General procedure for debenzylation of 1-benzyl-3-pyrrol-3-yl-1H-imidazol-3-ium bromides 1j,k,n. 1-Benzyl-1H-imidazol-3-ium bromide 1 (100 mg) was dissolved in MeOH (10 mL), Pd/C  and ammonium formate (10 equiv) were added. The suspension was stirred under reflux for 1 h (monitored by TLC). The reaction mixture was filtered to remove Pd/C, MeOH was evaporated under reduced pressure, water was added to the residue and the product was filtered, washed with water and dried to give the analytically pure compound.Methyl 4-(1H-imidazol-1-yl)-3,5-diphenyl-1H-pyrrole-2-carboxylate (12a): colorless solid, mp 239–241 °C , yield 128 mg, 90%, obtained from 1-benzyl-3-(5-(methoxycarbonyl)-2,4-diphenyl-1H-pyrrol-3-yl)-1H-imidazol-3-ium bromide , Pd/C  and ammonium formate  according to the general procedure. 1Н NMR (DMSO-d6) δ 3.66 , 6.89–6.90 , 7.10–7.11 , 7.21–7.26 , 7.30–7.33 , 7.51–7.52 , 12.49 ; 13С NMR (DMSO-d6) δ 51.2 (CH3), 117.3 (C), 119.1 (C), 122.5 (CH), 126.9 (CH), 127.1 (CH), 127.4 (CH), 128.2 (CH), 128.5 (CH), 128.69 (CH), 128.70 (C), 128.9 (C), 129.7 (CH), 131.1 (C), 131.8 (C), 139.1 (CH), 160.5 (C); HRMS–ESI (m/z): [M + H]+ calcd for C21Н18N3O2, 344.1394; found, 344.1401; IR  ν: 3124, 2951, 1688.General procedure for the synthesis of pyrrolydes 2 from 5-methoxycarbonylpyrrol-3-ylimidazolium bromides 1. A suspension of 3-(1H-pyrrol)-1H-imidazol-3-ium bromide 1 (1 mmol) in aqueous solution of KOH  was sonicated for 5 min and then vigorously stirred for 12 h. The precipitate was filtered, washed with water (2–3 mL) and dried to give analytically pure compound.2-Methoxycarbonyl-4-(1-methyl-1H-imidazol-3-ium-3-yl)-3,5-diphenylpyrrol-1-ide (2a): colorless solid, mp 237–238 °C (dec.), yield 188 mg, 71%, obtained from 3-(5-(methoxycarbonyl)-2,4-diphenyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazol-3-ium bromide  and a aqueous solution of KOH  according to the general procedure. 1Н NMR (DMSO-d6) δ 3.51 , 3.81 , 7.03–7.21 , 7.69–7.72 (m. 2H), 9.11 ; 13С NMR (DMSO-d6) δ 35.7 (CH3), 49.4 (CH3), 115.2 (C), 123.3 (CH), 124.6 (CH), 124.9 (CH), 125.2 (CH), 125.8 (C), 126.3 (CH), 127.0 (CH), 128.0 (CH), 128.0 (C), 129.6 (CH), 135.5 (C), 135.8 (C), 137.1 (C), 137.9 (CH), 165.0 (C); HRMS–ESI (m/z): [M + H]+ calcd for C22H20N3O2, 358.1550; found, 358.1566; IR  ν: 3528, 3144, 3059, 1676.General procedure for the synthesis of 4--1H-pyrrol-2-carboxylates 13 from pyrrolides 2. A suspension of 3-(1H-imidazol-3-ium-3-yl)-5-(methoxycarbonyl)-pyrrol-1-ide  and sulfur  in dry THF was stirred at rt for 1–2 hours (monitored by TLC). Then the reaction mixture was evaporated to dryness and the residue was purified by column chromatography on silica gel (hexane/ethyl acetate from 1:1 to 0:1) to give the analytically pure compound.Methyl 4--3,5-diphenyl-1H-pyrrole-2-carboxylate (13a): colorless solid, mp 261–262 °C, yield 43 mg, 80%, obtained from 2-(methoxycarbonyl)-4-(1-methyl-1H-imidazol-3-ium-3-yl)-3,5-diphenylpyrrol-1-ide  and sulfur  according to the general procedure. 