Bedaquiline is the first FDA-approved new chemical entity to fight multidrug-resistant tuberculosis in the last forty years. Our group replaced the quinoline ring with a naphthalene ring, leading to a new type of triarylbutanol skeleton. An asymmetric synthetic route was established for our bedaquiline analogues, and the goal of assigning their absolute configurations was achieved by comparison of experimental and calculated electronic circular dichroism spectra, and was confirmed by the combined use of circular dichroism and NMR spectroscopy. Tuberculosis is a widespread infectious disease that threatens one-third of the World population . Most ofM. tuberculosis . In. In90, aTo continue our research on determining which type of optical isomer of our analogues played a critical role in antituberculosis activity, we need to build a general asymmetric synthetic route to generate the four respective optical isomers and assign their absolute configurations (ACs).TM-05 as a starting material  was first brominated by excess bromine at positions 4 and 7, followed by selective debromination with tin powder in an acidic environment to generate the 7-monobromo-substituted naphthoic acid 3 −, found 344.8591. C11H5Br2O3− requires 344.8585.7-Bromo-3-hydroxy-2-naphthoic acid (3). Compound 2 , tin powder  and 250 mL of concentrated HCl were successively added into acetic acid (800 mL). The mixture was heated to reflux for 3 h. After cooling to room temperature, the reaction was quenched with ice water (1000 mL) and the solid was filtered and dried to afford 3  as a yellow solid. 1H-NMR : δ = 3.37 (br. s), 7.37 , 7.64 , 7.76 , 8.28 , 8.54  ppm. HRMS (ESI): [M − H]−, found 264.9502. C11H6BrO3− requires 264.9500. Methyl 7-bromo-3-methoxy-2-naphthoate (4). To a solution of 3  in acetone (2000 mL), anhydrous K2CO3  and dimethyl sulfate  were added at room temperature with mechanical stirring. After refluxing for 10 h, the reaction was cooled to room temperature. The insoluble K2CO3 was filtered and washed with EtOAc. The combined filtrate was concentrated under reduced pressure and extracted with EtOAc and water. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated to give 4  as a pale yellow solid. 1H-NMR : δ = 3.95 , 3.99 , 7.17 , 7.57 , 7.61 , 7.96 , 8.18  ppm. HRMS (ESI): calcd. for C13H11BrO3Na+ [M + Na]+ 316.9789, found 316.9785.7-Bromo-3-methoxy-2-naphthalenemethanol (5). To a suspension of LiAlH4  in THF (100 mL), a solution of 4  in THF (500 mL) was added dropwise via a dropping funnel at room temperature. After stirring for 8 h, water (20 mL) was added dropwise with vigorous stirring to quench the reaction. The mixture was filtrated through a plug of Celite and rinsed with EtOAc. The combined filtrate was dried with Na2SO4 and concentrated to give 5  as a white solid. 1H-NMR : δ = 3.97 , 4.82 , 7.09 , 7.50 , 7.60 , 7.65 , 7.9  ppm. GC-MS m/z: 266 (M+).7-Bromo-3-methoxy-2-naphthaldehyde (6). To a solution of 5  in CH2Cl2 (150 mL) at 0 °C, Dess-Martin periodinane  was added slowly. After stirring for 1 h at room temperature, the reaction mixture was quenched with saturated aqueous Na2S2O3 (50 mL) and saturated aqueous NaHCO3 (50 mL). The aqueous phase was extracted with CH2Cl2. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:10) to yield 6  as a yellow solid. 1H-NMR : δ = 4.01 , 7.58 , 7.72 , 7.86 , 8.35 , 10.46  ppm. GC-MS m/z: 264 (M+).Ethyl (E)-3-acrylate (7). To a solution of triethyl phosphonoacetate  in THF (10 mL) at 0 °C, NaHMDS  was added slowly and stirred for 30 min at room temperature. A solution of 6  in THF (10 mL) was added dropwise to the above reaction mixture and then stirred for another 2 h. The reaction mixture was quenched with water, concentrated under reduced pressure and extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4, rotary evaporated and crystallized to give 7  as a white solid. 1H-NMR : δ = 1.28 , 3.98 , 4.22 , 6.78 , 7.47 , 7.62 , 7.81 , 7.94 , 8.10 , 8.33  ppm. HRMS (ESI): [M + Na]+, found 357.0096. C16H15BrO3Na+ requires 357.0102.(E)-3-prop-2-en-1-ol (8). DIBAL-H  was added to a solution of 7  in CH2Cl2 (400 mL) at 0 °C. After being stirred for 2 h at room temperature, the reaction mixture was quenched with saturated aqueous Rochelle’s salt (300 mL) at 0 °C and the resulting suspension was stirred vigorously for 4 h. The reaction mixture was filtrated and the aqueous phase was extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:5) to afford 8  as a pale yellow solid. 1H-NMR : δ = 3.92 , 4.18 , 4.96 , 6.53 , 6.91, 7.36 , 7.51 , 7.75 , 7.99, 8.08 ppm. -3-oxiran-2-yl]methanol (9a). -3-oxiran-2-yl]methanol (9b) Ti(OiPr)4  was added to a suspension of powdered molecular sieves  in CH2Cl2 (30 mL). After stirring for 5 min, l-(+)-DIPT  in CH2Cl2 (10 mL) was added, and the reaction mixture was then stirred for 30 min, followed by slow addition of a solution of 8  in 120 mL of CH2Cl2. The resulting suspension was stirred for another 40 min and anhydrous tert-butyl hydroperoxide  was added at the same temperature and stirred for 4 h. After warming to 0 °C, 20% aqueous NaOH saturated with NaCl (30 mL) was added and the mixture was stirred for 1 h. The resulting slurry was filtered through a plug of Celite and rinsed thoroughly with CH2Cl2. The organic layer was separated and the aqueous layer was extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:5) to produce 9a (87% e.e.) as a white solid . 1H-NMR : δ = 3.10 , 3.58 , 3.80 , 3.95 , 4.16 , 5.05 , 7.40 , 7.56 , 7.63 , 7.79 , 8.11  ppm. HRMS (ESI): [2 M + Na]+ , found 640.9968. C28H26Br2O6Na+ requires 640.9973.9b (97% e.e.) was obtained with the same procedure as 9a using d-(−)-DIPT. HPLC .