The structures of the synthesized compounds were characterized by means of their IR and 1H-NMR spectral data. Synthesized compounds were screened for antiviral activity against influenza A  and influenza B viruses in MDCK cell culture. The antiHIV activities of the new compounds were screened for antiviral activity against replication of HIV-1(IIIB) in MT-4 cells. Cytotoxicity of the synthesized compounds was also tested in mock-infected MDCK and MT-4 cells. Compound CF-SD and CF-SDM inhibits the influenza A (H1N1) and compound GF-SDM inhibit the replication of influenza A (H5N1) and B in MDCK cells. All compounds displayed cytostatic propertity in MT-4 cells. Among the compounds tested, GF-SDM (CC50=39.44 μM) most toxic compound in this series.A series of novel N-Sulphonamidomethyl piperzinyl fluoroquinolones were synthesized and screened antiviral activity. Eight compounds were synthesized through modifying the N Quinolone is a versatile lead molecule for designing potential bioactive agents and its derivatives were reported to possess board spectrum antimicrobial activity. A new fluoroquinolone K12, bearing o-methoxyphenylpiperazinyl group and a difluoromethoxyl group at positions 7 and 8, respectively, was reported to have strong and selective antiHIV-1 activity–6 and so1H-NMR spectra were determined on Bruker AMX 400 MHZ with tetramethylsilane as an internal standard. The sample was dissolved in DMSO-d6 and the value was measured in δ ppm.Melting points were determined in open capillary tubes on a Thomas-Hoover melting point apparatus and are uncorrected. IR spectra were recorded using KBr pellets on a Jasco-410 infrared spectrophotometer, Synthesis of N-sulphonamidomethyl fluoroquinolones derivatives was achieved by stirring on equimolar (0.01 mol) mixture of sulphonamides , formaldehyde  and fluoroquinolone  with ethanol using a magnetic stirrer for 3 h . The mix−1: 3349 (NH), 2365 (C-Alkyl), 1763 (C=O), 1622 (C=N), 1440 (SO2), 670 (C-F); 1H-NMR (DMSO-d6) δ ppm:1.13 , 2.0 , 2.6 , 3.1 , 3.5 , 4.0 , 4.13 , 6-7.7 , 11.0 ; EI-MS (m/z):503. NF-SD yield:75.52%, mp:190°; IR (KBr) cm−1: 3352 (NH), 2365 (C-Alkyl), 1747 (C=O), 1622 (C=N), 1440 (SO2), 673 (C-F); 1H-NMR (DMSO-d6) δ ppm: 1.13 , 2.6 , 3.10 , 3.5 , 4.0 , 4.13 , 6-7.7 , 8.4 , 11.0 ; EI-MS (m/z): 581.NF-SDM yield: 69.52 %, mp: 210°, IR (KBr) cm−1:1745 (C=O), 1620 (C=N), 1440 (SO2), 3356 (NH), 2365 (C-Alkyl), 659 (C-F); 1 H-NMR (DMSO-d6) δ ppm: 1.13 , 2.3 , 2.6 , 3.10 , 3.5 , 4.0 , 4.13 , 6-7.7 , 11.0 ; EI-MS (m/z):609. CF-SD yield: 76.29%, mp: 200°; IR (KBr) cm−1: 3349 (NH), 2360 (C-Alkyl), 1748 (C=O), 1622 (C=N), 1440 (SO2), 667 (C-F); 1 H-NMR (DMSO-d6) δ ppm: 0.42 , 1.36 , 2.6 , 3.5 , 4.0 , 4.13 , 6-7.7 , 8.4 , 11.0 ; EI-MS (m/z): 593. CF-SDM yield: 89.25%, mp: 258°, IR (KBr) cm−1: 3350 (NH), 2365 (C-Alkyl), 1736 (C=O), 1622 (C=N), 1440 (SO2), 658 (C-F); 1 H-NMR (DMSO-d6) δ ppm: 0.42 , 1.36 , 2.6 , 3.10 , 3.5 , 4.0 , 4.13 , 2.2  6-7.7 , 11.0 ; EI-MS (m/z): 621. GF-SA yield: 82.48%, mp: 240°, IR (KBr) cm−1: 3352 (NH), 2367 (C-Alkyl), 1732 (C=O), 1624 (C=N), 1440 (SO2), 671(C-F); 1 H-NMR (DMSO-d6) δ ppm: 0.42 , 1.36 , 2.0 , 2.6 , 3.5 , 3.23 , 4.0 , 4.13 , 6-7.7 , 11.0 ); EI-MS (m/z): 545. GF-SD yield 80.34%, mp 255°, IR (KBr) cm−1: 3349 (NH), 2362 (C-Alkyl), 1750 (C=O), 1628 (C=N), 1440 (SO2), 659 (C-F); 1 H-NMR (DMSO-d6) δppm: 0.42 , 1.36 , 2.6 , 3.5 , 3.23 , 4.0 , 4.13 , 6-7.7 , 8.4 , 11.0 ; EI-MS (m/z): 623. GF-SDM yield 67.41%, mp 265°, IR (KBr) cm−1: 3352 (NH), 2362 (C-Alkyl), 1742 (C=O), 1624 (C=N), 1440 (SO2), 668 (C-F). 1H-NMR (DMSO-d6) δ ppm: 0.42 , 1.36 , 2.2 , 2.6 , 3.5 , 3.23 , 4.0 , 4.12 , 6-7.7 , 11.0 ; EI-MS (m/z): 651NF-SA yield: 65.24%, mp: 268°; IR (KBr) cm50) of virus were then added, and the plates were incubated for 3 d when inhibitor-free cell cultures were completely destroyed by virus. At this time, the mean percentage of cell viability in each set of three infected wells or set of two toxicity control wells was quantified by a neutral red dye uptake method, using 0.011% final concentration of the dye for 2 h. An Excel spreadsheet was developed for converting optical density readings to percentages of untreated control values. Concentrations of compounds reducing viral CPE by 50%  were calculated by plotting concentration versus percent inhibition on a semi log10 graph paper. Antiviral activity and cytotoxicity of standard ribavirin were also performed by a similar method.The inhibitory effects of the compounds on influenza A and B viruses replication were determined by cytopathic effect inhibition (CPE) assays in MDCK cell monolayers conducted in 96-well microplates9. CompouB) in MT-4 cells[50) were added to a flat-bottomed microtiter tray with 50 μl of medium containing various concentrations of compounds. MT-4 cells were added at a final concentration of 6×105 cells/ml. After 5 d of incubation at 37°, the number of viable cells were determined by the 3--2,5-diphenyl tetrazolium bromide (MTT) method. Cytotoxicity of test compounds against mock-infected MT-4 cells was also assessed by the MTT method. AntiHIV activity and cytotoxicity of standard AZT were also performed by a similar method in MT-4 cells.Compounds were tested for their inhibitory effects against replication of HIV-1  was 16 and 18 μg/ml, respectively, whereas the cytotoxic concentration (CC50) was found to be more than 100 μg/ml . Their inhibitory concentration (EC00 μg/ml . The com50) values of the compounds against the replication of HIV-1 were higher than the cytotoxic concentrations (CC50). Whereas the standard AZT (EC50) had 0.0064 μM against HIV-1 in MT-4 cells. Cytotoxic concentrations of test compounds were found to be 39-75 μM, whereas the standard AZT showed 65.9 μM in mock infected MT-4 cells Compound GF-SDM (CC50=39.44 μM)displayed marked cytostatic activity in the MT-4 cells. Presence of 2-pyrimidinyl/4,6-dimethyl-2-pyrimidinyl group in sulphonamide (SO2NH2) of N-sulphonamidomethyl piperzinyl flouroquinolone lead molecule is essential for antiviral activity against influenza viruses. Free SO2NH2 of N-sulphonamidomethyl piperizinyl flouroquinolone substitution will reduce the antiviral activity (The synthesized compounds were evaluated for antiHIV activity against HIV-1 replication in acutely infected MT-4 cells . The 50%activity . These l


Lobophytum pauciflorum resulted in the isolation of five new biscembranoids named lobophytones O–S (1–5) and a new “monomeric” cembrane lobophytone T (6). The structures of the new compounds were elucidated by interpretation of 1D and 2D NMR  spectroscopic data in association with MS and IR data. Lobophytone Q showed significant inhibition against lipopolysaccharide (LPS)-induced nitric oxide (NO) release in mouse peritoneal macrophages, while lobophytones Q and T showed inhibitory activities against the bacteria S. aureus and S. pneumoniae.Chemical examination of a Chinese soft coral  Sarcophyton . They are supposed to be biogenetically derived by a Diels–Alder cycloaddition of cembranoid-diene and cembranoid-dienophile +  and NMR data, requiring 10 degrees of unsaturation. The 1H NMR spectrum exhibited the resonances for nine methyls including two olefinic methyl singlets  and an OMe (δH 3.42), while the 13C NMR spectrum presented 41 carbon resonances including three ketone and one ester carbonyl carbon, four olefinic carbons, and five oxygen-bearing sp3 carbons. Six degrees of unsaturation, as accounted by the functional groups out of 10 in the molecule, suggested the presence of a tetracyclic nucleus. The 1H and 13C NMR data of 1 featured a biscembrane skeleton, closely related to that of ximaolide C as formerly isolated from Sarcophyton tortuosum +), indicating 42 amu more than that of 1. Comparison of NMR data revealed that the structure of 2 was closely related to that of 1, except for the presence of an acetyl group as evident from the 13C NMR signals at δC 171.3 (Cq) and 21.4 (CH3) and the corresponding 1H NMR signal at δH 2.06 . The HMBC correlation from H-26  to the acetyl carbonyl carbon ascertained the location of the acetoxy group at C-26. The similar NOE and NMR data of 2 and 1 indicated that 2 shared the same configurations of the stereogenic centers of 1. Thus, compound 2 was determined to be a C-26 acetylated analogue of 1.Lobophytone P (3) has a molecular formula of C41H63O8Cl as determined by HR-ESIMS , indicating 10 degrees of unsaturation and the presence of a chlorine atom. The 1D and 2D NMR spectroscopic analysis revealed that the structure of 3 was also closely related to that of 1. The presence of two D2O exchangeable protons at δH 4.18 (s) and 4.46 (br) was attributed to two hydroxy groups, which were deduced to be attached at C-31 and C-33, based on the HMBC interactions from OH-31  to C-30 (δC 90.1), C-31 (δC 73.2) and C-32 (δC 40.7), and the COSY correlation between OH-33  and H-33 (δH 4.82). The chlorine atom of 3 was assumed to be substituted at C-26 according to the upfield-shifted C-26 (δC 65.1), which showed around Δδ 5 less than the same carbon of 1 linked by hydroxy group. The relative configurations of the chiral centers in 3 were in agreement with those of 1 based on the similar NMR and NOE data.Lobophytone Q (4) was determined as C41H64O8 on the basis of HR-ESIMS (m/z 685.4670 [M + H]+), implying 10 degrees of unsaturation. Comparison of NMR data indicated that 4 is a biscembranoid possessing the same partial structure, with respect to rings A and B, of 1. However, the obvious downfield shifted C-26 (δC 77.8) and C-31 (δC 80.5), and the upfield shifted C-27 (δC 74.7) and C-30 (δC 80.0), in comparison with the corresponding carbons of 1, suggested that 4 contains an ether bridge at C-26/C-31 in ring C rather than C-27/C-30 ether bond of 1. The HMBC correlation from H-26 (δH 3.39) to C-31 confirmed this assignment. The COSY correlation between OH /H-30 (δH 2.91) and the HMBC relationships from OH (δH 4.18) to C-26 and C-27 disclosed two hydroxy groups at C-27 and C-30, respectively. The NOE relationships observed between H-2 /H-22 , H-22/H-26, and H-26/H3-40 (δH 0.92) indicated H-26 and H3-40 to be β-oriented, while the NOE correlations between OH-27 (δH 4.18)/H-26, H-30 /H-29a , and H3-39 (δH 0.98)/H-29a assigned to H3-39α and H-30α.The molecular formula of lobophytone R (5) has a molecular formula of C41H62O7, as determined by HR-ESIMS (m/z 689.4384 [M + Na]+). The NMR spectroscopic data of 5  in ring C of 5, in addition to the double bond located at C-22/C-23. This double bond was deduced to be located at C-26/C-27 based on the COSY and HMBC correlations. The