1Н NMR (DMSO-d6) δ 3.46 , 3.67 , 6.84 , 7.03 , 7.17–7.42 , 7.52 , 12.41 ; 13С NMR (DMSO-d6) δ 34.9 (CH3), 51.1 (CH3), 117.1 (C), 118.9 (CH), 119.6 (CH), 119.7 (C), 127.0 (CH), 127.3 (CH), 127.5 (CH), 128.1 (CH), 128.3 (CH), 129.4 (C), 129.7 (C), 130.1 (CH), 132.1 (C), 132.4 (C), 160.6 (C), 165.4 (C); HRMS–ESI (m/z): [M + Na]+ calcd for C22H19N3O2SNa, 412.1090; found 412.1112; IR  ν: 3304, 1662, 1454, 1375.5-(4-Chlorophenyl)-4-(1-methyl-1H-imidazol-3-ium-3-yl)-3-phenyl-1H-pyrrole-2-carboxylate (6b). A suspension of 3-(2-(4-chlorophenyl)-5-(methoxycarbonyl)-4-phenyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazol-3-ium bromide 1b  and LiOH  in dioxane (30 mL) and water (3 mL) was stirred at 110 °C for 24 h. Then reaction mixture was evaporated to dryness and water (5 mL) was added. The suspension was filtered, the solid was washed with water (2 × 5 mL) and thoroughly dried to obtain lithium salt 14b in quantitive yield. To convert the lithium salt 14b into 6b trichloroacetic acid  was added to a suspension of the lithium salt in water (5 mL). The suspension was sonicated for 5 min, stirred for 1 h and filtered. The solid was washed with water and dried to obtain 6b as a colorless solid, mp 205–207 °C, yield 79 mg, 98%. 1Н NMR (DMSO-d6) δ 3.82 , 7.11–7.26 , 7.29 , 7.36 , 7.77 , 7.80 , 9.26 ; 13С NMR (DMSO-d6) δ 36.0 (CH3), 115.1 (C), 120.8 (C), 123.9 (CH), 124.0 (C), 124.2 (C), 125.9 (CH), 126.0 (CH), 127.1 (CH), 128.4 (CH), 128.42 (C), 128.6 (CH), 130.1 (CH), 131.8 (C), 133.1 (C), 138.6 (CH), 162.2 (C); HRMS–ESI (m/z): [M + H]+ calcd for C21H17ClN3O2, 378.1004; found, 378.1009; IR  ν: 3498, 3033, 1694. Carboxylate 6a (CCDC 1406417) was analyzed by single X-ray diffraction. It is triclinic P−1, a = 9.2999(5) Å, b = 9.5698(4) Å, c = 15.3875(5) Å, α = 72.782(3)°, β = 78.712(4)°, γ = 66.401(4)°, V = 1194.02(9) Å3, Z = 2, 4960 unique reflections were measured, which were used in all calculations. The final R1 was 0.0561 and wR2 was 0.1681  (>2sigma(I)) (see File 11H and 13C NMR spectra for all new compounds. Computational details: energies of molecules, transition states and their Cartesian coordinates of atoms. X-ray details.Detailed experimental procedures including characterization data for all synthesized compounds,


AbstractTheraphosidae genus, Kankuamo Perafán, Galvis & Pérez-Miles, gen. n., is described from Colombia, with a new type of urticating setae. These setae differ from others principally by having a small distal oval patch of lanceolate reversed barbs. Males of Kankuamogen. n. additionally differ by having a palpal bulb organ very divergent from all known species, with many conspicuous keels dispersed across the median tegulum to the tip, mostly with serrated edges. Females differ by having spermathecae with a single notched receptacle, with two granulated lobes and several irregular sclerotized longitudinal striations. The new urticating setae, type VII, is characterized, illustrated and its releasing mechanism is discussed. It is hypothesized that these setae are the first in Theraphosinae subfamily whose release mechanism is by direct contact. Kankuamogen. n. is described and illustrated on the basis of the type species Kankuamomarquezi Perafán, Galvis & Gutiérrez, sp. n., and their remarkable characteristics, morphological affinities and cladistic relationship are analyzed.A new monotypic  Theraphosidae Thorell, 1869 is the most speciose of the Mygalomorphae with more than 130 genera and 980 species , mainly distributed in the tropical and subtropical regions, and currently divided into 11 subfamilies , and another four in Theraphosinae    (see Ber and VI) . Excepti and VI) , all othsensuTheraphosidae, with a large number of conspicuous keels on tegulum and embolus. The morphology and arrangement of these keels do not easily fit with the palpal bulb homologies proposed by Theraphosinae. However, this taxon shares the main general