-3--3-phenylpropane-1,2-diol (10a) -3--3-phenylpropane-1,2-diol (10b). A suspension of freshly prepared CuI  in THF (34 mL) was cooled to −20 °C, and PhMgBr  was added via syringe and stirred for 1 h. A solution of 9a  in THF (10 mL) was added and stirred. After 3 h, the reaction mixture was warmed to 0 °C and quenched by a saturated aqueous solution of NH4Cl (50 mL). After the reaction mixture was concentrated under reduced pressure and extracted with EtOAc, the combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:5) to obtain 10a as a white solid . 1H-NMR : δ = 3.20 , 3.32 , 3.83 , 4.38 , 4.53 , 7.10–7.33 , 7.50 , 7.71 , 8.08 , 8.12  ppm. HRMS (ESI): [M + Na]+, found 409.0425. C20H19BrO3Na+ requires 409.0415.10b was obtained with the same procedure as 10a using 9b.(2R)-2--1--2-phenylethanol (12a) (2S)-2--1--2-phenylethanol (12b). To a solution of 10a  in CH2Cl2 (100 mL), NaIO4 impregnated over silica  was added and stirred for 3 h at room temperature. The reaction mixture was filtrated and rotary evaporated to give the crude product 11a which was used immediately for the next reaction. All of the above 11a was dissolved in THF (22 mL), and freshly prepared naphthyl Grignard reagent  was added slowly at 0 °C and stirred for 2 h. A saturated aqueous solution of NH4Cl (50 mL) was added to quench the reaction, and the aqueous layer was extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:10) to obtain 12a as a white solid . 12b was obtained with the same procedure as 12a using 10b.(R)-2--1--2-phenylethanone (13a) (S)-2--1--2-phenylethanone (13b). To a solution of 12a  in CH2Cl2 (100 mL), Dess-Martin periodinane  was added slowly and stirred for 2 h. After the reaction finished, 30 mL of asaturated aqueous solution of Na2S2O3 and 30 mL of a saturated aqueous solution of NaHCO3 were added and stirred for 1 h. After the reaction mixture became clear, it was extracted by CH2Cl2 three times. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. 13a  was obtained without column chromatography. 1H-NMR : δ = 3.80 , 6.33 , 7.10 , 7.19 , 7.33–7.60 , 7.78 , 7.84 , 7.94 , 8.05 , 8.58 . HRMS (ESI): [M + Na]+, found 505.0616. C29H21BrO2Na+ requires 505.0602.13b was obtained with the same procedure as 13a using 12b.(1R)-1--2--1-phenylpent-4-en-2-ol (14a), (1S)-1--2--1-phenylpent-4-en-2-ol (14b). After the freshly prepared allylzinc bromide (83 mmol) was concentrated under reduced pressure, a solution of 13a  in THF (as little as possible) was poured into the allylzinc bromide’s bottle and stirred for 5 min. A saturated aqueous solution of NH4Cl (50 mL) was added, and the aqueous layer was extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:20) to obtain 14a as a white solid .14b was obtained with the same procedure as 14a using 13b.(4R)-4--3-hydroxy-3--4-phenylbutanal (16a), (4S)-4--3-hydroxy-3--4-phenylbutanal (16b). To a solution of 14a  and N-methyl morpholine-N-oxide  in THF/water/acetone  at 0 °C, OsO4  was added. The reaction was warmed to room temperature and stirred for 8 h. A saturated aqueous solution of Na2S2O3 (10 mL) was added and stirred for 30 min. The reaction mixture was concentrated under reduced pressure and extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated to give crude product 15a which was used immediately for the next reaction.15a was dissolved in CH2Cl2, and NaIO4 impregnated over silica  was added and stirred for 3 h at room temperature. The reaction mixture was filtrated and rotary evaporated to give 16a without column chromatography.All of the above 16b was obtained with the same procedure as 16a using 14b.(4R)-4--3--4-phenylbutane-1,3-diol (17a), (4S)-4--3--4-phenylbutane-1,3-diol (17b). All of the above 16a was dissolved in 30 mL of methanol, and NaBH4  was added slowly at 0 °C. After stirring at room temperature for 3 h, the reaction mixture was filtrated, rotary evaporated and passed through silica gel chromatography column (eluent: EtOAc/hexane = 1:10) to give 17a . ESI-MS m/z: 526 [M]+.17b was obtained with the same procedure as 17a using 16b.(4R)-4--3-hydroxy-3--4-phenylbutyl 4-methyl-benzenesulfonate (18a), (4S)-4--3-hydroxy-3--4-phenylbutyl 4-methylbenzenesulfonate (18b). 17a  was dissolved in 38 mL of pyridine, and 4-dimethylaminopyridine  and TsCl  were added slowly at 0 °C. After stirring for 16 h at room temperature, the reaction mixture was quenched by 5 mL of water. Most of the pyridine was rotary evaporated and the rest of the reaction mixture was extracted with EtOAc. The combined organic layer was washed with water and brine, dried with Na2SO4 and rotary evaporated. The crude product underwent silica gel chromatography (eluent: EtOAc/hexane = 1:10) to obtain 18a as a white solid . ESI-MS m/z: 698 [M + NH4]+.18b was obtained with the same procedure as 18a using 17b.-1--4-(dimethylamino)-2--1- phenylbutan-2-ol (TM-05-a) and -1--4-(dimethylamino)-2--1-phenylbutan-2-ol (TM-05-b). A solution of 18a  in Me2NH  was stirred at 45 °C for 24 h. The reaction mixture was rotary evaporated and passes through a silica gel chromatography column (eluent: EtOAc t/hexane = 1:5) to obtain the mixture of TM-05-a and TM-05-b, namely 19a . Further purification of 19a through a silica gel chromatography column (eluent: CH2Cl2) yielded the final products TM-05-a  and TM-05-b . The mixture of TM-05-c and TM-05-d, namely 19b  was obtained with the same procedure as 19a. -1--4-(dimethylamino)-2--1- phenylbutan-2-ol (TM-05-c). TM-05-c  was obtained after purification of 19b through a silica gel chromatography column (eluent: CH2Cl2). 1H-NMR : δ = 1.88–2.04 , 2.17 , 2.47 , 4.05 , 6.04 , 6.86–6.88 , 7.12–7.14 , 7.30–7.33 , 7.45–7.50 , 7.57-7.61 , 7.67 , 7.87–7.92 , 8.03 , 8.65 , 8.75 . 