carbon resonances of C-30  and C-31  were attributed to epoxy carbons, while a hydroxy group linked to C-33 was confirmed by the HMBC correlations from H-33  to C-21 and C-35 and the COSY relationship between OH /H-33. The NOE correlations between H3-39 (δH 1.52)/H2-25  and H-22 (δH 4.70)/H2-24  were in accordance with 22E and 26E geometries , H-33/H-22, and H-22/H-2 led to the assignment of β-faces of H-30 and H-33 and the trans-configuration of the epoxide group.Lobophytone S (ata of 5  and 2 inometries . The NOE6) has a molecular formula of C22H34O5 as determined by HR-ESIMS (m/z 401.2292 [M + Na]+), implying six degrees of unsaturation. The 1H MMR displayed five methyl singlets including two olefinic methyls at δH 1.65  and 1.80  and an acetyl methyl group at δH 1.87 , in addition to four olefinic protons for an AB spin system at δH 6.02  and 6.03  and for the exo-methylene at δH 4.86 (brs) and 4.82 (brs). The COSY correlations established the segments from C-2 to C-3, C-5 to C-7, C-9 to C-11, and C-13 to C-14, while 13C NMR and HMQC data indicated the methines of C-7 (δC 85.0), C-11 (δC 70.5) and C-14 (δC 69.7) bearing oxygen atom. The HMBC relationships connected each segment to form a backbone that featured a cembrane with 14-membered ring. An isopropene was deduced to position at C-1 based on the HMBC cross-peaks from H2-16  and H3-17 (δH 1.80) to C-1 (δC 143.5), while additional HMBC correlations indicated two vinyl groups to be conjugated at C-1/C-2 and C-3/C-4. The HMBC correlations from H3-19 (δH 1.01) to C-7, C-8 , and C-9 (δC 32.2), and from H3-20 (δH 1.05) to C-11, C-12 , and C-13 (δC 44.3), in addition to the HMBC relationship from H-7 (δH 3.26) to C-11 established an ether bridge which linked C-7 and C-11. The acetoxy group was deduced to position at C-14 on the basis of the HMBC correlations from H-14 (δH 6.01) to the acetyl carbonyl carbon and to C-1, C-2, and also C-15 . Thus, C-8 and C-12 were hydroxylated, and the gross structure of 6 was established as shown in 6 were determined through J values and NOESY analysis. The coupling constants of H-11  suggested its axial orientation in the chair-conformation of the perhydropyrano ring, while the NOE relationship between H-11/H-14 indicated the same face (β-orientation) of both protons. The NOE interactions of OH-12  with H-11 and H-14 assigned to OH-12β, whereas the NOESY cross-peaks between H-11/H-9a , H3-19/H-9a, and H-7/OH-8  suggested α-orientation for H-7 and OH-8.Lobophytone T (6) is likely a precursor (diene) to form the right part of biscembranes when it reacted with methyl tortuosoate + .Amorphous powder; [α]D25 + 133.7 ; IR (KBr) νmax 3467, 2959, 2928, 1741, 1708, 1459, 1373, 1237, 1028 cm−1; 1H and 13C NMR data, see m/z 765.4570 [M + Na]+ .Amorphous powder; [α]D25 + 121.2 ; IR (KBr) νmax 3440, 2958, 2930, 1741, 1706, 1459, 1374, 1206, 1027 cm−1; 1H and 13C NMR data, see m/z 741.4096 [M + Na]+ .Amorphous powder; [α]D25 + 151.1 ; IR (KBr) νmax 3423, 2958, 2928, 1709, 1460, 1376, 1203, 1050 cm−1; 1H and 13C NMR data, see m/z 685.4670 [M + H]+ .Amorphous powder; [α]D25 + 101.1 ; IR (KBr) νmax 3399, 2958, 2927, 1742, 1706, 1459, 1435, 1387, 1204, 1074 cm−1; 1H and 13C NMR data, see m/z 689.4384 [M + Na]+ .Amorphous powder; [α]D25 + 36.5 ; IR (KBr) νmax 3423, 2969, 2935, 1734, 1453, 1375, 1246, 1071, 1026 cm−1; 1H NMR δ : 6.02 , 6.03 , 2.47 , 1.96 , 1.44 , 2.00 , 3.26 , 1.42 , 1.50 , 1.43 , 1.77 , 3.71 , 1.85 , 1.97 , 6.01 , 4.82 , 4.86 , 1.80 , 1.65 , 1.01 , 1.05 , 1.87 ; 13C NMR δ : 143.5 , 120.5 , 119.6 , 137.8 , 39.4 , 21.6 , 85.0 , 69.2 , 32.2 , 19.9 , 70.5 , 73.7 , 44.3 , 69.7 , 144.2 , 114 , 23.0 , 18.1 , 27.8 , 25.0 , 169.9 , 21.5 ; HR-ESIMS m/z 401.2292 [M + Na]+ .Colorless oil; [α]2 in 37 °C. Mouse PEMΦ were incubated with compounds for 1 h at 37 °C before stimulation with 1 μg/mL of lipopolysaccharide (LPS) for 24 h. Cells (5 × 105 cells) were preincubated at 37 °C for 24 h in serum-free medium and NO production was monitored by measuring nitrite levels in culture media using Griess reagent. Absorbance was measured at 548 nm in incubated media with Griess reagent for 10 min. Viable adherent cells were stained with MTT (2 μg/mL) for 4 h. The medium was then removed and the produced formazan crystals were dissolved in DMSO (200 μL). Absorbance was measured at 540 nm.Dexamethasone  and each compound (20 mM in DMSO) were diluted to 1–20 μM range at r.t. before experiment. The final percentage of DMSO in the reaction mixture was less than 0.5% (v/v). LPS (1 μg/mL), 4% sodium thioglycolate, RPMI1640, FBS, PBS, MTT and Griess reagents were purchased from Sigma . Mouse peritoneal macrophages (PEMΦ) were obtained from C57BL6J male mice, and then plated onto 48 well plates and cultured for 2 hrs in DMEM containing 5% FBS at 5% COOne-way analysis of variance was applied for all statistical analyses by independent experiments, and data were represented as means ± standard error of the measurement. Individual values were compared by t-test and a P-value < 0.01 to evaluate the significant.Staphylococcus aureus, S. pneumoniae, Escherichia coli, and Pseudomonas aeruginosa were grown on Mueller-Hinton agar. The yeasts, Candida albicans and Saccharomyces cerevisiae, were grown on Sabouraud dextrose agar, and the fungus, Aspergillus fumigatus, was grown on potato dextrose agar. Targeted microbes (3–4 colonies) were prepared from broth culture , and the final spore suspensions of bacteria (in MHB medium), yeasts (in SDB medium), and fungus (in PDB medium) were 106 and 105 cells/mL and 104 mycelial fragments/mL, respectively. Testing compounds  were transferred to a 96-well clear plate in triplicate, and the suspension of the test microorganisms were added to each well (200 μL) . After incubation, the absorbance at 595 nm was measured with a microplate reader (TECAN), and the inhibition rate was calculated and plotted versus test concentrations.Antimicrobial and antifungal bioassays were conducted in triplicate following the method as recommended by the National Center for Clinical Laboratory Standards (NCCLS) . The bac


DSS-18. Their structures were established on the basis of detailed spectroscopic analyses, including 1D-, 2D-NMR and HR-ESI MS techniques.Three new polyketides, phaeochromycins F ( Streptomyces was isolated from deep sediment collected from the west Pacific.Marine microorganisms are widely recognized as rich sources of novel natural products –2. In re1), G (2) and H (3), from Streptomyces sp. DSS-18  under constant agitation (240 rpm). After filtration of the harvested culture broth, the culture filtrate was extracted exhaustively with ethyl acetate. The ethyl acetate extract was purified by column chromatography  to afford three new polyketides.1) was obtained as a brown powder. The IR absorption at 3424 and 1653 cm−1 indicated the presence of OH groups and aryl ketone. The 1H NMR spectra of 1 (2 moieties , five aromatic CH groups . The 13C NMR (DEPT) spectrum of 1 , 7.57 , 7.39  suggested the presence of a trisubstituted aromatic ring. The HMBC correlations from H-C(8) to C(5), C(6), C(7), C(9), C(10), C(11), C(12) and C(13), and H-C(9) to C(7), C(8), C(10), C(11) and C(12), and from H-C(10) to C(7), C(8), C(9), C(11), C(12) and C(13), and from H-C(14) to C(12), C(13), C(15) and C(16), along with 1H,1H-COSY correlations, and the connection from C(11) to C(15) via oxygen was confirmed by downshift of their chemical shift, established the presence of fragment 1a (benzopyranone). The assignments of the benzopyranone carbons are in good agreement with those of SEK34b + peak at m/z 603.1282, establishing the molecular formula C33H24O10, which further supported the symmetrical structure. Therefore, from those data, the structure of 1 was elucidated as 5-{[4-hydroxy-3-({4-hydroxy-6-[(2-methyl-4-oxo-4H-chromen-5-yl)methyl]-2-oxo-2H-pyran-3-yl}methyl)-2-oxo-2H-pyran-6-yl]methyl}-2-methyl-4H-chromen-4-one, named phaeochromycin F.Phaeochromycin F  was isolated as an amorphous powder. Its molecular formula was determined as C13H12O3 based on HR-ESI mass spectra and NMR data. The IR absorption at 1660 cm−1 indicated the presence of aryl ketone. Inspection of the NMR spectral data  (2 had the same fragment 1a. The long-range correlations from H-C(2) to C(3) and C(4) and H-C(4) to C(3), C(5), C(6) and C(10) suggested the presence of the acetonyl group connected to C(5). Again, NMR assignments were in excellent agreement with the literature -2-oxo-2H-pyran-3-yl}methyl)-2-oxo-2H-pyran-6-yl]methyl}-2-methyl-4H-chromen-4-one 1), brown, amorphous powder. UV (MeOH): 301.5; [α]D20 = 0 ; IR (KBr): 3424, 2974, 1653, 1391, 1089, 1050; 1H and 13C NMR: see m/z 603.1282 .: 248, 300; [α]D20 = 0 ; IR (KBr): 1660, 1603, 1391, 1118; 1H and 13C NMR: see m/z 217.0897 ., white powder; UV: 230, 249.5, 302; [α]D20 = +13.2 ; IR: 3438, 2971, 1712, 1644, 1605, 1392, 1366, 1118; 1H and 13C NMR: see m/z 283.2717 [M + Na]+ calc. 283.2749).[5-(4-hydroxy-2-oxopentyl)-2-methyl-4File 11–3NMR spectra of compounds


GS-17 along with three known compounds, 5-hydroxyl-3-[(1S)-1-hydroxyethyl]-4-methylfuran-2(5H)-one (3), 3,5-dimethyl-4- hydroxylmethyl-5-methoxyfuran-2(5H)-one (4), and 3,5-dimethyl-4-hydroxymethyl-5- hydroxyfuran-2(5H)-one (5). Compound 1 is the first natural occurring polyketide with a unique furylisobenzofuran skeleton. The structures of these compounds were established via extensive spectroscopic analyses including 1D-, 2D-NMR, HRESI-MS, and crystal X-ray diffraction analysis.Two new polyketides, myrothecol ( Myrothecium sp., was isolated from a soil sample taken from a saline field in Gansu Province in China. The fermentation broth of Myrothecium sp. GS-17 exhibited inhibitory activity against the human promyelocytic leukemia HL-60 cell line. Previous investigation of this fungus had led to the isolation of eight trichothecenes 20D = −17.65) [3 was identified as 5-hydroxy-3-[(1S)-1-hydroxyethyl]-4-methylfuran-2(5H)-one 20D = 0). By comparison of its spectroscopic data with those reported previously, the structure of 4 was elucidated as 3,5-dimethyl-4-hydroxymethyl-5-methoxy- furan-2(5H)-one 20D = 0). By comparison of its spectroscopic data with those reported previously, the structure of 5 was elucidated as 3,5-dimethyl-4-hydroxymethyl-5-hydroxyfuran-2(5H)-one 20D = –17.65 . UV (MeOH) λmax: 205 nm. IR (KBr) νmax: 3401, 2983, 1749, 1050, 1029 cm−1. 1H-NMR  and 13C-NMR : see −. HRESI-MS: 181.0474 .Colorless oil. [α]20D = 0 . UV (MeOH) λmax: 210 nm. IR (KBr) νmax: 3399, 2938, 1758, 1071, 1026 cm−1. 1H-NMR  and 13C-NMR : see +. HRESI-MS: 195.0623 .Colorless oil. [α]20D = 0 . UV (MeOH) λmax: 207 nm. IR (KBr) νmax:3368, 2993, 2933, 1749, 1199, 1044 cm−1. 1H-NMR  and 13C-NMR : see −. HRESI-MS: 181.0466 . Colorless oil. [α]1 was determined using data collected on a SMART APEX II CCD diffractometer (Mo Kα radiation). A suitable colorless crystal (0.18 × 0.14 × 0.12 mm) of 1 for diffraction was obtained from methanol solution. Crystal data: C16H18O5 orthorhombic, a = 7.180(5) Å, b = 9.723(5) Å, c = 10.924(5) Å, V = 722.6(7) Å3, Z = 2, Dcalcd 1.334 mg/m3, λ = 0.71069 Å, F(000) = 308, T = 293(2) K. A total of 2433 reflections were collected, of which 1545 unique reflections (Rint = 0.0532) with I > 2σ (I) were used for the analysis. The data was solved using direct method SHELXL-97, and the structure was refined full-matrix least-squares on F2 values. The refined structure model converged to a final R1 0.0476, wR2 = 0.1205 with Goodness-of-fit = 0.951. CCDC 969517 contains the supplementary crystallographic data for 1. These data can be obtained free of charge via http://www.ccdc.cam.ac.uk/conts/retrieving.html  1223 336033; E-mail: deposit@ccdc.cam.ac.uk).The crystal structure of Myrothecium sp. GS-17 has resulted in the isolation of five polyketides, including two new ones, myrothecol and 5-hydroxy-3-methyl- 4-(1-hydroxyethyl)-furan-2(5H)-one. Myrothecol (1) represents a novel class of natural polyketides.Our investigation on the metabolites of the halotolerant fungus