characteristics of the Theraphosinae subfamily, extended subtegulum, keels on palpal bulb, and urticating setae . Photographs were taken with an Infinity Lite camera adapted to the stereoscope lens (Nikon SMZ-10), SEM with JEOL JSM-5900 LV. The geographic coordinates are taken by GPS, Datum WGS84. The distribution map was produced using DIVA-GIS version 7.5.0 (http://www.diva-gis.org/). The type material is stored in 70% ethanol, and deposited in the Instituto de Ciencias Naturales Arachnological Collection (ICN-Ar), at the Universidad Nacional de Colombia, Bogotá, Colombia.Urticating setae terminology follows Abbreviations are listed below: accessory keels AcK apical keel AK anterior lateral eyes ALE anterior median eyes AME apical ap dorsal D dorsal keels DKsUniversidad Nacional de Colombia, Bogotá, Colombia  Instituto de Ciencias Naturales, ICN concavity constant k ocular quadrangle  OQ prolateral P prolateral inferior keel PIK posterior median eyes PME posterior median spinnerets PMS posterior lateral eyes PLE posterior lateral spinnerets PLS prolateral superior keel PSK subapical keel SAK supra-accessory keels SpAcK retrolateral R ventral VPageBreakCladistic analysis. Cladistic analysis was based on the previous matrix of Theraphosinae genera used by Schismatothelinae taxon Guyruita Guadanucci et al., 2007 . Character optimization was performed in Winclada 1.00.08  coxal stridulatory setae: absent = 0; present = 1. (21) Coxal spinules: absent = 0; present = 1. (22) Labial cuspules: numerous (more than 15) = 0; few or none = 1. (23) Fovea: normal = 0; with spheroid process = 1. (24) Metatarsus I of males: normal = 0; strongly curved = 1. (25) urticating hairs on prolateral palpal femur: absent = 0; PageBreakpresent = 1. (26) Urticating setae type VI: absent = 0; present = 1. (27) Coxae: normal = 1; retrolaterally extend = 1.PIK: absent = 0; present = 1. (34) Prolateral superior keel: absent = 0; presente = 1.(36) Urticating setae type VII: absent = 0; present = 1.Taxon classificationAnimaliaAraneaeTheraphosidaePerafán, Galvis & Pérez-Milesgen. n.http://zoobank.org/622CB9E5-59D1-4E45-A5CE-21F5248CF9EFKankuamomarquezi Perafán, Galvis & Gutiérrez, sp. n.Theraphosidae by having a distinct type of urticating setae  and supra-accessory keels (SpAcK). Tibial apophysis on leg I absent : male palpal bulb with large and extended subtegulum, and numerous developed keels; legs spinose, with normal scopulae on tarsi and presence of abdominal urticating setae , but with the subtegulum more elongated in Kankuamo gen n.  for those placed on proximal subtegulum  for those placed principally on dorsal face , but notched and longitudinally striated  found between 3 and 10 shortest trees. The strict consensus of each of these tress recovered different topologies, but between k = 8 and k = 12 the topology of strict consensus did not vary, for this reason we have selected this topology to test Kankuamo gen. n. affinities  on character 16 “absence the urticating setae type I”, a parallelism of the state (1) on character 31 “presence the accessory keels under prolateral inferior keel” and an autapomorphic character “presence of urticating setae type VII”.Kankuamo gen. n. and Metriopelma were resolved as sister genera within the largest clade of our selected topology. The relationship of Kankuamo gen. n. + Metriopelma as the sister group of the clade (Theraphosa (Sericopelma (Brachypelma (Megaphobema (XenesthisPamphobeteus) (Sphaerobothria (Phormictopus (CyrtopholisAcanthoscurria))))))) is supported by “absence of retrolateral keel” and “absence of sub-apical keel”, with some homoplasies. The Theraphosa clade is supported by two synapomorphies “apical region of palpal bulb with cancave-convex aspect” and “apical keel very long”.Taxon classificationAnimaliaAraneaeTheraphosidaePerafán, Galvis & Gutiérrezsp. n.http://zoobank.org/7EF097DE-5147-4995-985C-35504F164C6111.11 N, -74.058 W, PageBreak30-Aug-2014, leg. W. Galvis and J. Moreno (ICN-Ar 7983). Allotype female, same data as the holotype (ICN-Ar 7983). Paratypes: one female, same data as the holotype (ICN-Ar 7984); one male from the same locality as the holotype, 11.1 N, 74.05 W, 9-10-Sept-2014, leg. Miguel Gutierrez (ICN-Ar 7985).Holotype male from Colombia, Magdalena, Santa Marta, Corregimiento Minca, Sector San Lorenzo, 2200m above sea level, The specific epithet is a noun in genitive in honor to Gabriel García Márquez , who was a renowned Colombian writer, considered one of the most significant authors of the 20th century, and awarded the 1982 Nobel Prize in Literature for “One hundred years of solitude”.See diagnosis of the genus.Male (holotype ICN-Ar 7982) , most of them with serrated edge   Figs  and 4. Tous Fig. . Basal sous Fig. , length ous Fig. , with 56ous Fig.  and 5F; ous Fig. . Palpal ous Fig. . Cymbiumous Fig. , curved,dge Fig. . PresenccK) Fig. . Colour PageBreakSpination : Femur: palp: 0V, 0D, 0-0-1P, 0R; I: 0V, 1-1-0D, 0-1-1P, 0-1-2R; II: 0V, 1-1-0D, 1-1-3P, 1-5-2R; III 0V, 1-1-0D, 0-2-1P, 0-2-3R; IV: 0V, 3-2-0D, 1-2-1P, 0-1-2R. Patella: palp: 0V, 0D, 0-2-0P, 0R; I: 0-2-2V, 0D, 0-2-0P, 0R; II: 0-0-2V, 0D, 0-2-0P, 0R; III: 0V, 0D, 0-3-0P, 0-1-0R; IV: 0V, 0D, 0-3-0P, 0-1-0R.Tibia: palp: 2-1-2V, 0D, 2-2-2P, 0R; I: 4-6-2apV, 0D, 0-2-2P, 0-1-1R; II: 3-5-2apV, 0D , 0-1-1P, 0-2-0R; III: 3-3-2apV, 0D, 2-2-1P, 2-2-1R; IV: 3-3-2-1-2apV, 0D, 2-2-2P, 1-2-2-2apR. Metatarsus: I: 2-3-1V, 0D, 0-1-1apP, 0-1-1apR; II: 3-3-1V, 0D, 0-1-1-1apP, 0-1-1apR; III: 4-4-2-1apV, 0-0-2D, 2-2-1-1apP, 1-2-1-1apR; IV: 5-5-3-1apV, 0-0-2D, 2-3-1-1apP, 2-2-1-1apR. Tarsus: palp and legs: 0. Legs and palpal segments lengths in Table ICN-Ar 7983)  Figs  and 6. Ttae Fig. . Basal stae Fig. , length lar Fig. , with 15lar Fig.  length 7lar Fig. ; palp anlar Fig. . Color : Femur: palp: 0V, 0-1-0D, 0-0-3P, 0-0-1R; I: 0V, 0-1-0D, 0-0-3P, 0R; II: 0V, 0-2-0D, 0-0-3P, 0-1-0R; III: 0V, 0D, 0-2-1P, 1-3-1R; IV: 0V, 1-0-0D, 0-0-3P, 0-0-1R. Patella: palp: 0V, 0D, 0-2-0P, 0R; I: 0V, 0D, 0-1-0P, 0R; II: 0V, 0D, 0-2-0P, 0R; III: 0V, 0D, 0-2-0P, 0R; IV: 0V, 0D, 0-1-0P, 0R. Tibia: palp: 1-4-4V, 0D, 0-3-0P, 0-1-0R; I: 0-1-2V, 0D, 1-1-0P, 0R; II: 0-2-2V, 0D, 0-2-0P, 0R; III: 3-3-3V, 0D, 2-1-2P, 1-2-1R; IV: 2-2-2V, 0D, 2-2-1P, 2-2-1R. Metatarsus: I: 1-5-1V, 0D, 0-1-1P, 0R; II: 1-5-1V, 0D, 0-0-1P, 0R; III: 4-3-5V, 0-0-2D, 2-2-2P, 1-3-1R; IV: 5-4-5V, 0-0-2D, 3-3-2P, 1-2-2R. Legs and palpal segments lengths in Table PageBreakSee distribution of the genus Fig. .Kankuamomarquezi sp. n. inhabits Cuchilla San Lorenzo from Sierra Nevada de Santa Marta National Natural Park. Cuchilla San Lorenzo is located in its northwestern flank, in a gradient of altitude from 2000–2300 meters above sea level, in life zone of lower montane wet forest .t forest . The Sielanaceae , and inhPageBreakPageBreakMorphology. Urticating setae differ from body covering setae by the insertion feature through a stalk  or attached into a specialized socket (types V