13C-NMR : δ = 33.4, 44.7, 48.9, 55.9, 56.3, 82.4, 104.7, 116.8, 124.4, 125.1, 125.4, 125.5, 127.0, 127.4, 127.9, 128.0, 128.9, 129.8, 129.9, 130.1, 130.5, 131.6, 133.4, 134.7, 141.1, 142.2, 156.7 ppm. HRMS (ESI): [M + H]+, found 555.1769. C33H33BrNO2+ requires 555.1728. .-1--4-(dimethylamino)-2--1-phenylbutan-2-ol (TM-05-d). TM-05-d  was obtained after purification of 19b through a silica gel chromatography column (eluent: CH2Cl2). 1H-NMR : δ = 1.93–2.06 , 2.32 , 2.49 , 3.00 , 5.86 , 6.46 , 7.20–7.25 , 7.34–7.42 , 7.51–7.55 , 7.77 , 7.86 , 7.91-7.97 , 8.34 , 8.52 . 13C-NMR : δ = 35.0, 42.0, 43.7, 54.4, 54.9, 104.6, 116.6, 124.0, 124.9, 125.7, 126.5, 127.4, 127.5, 128.6, 128.7, 128.9, 129.1, 129.2, 129.4, 129.6, 129.7, 131.1, 131.2, 131.4, 134.3, 138.1, 139.1, 155.2 ppm.R, 2R) isomer of TM-05 strongly indicate that the replacement of naphthalene did not alter the conformation of bedaquiline too much, which is a reasonable explanation for our early bioactivity results. Further research to find the most bioactive isomers of our analogues is ongoing.Asymmetric synthesis and AC assignment were achieved for our bedaquiline analogues with a TARB skeleton. Comparison of theoretical and experimental ECD spectra is a very useful and reliable tool to determine the AC and is especially suitable for our analogues, which are not very flexible and have many chromophoric groups. The vicinal diol intermediate revealed its critical role for our AC confirmation due to the special reactivity of the vicinal diol moiety. Moreover, our results consolidated the generality of Sharpless epoxidation. Our NOESY spectra and existing crystallographic data on the (1


In addition, seven known phenolic compounds 4–10 were isolated. These structures were determined on the basis of NMR experiments as well as chemical evidence. Furthermore, all the isolated compounds 1–10 were tested for antioxidative activity by the superoxide dismutase (SOD)-like assay.One new phenolic compound, sapnol A ( Acer saccharum (Aceraceae), which is a tree native to North America -2,6-dimethoxyphenoxy]-1-(4-hydroxy-3-methoxyphenyl)-1,3-propanediol (5) , (+)panediol  [10] (Fi1) was obtained as an amorphous powder. HREIMS of 1 established the molecular formula C18H22O7, with an [M]+ peak at m/z 350.1349, indicating eight degrees of unsaturation. The UV spectrum of 1 showed absorption bands at 226, 280, and 307 nm, indicating the presence of a conjugated system. The IR spectrum of 1 suggested the presences of hydroxyl  and aromatic ring groups . In the aromatic region, the 1H-NMR spectrum of 1 showed one set of ABC-type signals at δ 7.23 , δ 7.24 , and δ 7.31 , an AA′-type signal at δ 7.03 , and in the aliphatic region, three methoxy signals at δ 3.64  and δ 3.72 , and one each of methine at δ 3.63 , oxymethylene at δ 4.37 , δ 4.58 , and oxymethine at δ 5.73  , one oxymethine carbon (δ 74.8), and five tertiary and seven quaternary carbons due to two aromatic rings, and the presence of 1,3-propanediol could be revealed by the COSY correlations (1 is an erythro isomer + peak at m/z 707.2530 in HRFABMS, which corresponded to the molecular formula C32H44O16, requiring 11 degrees of unsaturation. The IR absorption bands at 3,530, 1,605 and 1,590 cm−1 were characteristic of hydroxyl and aromatic groups. The 1H-NMR and 13C-NMR spectra of 2 indicated the presence of two β-glucopyranosyl units  -2,6-dimethoxy xyphenoxy]-1-(4-hydroxy-3-methoxyphenyl)-1,3-propanediol . Fraction 3 (2.0 g) was passed through silica gel with CHCl3-MeOH-H2O (25:8:0.1) and purified by preparative HPLC  to afford 1 (20.6 mg), acernikol , erythro-1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol , threo-1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol , and (+)-sakuraresinol . Fractions 4 (2.13 g), and 5 (1.7 g) were successively purified by preparative HPLC  to afford erythro-1,2-bis-(4-hydroxy-3-methoxyphenyl)-1, 3-propanediol  from fraction 4, and 2 (11.5 mg), and 3 (56 mg) from fraction 5, respectively.The maple sap (400 L) was passed through an Amberlite XAD-1180N column and eluted with 20% MeOH, 50% MeOH, and 100% MeOH. The MeOH fraction (19.5 g) was subjected to silica gel column chromatography with CHClSapnol A (1): An amorphous powder; [α]D22 −12.2° ; UV (MeOH) λmax nm (log ε): 226 (4.01), 280 (3.61), 307 (3.43); FT-IR (dry film) νmax cm−1: 3,395 (OH), 1,605, 1,520 (C=C), 1,070 (OH); 1H-NMR and 13C-NMR data (C5D5N) see m/z 350.1349 .Saposide A (2): An amorphous powder; [α]D22 +9.2° ; UV (MeOH) λmax nm (log ε): 230 (4.06), 283 (3.76); CD (MeOH) nm (Δε): 292 (−6.30), 276 (+4.54), 239 (+4.55); FT-IR (dry film) cm−1: 3,530 (OH), 1,605, 1,590 (C=C); 1H-NMR and 13C-NMR data (C5D5N) see m/z 707.2530 .Saposide B (3): An amorphous powder; [α]D22 −6.0° ; UV (MeOH) λmax nm (log ε): 222 (4.09), 228 (4.10), 282 (3.75); CD (MeOH) nm (Δε): 294 (+0.67), 244 (+0.88), 226 (−1.67); FT-IR (dry film) λmax cm−1: 3,380 (OH), 1,612, 1,520 (C=C), 1,035 (OH); 1H-NMR and 13C-NMR data (C5D5N) see m/z 707.2522 .2SO4-dioxane (1:1) was heated at 100 °C for 2 h. The reaction mixture was diluted with H2O, neutralized with Amberlite IRA-35 and evaporated in vacuo to dryness. The identification and the d or l configuration of glucose were determined using RI detection (Shimadzu RID-10A) and chiral detection (Shodex OR-1) by HPLC , by comparison with an authentic sugar (10 mmol d-glc). The sugar portion gave the following peak of d-(+)-glucose at 20.6 min.Each sample (1 mg) in 5% HSOD-like activity was determined according to the method of Ukeda  using a 1), lignan sophoroside saposide A (2), and neolignan sophoroside saposide B (3) were isolated from sugar maple sap collected from A. saccharum (Aceraceae), together with seven known compounds, which included phenylpropanoids 4, 10 and neolignans 5–9. Additionally, all known compounds, 4–10 are reported for the first time from sugar maple sap. Some phenolics in maple syrup are possibly formed under the conditions of intensive heating involved in transforming sap to syrup. Here it should be noted that compounds 1–4, 7 were not present in maple syrup. All isolated compounds showed moderate antioxidative activities . It was noted that compound 1 showed activity at IC50 11.7 μM in this study. These results provide a potential explanation for the use of maple sap as a herbal medicine in the prevention of aging. Moreover, it should be noted that a number of compounds in maple sap remain unidentified.The phenylpropanoid sapnol A (