The obtained compounds were identified by their spectral  data. Moreover, DFT quantum chemical calculations were used to study the mechanism of the cycloaddition reaction. The regioselectivity was explained using transition state calculations, where the calculations agreed with the formation of products 4 and 5 in almost the same ratio. The reaction was also extended for diphenylaceylene as dipolarophile to give only two products instead of three.Regioselectively, ethyl propiolate reacted with 1-(propergyl)-pyridinium-3-olate to give two regioisomers; ethyl 4-oxo-8-(prop-2-ynyl)-8-aza-bicyclo(3.2.1)octa-2,6-diene-6-carboxylate  Nowadays, improving the traditional methods for organic synthesis is an important issue. Ultrasound is widely used to avoid using expensive reagents, highly acidic solutions, long reaction times and high temperatures as well as to improve reaction yields and incompatibility with other functional groups .The use of ultrasound irradiation in the organic synthesis offers many benefits, which include milder reaction conditions, decrease in the reaction times, and increase in selectivity, thus enhancing the purity and reaction efficiency ,3,4. UltIa in polar solvents exists to a considerable extent in the unstable mesomeric form, pyridinium-3-olate Ib, via prototropic rearrangement , base peak at m/z = 95 [M+ − Cl-CH2CCH].1) with ethyl propiolateReaction of 1-(propergyl)-pyridinium-3-olate ( (2): To a mixture of 3-hydroxy prop-2-yne-1-yl-pyridinium chloride  and ethyl propiolate  in the presence of hydroquinone (0.01 g), triethylamine (1 mL) was added dropwise. The reaction mixture was heated in ultrasonic bath, for 6 h at 40 °C. At the end of the reaction, the brown residue obtained was triturated with distilled water and then extracted with chloroform that was separated, dried over anhydrous sodium sulfate, filtered off, and evaporated under reduced pressure to give a brown sticky residue, which gave three spots on TLC. The residue obtained was eluted through a column of alumina with a mixture of ethyl acetate-light petroleum (1:2). Three products 4–6 were obtained in a combined yield of 80% (1.84 g).Ethyl 4-oxo-8-(prop-2-ynyl)-8-aza-bicyclo(3.2.1)octa-2,6-diene-6-carboxylate (4): Yellow crystals , mp 176–177 °C, IR ν = 2929 (CH), 1698 (conjugated CO), 1735 cm−1 (CO ester), C13H13NO3 (231.25), [M+] at m/z = 231, base peak at m/z = 80.Ethyl 2-oxo-8-(prop-2-ynyl)-8-aza-bicyclo[3.2.1]octa-3,6-diene-6-carboxylate (5): Orange crystals , mp 180–182 °C, IR ν = 2931 (CH), 1686 (conjugated CO), 1726 cm−1 (CO ester), C13H13NO3 (231.25), [M+] at m/z = 231, base peak at m/z = 80.Ethyl 2,6-dihydro-6-(prop-2-ynyl)furopyridine-3-carboxylate (6): Yellow oil , IR ν = 2924 (CH), 1723 (CO ester), 1616 (C=C-N), 1348 cm−1 (furan ring). C13H13NO3 (231.24), [M+] at m/z = 231, base peak at m/z = 180.1)  and diphenyl acetylene  in the presence of hydroquinone (0.01 g), triethylamine (1 mL) was added dropwise. The reaction mixture was heated in ultrasonic bath, for 6 h at 40 °C. At the end of the reaction, the brown residue obtained was triturated with distilled water and then extracted with chloroform that was separated, dried over anhydrous sodium sulfate, filtered off, and evaporated under reduced pressure to give a brown sticky residue. The residue obtained was eluted through a column of alumina with a mixture of chloroform–light petroleum (3:1). The solvent was removed under reduced pressure to give two products, 10–11 .To a mixture of 1-(propergyl)-pyridinium-3-olate -8-aza-bicyclo(3.2.1)octa-3,6-dien-2-one (10): A brown oil, , IR ν = 2920 (CH), 1690 cm−1 (conjugated CO). C22H17NO (311.37), [M+] at m/z = 311 a.m.u., base peak at m/z = 95 a.m.u 1H-NMR (DMSO) at δ = 2.18 ; 4.05 ; 4.93 ; 5.22 ; 6.00 ; 6.58 ; 7.20–7.91 .2,6-Dihydro-2,3-diphenyl-6-(prop-2-ynyl)furopyridine (11): orange oil, , IR ν = 2924 (CH), 1628–1348 (furan ring), 1525 cm−1 (C=C-N). C22H17NO (311.37) C (84.84%), H (5.50%), N (4.50%) required C (84.65%), H (5.33%), N (4.42%). 1H-NMR (DMSO) at δ = 2.1 ; 3.55 ; 4.92; 5.2; 6.35 ; 7.90–8.07 .1 with either symmetric or asymmetric 2π-acetylene 1,3-dipolarophiles proceeded effectively in a regioselective manner to give bicyclic compounds (8-(prop-2-ynyl)-8-aza-bicyclo(3.2.1)octa-3,6-diene derivative) and 2,6-dihydro-6-(prop-2-ynyl)furopyridine derivative. The results of the DFT calculations showed that in the cycloaddition reaction between 1-(propergyl)pyridinium-3-olate 1 and ethyl propiolate 2, the reactant 1 is the nucleophile while 2 is the electrophile. The global and local reactivity descriptors were used to study the mechanism of the studied CA reaction. Transition state calculations were used to explain the regioselectivity of the CA reaction. Product 7 could not be formed as it has the highest activation energy. Products 4–6 have lower activation energy where 4 and 5 have very close energy barrier so they are formed almost in the same ratio. Product 6 has higher activation energy; hence, it is formed in a lower amount than the others.The cycloaddition reaction between 1-(propergyl)pyridinium-3-olate


Pennington. From wood the coumarin scopoletin (9) was isolated. The structures were established by NMR , mass spectroscopy and infrared (IR) spectral data. The hexane and methanol extracts of the leaves, the protolimonoids lepidotrichilins A (1) and B (2) (IC50 42.7 µg mL−1) and the protolimonoid deoxy-flindissone  exhibited significant cytotoxic activity against the MOLT-4 and U937 leukemic cell lines.Two novel protolimonoids, named lepidotrichilins A ( Trichilia lepidota subsp. schumanniana (Harms) T.D. Pennington  is a tree of 3-6 meters in height. This wood of this species is used to make furniture , ten to 1 (four sp2 and six sp3 including two linked to oxygen atom) and eight to 2 (two sp2 and six sp3 including two linked to oxygen atom) methine, five to 1  and seven to 2  methylene and seven methyl (1 and 2) carbon atoms, corresponding to 30 carbon signals for each compound (2(C)4(=CH)4(CH-O)(CH-OH)(CH)4(CH2)5(CH3)7 = C30H42O4 for 1  and (C=O)(COO)(=C)2(C)4(=CH)2(CH-O)(CH-OH)(CH)6(CH2)5(CH3)7 = C30H42O4 for 2 . Thus, seven methyl groups and eight non-hydrogenated carbon atoms (four sp2 and four sp3) on each compound 1 and 2, revealed clearly the principal differences anticipated by the presence of a double bond between the carbon atoms 1 and 2 of 1 [δC 158.9/δH 7.14  and δC 125.4/δH 5.83 ] and their corresponding absence in 2 suggested by two additional methylene signals . The signals corresponding to methine carbons linked to an oxygen atom HOCH-7 were observed at δC 72.1/δH 3.98  and 71.6/δH 3.98 , using a value of J = 11.7 Hz  to locate the hydroxyl group at OH as equatorial (1a and 2a) and HMQC spectrum to confirm these deductions by direct heteronuclear (1JHC) correlations. The signals (δC/δH) at δC 75.3/δH 5.19  were attributed to CH-23 of 1 and 2. Analogous results for 1 and 2 were attributed to carbon signals of the double bonds located between C-14 (δC 161.0) and CH-15 [δC 119.6/δH 5.51 (m)] and between CH-24 [δC 122.8/δH 5.05 (m)] and C-25 (δC 140.0). The proposed structures of 1 and 2 were confirmed by analyses of the 2D NMR spectra , shown in Comparative analysis of the compound : (C=O) and 2 . The value from the coupling constants (J = 11 7 Hz) corresponding to a vicinal interaction  between the hydrogens H-7  with H-6ax is consistent with the axial-axial interaction shown in 1/1a and 2/2a. The significant differences between 1 and 2 can be attributed to the presence of the one more endocyclic  double bond in the ring-A (CH-1/CH-2) of 1. The protolimonoids 1 and 2 were named lepidotrichilin A (1) and lepidotrichilin B .The location of a hydroxyl group at CH-7 in both compounds was confirmed by HMBC correlations of the signals corresponding to CH-7 of 3, 4 and 5 were obtained as colorless crystals, their 13C-NMR spectra showed 30 carbon signals. The compounds 3, 4 and 5 were identified as 21,23-epoxy-7α-21α-dihydroxy-apotirucalla-14,24-dien-3-one, and 21,23-epoxy-7α-21β-dihydroxyapotiru-calla-14,24-dien-3-one previously isolated from Luvunga sarmentosa ; ; ; ; ; ; ; ; .Deoxyflindissone (6): Purple crystals; m.p. 123 ºC; 13C-NMR δ (ppm): 217.1 (C-3); 48.1 (C-4); 145.7 (C-8); 35.3 (C-10); 43.9 (C-13); 50.9 (C-14); 135.3 (C-25); 51.1 (CH-5); 118.3 (CH-7); 48.5 (CH-9); 52.6 (CH-17); 42.4 (CH-20); 74.7 (CH-23); 127.0 (CH-24); 38.9 (CH2-1); 35.3 (CH2-2); 24.5 (CH2-6); 17.9 (CH2-11); 34.5 (CH2-12); 32.1 (CH2-15); 28.0 (CH2-16); 72.3 (CH2-21); 38.9 (CH2-22); 12.9 (CH3-18); 22.8 (CH3-19); 18.3 (CH3-26); 26.0 (CH3-27); 24.8 (CH3-28); 21.8 (CH3-29); 27.5 (CH3-30). 1H-NMR δ (ppm): ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; . HR-ESIMS (positive mode) (m/z) 439.3402 [M+H]+ ; 421.3313 [M−H2O+H]+ ; 313.2400 [M−H2O−C8H12+H]+ ; 295.2288 [M−2H2O−C8H12+H]+ ; 403.3233 [M−2H2O+H]+ .