and VI) that facilitates detachment, plus presence of a penetrating acute tip, and barbs or scales that aid embedding them into targets  are narrow, mean width ranging from 6 to 7 µm, and length/width ratio ranging from 98.3 to 208.3. In contrast, for urticating setae released by direct contact with the target, the width was greater, ranging from 15 to 22 µm and the length/width ratio of 37.3-46.8  involved in their urticating effects, presumably often contribute their ability to float in the air. Conversely, in both urticating setae types II and VII, which presumably do not float by air, the main barbs are only residual denticles, being much smaller than those of other types with known air dispersal.Furthermore, the larger size, broader shape and often dispersed arrangement of many of the main barbs of The differences in the position of the penetrating tip between type VII and II also suggest a different penetration mechanism. On first contact with the target, the urticating setae type II pivots on its stalk so the apical end moves away from the target, while the basal penetration tip  instead lodges into the target as the stalk releases see , fig. 3.PageBreakthird of their length. When we attempted to remove them from the skin, these setae were easily broken at their distal end. Based on our observations together, our assumption is that the patch of the lanceolate reverse barbs in the distal area can serve as a breakpoint into the skin, ensuring that the apex of the seta remains within the tissues.Experimentally, the possible mechanism of action of the urticating setae type VII was observed while handling specimens in alcohol. These urticating setae easily perforated the skin of human fingers perpendicularly. The dorsum of the tarantula’s abdomen was touched intentionally, and on further examination of the affected fingers with a stereoscope microscope, many of these setae were found embedded in the skin Fig. . These sKankuamo gen. n. therefore represents a remarkable exception to this suggested character association because, in contrast, they do have legs with some spines, mainly conspicuous on metatarsi and tarsi of hind-leg pair IV. Leg spines in other theraphosids have been functionally associated with releasing of airborne urticating setae by facilitating rubbing, with the exception of Aviculariaversicolor which use claw tufts for rubbing  could have facilitated the evolutionary transformation of contact setae into derived airborne setae, which are now widespread among Theraphosinae. However, if contact setae are interpreted as derived, the posterior leg spines present in Kankuamo gen n. could be regarded as a plesiomorphic retention.Another aspect to consider is that all previously known species with contact urticating setae have leg spines absent or reduced . Kankuam rubbing . One posPageBreakKankuamo gen. n. fits the diagnostic characters of Theraphosinae, but also shows a very divergent palpal bulb morphology and the presence of a new abdominal urticating setae type. These setae are unique, and here are proposed to be the only contact released urticating setae yet known within the Theraphosinae, although this release mechanism was previously well known only for Aviculariinae. Also, the supernumerary keels on the male palpal bulb clearly distinguish it from all known theraphosid species. Kankuamo gen. n. was resolved as the sister group of Metriopelma on our preferred phylogeny of Theraphosinae.