Garcinia xanthochymus has been widely used in traditional Chinese medicine for expelling worms and removing food toxins. Bioassay-guided fractionation of an EtOAc-soluble extract of G. xanthochymus stem bark led to the isolation of six new xanthones. Their structures were elucidated by spectroscopic methods, especially 2D-NMR techniques. Free-radical-scavenging activities of the isolated compounds were elucidated through DPPH method. Most of the isolated compounds showed considerable free radical scavenging activity on DPPH assay. Compound 1 exhibited effective antioxidant scavenging activity against DPPH radical with an IC50 value of 19.64 μM, and compound 6 showed the lowest activity among all the tested molecules, with an IC50 value of 66.88 μM. These findings support the notion that the plant genus Garcinia is a good source of bioactive compounds.  The presence of the fused furan ring was substantiated by the methine carbons (δc 104.9 and 144.8) in the 13C-NMR spectrum. The HMBC correlations of the hydrogen-bonded proton (1-OH) with an oxygenated aromatic carbon at δc 160.9, a quaternary aromatic carbon at δc 105.8 and a methine aromatic carbon at δc 94.1 corresponding to an aromatic proton [δH 6.85 ] in HSQC spectrum. It suggested that this proton may be attributed to H-2. The position of the furan ring was determined as follow. In the HMBC spectrum, one proton signal at δH 7.39  of the furan ring showed correlations with a quaternary aromatic carbon at δc 108.5 (C-4) and an oxygenated aromatic carbon at δc 160.6 (C-3). The signals at δc 108.5 and 160.6 also correlated with the aromatic proton signal at δH 6.85 . Therefore, the furan ring was fused at C-4 through an oxygen at C-3. The locations of other substituents were determined as follows. In the 13C-NMR spectrum, the aromatic carbons with an oxygen function were observed at δc 149.4, 130.3 and 150.5, which suggested the presence of a 1, 2, 3-trioxygenated benzene ring in partial structure B. In HMBC spectrum (2-1′′/C-6 (δC 150.5), C-7 (δC 126.1) and C-8 (δC 136.4) indicated that one 3-methyl-2-butenyl group was located at C-7. Thus, the remaining 3-hydroxy-3-methylbutyl group should be located at C-8. Compound 1 was thus identified to be 1,5,6-trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)furano xanthone.Compound spectrum , the cor2 was obtained as a yellow powder. The molecular formula was determined as C28H32O7 (m/z 480.2118) by HREIMS. Comparing its 13C-NMR and DEPT data with those of 1, it was found that compound 2 had almost the same chemical shifts as those of 1, except for the dimethylpyran ring carbon signals at δc 115.9 (d), 127.2 (d), 78.3 (s), 28.0 (q) and 28.0 (q) in 2 instead of furan ring carbon signals at δc 104.9 (d) and 144.8 (d) in 1. These facts suggested that dimethylpyran ring in the structure of 2 replaced furan ring found in 1. Thus, 2 was identified to 1,5,6-trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)–6′, 6′-dimethylpyrano  xanthone.Compound 3 was obtained as a yellow powder, whose molecular formula was determined as C28H34O8 by the HREIMS . Comparison of NMR data of 3 with those of 1 indicated that the two compounds were closely related. The obvious spectroscopic differences between them resulted from the presence of a 2-(1-hydroxy-1-methylethyl)-2,3-dihydrofuran ring in 3, instead of furan ring in 1. The location of 2-(1-hydroxy-1-methylethyl)-2,3-dihydrofuran ring was fused at C-3 and C-4 of the xanthone nucleus with an ether linkage at C-3 by the HMBC correlations (H 6.19 (H-2) to δC 163.7(C-1), 166.7(C-3) and δH 3.27(H2-1') to δC 166.7 (C-3), 102.5 (C-4). Therefore, the structure of 3 was determined as 1,5,6-trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)–5′-(1-hydroxy-1-methylethyl)-4′, 5′-dihydrofurano  xanthone.Compound elations  between 4 was obtained as a yellow amorphous powder. The [M+] at m/z 428.1464 in the HREIMS corresponds to C23H24O8 . The 1H-NMR spectrum of 4 exhibited one chelated hydroxy group [δH 13.0 ], two aromatic proton as a singlet at δH 7.34  and δH 7.79 , a typical signal of a 1,1-dimethylallyl group at δH 6.36 ; 5.16 ; 5.02  and 1.65 , as well as a 2-(1-hydroxy-1-methylethyl)-2, 3-dihydrofuran-3-ol moiety from resonances at δH 5.53 ; 4.49 ; 1.32  and 1.30 . The 13C-NMR and DEPT experiments displayed the presence of four methyl, five methine, one methylene, 12 quaternary carbons and one carbonyl. A partial structure A of 4, 1, 2-dihydroxy-4-xanthone, was deduced by comparison of the 1H-NMR and 13C-NMR data of 4 with those of subelliptenone H , two meta-aromatic proton at δH 6.20  and δH 6.40 . Therefore, a coupling of meta-aromatic protons were assigned to C-2 and C-4 respectively, which was further supported by the HMBC correlations of H-2 with δC 165.5(C-3), 94.3(C-4) and 102.8(C-9a). Based on the above observation, the structure of 5 was established as 1, 3, 5, 6-tetrahydroxy-7-geranylxanthone.Compound xanthone . The ger6 was obtained as a yellow powder. The molecular formula