β-Sitosterol (7): 13C-NMR δ (ppm): 140.8 (C-5); 36.5 (C-10); 42.4 (C-13); 71.8 (CH-3); 121.7 (CH-6); 31.9 (CH-8); 50.2 (CH-9); 56.8 (CH-14); 56.1 (CH-17); 39.8 (CH-20); 45.9 (CH-24); 29.7 (CH-25); 37.3 (CH2-1); 31.9 (CH2-2); 42.4 (CH2-4); 34.0 (CH2-7); 21.1 (CH2-11); 39.8 (CH2-12); 24.3 (CH2-15); 28.9 (CH2-16); 31.9 (CH2-22); 26.2 (CH2-23); 23.1 (CH2-28); 12.0 (CH3-18); 19.4 (CH3-19); 18.8 (CH3-21); 19.8 (CH3-26); 19.8 (CH3-27); 11.9 (CH3-29). LRMS m/z (rel. int.): 414 (100%); 399; 381; 329; 273; 255; 231; 213.Stigmasterol (8): 13C-NMR δ (ppm): 140.8 (C-5); 36.2 (C-10); 42.4 (C-13); 71.8 (CH-3); 121.7 (CH-6); 31.9 (CH-8); 50.2 (CH-9); 56.8 (CH-14); 56.1 (CH-17); 39.8 (CH-20); 138.3 (CH-22); 129.3 (CH-23); 50.2 (CH-24); 31.7 (CH-25); 37.3 (CH2-1); 31.9 (CH2-2); 42.4 (CH2-4); 34.0 (CH2-7); 21.1 (CH2-11); 39.8 (CH2-12); 24.3 (CH2-15); 28.9 (CH2-16); 25.4 (CH2-28); 12.0 (CH3-18); 19.4 (CH3-19); 19.1 (CH3-21); 21.2 (CH3-26); 19.8 (CH3-27); 12.0 (CH3-29). LRMS m/z (rel. int.): 412 (100%); 394; 273; 255; 231; 213.Scopoletin (9): white amorphous powder; m.p. 206-207 ºC; 13C-NMR δ (ppm): 161.4 (C-2); 144.0 (C-6); 149.7 (C-7); 150.0 (C-9); 111.5 (C-10); 113.4 (CH-3); 143.3 (CH-4); 107.5 (CH-5); 103.2 (CH-8); 56.4 (OCH3). 1H-NMR δ (ppm): ; ; ; ; . LRMS m/z (rel. int.): 192 (100%); 177 (63%); 164 (30%); 149 (59%); 121 (30%).−1 of gentamicin. The cultures were split every two or three days in 25 cm2 culture flask and maintained in incubator at 37 °C with 5% of CO2 and controlled humidity.Human Leukemia cells lineages U937 and MOLT-4 were grown in DMEM-12  supplemented with 10% of bovine fetal serum and 20 μgmL1) and B (2) were dissolved in DMSO and 500.0 μg·mL−1 was the highest concentration used for cell culture treatments. Due to the low solubility of the isolated protolimonoid desoxyflindissone (6) and the viscosity of the methanol and hexane extracts, 300 μL of DMSO was added to the each sample, which was homogenized and left standing for 24 h at room temperature. Samples were centrifuged at 11,000 × g for 10 min., then 200 µL of supernatant were collected and submitted to lyophilization. Freeze-dried samples were weighed and diluted in DMSO again. The starting dilution of protolimonoid desoxyflindissone (6) was 150 μg·mL−1, methanol extract 512.5 μg·mL−1 and hexane extract 1000.0 μg·mL−1. Cell viability assay was performed in 96 well plates with 5.0 × 105 cellmL−1. As controls DMSO and cisplatin were included in all experiments.The protolimonoids lepidotrichilin A (Quantification of the enzyme lactate dehydrogenase (LDH) was performed in cell culture supernatants after treatment with the isolated compounds and plant extracts after the period of 48 hours of incubation at 37 °C. The enzyme LDH release was measured by the colorimetric assay from Doles  according to the manufacturer's recommendation. For negative control, cells were treated with DMSO and positive control Triton was added to the culture. Optical density of the samples was measured in multichannel spectrophotometer with wavelength adjusted for 492 nm.−1, 20 μL·well−1) was added to the cell culture after 48 h of treatments with different samples and incubated for 4 h at 37 °C. After incubation, 150 μL of medium from each one well was discarded and the purple crystals of MTT were dissolved by adding 100 μL of isopropanol solution with 0.0014% of HCl. After vigorous agitation the optical density of the samples was measured in multichannel spectrophotometer with wavelength adjusted for 570 nm. For negative and positive control cells were treated with DMSO and Triton, respectively.Cell viability was determined by a 3--2,5-diphenyltetrazolium bromide (MTT) assay. MTT  and B (2) are described for the first time in the literature. The hexane and methanol extracts of the leaves, the protolimonoids lepidotrichilins A (1) and B (2) (IC50 42.7 µg·mL−1) and the protolimonoid deoxyflindissone  exhibited significant cytotoxic activity against the MOLT-4 and U937 leukemic cell lines.The hexane extract from the fruits of


The good performance of this triolborate for metal-catalyzed bond-forming reactions was demonstrated in palladium-catalyzed cross-coupling reactions with haloarenes. Cross-coupling reaction of [MeB(OCH B-alkyl cross-coupling has been frequently used in organic synthesis. Classically, alkylboron reagents have been synthesized from the corresponding alkyllithium or alkylmagnesium compounds by transmetalation with trialkoxyboranes [B-MeO-9-BBN) to produce the corresponding alkylborinate complexes [B-methyl-9-borabicylco[3.3.1]nonane (B-Me-9-BBN) M , that have good stability in air and water and high solubility in organic solvents and that undergo very smooth transmetalation to various transition metal complexes Li was obtained in high yield as an air-stable white solid (97%). Triolborate is a bench-stable ate-complex that can be handled and stored without special precautions.We developed a method for synthesis of lithium methyltriolborate. It was synthesized by methylation of B(Ol)ethane . By usinNext, we chose 4-bromobiphenyl to examine its efficiency toward cross-coupling reaction. The yields were highly sensitive to palladium complexes and phosphine ligands in the cross-coupling reaction between 4-bromobiphenyl and lithium methyltriolborate .2 was used, the yield was 35% (entry 1). The use of Pd(OAc)2 gave the best results (entry 6), but palladium chloride complexes such as PdCl2, PdCl2(PhCN)2, PdCl2(PPh3)2 and PdCl2(cod) resulted in yields of 59%, 66%, 67% and 49%, respectively (entries 2–5).When Pd(dba)3 resulted in low yields (entries 9–12). Next, we screened bidentate ligands, and coupling products were obtained in moderate yields (entries 13–17). Furthermore, we optimized the reaction conditions  and temperature (entries 1 and 14), a methylated product was finally obtained in 94% yield using 1 mol% Pd(OAc)2/2 mol% RuPhos with MeOH/H2O as a solvent at 80 °C for 12 h (entry 13). The yields were low when 1.1 or 1.3 equivalents of boronic acid were used (52% or 83%), but they were increased to practical levels in the presence of 1.5–2.0 equivalents of boronic acid.Among phosphine ligands screened for optimizations, RuPhos was found to be best ligand to achieve quantitative yield (entry 7). The use of Brettphos gave a methylation product as in the case of SPhos (entry 8). Other monodentate ligands such as Johnphos, XPhos and PCynditions . The rea2 (1 mol%) and RuPhos (2 mol%). 2-Naphthyliodide showed a slight decrease in reactivity compared to the corresponding bromide and chloride (entries 6–8). It was also interesting that the steric hindrance of ortho-substituents did not affect the yields (entries 10–12). Under the optimized reaction conditions, the scope for representative aryl halides is summarized in The use of 1-bromo-2-methoxynaphthalene, 1-bromo-2-methylnaphthalene and 2-bromo-1,3,5-trimethylbenzene resulted in yields of 96%, 88% and 86%, respectively. The reaction is highly sensitive to electron density of halides. For example, the methylation of furyl and thienyl halides results in low yields (entries 14 and 15).1H-NMR spectra were recorded on a JEOL ECX-400 (400 MHz) in CDCl3 with tetramethylsilane as an internal standard. Chemical shifts are reported in part per million (ppm), and signal are expressed as singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m), and broad (br). 13C-NMR spectra were recorded on a JEOL ECX-400 (100 MHz) in CDCl3 (δC = 77.0) with tetramethylsilane as an internal standard. 11B NMR spectra was recorded on a JEOL ECX-400 (128 MHz) with BF3·OEt2 as an external standard. Chemical shifts are reported in part per million (ppm). Kanto Chemical silica gel 60N (particle size 0.063–0.210 mm) was used for flash column chromatography. All reactions were conducted under an atmosphere of nitrogen. Glassware was oven dried at 130 °C and allowed to cool under a stream of dry nitrogen. All chemicals were purchased from Aldrich, Wako, TCI, or Kanto Chemicals and used as received.1H-NMR (DMSO-6d) δ = 3.40 , 0.37 , −0.75 ; 13C-NMR (DMSO-6d) δ = 73.5, 34.5, 16.9, 6.54; 11B NMR (DMSO-6d) δ = 1.44.MeLi (50 mmol) in ether was added to a solution of triisopropoxyborane (50 mmol) in ether (100 mL) at −78 °C. The resulting mixture was stirred for 30 min at −78 °C, then allowed to warm to room temperature and was stirred for 8 h. 1,1,1-tris(hydroxymethyl)ethane (50 mmol) was then added in one portion, and the resulting mixture was stirred at 60 °C for 1 h. The mixture is poured into 1 L of acetone. The solid product is isolated by filtration, washed with acetone and dried under vacuum to afford 7.3 g (97%) of lithium methyltriolborate as a white solid. 2O (2.5 mL/0.5 mL) was added, and then the mixture was stirred for 30 min at room temperature. After addition of lithium methyltriolborate (1 mmol) and aryl halide (0.5 mmol), the mixture was heated at 80 °C for 12 h. After cooling to room temperature, the product was extracted with benzene, and dried over anhydrous MgSO4. The desired product was purified by column chromatography on silica gel.Palladium acetate (1 mol%) and RuPhos (2 mol%) were placed in a flask under an atmosphere of nitrogen. MeOH/H4-Methylbiphenyl (entry 1): 1H-NMR (CDCl3) δ = 7.58 , 7.49 , 7.42 , 7.42 , 7.32 , 7.25 , 2.40 .p-Methylacetophenone (entry 2): 1H-NMR (CDCl3) δ = 7.82 , 7.22 , 2.54 , 2.37 .1-Methyl-4-phenoxybenzene (entry 3): 1H-NMR (CDCl3) δ = 7.33 , 7.16 , 7.09 , 7.01 , 6.95 , 2.36 .2-Methoxy-6-methylnaphthalene (entry 4): 1H-NMR (CDCl3) δ = 7.70 , 7.59 , 7.34 , 7.19 , 7.15 , 3.94 , 2.53 .2,7-Dimethylnaphthalene (entry 5): 1H-NMR (CDCl3) δ = 7.76 , 7.58 , 7.31 , 2.56 .2-Methylnaphthalene (entries 6–8): 1H-NMR (CDCl3) δ = 7.82–7.72 , 7.67 , 7.51–7.40 , 7.33 , 2.57 .1-Methyl-4-nitrobenzene (entry 9): 1H-NMR (CDCl3) δ = 8.10 , 7.31 , 2.45 .2-Methoxy-1-methylnaphthalene (entry 10): 1H-NMR (CDCl3) δ = 8.03 , 7.86 , 7.78 , 7.56 , 7.43 , 7.32 , 3.99 , 2.65 .1,2-Dimethylnaphthalene (entry 11): 1H-NMR (CDCl3) δ = 8.08 , 7.85 , 7.67 , 7.54 , 7.46 , 7.35 , 2.65 , 2.54 .1,2,3,5-Tetramethylbenzene (entry 12): 1H-NMR (CDCl3) δ = 6.88 , 2.29 , 2.18 .2′-Methylbiphenyl-4-carbonitrile (entry 13): 1H-NMR (CDCl3) δ = 7.72–7.68 , 7.44–7.41 , 7.32–7.25 , 7.19 , 2.26 .Methyl 3-methylbenzoate (entry 14): 1H-NMR (CDCl3) δ = 7.85–7.82 , 7.36–7.29 , 3.89 , 2.38 .Methyl 5-methylfuran-2-carboxylate (entry 15): 1H-NMR (CDCl3) δ = 7.05 , 6.08 , 2.34 .2-Acetyl-5-methylthiophene (entry 16): 1H-NMR (CDCl3) δ = 7.48 , 6.76 , 2.50 , 2.41 .In summary, we have demonstrated the efficiency of lithium methyltriolborate for methylation of aryl halides. This borate showed several advantages over boronic acid, including high nucleophilicity of methyl groups for smooth transmetalation to a palladium catalyst. Since the use of a base is avoided, a variety of functional groups may be accommodated in this reaction system.