Page 1: In the Funding Information section, grant number “09-546-2-206” should read “6-059-2-023.”Volume 4, no. 2,


S-isopropanide. In the absence of the starting thioketone, the corresponding thiiranes and/or ethene derivatives, formed from them via spontaneous desulfurization, are the main products. In contrast, reactions with diazoethane occurred predominantly via initially formed diradicals, which in cascade processes gave sterically crowded 4,4,5,5-tetrahetaryl-1,3-dithiolanes as major products. Finally, the reaction of dihetaryl thioketones with (trimethylsilyl)diazomethane occur smoothly at −75 °C leading to the corresponding 4,4,5,5-tetrahetaryl-1,3-dithiolanes as the exclusive [3 + 2]-cycloadducts formed via a cascade of postulated diradicals. The presence of S or Se atoms in the hetaryl rings is of importance for stabilizing diradical intermediates. Remarkably, in no single case, the ‘head-to-head dimerization’ of aryl/hetaryl and dihetaryl substituted thiocarbonyl ylides was observed.Reactions of dihetaryl and aryl/hetaryl thioketones with 2-diazopropane, diazoethane, and (trimethylsilyl)diazomethane were studied at variable temperature. The experiments showed that reactions with 2-diazopropane carried out at –75 °C occur mainly via the initially formed, relatively stable 1,3,4-thiadiazolines as products of the [3 + 2]-cycloaddition of the diazo dipole onto the C=S bond. The latter decompose only at higher temperature (ca. −40 °C) to generate thiocarbonyl  Cycloaddition reactions belong to the most important classes of organic reactions, and 3 + 2]- cycloadditions, also known as 1,3-dipolar cycloadditions or Huisgen reactions, offer a universal tool for the preparation of five-membered heterocycles with a variable number of heteroatoms in the ring - cycload. The fir1a), the reaction performed at –65 ºC occurs without evolution of N2 and the in situ formed 2,2-diphenyl-1,3,4-thiadiazoline 2a can be subsequently used as a precursor of the reactive thiobenzophenone S-methanide (a thiocarbonyl ylide) 3a at ca. −45 °C, when the evolution of N2 takes place -cycloaddition reactions with diverse dipolarophiles leading to five-membered S-heterocycles, such as di- and tetrahydrothiophenes, 1,3-oxathiolanes, 1,3-dithiolanes, etc.Prompted by these observations, we decided to examine reactions of aryl/hetaryl and dihetaryl thioketones omethane . An impo1a) and 2-diazopropane (7a) at ca. −75 °C in THF led to a change of the blue color of the solution without evolution of N2. The addition of the solution of 7a was continued until the color of the mixture changed to pale red. The mixture was warmed up, and around 0 °C the evolution of N2 was observed. The 1H NMR analysis of the crude product revealed the presence of only one singlet for two Me groups located at 1.62 ppm. After chromatographic work-up, the only product isolated as colorless crystals was identified as 2,2-dimethyl-3,3-diphenylthiirane  and 7a. Whereas at −75 °C and −60 °C the corresponding thiirane 8b and ethene 9b, formed via spontaneous desulfurization of 8b, were obtained as the sole products, in the experiment carried out at –15 ºC, the sterically crowded 1,3-dithiolane 10b was also detected as a minor product  with diazoethane (7b) performed at –75 °C led to 5-methyl-2,2-diphenyl-1,3,4-thiadiazoline as the exclusive product -cycloadducts. Elimination of N2 from the initially formed diradical of type 11 leads