was determined as C18H14O5 (m/z 310.0848) by HREIMS. The 13C NMR spectrum showed 18 carbon signals, which were classified into 2 methyl, 6 methine and 10 quaternary carbons by analysis of the DEPT spectra. The 1H NMR data showed two sets of ortho-aromatic protons at δH 6.64  and 7.32 ; 7.18  and 7.72 , one chelated hydroxyl group at δH 12.02 , one cis olefinic group  as well as two tertiary methyls attached to an oxygenated carbon [δH 1.54 ] indicating the presence of a dimethylpyran ring system. The positions of the substituents were deduced by analysis of HMBC. In the HMBC spectrum, there were correlations between the chelated hydroxyl group at δH 12.02 and the carbon signal at δc 154.2 (C-1) and 109.6 (C-2) corresponding to one of the ortho-coupled proton δH 6.64  in the HSQC spectrum. These results indicated that 6 was a 1, 4-dihydroxyxanthone derivative. The other coupling of ortho-aromatic protons at δH 7.18  and 7.72  were assigned as H-7 and H-8, respectively, by the HMBC correlation of δH 7.72 with carbonyl carbon C-9 (182.4). The dimethylpyran ring was fused with the xanthone in an angular form which was further supported by HMBC correlation of δH 6.59  with δc 141.7 , 127.6  and 122.3 and δH 6.02  with δc 127.6 . Thus, the structure of compound 6 was established as 1, 4-dihydroxy-6′, 6′-dimethylpyrano  xanthone.Compound 1 > 3 > 2 > 5> 4 > 6. The six xanthones were evaluated for their antioxidant activities by DPPH free radical scavenging method . Most of1 exhibited effective antioxidant scavenging activity against DPPH radical, with an IC50 value of 19.64 μM, and compound 6 showed the lowest activity with an IC50 value of 66.88μM among all the tested molecules. The DPPH radical scavenging activities of these compounds seemed to be related to the number of phenol-like OH groups at the xanthone skeleton. It was reported previously that the radical scavenging activity was increased in the presence of an increasing number of phenol like OH groups in a molecule [5, having four phenol-like OH groups, showed a lower radical scavenging activity compared to that of compound 1 having three phenol-like OH groups. This was because the presence of furan ring in compound 1 extended the conjugation system to participate in stabilizing the phenoxy radical by resonances, therefore increasing the radical-scavenging activity of compound 1 [Garcinia xanthochymus were the phenolic compounds, such as xanthone derivatives.Compound molecule . Howevermpound 1 . From ab6d or Me2CO-6d on a Bruker AM-400 (1D) or Varian Inova-600 (2D) spectrometer with TMS as an internal standard. EIMS and HREIMS measurements were conducted with a Finnigan MAT 95 instrument. Thin-layer chromatography (TLC) was performed on silica gel 60 GF254, while column chromatography was carried out using silica gel (200-300 mesh) from Qingdao Haiyang Chemical Group Co., P. R. China and C18 reversed-phase silica gel from YMC CO., LTD., Japan. UV spectra were measured on an SP-2102UVPC spectrometer using MeOH as the solvent. NMR spectra were run in DMSO-Garcinia xanthochymus was collected from Xishuangbanna Prefecture, Yunnan Province, P.R. China and identified by Xishuangbanna Prefecture National Medicine Research Institute. The voucher specimen (06061201) was deposited with the Herbarium of College of Pharmacy, South Central University for Nationalities.The bark of G. xanthochymus (6.5 kg) was extracted with 95% EtOH (25 L × 3) and then successively partitioned with petroleum ether (P.E.) (3.0 L × 3), EtOAc (3.0 L × 3) and n-BuOH (3.0 L × 3). The combined EtOAc extract (590 g) was chromatographed on silica gel with P.E-Me2CO  to give thirteen fractions (fr.1−fr.13). Fr.6 (17.0 g) was separated on a silica column (toluene/Me2CO 95:5→3:7 gradient system), and then purified by chromatography on a silica gel  and RP-18  to afford compound 6 (2.8 mg). Fr.7 (33.8 g) was extensively separated over a silica column (toluene/Me2CO 95:5→3:7 gradient system) and RP-18  to afford 1 (3.2 mg), 2 (9.8 mg) and 5 (4.8 mg). Fr. 9 (10.8 g) was also subjected to silica gel with a gradient elution  and RP-18  to afford compounds 3 (7.4 mg) and 4 (8.8 mg). The powdered bark of 1,5,6-Trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)–furano xanthone (1). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 231 (3.50), 265 (3.49), 350 (3.58); For 1H-NMR and 13C-NMR spectroscopic data (in Me2CO-6d), see m/z (%): 438 , 420 (36), 377 (76), 364 (48), 349 (100), 323 (80); HREIMS m/z 438.1698 .1,5,6-Trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)–6′,6′-dimethylpyrano  xanthone (2). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 229 (3.52), 263 (3.52), 350 (3.63); For 1H-NMR and 13C-NMR spectroscopic data (in Me2CO-6d), see m/z (%): 480 , 463 (76), 447 (100), 419 (72), 391(84), 365 (64), 349 (56); HREIMS m/z 480.2118 .1,5,6-Trihydroxy-7-(3-methyl-2-butenyl)-8-(3-hydroxy-3-methylbutyl)–5′-(1-hydroxy-1-methyl-ethyl)-4′,5′-dihydrofurano xanthone (3). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 250 (4.21), 285 (4.00), 334 (4.11); For 1H-NMR and 13C-NMR spectroscopic data (in DMSO-6d), see m/z (%): 498 , 480 (40), 437 (100), 424 (44), 409 (52), 383 (100), 365 (32); HREIMS m/z 498.2256 .1,2,5,4′-Tetrahydroxy-4-–5′-(2-hydroxypropan-2-yl)-4′,5′-dihydro furano- xanthone (4). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 230 (3.46), 264 (3.47), 337 (3.46), 389(sh)(3.09); For 1H-NMR and 13C-NMR spectroscopic data (in Me2CO-6d), see m/z (%): 428 , 410 (6), 392 (16), 352 (56), 319 (56), 319 (100); HREIMS m/z 428.1464 .1,3,5,6-Tetrahydroxy-7-geranylxanthone (5). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 260 (4.00), 341(4.01); For 1H-NMR and 13C-NMR spectroscopic data (in Me2CO-6d), see m/z (%): 396 , 327 (32), 311 (100), 274 (56), 123 (181), 69 (24); HREIMS m/z 396.1574 .1,4-Dihydroxy-6′,6′-dimethylpyrano  xanthone (6). Yellow amorphous powder; UV λmax (MeOH) nm (logε): 230 (3.33), 264 (3.33), 354 (3.45), 400 (sh) (3.26); For 1H-NMR and 13C-NMR spectroscopic data (in Me2CO-6d), see m/z (%): 310 , 295 (100), 148 (15); HREIMS m/z 310.0848 .G. xanthochymus towards DPPH radical were assessed by using the method described by Scherer and Godoy with a slight modification [0 was the absorbance of control and A1 was the absorbance in the presence of the standard, solvent extracts or purified compounds at different concentrations. Ascorbic acid (VC) and gallic acid were used as positive controls, respectively. All the tests were performed in triplicate. The scavenging activities of the purified compounds towards DPPH radical were expressed as IC50, which was determined to be the effective concentration at which DPPH radical was scavenged by 50%. The IC50 value was obtained by interpolation from linear regression analysis.Scavenging activities of the purified compounds from fication ,26. Brie50 values were calculated by linear regression analysis. The results were performed as mean ± standard deviation (SD) of three determinations. Analysis of significance differences among means were tested by one-way analysis of variance. The ICG. xanthochymus, an EtOAc-soluble partition of the EtOH extract of the bark of G. xanthochymus displayed significant antioxidant activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging bioassay. This prompted us to perform a detailed bioassay-guided isolation from this plant, which led to the isolation of six new xanthones. Notably, most of the isolated compounds showed considerable free radical scavenging activity in the DPPH assay. Compound 1 exhibited effective antioxidant scavenging activity against DPPH radicals with an IC50 value of 19.64 μM, and compound 6 showed the lowest activity among all the tested molecules, with an IC50 value of 66.88 μM. These findings support that plant genus Garcinia is a good source of bioactive compounds.In the course of our ongoing research project on bioactive natural products from


Moorea producens was investigated. The organic extract of the Red Sea cyanobacterium afforded one new cerebroside, mooreaside A (1), two new nucleoside derivatives, 3-acetyl-2′-deoxyuridine (2) and 3-phenylethyl-2′-deoxyuridine (3), along with the previously reported compounds thymidine (4) and 2,3-dihydroxypropyl heptacosanoate (5). The structures of the compounds were determined by different spectroscopic studies , as well as comparison with the literature data. Compounds 1–5 showed variable cytotoxic activity against three cancer cell lines.In the course of our ongoing efforts to identify marine-derived bioactive compounds, the marine cyanobacterium  Nucleosides and cerebrosides are found in both terrestrial and marine organisms. Cerebrosides are composed of a hydrophobic part named ceramide, which is linked to one sugar moiety . CerebroMoorea (formerly Lygnbya) +. The attachment of the glucose moiety at C-1 was established by the downfield shift of C-1 (δC 68.5) and HMBC correlation of H-1′′ (δH 4.28) with C-1. Moreover, the 1H-NMR spectrum of 1 showed signals at δH 3.61 , 3.97 , 3.91 and 3.74  attributable to oxymethine, NH-bonded methine, and oxymethylene groups, respectively. They correlated with the carbon signals at 73.4 (C-3), 59.3 (C-2), and 68.5 (C-1) in the HSQC spectrum. 1H-1H COSY cross peaks were observed between the NH proton (δH 7.51) and H-2, which coupled to H-1 and H-3, suggesting the presence of hydroxyl group in the long chain base. This was further confirmed by the HMBC correlations of H-1 to C-2 and C-3, H-2 to C-1, C-3, and C-4, H-3 to C-1 and C-2, and 2-NH to C-2 and C-3 23CONH]+, 365 [CH3(CH2)23CO]+, 337 [CH3(CH2)23]+, 225 [CH3(CH2)10-CH=CH-CH2(CHOH)]+, 181 [CH3(CH2)10-CH=CH]+, and 155 [CH3(CH2)10]+ (1. Methanolysis of 1 gave long chain base (LCB) and fatty methyl ester (FAME). The FAME in the n-hexane layer was identified as pentacosanoic acid methyl ester based on the GCMS molecular ion peak at m/z 396 [M]+. The LCB showed an EIMS molecular ion peak at m/z 285 [M]+, corresponding to -2-aminoheptadec-5-ene-1,3-diol. Based on the above evidence and discussion, compound 1 was assigned as N--3-hydroxy-1-O-((β-d-glucopyranosyl)heptadec-5-en-2-yl)pentacosanamide. Compound 1 was generically named mooreaside A.Compound 1  was obta and 1H-  and 13C- ,25,26. T and C-3 . Compariart of 1 ,31,32. TCH2)10]+ , support2  and 1693 (amide carbonyl) cm−1. These data in conjunction with characteristic UV absorption bands at λmax 253 and 268 nm along with 1H NMR spectrum , 162.8 (C-4), and 183.4 (C-8). The 1H-1H COSY spectrum  and 7.98 . These protons correlated to the carbon signals at δC 102.6 and 142.5, respectively in the HSQC, indicating the presence of uracil moiety in 2. This was confirmed by the HMBC /δC 86.6 (C-1′), 2.28 /41.4 (C-1′), 4.38 /72.2 (C-3′), 3.89 /89.0 (C-4′), and 3.75  and 3.71 /62.8 (C-5′) supported the presence of 2′-deoxyribose moiety +. Based on the 13C chemical shifts, the acetyl group was assigned at N-3 of the uracil moiety, completing the molecular formula of 2 and the degrees of unsaturation. Consequently, 2 was assigned as 3-acetyl-2′-deoxyuridine and is reported here as a new natural product.Compound 2  was obtaspectrum  suggeste The 13C  and mult The 13C  spectra spectrum  showed tthe HMBC  cross peobserved . This wa3 /130.0 , 7.27 /129.8 , 7.25 /128.3 (C-6′′), 2.95 /34.7 (C-2′′), and 3.16 /34.7 (C-1′′) were observed. The signals suggested the presence of a N-bonded phenylethyl moiety. This was