AbstractA total of 162 species and subspecies of marine macroinvertebrates were recorded in the submerged soft and hard substrates around the PERU LNG marine terminal and surrounding area, in the central coast of Peru, 167 km south of Lima, Peru. The collection of specimens was carried out from June 2011 to June 2015 as part of the research studies conducted by the Biodiversity Monitoring and Assessment Program (BMAP) around the marine terminal. The area is part of the Humboldt Current Large Marine Ecosystem, one of the most important upwelling systems in the world.Annelida, Arthropoda, Brachiopoda, Cnidaria, Echinodermata, Bryozoa and Mollusca. Phyla Annelida (60 spp.), Arthropoda (47 spp.)and Mollusca (45 spp.) exhibited the largest number of species.We identified specimens belonging to 83 families and seven phyla. The list was assembled from the taxonomic identifications made by the BMAP. We identified species and subspecies belonging to phyla  Marine studies along the coastline of Peru are largely focused on species of economic importance . SpeciesHere, we have assembled a taxonomic list of macrobenthic species present in the shallow coastal area near the international marine terminal of company PERU LNG (PLNG), in Pampa Melchorita, on the central coast of Peru, in the southeast Pacific. The area where the terminal is located is part of the Humboldt Current Large Marine Ecosystem (HCLME), an important upwelling system encompassing submerged habitats from the southern tip of Chile to northern Peru. The HCLME is considered amongst the most productive marine ecosystems in the world and knowledge about the diversity and natural processes characterising it are in great need, given its importance in global fisheries economy . This ta2 Van Veen grab was used. Samples were sieved through a bag of 0.5 mm2 mesh size and the retained material was fixed with 4% formaldehyde in seawater. To facilitate later triage work and taxonomic analysis, the samples were stained with 1% rose bengal. To collect samples from hard substrate, a 25 cm x 25 cm quadrat was used and samples were obtained by clearing all specimens within the quadrat using a chisel and hammer.The checklist of benthic macrofauna species was assembled using data collected by the Biodiversity Monitoring Assessment Program (BMAP). This programme is carried out in collaboration with the Smithsonian Conservation Biology Institute and PLNG. The area of study is the area of influence of PLNG marine terminal , situated 167 km south of Lima, Peru. The submerged area is characterised by sediment flats with scattered patches of hard bottom and new artificial hard bottom created after establishment of an 800 m-long breakwater. Sampling was carried out close to the PLNG marine terminal and surroundings from June 2011 to June 2015, with a biennial periodicity. Samples were taken in three replicates both from soft and hard substrates . In this study, we only considered taxa identified at species or subspecies levels. Names of higher taxa as well as names of species and subspecies within them are listed alphabetically. For each of the species, we provide information about its original combination, the type of substrate (soft or hard), depth or bathymetric range, code of station where it was reported (with the name of transect and the depths in parentheses). We include remarks when necessary, particularly in the case of identified non-native species. Voucher specimens were deposited at the scientific collection of Laboratorio de Biología y Sistemática de Invertebrados Marinos (LabSIM) at Universidad Nacional Mayor de San Marcos (UNMSM).We estimated species richness to test sampling effort using the non-parametric estimator that better fits our occurrence-data from multiple samples Chao2, , implemeData package title: Macrobenthos_bmapperuResource link: http://ipt.pensoft.net/resource?r=macrobenthos_bmapperu&v=1.2Number of datasets: 1Data set name: Macrobenthos from upwelling coastal area of PeruData format: Darwin Core Archive (DwC-A)Arenicolaassimilisaffinis Ashworth, 1902Types of substrate: soft bottom. Depth / bathymetric range: 0-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N(15); BT3S;BT4N.Hartmann-Schröder, 1962Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N ; BT1S; BT2N; BT2S; BT3S(8); BT4N(8); D1; D2; D3; D4(0).Clymenellarubrocincta Johnson, 1901Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D2.Hartmann-Schröder, 1965Material examined: Fig. Stauronereisannulatus Moore, 1906Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2; D3; D4.Hartmann-Schröder, 1960Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(5); D4.Hartmann-Schröder, 1960Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-10 m. Station code: BT2S(0); D1; D2; D3; D4; D5(0).Quatrefages, 1866Material examined: Fig. Hartman, 1944Types of substrate: hard and soft bottoms. Depth / bathymetric range: 8-15 m. Station code: BT1N(10); BT1S; BT2N; BT2S; BT3S; MU(17).Haploscoloploskerguelensischilensis Hartmann-Schröder, 1965Types of substrate: soft bottom. Depth / bathymetric range: 8-12 m. Station code: BT1N; BT1S(10); BT2N(12); BT2S; BT3N; BT3S(8).Scoloploskerguelensis McIntosh, 1885Types of substrate: soft bottom. Depth / bathymetric range: 8-12 m. Station code: BT1N(10); BT1S; BT2S(12); BT4N(10).Hartmann-Schröder, 1960Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2(5); D3; D4.Hartmann-Schröder, 1962Material examined: Fig. Ariciarubra Webster, 1879Types of substrate: soft bottom. Depth / bathymetric range: 0-12 m. Station code: BT1N; BT1S; BT2N(0).Hartman, 1955Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N; BT3S; BT4N.Leidy, 1855Types of substrate: hard and soft bottoms. Depth / bathymetric range: 5-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N; BT3S(8); BT4N; D3Hemipoduscaliforniensis Hartman, 1938Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-12 m. Station code: BT1N; BT1S(0); BT2N; BT2S; BT3S(0); BT4N(0); D2.Grube, 1870Types of substrate: soft bottom. Depth / bathymetric range: 0-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N(10); BT3S; BT4N.Hartmann-Schröder, 1962Material examined: Fig. Hartmann-Schröder, 1962Types of substrate: soft bottom. Depth / bathymetric range: 8-10 m. Station code: BT2N.Podarkefurcata Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2; D3; D4; D5(0).Hartman, 1940Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N(10); BT1S; BT2N; BT3N(8); BT4N(8).Baird, 1873Material examined: Fig. Nereilepascallaona Grube, 1857Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N(15); BT1S(10); BT4N(8); D1; D2; D3; D4; D5(0).Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D3(0).Kinberg, 1865Material examined: Fig. Gravier, 1907Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D3; D4(0).Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(10); D2; D3; D4(0).Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2; D3; D4; D5(5).Hartmann-Schröder, 1962Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-12 m. Station code: BT1N(4); BT1S; BT2N; BT2S; BT3S; BT4N(8); D1(0); D2(0); D3.Hartmann-Schröder, 1960Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5(0).Loandaliafauveli Berkeley & Berkeley, 1941Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N(12); BT3S; BT4N; D3(0).Ancistrosyllisbassi Hartman, 1945Types of substrate: soft bottom. Depth / bathymetric range: 10 m. Station code: BT1N(10); BT1S(10); BT4N(10).Polynoefuscomarmorata Grube, 1876Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(0); D2; D3; D4(0); D5(0).Lepidonotusjohnsoni Darboux, 1899Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5(0).Kinberg, 1856Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D3; D4(0).Johnson, 1897Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D3(10).Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2(10); D3; D4(0).Peisidiceaspera Johnson, 1897Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D2(10).Siewing, 1955Types of substrate: soft bottom. Depth / bathymetric range: 0 m. Station code: BT1N(0); BT1S(0); BT2N(0); BT2S(0); BT3S(0); BT4N(0).Kinberg, 1856Types of substrate: soft bottom. Depth / bathymetric range: 10-15 m. Station code: BT1S(10); BT4N(15).Odontosyllisliniata Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4(0).Aguirre, San Martín & Álvarez-campos, 2015Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D1(10); D2(10); D3(10). Remarks: This species was described recently from the study area Odontosyllismicropedata Hartmann-Schröder, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(10); D2; D3; D4(10).Syllisprolixa Ehlers, 1901Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(10); D2(10); D3; D4(0).Choneparacincta Hartmann-Schröder, 1962Types of substrate: hard and soft bottoms. Depth / bathymetric range: 8- 10 m. Station code:BT1N(8); BT2N(10); D3(10).Demonaxleucaspis Kinberg, 1867Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D1(10); D2(10); D3(10).Polydorapolybranchia Haswell, 1885Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4.Blake, 1979Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D3(0); D4(0).Leucodoresociales Schmarda, 1861Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D2; D3.Prionospio (Paraprionospio) pinnata Ehlers, 1901Material examined: Fig. Blake, 1969Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-12 m. Station code: BT1N(10); BT1S; D1(5); D2(0); D3; D4(0).Hartman in Loosanoff & Engle, 1943Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2 D3; D4. Remarks: considered as non-native species, with North American Pacific coast being its native distribution area (Cinar 2012). Considered as an invasive species in Hawaii Islands (Cinar 2012), Venezuela and Australia (Díaz and Liñero-Arana 2009), which has caused extensive damage to oysters.Hartmann-Schröder, 1962Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N; BT3S; BT4N.Scolecolepisglutaea Ehlers, 1897Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-10 m. Station code: BT1N(10); BT1S(10); BT2N(10); BT2S(10); D3(5); D4(0).Nerinecirratuluschilensis Hartmann-Schröder, 1962Material examined: Fig. Lumbricussquamatus O.F. Müller, 1806Types of substrate: soft bottom. Depth / bathymetric range: 8-112 m. Station code: BT1N(10); BT1S; BT2N(10); BT2S; BT3S(8); BT4N(8).Nilsson, 1928Types of substrate: hard and soft bottom. Depth / bathymetric range: 10-15 m. Station code: BT1N(12); BT1S; BT2N(15); BT2S(15); BT3S; D1(10).Kinberg, 1866Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-10 m. Station code: BT1N(10); D1; D2; D3; D4.McIntosh, 1885Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D3(10).Templeton, 1836Types of substrate: hard and soft bottom. Depth / bathymetric range: 5-12 m. Station code: BT1N(12); BT2N(10); BT1S(12); D2(5). Remarks: considered as non-native species, being native to the western Indian .Corophiumacherusicum Costa, 1853Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2; D3. Remarks: considered as non-native species in Chile . Its natCorophiuminsidiosum Crawford, 1937Types of substrate: hard and soft bottom. Depth / bathymetric range: 0-15 m. Station code: BT1N(15); BT2N(15); BT3S(15), D1; D2; D3; D4. Remarks: considered as non-native species in Chile . After FPodoceruspunctatus Bate, 1857Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1 ; D2 ; D3; D4.Costa, 1853Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1S; BT2S(10), BT3N(15); BT3S(8); D1; D2; D3; D4; D5(0). Remarks: Considered as non-native species from the Pacific . After HDana, 1852Types of substrate: Hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N(15); D2(5); D3.Rathbun, 1898Types of substrate: soft bottom. Depth / bathymetric range: 10-12 m. Station code: BT1S(10); BT2S(12). Remarks: This species has a North American distribution  and for Benedict, 1903Types of substrate: soft bottom. Depth / bathymetric range: 0-8 m. Station code: BT1S(0); BT2S(8).Lenz, 1902Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D3(5).Wicksten & Méndez, 1981Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D3.Alpheusspinifrons Milne Edwards, 1837 :Material examined: Fig. Cancerpolyodon Poeppig, 1836Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D3(0).Milne Edwards, 1834Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D3(0).Grapsustransversus Gibbes, 1850Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D1(0); D3(0); D5(0).Hippaanaloga Stimpson, 1857Types of substrate: soft bottom. Depth / bathymetric range: 0-8 m. Station code: BT1N(0); BT1S(0); BT2N(0); BT2S; BT3S(0); BT4N(0).Milne Edwards, 1848Types of substrate: soft bottom. Depth / bathymetric range: 8-10 m. Station code: BT1S(8); BT2N(8); BT2S(10); BT3S(8).Nicolet, 1849Material examined: Fig. Panopeustransversus Stimpson, 