immediately to the new diradical species 3, which adds regioselectively to the C=S group of the non-converted thioketone 1.Based on these results, the reaction mechanism can be proposed as formulated in 1 with diazomethane, in which the formation of sterically crowded 1,3-dithiolanes side-by-side with 12-membered dimers of the thiocarbonyl ylide was observed [1 with the in situ generated thiocarbonyl S-methanides [3 derived from hetaryl thioketones can be explained by steric hindrance resulting from the presence of the Me or Me3Si groups in the S-methanide moiety. However, electronic effects resulting from the diradical nature of the intermediate thiocarbonyl ylides, can also play certain role. The same effect was observed in the case of diaryldiazomethanes used in reactions with aryl/hetaryl thioketones [This interpretation is consistent with the course of the reactions of aryl/hetaryl thioketones observed . In addithanides . The misoketones –30.General information: Melting points were determined in capillaries using a MEL-TEMP II apparatus (Aldrich) and are uncorrected. IR spectra were recorded with a FTIR NEXUS spectrophotometer as KBr pellets or as film; absorptions (ν) in cm−1. 1H and 13C NMR spectra were measured on a Bruker Avance III (1H at 600 and 13C at 150 MHz) instrument in CDCl3; chemical shifts (δ) are given in ppm, solvent signals as reference, coupling constants (J) in Hz. The multiplicity of the 13C signals was deduced from DEPT, supported by 1H,13C HMQC spectra. 1H NMR data are presented as follows: chemical shift, multiplicity , coupling constant, integration. The mass spectra were recorded on a Finnigan MAT-95 (ESI). Elemental analyses were performed in the Microanalytical Laboratory of the Chemistry Faculty of the University of Łódź. Applied reagents such as 2-diazopropane (7a) and diazoethane (7b) were prepared by known methods according to the literature protocols [1a), fluorene-9-thione , symmetrical dihetaryl thioketones 1d,e, and nonsymmetrical aryl/hetaryl thioketones 1c, 1f–h were obtained from the corresponding ketones using the known procedures [rotocols –32. Thioocedures . Other rReaction of thioketones 1a–h with 2-diazopropane (7a) and diazoethane (7b) – General procedure: Corresponding thioketones 1a–h (1 mmol) were dissolved in freshly distilled THF (2.5 mL) and the solution was cooled to the corresponding temperature . Then, the mixture was treated with small portions of ethereal 2-diazopropane (7a) or diazoethane (7b) solution, until the intense color of the thioketone vanished. Then, the mixture was allowed to warm slowly to rt (ca. 2–4 h). After removal of the solvent under vacuum, the residue was subjected to 1H NMR analysis in CDCl3 solution with a weighed amount of 1,1,2,2-tetrachloroethane as a standard. Crude products were purified by CC (CHCl3/hexane 2:8).7a) and thioketones 1c,e,g,h and from diazoethane (7b) and thioketones 1b–f, the presence of thiirane and the corresponding ethylene was evidenced based on the 1H NMR spectra. In these cases, no isolation of the thiirane was performed; after addition of tris(dimethylamino)phosphine desulfurization leading to the ethane derivative was carried out.In crude mixtures obtained from 2-diazopropane (Reaction of thioketones 1d–e with (trimethylsilyl)diazomethane (7c) – General procedure: The corresponding thioketones 1d–e (1 mmol) were dissolved in freshly distilled THF (2.5 mL) and the