established by 1H-1H COSY cross peaks, and further confirmed by HMBC correlations  + ; NMR spectral data, see 3-Acetyl-2′-deoxyuridine (3): White amorphous powder; c 0.2, MeOH); UV (MeOH) λmax (log ε) 257 (2.42), 269 (2.87) nm; IR (film) νmax 3289, 2959, 1694, 970, 746 cm−1; HRESIMS m/z 333.1446 [M + H]+ ; NMR spectral data, see 3-Phenylethyl-2′-deoxyuridine (1–5) were evaluated for their cytotoxic activity against colorectal carcinoma (HCT-116), hepatocellular carcinoma (HepG2), and breast cancer (MCF-7). The cells were obtained commercially from ATCC. The cytotoxicity was evaluated by the sulforhodamine B (SRB) assay, as described previously [The isolated compounds (eviously . Doxorub1 (4.5 mg) was treated with 6 mL of 1N HCl in MeOH at 90 °C for 15 h in a sealed ampoule. The reaction mixture was diluted by adding 20 mL of distilled water, then extracted with n-hexane (3 × 15 mL) to give a corresponding FAME, which was identified by GCMS. The aqueous layer was evaporated to dryness and subjected to Sephadex LH-20 using CHCl3:MeOH (10:90) to give LCB and sugar. The base was analyzed by EIMS [Compound  by EIMS .Moorea producens led to the isolation of a new cerebroside (1) and two new nucleoside derivatives (2 and 3), along with two known compounds (4 and 5). Their structures were determined using extensive spectroscopic studies. Compounds 1–3 showed moderate cytotoxic activity against breast cancer cell lines. In conclusion, the investigation of the Red Sea strain of the marine cyanobacterium


The first detectable intermediates are the (RP)- and (SP)-diastereomers of the monodeacetylated triester 14, which subsequently undergo concurrent retro-aldol condensation to diester 4 and enzyme-catalyzed hydrolysis to the fully deacetylated triester 15. The former pathway predominates, representing 90% of the overall breakdown of 14. The diester 4 undergoes the enzymatic deacetylation 700 times less readily than the triester, but gives finally thymidine 5´-monophosphate as the desired main product. To elucidate the potential toxicity of the electrophilic 2-cyano-N-(2-phenylethyl)acrylamideby-product 17 released upon the deprotection, the hydrolysis of 1 has also been studied in the presence of glutathione (GSH).Thymidine 5´-bis[3-acetyloxy-2-cyano-2-(2-phenylethylcarbamoyl)propyl]phosphate ( Al. Al-N-(2ite (12)  . At pH 7.5-10, hydrolysis of the first acetic ester linkages of 1 (Reaction A in k = 7.0 × 10-6 s-1). The subsequent departure of the remnants of this protecting group by retro-aldol condensation gave diester 4 (Route C) without accumulation of the monodeacetylated triester 14 as an intermediate. Hydrolysis of the second ester linkage (Reaction D) was 16 times slower, the half-live being 460 h. Accordingly, the compound is slightly more stable than the corresponding 3-acetyloxy-methoxy-2,2-bis(ethoxycarbonyl)propyl derived triester 2. The half-lives for the consecutive deacetylations of 2 have been reported to be 32 h and 148 h under these conditions ethanesulfonic acid (HEPES) buffer (3.5 mL) at pH 7.5 and 37 °C (I = 0.1 mol L-1 with NaCl) resulted in an intermediary accumulation of the RP- and SP-distereomers of monodeacetylated triester 14  being 3.8 min. The subsequent retro-aldol condensation of 14 to diester 4  was, however, so fast that the amount of 14 remained less than 5% of the initial concentration of 1. Treatment of triester 14 to the dideacetylated triester 15  competed with the retro-aldol condensation, representing approximately 10% of the total disappearance of 14. Both, 4 and 15 were subsequently decomposed to diester 16  that was rapidly converted to 5´-TMP . The half-life for the disappearance of 4 was 47 h. The level of accumulation of the deacetylated intermediate 16 remained below 5%. Prolonged treatment with the esterase resulted in dephosphorylation of 5´-TMP to thymidine. 1.Deacetylation of 1 and its 3-acetyloxy-2,2-bis(ethoxycarbonyl) and 3-acetyloxymethoxy-2,2-bis(ethoxycarbonyl) counterparts 3  and 2, respectively. Although at the high HLE concentration employed the rates of Reactions B and C are comparable, Reaction C predominates at low enzyme concentrations. Comparison of the half-lives of Reaction D reveals that 1 is under such conditions converted to 5´-TMP 3 times as fast as 3, but still one order of magnitude more slowly than 2. The advantage of faster retro-aldol condensation of the intermediated derived from 1 is overcompensated by the slower enzymatic deacetylation of diester 4. In other words, the desired overall acceleration of the exposure of 5´-TMP was not achieved.14, 15 and 16, evidently produces 2-cyano-N-(2-phenyl)ethylacrylamide  or nitrogen (Route G) on the β-carbon of 17. In fact, the signals at m/z 199.2 could be observed at different elution times. The mass spectra of these compounds additionally exhibited an unidentified m/z peak of 187.2. The m/z values ([M+H]+ = 219.5 and [M-H]- = 217.2) referring to the molecular ion of 2-cyano-3-hydroxy-N-(2-phenyl)ethylpropanamide 19 (see 17 may take place (Route F).As discussed above, retro-aldol condensation of the deacetylated intermediates, mide 17; . Consiste 19 see  was visi1 was also carried out in the presence of glutathione ([GSH] = 5.4 mM) and carboxyesterase ([HLE] = 28.0 units mL-1) in a 2-(N-morpholino)ethanesulfonic acid (MES) buffer (0.005/0.005 mol L-1) at pH 6.2 and 37 °C. Under these conditions, the half-live for the deacetylation of 1 was 18 h. In 2 days, the starting material was converted almost quantitatively to a mixture of acetylated and deacetylated phosphodiester 4 and 16 ([4]/[16] is 1:1). At this stage, 2-cyano-N-(2-phenyl)ethylacrylamide (17) was captured as the glutathione conjugate . Product 18 was