1860Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2, D3.Hepatusperlatus Poeppig, 1836Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5(0).Ostracotherespolitus Smith, 1870Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(10); D2(10); D3.Rathbun, 1907Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N(10); BT1S(15); BT2N.Xanthoorbignyi Milne Edwards & Lucas, 1843Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2(10); D3.Porcellanaangulosa Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(5); D3(0).Porcellanapunctata Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D3(0); D5(0).Porcellanaspinifrons Milne Edwards, 1837Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D4(5).Porcellanamitra Dana, 1852Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2(5); D3; D4; D5.Haig, 1960Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2(0); D3; D4(0); D5(5).Porcellanagrossimana Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D3(10).Gibbes, 1850Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D1(5); D2(5)Porcellanadesmarestii Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D3(10).Porcellanagranulosa Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D2(5); D3.Porcellanalaevigata Guérin, 1835Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D3.Euctenotamexicanus Gerstaecker, 1856Material examined: Fig. Portunusruber Lamarck, 1818Material examined: Fig. Cancersetosus Fabricius, 1798Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D1(10); D2(10); D3(5).Xanthogaudichaudii Milne Edwards, 1834Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(5); D2(5); D3.Leptodiustridentatus Lenz, 1902Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2(5); D3.Lemos de Castro, 1959Types of substrate: soft bottom. Depth / bathymetric range: 10 m. Station code: BT1N(10). Remarks: It is a species from Western Atlantic  probablyGlynn & Glynn, 1974Material examined: Fig. Richardson, 1912Types of substrate: soft bottom. Depth / bathymetric range: 0-10 m. Station code: BT1N(0); BT1S(0); BT2N(0); BT2S; BT3S(0); BT4N(0).Menzies, 1962Types of substrate: soft bottom. Depth / bathymetric range: 10-12 m. Station code: BT1N.Dynamenopsisbakeri Menzies, 1962Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2(0); D3; D4.Lepaspsittacus Molina, 1782Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5(0).Bruguière, 1789Material examined: Fig. Balanustintinnabulum Linnaeus, 1758Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(10) D2(0); D3.Darwin, 1854Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(0); D2(0); D3; D4(0).Chthamalusscabrosus Darwin, 1854Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2; D3(0); D4(0); D5(0).Orbiculalamellosa Broderip, 1833Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-10 m. Station code: BT4N(8); D1; D2; D3; D4.Sertularianeritina Linnaeus, 1758Types of substrate: soft bottom. Depth / bathymetric range: 10-15 m. Station code: BT1N; BT3N(15). Remarks: It is a biofouling species and considered as non-native in Australia and Europe .Metridiumconcinnatum Drayton in Dana, 1846Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(5); D3.Actiniachilensis Lesson, 1830Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-10 m. Station code: BT1S(10); BT2N(10); BT2S; BT4N(0); D1; D2(10); D3; D4(0); D5(0).Echinusspatuliger Valenciennes, 1846Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D4; D5(5).Echinusniger Molina, 1782Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5.Echinus (Toxopneustes) gibbosus L. Agassiz & Desor, 1846Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3(0); D5(5).Echinusalbus Molina, 1782Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D1; D2(5).Lütken, 1856Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-12 m. Station code: BT1S(10); BT2S(12); BT4N; D1; D2; D3; D4; D5(0).Mytilusrugosus Linnaeus, 1767Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D1(10); D3(10).Solenscalprum King, 1832Material examined: Fig. Pholassubtruncata G.B. Sowerby I, 1834Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D3(10).Mytilusater Molina, 1782Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D2(5).Mytilusgranulatus Hanley, 1843Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1; D2(0); D3(0).Modiolapurpurata Lamarck, 1819Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2(0); D3; D4; D5(0).Mytilusalgosus Gould, 1850Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N(10); BT3S; BT4N; D1; D2; D3; D4; D5(0).Reeve, 1854Types of substrate: soft bottom. Depth / bathymetric range: 8 m. Station code: BT1N(8); BT1S(8); BT2N(8); BT2S(8); BT3S(8).Poroniapetitiana Récluz, 1843Types of substrate: hard and soft bottoms. Depth / bathymetric range: 5-10 m. Station code: BT1N(10); BT2N(15); D2(10); D3.Mactravelata Philippi, 1849Types of substrate: soft bottom. Depth / bathymetric range: 10 m. Station code: BT3S(10).Mactraedulis King, 1832Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N ; BT3S ; BT4N.Bernard, 1983Types of substrate: hard and soft bottoms. Depth / bathymetric range: 0-15 m. Station code: BT1N; BT1S(8); BT3N(15); BT3S; D1; D2; D3; D4.Lamarck, 1822Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(0); D2(0); D3(0); D4.Sowerby, 1835Material examined: Fig. Sowerby, 1835Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4(0); D5(0).Sowerby, 1834Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2; D3; D4(0).Lamarck, 1822Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(0); D2; D3(0) D4(0).Acmaeaorbignyi Dall, 1909Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D1(0); D4(0).Patellaceciliana Orbigny, 1841Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D4(0).Lottiavariabilis Sowerby, 1839Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D1(0); D4(0).Patellaviridula Lamarck, 1819Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D4(0).Blainville, 1827Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D4(0).Trochusater Lesson, 1830Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D4; D5.Trochuseuryomphalus Jonas, 1844Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D1(5); D2(5).Trochusluctuosus Orbigny, 1841Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D1; D2(5).Monodontatridentata Potiez & Michaud, 1838Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1(0); D2(5).Turboniger W. Wood, 1828Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3(5); D4(5); D5.Stuardo, 1962Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D4(5).Crepiduladilatata Lamarck, 1822Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4; D5(0).Turbotrochiformis Born, 1778Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D1(10).Phasianellaperuviana Lamarck, 1822Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D5(0).Albuladidyma Röding, 1798Types of substrate: soft bottom. Depth / bathymetric range: 8-15 m. Station code: BT1N; BT1S; BT2N; BT2S; BT3N(15); BT3S; BT4N.Naticacymba Menke, 1828Types of substrate: soft bottom. Depth / bathymetric range: 8-12 m. Station code: BT1S; BT3S(8).Columbellaunifasciata G. B. Sowerby I, 1832Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D1(5).Buccinumconcholepas Bruguière, 1789Types of substrate: hard bottom. Depth / bathymetric range: 0-5 m. Station code: D1; D2.Murexcrassilabrum Sowerby, 1834Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D1(5); D2(10).Buccinumhaemastoma Linnaeus, 1767Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4(0); D5(0).Purpurachocolata Duclos, 1832Types of substrate: hard bottom.Depth / bathymetric range: 0-10 m. Station code: D1(0); D2; D3.Murexbuxeus Broderip in Broderip & Sowerby, 1833Types of substrate: hard bottom. Depth / bathymetric range: 5-10 m. Station code: D1.Nassadentifera Powys, 1835Types of substrate: hard and soft bottoms. Depth / bathymetric range: 8-12 m. Station code: BT1S(10); BT2N(10), BT2S; D2(10).Millen, Schrödl, Vargas & Indacochea, 1994Types of substrate: hard bottom. Depth / bathymetric range: 10 m. Station code: D2(10).Marcus, 1955Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D2(5); D3.Chitonhennahi Gray, 1828Types of substrate: hard bottom. Depth / bathymetric range: 5 m. Station code: D4(5).Chitonechinatus Barnes, 1824Types of substrate: hard bottom. Depth / bathymetric range: 0 m. Station code: D2(0).Frembly, 1827Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1; D2; D3; D4(0).Frembly, 1827Types of substrate: hard bottom. Depth / bathymetric range: 0-10 m. Station code: D1(0); D2; D3; D4(0).We recorded 162 species and subspecies of marine macroinvertebrates in the submerged soft and artificial hard substrates around the PERU LNG marine terminal and surrounding area between June 2011 and June 2015 , followed by Crustacea and Mollusca with 47 and 45 species, respectively. Less numerous in species, but present in the study area, were the phyla Brachiopoda, Bryozoa, Cnidaria and Echinodermata (one to five species). The photographs for some of the species listed in this study are presented in Fig. The This study reports the diversity of macrobenthonic species associated with the coastal soft and hard bottom habitats around PERU LNG marine terminal in central Peru. In general, species richness and taxonomic composition observed in our study area are similar to other upwelling areas, north of the terminal, like Ancon Bay  and ChanPolychaeta species, Myrianidaparedesi, described from specimens obtained from biofouling from main pier piles at PERU LNG marine terminal . The most likely vector of introduction may be the maritime traffic occurring along the coast of Peru.We highlight the report of a new terminal . Areas wAbarenicolaaffinischilensis, Capitellacapitata, Cirratulusmegalus, Dodecaceriaopulens, Eunicepelamidis, Hemipodiasimplex, Kinbergonuphismicrocephala, K.multidentata, Lumbrinerisannulata, Magelonaphyllisae, Paleanotuschrysolepis, Phymactisclematis, Pisioneoerstedii, Polydorapygidialis, Scoletomatetraura, Syllisgracilis, Thoracopheliamucronata and Spiophanesnorrisi. We recommend an increased sampling effort as well as an extensive review to confirm their presence in the area. Further, the application of molecular tools (i.e. barcode sequencing) could be integrated into the analyses to help improving biodiversity assessments (e.g. We detected species in our study area (but not listed here) that were challenging to identify because their presence was limited to individuals at early developmental stages (i.e. juveniles), they were present in low numbers or because of taxonomic complexity. These putative species include nts e.g.  and for nts e.g. . Moleculnts e.g.  and imprnts e.g. . ConsideSupplementary material 1Table 1. Sampling sites at central coast of Peru and influence area of PERU LNG marine terminal, including depth(s), type of substrate and geographic decimal coordinates.Data type: Formatted textBrief description: Word table with geographic information of sampling sitesFile: oo_221022.docxV. Tasso, M. El Haddad, C. Assadi, R. Canales, L. Aguirre, X. Velez-ZuazoSupplementary material 2Macrobenthos species ocurrence listData type: Darwin Core Archive (.zip) of occurrence data and associated metadataBrief description: Resource link: http://ipt.pensoft.net/resource?r=macrobenthos_bmapperu&v=1.2File: oo_225921.zipTasso V, El Haddad M, Assadi C, Canales R, Aguirre L, Velez-Zuazo XSupplementary material 3Species accumulation curve using rarefaction method for macrobenthos reported at soft-bottom sampling sites. Light blue shaded area indicates 95% confidence interval.Data type: imageFile: oo_226161.jpegTasso V, El Haddad M, Assadi C, Canales R, Aguirre L, Velez-Zuazo XSupplementary material 4Species accumulation curve using rarefaction method for macrobenthos reported at hard-bottom sampling sites. Light blue shaded area indicates 95% confidence interval.Data type: imapeFile: oo_226162.jpegTasso V, El Haddad M, Assadi C, Canales R, Aguirre L, Velez-Zuazo X