solution was cooled to −75 °C (acetone/dry ice). Then, the mixture was treated with small portions of an ethereal solution of 7c (1 mmol). The mixture was kept in a cold bath, and the intense color of the thioketone vanished after 15 min. Subsequently, the mixture was allowed to slowly warm to rt (ca. 2–4 h). After removal of the solvent under vacuum, the residue was subjected to the 1H NMR analysis in CDCl3 solution with a weighed amount of 1,1,2,2-tetrachloroethane as an internal standard. Crude products were purified by CC (CHCl3/hexane 2:8).2,2-Dimethyl-3,3-diphenylthiirane (8a): Yield: 182 mg (76%). White crystals; mp 66–67 °C (chromatographic purification); IR (KBr) ν: 2987 (w), 2921 (w), 1596 (w), 1490 (m), 1443 (m), 773 (m), 747 (m), 705 (s), 691 (m) cm−1; 1H NMR  δ 7.60–7.58 , 7.31–7.29 , 7.23–7.21 , 1.62  ppm; 13C NMR  δ 142.4 (for 2 Carom), 129.4, 127.9, 126.8 (for 10 CHarom), 67.8, 52.9 , 27.9 (2 CH3) ppm; ESIMS m/z (%): 241 ; anal. calcd for C16H16S (240.36): C, 79.95; H, 6.71; S, 13.34; found: C, 79.69; H, 6.50; S, 13.40.1,1-Di(thiophen-2-yl)-2-methylpropene (9d): Yield: 4 mg (2%). Yellow oil; IR (film) ν: 2927 (w), 2908 (w), 2850 (w), 1436 (m), 1369 (m), 1227 (m), 1016 (w), 827 (m), 693 (s) cm−1; 1H NMR  δ 7.29 , 7.04–7.02 , 6.91 , 2.03  ppm; 13C NMR  δ 144.9, 137.1 , 126.6, 126.4, 124.9 (for 6 CHarom), 122.9 (C=C(CH3)2), 23.3 (2 CH3) ppm; anal. calcd for C12H12S2 (220.35): C, 65.41; H, 5.49; S, 29.10; found: C, 65.31; H, 5.76; S, 28.81.2,2-Dimethyl-4,5-diphenyl-4,5-di(thiophen-2-yl)-1,3-dithiolane (10f): Isolated as a mixture of cis-, trans-isomers (crude product ratio 65:35). Yield: 142 mg (63%). White crystals; mp 153–154 °C (MeOH/CHCl3); IR (KBr) ν: 3062 (w), 2919 (w), 1595 (w), 1490 (m), 1443 (m), 1231 (m), 1156 (m), 854 (w), 697 (s) cm−1; 1H NMR  δ 7.59–6.68 , 1.86 , 1.70 , 1.39  ppm; 13C NMR  δ 151.2, 149.8, 144.0, 142.9 (for 8 Carom), 131.3, 131.1, 130.5, 127.5, 127.1, 127.0, 126.6, 126.1, 125.4, 125.2, 125.0 (for 32 CHarom), 78.8, 78.4 , 55.4, 55.3  33.1 (CH3-cis), 32.9 (for 2 CH3-trans), 32.5 (CH3-cis) ppm; anal. calcd for C25H22S4 (450.70): C, 66.62; H, 4.92; S, 28.46; found: C, 66.46; H, 4.91; S, 28.33.1,1-Di(thiophen-2-yl)-propene (9k): Yield: 52 mg (25%) – After desulfurization of tiirane 8k. Yellow oil; IR (film) ν: 3104 (w), 3071 (w), 2930 (w), 2908 (w), 2850 (w), 1438 (m), 1362 (w), 1249 (m), 1223 (m), 1036 (w), 850 (s), 836 (s), 818 (s), 695 (s) cm−1; 1H NMR  δ 7.23–7.22 , 7.01–7.00 , 6.96–6.94 , 6.90–6.89 , 6.80–6.79 , 6.71–6.70 , 6.18 , 1.74  ppm; 13C NMR  δ 129.4, 139.8, 146.8 , 127.6, 127.0, 126.6, 126.3, 125.5, 124.8, 123.4 , 15.6 (CH3) ppm; anal. calcd for C11H10S2 (206.33): C, 64.04; H, 4.89; S, 31.08; found: C, 63.83; H, 4.98; S, 31.14.2-Trimethylsilyl-4,4,5,5-tetra(thiophen-2-yl)-1,3-dithiolane (10r): Yield: 220 mg (87%). Yellow crystals: mp 152–154 °C (hexane/CH2Cl2); IR (KBr) ν: 2951 (w), 1618 (w), 1424 (m), 1250 (s), 1232 (m), 1122 (m), 1077 (m), 1050 (m), 842 (s), 752 (s), 699 (s), 632 (m) cm−1; 1H NMR  δ 7.23–7.22 , 7.20–7.19 , 6.96–6.95 , 6.89–6.87 , 6.83–6.81 , 3.89 3Si)HC), 0.34 3Si) ppm; 13C NMR  δ 148.1, 146.1 (for 4 Carom), 130.2, 129.4, 127.0, 125.6, 125.5, 125.4 (for 12 CHarom), 74.9 , 36.5 (C-2), −1.53 ((CH3)3Si) ppm; anal. cald for C22H22S6Si (506.89): C, 52.13; H, 4.37; S, 37.96; found: C, 52.44; H, 4.55; S,37.71.File 18–10, and the original 1H and 13C NMR spectra for all products.Experimental data for selected compounds