Interaction of 3 with DMF/POCl3 gave β-chloroacroline derivative 8. Treatment of 3 with malononitrile gave benzo[c]chromone and 2-aminobenzonitrile derivatives 9 and 10, respectively with respect to the reaction conditions. Condensation of 3-(2-bromoacetyl)-8-ethoxycoumarin (4) with o-phenylenediamine gave 3--8-ethoxycoumarin hydrobromide (11), while 4 reacted with 2-aminopyridine to give chromenopyridopyrimidine derivative 12. Condensation of 4 with potassium thio-cyanate/methanol gave an unexpected derivative, 2H-chromeno-3-carboxy(methyl-carbonimidic)thioanhydride 16, which upon treatment with (NH2)2·H2O gave 3-ethoxy-2-hydroxybenzaldehyde azine 19. Interaction of 4 with thiourea derivatives gave thiazole derivatives 20a–c. The structures of the newly synthesized compounds were confirmed by their spectra data. The newly synthesized compounds were also screened for their antimicrobial activity.Condensation of 3-acetyl-8-ethoxycoumarin ( Coumarin and its derivatives are used as additives in food, perfumes, cosmetics, pharmaceuticals, agrochemicals ,2, for t1) with ethyl acetoacetate (2) in boiling ethanol containing few drops of piperidine afforded 3-acetyl-8-ethoxycoumarin (3) [3 in acetic acidgave the corresponding ω-bromo-8-ethoxy-3-acetylcoumarin (4) [Treatment of 3-ethoxysalicylaldehyde (arin (3) . Brominaarin (4)  Scheme  with eth3 with thiosemicarbazide afforded 2-[1-(8-ethoxycoumarin-3-yl)ethylidene]-hydrazinecarbothioamide (5). Reaction of 5 with chloroacetic acid or ethyl chloroacetate afforded 2-[(1-(8-ethoxycoumarin-3-yl)ethylidene]hydrazonothiazolidin-4-one (6). Treatment of the thiazolidin-4-one derivative 6 with p-methoxybenzaldehyde or α-cyano-p-methoxycinnamonitrile in ethanolic piperidine afforded the corresponding 2-[(1-(8-ethoxycoumarin-3-yl)ethylidene)hydrazono]-5-(4-methoxybenzylidene)thiazolidin-4-one (7) (Condensation of -one (7) .5–7 were established on the basis of spectral data. IR spectra showed the presence of NH2 and NH absorptions at υ 3,404, 3,254, 3,174 cm−1 and CO at υ 1,720 cm−1 for 5, NH at υ 3,147 cm−1, υ 2,977 C-H , CO at υ 1,720, 1,648 cm−1 for 6 and a CO at υ 1,705, 1,655 cm−1 and υ 1,605 cm−1 for N=C and 2,939.3 cm−1 for 7. The 1H-NMR spectrum of 6 showed signals at δ 12.05 ppm , 3.87 ppm , 2.49 ppm  and that of 7 signals atδ 12.40 , 8.44 ppm , 8.25 ppm , 3.84 ppm . The mass spectra of compounds 5,6 provided additional evidence in support of the proposed structures.Structures 3) with DMF/POCl3 gave 3-chloro-3-(8-ethoxycoumarin-3-yl)acrylaldehyde(8)  . The forvative 8 .3 with malononitrile or ethyl cyanoacetate in ethanolic triethylamine solution gave the same product, the benzo[c]chromene-6-one derivative 9  as intermediate which undergoes an intramolecular cyclization through the addition of an active methylene group to the activated 3,4-double bond forming the intermediate (B), which then underges spontaneous oxidation to the final product 9 pyridine-2-yl-8-ethoxychrome-2-one 13 , 8.74  for 22. The mass spectrum of 20c gave a molecular ion peak at m/z (%) 394  and 392  with a base peak at 284 (M+-(HBr+CO), 100); for 21: m/z (%) 448 , 450  and a base peak at 368 ; for 22: m/z: 449 , 447 , 367 .The structures of 20c with various aromatic aldehydes 23 afforded the corresponding dicoumarin-3-ylthiazole derivatives 24, 25 and iminocoumarin derivative 26, while condensation of 20c with aromatic aldehydes in methanolic piperidine solution give 27a,b. Compound 20c was also readily coupled with p-methoxybenzenediazonium chloride to afford 28 (28 was supported by its independent synthesis from 4 and (4-methoxyphenylazo)-2-cyanoethanethioamide (29) (Condensation of fford 28 . The stride (29) .24–28 were established on the basis of spectral data. The IR spectra showed the presence of a CO bands at υ 1,720 cm−1 for 24–26, while a NH band appeared at υ 3,449, 3,209 cm−1 for 26; a CN and at υ 2,207 cm−1 a CO band at υ 1,728 cm−1 for 27a; a CN band at υ 2,214 cm−1 and a CO band at υ 1,720 cm−1 for 27b and a NH band at υ 3,156 cm−1 a CN band at υ 2,214 cm−1 CO at υ 1,736 cm−1 for 28. The 1H-NMR spectra showed two signals at δ 8.95, 8.54 ppm  9.14  for 24; at δ 8.39  and 3.31 ppm  for 26 and δ 11.93 , 8.39  and 3.76 ppm  for 28. The mass spectra of compounds 24–28 provided additional evidence for the proposed structures.Structures 1H-NMR spectra were recorded using a Varian Mercury (300 MHz) spectrometer. The mass spectra recorded on a Shimadzu GC-MS QP 1000 EX spectrometer. Elemental analyses were performed on a Perkin-Elmer 240 microanalyser in the Cairo University Faculty of Science.Melting points were determined with a Stuart Scientific Co. Ltd. apparatus. IR spectra were determined as KBr pellets on a Jasco FT/IR 5300 spectrophotometer. Synthesis of 3-acetyl-8-ethoxycoumarin (3). 3-Ethoxysalicylaldehyde 1 (10 mmol) was refluxed with ethyl acetoacetate 2 (10 mmol) in ethanolic/piperidine solution  for two hours. The precipitate formed on cooling was collected by filtration, washed with cold ethanol and dried. The solid formed was recrystallized from diluted ethanol to give pale yellow crystals; yield 88%; m.p. 130–132 °C, lit. (135–7 °C) chromen-6-one (9). Method A: A solution of 1 (10 mmol), malononitrile (10 mmol) in absolute ethanol (30 mL) was refluxed in the presence of TEA (0.2 mL). A solid formed after 30 min. boiling and the refluxing were continued for 1.5 hours more. The crude product was recrystallized from an ethanol/benzene mixture to give colourless crystals; yield 77%; m.p. 236–8 °C; IR (KBr) υ (cm−1): 3,425 (OH), 3,094, 2,978, 2,928 (CH stretching), 1,704, 1,694 (CO); 1H-NMR (DMSO-d6) δ: 1.45 , 4.20 , 7.30–8.16 , 8.55  and 11.22 ; MS m/z (%): 444 , 388 (100), 360 (28), 304 (21), 189 (15); Anal. calcd. for C26H20O7: C, 70.25; H, 4.50. Found: C, 70.27; H, 4.52%. Method B: The reaction was carried out as described in Method A, but using ethyl cyanoacetate (1.13 mL) instead of malononitrile; yield (82%) (identity confirmed by m.p. and mixed m.p.).Synthesis of 2-amino-4,6-bis(8-ethoxycoumarin-3-yl)benzonitrile (10). The reaction was carried out with the same procedure described for compound 9 (Method A) but using absolute methanol as a solvent and piperidine as a catalyst; m.p. 330–332 °C, 75%; IR (KBr) υ (cm−1): 3,474, 3,320 (NH2), 3,092, 3,042, 2,980 (CH stretching); 2,206 (CN), 1,720, 1,696 (CO); 1H-NMR (DMSO-d6) δ: 1.43 , 4.23 , 7.31 , 7.69 , 8.37 ; MS m/z (%): 494 , 412 (100), 384 (19), 344 (28), 316 (44), 243 (19), 242 (10), 206 (22); Anal. calcd. for C29H22N2O6: C, 70.44; H, 4.48; N, 5.67; found: C, 70.40; H, 4.47; N, 5.68%.Synthesis of 3--8-ethoxycoumarin hydrobromide (11). A solution of 4 (5 mmol) and o-phenylenediamine (5 mmol) in absolute methanol (20 mL) was refluxed for 3 hours. The solid obtained was filtered, washed with ethanol and dried under vacuum. The crude product was recrystallized from ethanol/benzene mixture to give compound 11 as pale brown crystals; yield 77%; m.p. 208–209 °C; IR (KBr) υ (cm−1): 3,417 broad (NH), 3,098, 3,022, 2,987, 2,896 (CH stretching); 1,720 (CO); 1H-NMR (DMSO-d6) δ: 1.43 , 4.21 , 7.37–8.15 , 8.88 , 9.58 ; MS m/z (%): 400 , 398 , 370 (98), 368 (100), 289 (69), 205 (96), 102 (53), 76 (57); Anal. calcd. for C19H15BrN2O3: C, 57.16; H, 3.79; N, 7.02; found: C, 57.26; H, 3.69; N, 6.98%.7-(Bromo-(8-ethoxycoumarin-4-yl)methylene)-4-ethoxychromenopyrido-pyrimidin-6(7H)-one (12). Compound 12 was prepared from 4 (5 mmol) and 2-aminopyridine (5 mmol) according to the procedure described for 11 to give 12 from acetic acid as pale brown crystals; yield 63%; m.p. 231–233 °C; IR (KBr) υ (cm−1): 2,924 (CH stretching), 1,713, 1,692 (CO); 1H-NMR (DMSO-d6) δ: 1.18 , 1.37 , 4.02 , 4.18 , 6.46 , 7.04–8.36 ; MS m/z (%): 574 , 572 , 493 (100), 465 (97), 304 (49), 280 (30), 224 (51), 196 (41), 134 (17); Anal. calcd. for C29H21BrN2O6: C, 60.75; H, 3.69; N, 4.89; found: C, 60.85; H, 3.68; N, 4.91%.Synthesis of 2-bromomethylene-8-ethoxy-2H-chromene-3-carboxylic (methyl-carbonimidic)thioanhydride (16). A solution of compound 4 (10 mmol) in absolute methanol (40 mL) was refluxed with potassium thiocyanate (10 mmol) for 2 hours. The solid formed on cooling filtered off, washed with ethanol and dried under vacuum. The product was then recrystallized from acetic acid to give 16 as brown needles; yield 83%; m.p. 165–167 °C; IR (KBr) υ (cm−1): 3141 (NH), 3093, 3021, 2985, 2890 (CH stretching); 1717 (CO); 1H-NMR (DMSO-d6) δ: 1.41 , 4.10 , 4.17 , 7.16–7.54 , 7.88 , 8.58 ; MS m/z (%): 385 , 383 , 303 (100), 355 (37), 276 (67), 232 (44), 218 (28); Anal. calcd. for C15H14BrNO4S: C, 46.89; H, 3.67; N, 3.65; found: C, 47.01; H, 3.67; N, 3.67%.4-Bromo-2-(3-ethoxy-2-hyroxybenzylidine)-3-hydroxybut-3-enoic cyanic thioanhydride (18). A solution of 16 (10 mmol) was boiled in glacial acetic acid/methanol mixture . HCl gas stream was bubbled into the hot solution for 2 hours. The reaction mixture was allowed to cool down and the solid formed was filtered off and recrystalized from methanol as yellow crystals; yield 89%; m.p. 235–237 °C; Anal. calcd. for C14H12BrNO4S (368.97): C, 45.53; H, 3.25; Br, 21.41; N, 3.79; S, 8.67; found: C, 45.55; H, 3.27; Br, 21.42; N, 3.81; S, 8.68%.Synthesis of 3-ethoxy-2-hydroxybenzaldehyde azine (19). A solution of compound 16 (5 mmol) in absolute methanol (20 mL) was stirred at room temperature with 85% hydrazine hydrate (10 mmol) for 4 hours. The solid obtained filtered, washed with ethanol  and dried under reduced pressure. The product was recrystallized from acetic acid to give 19 as yellow crystals; yield 93%; m.p. 275–277 °C; IR (KBr) υ (cm−1): 3,302 (OH), 3,083, 3,025, 2,965, 2,870 (CH stretching), 1,562, 1,597 (C=N); 1H-NMR (DMSO-d6) δ: 1.34 , 4.09 , 6.87–7.35 , 9.20 , 12.10, 12.45 ; MS m/z (%): 328 , 164 (57), 136 (46), 121 (51), 80 (65); Anal. calcd. for C18H20N2O4: C, 65.84; H, 6.14; N, 8.53; found: C, 65.81; H, 6.10; N, 5.55%.Synthesis of 3-(2-aminothiazol-4-yl)-8-ethoxycoumarin (20a). Compound 4 (10 mmol) and thiourea (10 mmol) were disolved in absolute methanol (40 mL). The reaction mixture was refluxed for 1 hour. A precipitate formed on cooling, which was collected by filtration, washed with ethanol and dried under vacuum. The crude product was recrystallized from ethanol to give pale yellow crystals; yield 94%; m.p. 200–202 °C; IR (KBr) υ (cm−1): 3,387, 3,302, 3,148 (NH2), 3,020, 2,983, 2,887 (CH stretching), 1,705 (CO); MS m/z (%): 288 , 261 (26), 203 (21), 134 (22), 89 (40); Anal. calcd. for C14H12N2O3S: C, 58.32; H, 4.20; N, 9.72; found: C, 58.31; H, 4.18; N, 9.73%.Synthesis of 3-(2-(phenylamino)thiazol-4-yl)-8-ethoxycoumarin (20b). Compound 20b was prepared from 4 (10 mmol) and phenylthiourea (10 mmol) according to the procedure described for 20a to give 20b from ethanol/benzene mixture as yellow crystals; yield 69%; m.p. 216–218 °C; IR (KBr) υ (cm−1): 3,302, 3,148 (NH), 3,083, 3,011, 2,955, 2,850 (CH stretching), 1,697 (CO); 1H-NMR (DMSO-d6) δ: 1.41 , 4.13 , 7.00–7.76 , 8.61 , 10.31 ; MS m/z (%): 364 , 279 (27), 207 (12), 150 (39), 77 (41); Anal. calcd. for C20H16N2O3S: C, 65.92; H, 4.43; N, 7.69; found: C, 65.95; H, 4.40; N, 7.71%.Synthesis of 2-(4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)acetonitrile hydrobromide (20c). Compound 20c was prepared from 4 (10 mmol) and cyanothioacetamide (10 mmol) according to the procedure described for 20a to give 20c from benzene as yellow crystals; yield 96%; m.p. 195–197 °C; IR (KBr) υ (cm−1): 1,722 (CO), 3,021, 2,946, 2,890 (CH stretching), 2,262 (CN); 1H-NMR (DMSO-d6) δ: 1.43 , 4.16 , 4.55 , 7.29–7.32 , 8.42 ; 13C (75 MHz) (DMSO-d6) δ: 14.05 (CH3-ester), 22.50 (CH2-), 61.92 (CH2-ester), 115.47 (C-7), 116.43 (CN), 118.95 (C-5), 122.15 (C-4a), 124.26 (C-5 thiazole), 126.91 (C-6), 127.51 (C-3), 142.11 (C-4 thiazole), 144.92 (C-4), 151.32 (C-8a), 154.54 (C-8), 158.65 (C-2 thiazole), 159.89 (CO-lactone); MS m/z (%): 394 , 392 , 390 364 , 312 , 284 (M+-(HBr+CO), 100), 256 (43.6), 227 (18), 198 (13.7), 134 (11.2), 89 (27.1), 63 (23.1); Anal. calcd. for C16H13BrN2O3S: C, 48.87; H, 3.33; N, 7.12; found: C, 49.25; H, 2.84; N, 7.20%.Synthesis of 2-(4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)-3-ethoxyacrylonitrile hydrobromide (21). A solution of compound 20c (5 mmol) was refluxed with triethyl orthoformate (5 mL) for 3 hours. The excess triethyl orthoformate removed under reduced pressure. The residual solid was treated with ethanol, filtered off, washed with ethanol  and dried under vacuum. The crude product was recrystallized from benzene/petroleum ether 40–60 °C mixture to give compound 21 as green crystals; yield 83%; m.p. 145–147 °C; IR (KBr) υ (cm−1): 3,027, 2,966, 2,876 (CH stretching), 2,222 (CN), 1,728 (CO); 1H-NMR (DMSO-d6) δ: 1.39 , 1.42 , 4.17 , 4.45 , 7.28–7.45 , 8.15 , 8.43 ; MS m/z (%): 450 , 448 , 368 (100), 340 (50.1), 312 (67), 227 (29.3), 134 (35.7), 89 (49.9); Anal. calcd. for C19H17BrN2O4S: C, 50.79; H, 3.81; N, 6.23; found: C, 51.14; H, 3.35; N, 6.30%.Synthesis of 2-(4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)-3-(dimethylamino)acrylonitrile hydrobromide (22). A mixture of 21 (2.5 mmol) and dimethylamine (2.5 mmol) in absolute methanol (30 mL) was refluxed for 6 hours. The resulting solid was filtered off, washed with ethanol and dried. The crude product was then crystallized from acetic acid to give compound 22 as brown crystals; yield 89%; m.p. 206–208 °C; IR (KBr) υ (cm−1): 3,012, 2,944, 2,927, 2,890, 2,874 (CH stretching), 2,191 (CN), 1,720 (CO); 1H-NMR (DMSO-d6) δ: 1.41 , 3.28 2), 4.18 , 7.22–7.59 , 7.89 , 8.12 ; MS m/z (%); 449 , 447 , 367 , 334 (35.8), 230 (28.6), 170 (18.3), 132 (29.1), 89 (48.9); Anal. calcd. for C19H18BrN3O3S: C, 50.90; H, 4.05; N, 9.37; found: C, 51.24; H, 3.61; N, 9.46%.Synthesis of 3-(2-(coumarin-3-yl)thiazol-4-yl)-8-ethoxycoumarin hydrobromide (24). A solution of the acetonitrile derivative 20c (10 mmol) in absolute methanol (40 mL) was refluxed with salicyaldehyde (10 mmol) in the presence of piperidine for 2 hours. The solid formed was filtered off, washed with ethanol and dried under reduced pressure. The crude product was recrystallized from acetic acid to give compound 24 as pale yellow crystals; yield 91%; m.p. 265–7 °C; IR (KBr) υ (cm−1): 2,948, 2,927, 2,897, 2,874 (CH stretching), 1,720 (CO); 1H-NMR (DMSO-d6) δ: 1.43 , 4.19 , 7.31–8.00 , 8.54, 8.95 , 9.14 ; Anal. calcd. for C23H16BrNO5S: C, 55.34; H, 3.24; N, 2.81; found: C, 55.77; H, 2.85; N, 2.84%.Synthesis of 3-thiazol-4-yl)-8-ethoxycoumarine hydrobromide (25).Compound 25 was prepared from 20c (10 mmol) and 3,5-diiodosalicyaldehyde (10 mmol) according to the procedure described for 24 to give 25 from benzene as pale green crystals; yield 96%; m.p. 310–312 °C; IR (KBr) υ (cm−1): 2,938, 2,927, 2,898, 2,877 (CH stretching), 1,720 (CO); MS m/z (%): 749 , 669 (100), 668 (59.6), 641 (69), 613 (24), 585 (14), 321 (13), 190 (15), 134 (14), 89 (19). Anal. calcd. for C23H14BrI2NO5S: C, 36.83; H, 1.88; N, 1.87; found: C, 36.98; H, 1.63; N, 1.89%.Synthesis of 3-(2-(6-(dimethylamino)-2-iminocoumarin-3-yl)thiazol-4-yl)-8-ethoxycoumarin hydrobromide (26). Compound 26 was prepared from 20c (10 mmol) and 5-dimethylaminosalicyaldehyde (10 mmol) according to the procedure described for 24 to give 26 from benzene as brown crystals; yield: 90% m.p. 223–5 °C; IR (KBr) υ (cm−1): 3,449, 3,209, (NH), 3,015, 2,978, 2,921, 2,890, 2,873 (CH stretching), 1,597 (C=N), 1,720 (CO); 1H-NMR (DMSO-d6) δ: 1.43 , 3.31 , 4.19 , 6.63–7.67 , 8.39 ; Anal. calcd. for C25H22BrN3O4S: C, 55.56; H, 4.10; N, 7.78; found: C, 55.88; H, 3.74; N, 7.83%.20c (5 mmol) in absolute methanol (40 mL) was refluxed with 4-methoxybenzaldehyde and/or 4-chlorobenzaldehyde (5 mmol) in the presence of piperidine for 1 hour. The solid formed was filtered off, washed with ethanol and dried under reduced pressure. The crude product was recrystallized from a suitable solvent to give 27a,b. The physical and spectral data of compounds 27a,b are as follows:A solution of 3-(4-Methoxyphenyl)-2-(4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)acrylonitrile hydrobromide (27a). Yellow crystals; yield 91%; m.p. 215–7 °C; IR (KBr) υ (cm−1): 3,425, 3,139, 2,923 (NH and CH stretching), 2,207 (CN), 1,728 (CO); MS m/z (%): 510 [M+] (7.5), 508 (76.3), 430 (100), 265 (15), 216 (42), 134 (15), 89 (34); Anal. calcd. for C24H19BrN2O4S: C, 56.37; H, 3.74; N, 5.48; found: C, 56.71; H, 3.37; N, 5.53%.3-(4-Chlorophenyl)-2-(5-bromo-4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)acrylonitrile hydrobromide (27b). Yellow crystals; yield 86%; m.p. 196–7 °C; IR (KBr) υ (cm−1): 3,011, 3,332.8 and 3,224.8, 2,923 (NHBr and CH stretching), 2,214 (CN), 1,720 (CO); MS m/z (%): 516 , 514 , 512 , 433 (76.5), 295 (46), 215 (63), 125 (57), 89 (69); Anal. calcd. for C23H16BrClN2O3S: C, 53.56; H, 3.13; N, 5.43; found: C, 53.90; H, 2.71; N, 5.48%.Synthesis of 2-(2-(4-methoxyphenyl)hydrazono)-2-(4-(8-ethoxycoumarin-3-yl)thiazol-2-yl)acetonitrile hydrobromide (28). Method A: To a solution of compound 20c (10 mmol) in methanol (20 mL), a combination of 4-methoxyaniline (10 mmol), hydrochloric acid (5 mL) and sodium nitrite (10 mmol) in glacial acetic acid (20 mL) at 0 °C was added dropwise. The resulting mixture was stirred at 0 °C for 1 hour then the stirring was continued for 2 hours more at room temperature. The solid formed was filtered, washed with ethanol and dried under vacuum. The crude product was recrystallized from dioxane to give pale green crystals; yield 73%; m.p. 258–60 °C; IR (KBr) υ (cm−1): 3,157 (NH), 3,011, 2,883 (CH stretching), 2,214 (CN), 1,735.8 (CO); 1H-NMR (DMSO-d6) δ: 1.44 , 3.76 , 4.18 , 6.96–7.47 , 8.39 , 11.93 ; MS m/z (%): 526 , 445 (100). Anal. calcd. for C23H19BrN4O4S: C, 52.38; H, 3.63; N, 10.62; found: C, 52.69; H, 3.26; N, 10.69%. Method B: A solution of 4 (5 mmol) in absolute methanol (20 mL) was refluxed with (4-methoxyphenylazo)-2-cyanoethanethioamide 29 (5 mmol). The work up continued as previously mentioned in Method A to give 28; yield 79 %; (identified by m.p. and mixed m.p.).in vitro against two Gram-negative Bordetella bronchiseptica (ATCC 4617) and Escherichia coli (ATCC 14169) and four Gram-positive Bacillus pumilus (ATCC 14884), Bacillus subtilis (ATCC 6633), Staphylococcus aureus (ATCC 29737) and Staphylococcus epidermidis (ATCC 12228) pathogenic bacteria and two fungi Candida albicans (ATCC 10231) and Saccharomyces cervesia (ATCC 9080). The activities of these compounds were tested using the disc diffusion method [−1) as standard antibiotic Micostatin (25 μg mL−1) was used as a reference antifungal. The tested compounds were dissolved in N,N-dimethylformamide (DMF) to give a solution of 1 mg mL−1. The inhibition zones were measured in millimeters at the end of an incubation period of 48 hours at 28 °C. N,N-dimethylformamide (DMF) showed no inhibition zone. Test results are shown in The newly synthesized compounds were screened for their antimicrobial activities n method  for bactli ATCC 169 and fo1H-NMR, 13C-NMR and MS data. The screening results demonstrated that replacing the hydrogen atom attached to the coumarin nucleus at C-3 with a side chain as in compound 5 and thiazoles 7 and 28 results in wide spectrum antimicrobial activity against all tested bacteria and fungi compared to ampicillin and mycostatin, while the other compounds with other side chains showed moderate to weak activity.Our interest in the synthesis of the title compounds was to focus on their study as antimicrobial agents as a part of our program which is aimed at the development of new heterocyclic compounds as more potent antimicrobial agents. In this paper we reported the synthesis of some 8-ethoxycoumarin derivatives bearing side chains, thiazole derivatives and the antimicrobial evaluation of all the novel compounds. The structures of the novel compounds were elucidated on the basis of IR,


Arrabidaea pulchra (Cham.) Sandwith (EEAPL) that had shown in vitro activity in a previous screening using DNA and RNA viruses. The activity of EEPAL was evaluated against the DNA viruses Human herpesvirus 1 (HSV-1) and Vaccinia virus Western Reserve (VACV-WR) as well as against the RNA viruses Murine encephalomyocarditis virus (EMCV), and Dengue virus 2 (DENV-2) by the 3--2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Cytotoxicity was determined in LLCMK2 and Vero cells and the Selectivity Indexes (SI) were calculated. The most potent effect was observed against DENV-2 . For HSV-1 and VACV-WR EC50 values > 200 µg mL−1 were determined, while no inhibition of the cytopathic effect was observed with EMCV. Bioguided fractionation of EEAPL by partition between immiscible solvents followed by chromatography over a Sephadex LH20 column afforded two arylpropanoid glycosides, verbascoside (AP 1) and caffeoylcalleryanin (AP 2), along with a terpenoid, ursolic acid (AP 3). AP 1 and AP 3 exhibited similar anti-DENV-2 profiles, with SI values of 3.8 and 3.1, respectively, while AP 2 was the most effective anti-DENV-2 constituent, with a SI of 20.0. Our results show that A. pulchra leaves ethanol extract (EEAPL) affords compounds with antiviral activity, mainly against DENV-2.The aim of the present work was to carry out a bioguided isolation of antiviral chemical constituents from an ethanol extract of leaves from  Viral infections represent a current problem in industrialized and developing countries, accounting for severe damages to human health and economic losses in livestock. Plants afford an extensive chemical diversity and represent rich and renewable sources of natural products with promising biological activities. Antivirals of ethnomedicinal origin are of great interest and have been widely explored . AntivirSome viral diseases, such as dengue, which is considered the most important human arboviral disease, have a high impact in public health. It is estimated that 100 million cases occur every year around the World. The need for a safe and efficient approach either for treatment or prevention for virus infections has been a global priority, as neither an effective drug nor a vaccine for human use is available. However, investigations against dengue viruses have started in the previous decade and opportunities for the development of anti-dengue drugs have been pointed out by the WHO .Arrabidaea belongs to the tribe Bignonieae (family Bignoniaceae), a large clade of neotropical lianas occurring in Central America, Amazonia, the Atlantic forests of eastern Brazil, and the open dry forests and savannas of Argentina, Bolivia, Brazil, and Paraguay ; calculated for C29H35O15Na m/z 647.9–151 °C . UV . Light yellow powder. MP 167–171 °C. UV (MeOH): λmax (log ε) 217 (4.3), 250 (sh), 286.5 (4.1), 300 (sh), 329.5 (4.2) nm. IR: umax 3270, 1660, 1599, 1510, 1443, 1353, 1253, 1175, 1150, 1035, 803 cm−1. 1H-NMR (DMSO-6d): δ = 7.07 , 7.02 , 6.78 , 7.12 , 6.89 , 6.81 , 7.52 , 6.32 , 4.71 , 3.2–3,84 . 13C-NMR (DMSO-6d): δ = 131.0 (C1), 115.9 (C2), 145.1 (C3), 146.8 (C4), 116.7 (C5), 119.3 (C6), 65.1 (C7), 102.3 (C1′), 75.9 (C2′), 77.2 (C3′), 69.6 (C4′), 73.3 (C5′), 60.8 (C6′), 125.4 (C1′′), 114.8 (C2′′), 145.6 (C3′′), 148.6 (C4′′), 115.7 (C5′′), 121.4 (C6′′), 145.4 (C7′′), 113.7 (C8′′), 166.4 (C9′′). ESI-MS: found m/z 487.1 [M+Na]; calculated for C22H24O11Na m/z 487.0.Ursolic acid (AP 3). Colourless crystals. MP 282–284 °C, Lit 285–287 °C [max 3392, 2973, 2852, 1687, 1620, 1456, 1387, 1259, 1089, 867, 798; 1H-NMR (DMSO-6d): 5.23 , 3.0 , 2.11 , 0.69; 0.71; 0.87; 0.91; 1.08; 1.29 . 13C-NMR (DMSO-6d): δ 38.1 (C-1), 26.9 (C-2), 76.7 (C-3), 32.6 (C-4), 54.7 (C-5), 17.9 (C-6), 36.4 (C-7), 39.5 (C-8), 46.9 (C-9), 38.3 (C-10), 22.8 (C-11), 124.5 (C-12), 138.1 (C-13), 41.6 (C-14), 27.4 (C-15), 23.7 (16), 46.7 (C-17), 52.3 (C-18), 38.4 (C-19), 38.4 (C-20), 30.1 (C-21), 36.2 (C-22), 28.2 (C-23), 15.1 (C-22), 28.2 (C-23), 15.1 (C-24), 16.0 (C-25), 16.8 (C-26), 23.2 (C-27), 178.5 (C-28), 16.9 (C-29), 21.0 (C-30). EI-MS m/z (%): 43 (100), 55 (56), 67 (26), 68 (36), 79 (23), 81 (30), 91 (23), 93 (23), 95 (26), 105 (24), 107 (20), 109 (16), 133 (44), 147 (17), 163 (7), 175 (9), 189 (15), 203 (31), 207 (26), 248 (76), 249 (13), 456 (M+) (2).5–287 °C . IR: umaFingerprints were registered by RP-HPLC-DAD on a Waters 2695 apparatus equipped with a UV-DAD detector (Waters 2996). A LiChrospher 100 RP-18 column  was employed at 40 °C, flow rate of 1.0 mL/min and detection at wavelengths of 220, 280 and 350 nm. To an aliquot (10.0 mg) dried extract/fractions and (1.0 mg) of each of the isolated compounds , HPLC grade methanol was added and the mixture was dissolved by sonication in an ultrasound bath for 15 min, followed by centrifugation at 10,000 rpm for 10 min. The supernatant was filtered through a Millipore membrane (0.2 μm) and injected (10.0 μL) onto the equipment. Elution was carried out with a linear gradient of water (A) and acetonitrile (B) (from 5% to 95% of B in 60 min).2 cells were cultured in Dulbecco’s modified Eagle’s medium  at 37 °C, in 5% CO2 atmosphere, supplemented with 5% fetal bovine serum, 50 µg/mL gentamicin, 100 U/mL penicillin and 5 µg/mL amphotericin B. HSV-1 was obtained from the collection of Laboratório de Virus, UFMG, Belo Horizonte, Brazil. DENV-2, EMCV and VACV-WR were kindly donated by Dr. L. Figueiredo , Dr. I. Kerr  and Dr. C. Jungwirth , respectively. The viruses were titrated by TCID50 in Vero cells [6; 1.0 × 106; 1.0 × 106 and 1.0 × 104 TCID50/mL, respectively, for HSV-1, EMCV, VACV-WR and DENV-2 virus.Vero cells (ATCC CCL-81) and LLCMKro cells  and the 2 cells were exposed to different concentrations of extracts/fractions/compounds for 48 and 72 h. After incubation, cell viability was assessed by the 3--2,5-diphenyltetrazolium bromide  assay at a concentration of 2 mg mL−1 in PBS [−1. The cytotoxicity of each sample was expressed as CC50, i.e. the concentration of sample that inhibited cell growth by 50%.Vero and LLCMK1 in PBS . Each sa50) was evaluated by the MTT assay [50) for each extract, fractions, and constituents were calculated from concentration-effect-curves after no linear regression analysis  to be a promising source of the anti-dengue arylpropanoid glycoside derivatives verbascoside and caffeoylcallerianin, which might represent the main responsible constituents for its activity against DENV-2. Our findings are in line with the traditional use of Arrabidaea species as anti-infectious agents in different South American countries [Our results reveal the ethanol extract of ountries  and are ountries .