detected by both HPLC and LC-MS analysis .The hydrolytic reactions of N-(2-phenyl)ethylacrylamide (17) is largely trapped by glutathione, cyclization (Routes G and H) and hydration (Route F) of 17 seem to occur as a side reaction. As seen from - ion peak at m/z 217.3, referring most likely to 2-cyano-3-hydroxy-N-(2-phenyl)ethyl-propanamide (19), appears and undergoes fragmentation (loss of H2O) to yield an ion at m/z 199.2 (17). Similarly, the molecular ion signals of the cyclization products 20 and 21 (m/z 199.2) were detected as in the absence of GSH. By contrast, no sign of S-hydroxymethylglutathione, obtained by the reaction of GSH with formaldehyde, was observed.Although 2-cyano-2 and distilled before use. 1H-, 13C-, 31P-NMR spectra were recorded on a Bruker Avance 400  or 500  NMR spectrometer. HRMS spectra were recorded on a Bruker Daltonics micrOTOF-Q instrument and LC-MS spectra were recorded on a Perkin-Elmer Sciex-API-365 triple-quadrupole instrument. For column chromatography, Fluka silica gel 60 (230-400 mesh) was used. Hydrolytic reactions were followed by Merck Hitachi LaChrom D7000 HPLC.Chemicals were purchased from Sigma–Aldrich, Fluka and Merck. Dichloromethane, acetonitrile, and pyridine were dried over 4Å molecular sieves. Dioxane was dried over 3Å molecular sieves. Triethylamine was dried by refluxing over CaHO-Levulinoylthymidine (11) and 2-cyano-3-hydroxy-2-(hydroxymethyl)-N-(2-phenylethyl)propanamide (8) were prepared as described previously +, calcd. 341.1.eviously . 1H-NMR 2-cyano-3-hydroxy-2-(hydroxymethyl)-N-(2-phenylethyl)propanamide (8) was obtained as described previously + obsd. 271.2, calcd. 271.2.eviously . 1H-NMR 2-cyano-2-(hydroxymethyl)-3-oxo-3-(2-phenethylamino)propyl acetate (10). Concentrated H2SO4 was added to a mixture of compound 8  and triethyl orthoacetate  in dry THF (10 mL). The reaction was allowed to proceed overnight and the mixture was the poured into an ice-cold solution of 5% NaHCO3 (50 mL). The product was extracted with diethyl ether (3 × 50 mL), washed with saturated aqueous NaCl (3 × 50 mL) and dried over Na2SO4. The solvent was evaporated and the crude product, 5-cyano-2-ethoxy-2-methyl-N-(2-phenyl)-ethyl-1,3-dioxane-5-carboxamide (9), was subjected to silica gel chromatography, eluting with a mixture of DCM and MeOH . The product (9) was obtained as oil . 1H-NMR  δ = 8.55, 8.44, 8.26, 7.98 , 7.28 , 7.21 , 4.32, 4.02, 3.57, 3.45 , 5.82, 5.81, 5.54, 5.53, 4.28, 4.23, 4.07, 3.99 , 3.79-3.66 and 3.41-3.29 , 2.77, 2.74, 2.56 , 2.03, 1.99, 1.42 1.36 , 1.33, 1.17, 1.15, 1.06 . ESI+-MS: m/z [M + Na]+ obsd.341.3, calcd. 341.1.5-cyano-2-ethoxy-2-methyl-N-(2-phenyl)-ethyl-1,3-dioxane-5-carboxamide (9)  was dissolved in 80% aqueous acetic acid (50 mL) and left for 1 h at room temperature. The solution was evaporated to dryness and the residue was coevaporated three times with water. The product was purified by silica gel colum chromatogaphy eluting with a mixture of DCM and MeOH . The product (10) was obtained as solid foam . 1H-NMR : δ = 8.28 , 7.30 , 7.21 , 5.81 , 4.34 , 3.74 , 3.31 , 2.74 , 2.03 CH3). 13C-NMR : δ = 170.05 (C=O Ac), 164.73 (C(O)NH), 137.93 (Ph), 128.82 (Ph), 128.75 (Ph), 126.92 (Ph), 117.60 (CN), 63.41(CH2OAc), 62.96 (CH2OH), 51.01(C2), 41.55 (CH2NH), 35.27 (CH2Ph), 20.54 (CH3) ppm. ESI+-MS: m/z [M + Na]+ obsd. 313.1146, calcd. 313.1159.Thymidine 5´-bis[3-acetyloxy-2-cyano-2-(2-phenylethylcarbamoyl)propyl]phosphate (1). 3’-O-Levulinoylthymidine (11) was coevaporated three times from dry pyridine and dried on P2O5 overnight. To a solution of dried 11  in dry DCM (1.2 mL), anhydrous triethylamine  and bis(diethylamino)chlorophosphine  was added, and the reaction mixture was stirred for 2 h under nitrogen. The product 12 was filtered through a short silica gel column eluting with a mixture of anhydrous ethyl acetate and triethylamine in hexane . The solvent was removed under reduced pressure and the residue was coevaporated three times from dry MeCN to remove the traces of triethylamine.The residue (0.16 g) was dissolved in dry MeCN (0.5 mL) and compound 10  in dry MeCN (0.5 mL) and tetrazole  were added under nitrogen. The reaction mixture was stirred for 4 h at room temperature. The phosphite ester 13 formed was oxidized with I2 (0.1 mol L-1) in a mixture of THF, H2O and lutidine  by stirring overnight at room temperature. The crude product, 3´-O-levulinoylthymidine 5´-bis[3-acetyloxy-2-cyano-2-(2-phenylethylcarbamoyl)propyl]phosphate (13) was isolated by DCM/aq. NaHSO3 work up, and purified on a silica gel column eluted with ethylacetate, followed by a stepwise gradient from a mixture of DCM and MeOH  up to 30% MeOH. The purification was repeated using ethyl acetate and a mixture of DCM and MeOH  as an eluent. The product (13) obtained as solid foam was contaminated by some unidentified impurities. The product was characterizied by HPLC-MS eluting with a mixture of water and MeCN (linear gradient from 10% up to 40.0% MeCN in 40 min)  and used without further purification. Compound 13 was dissolved in dry DCM (2.0 mL) and hydrazine acetate  in dry MeOH (0.20 mL) was added. After 1 h, hydrazinium acetate  in a mixture of DCM (100 μL) and MeOH (20 μL) was added. The reaction was allowed to proceed for 1.5 h and the addition of hydrazinium acetate was repeated. The reaction was quenched with acetone and the mixture was evaporated to dryness. The crude product was purified by reversed phase chromatography on a Lobar RP-18 column , eluting with a mixture of water and acetonitrile . The product was obtained as solid foam . 1H-NMR : δ = 8.65 , 7.35-20 , 6.72 , 6.15 , 4.51-4.31 , 4.00 ,3.63 , 3.51 , 2.86 , 2.32 , 2.08 CH3) 2.01 . 13C-NMR : δ = 169.69 (C=OAc), 163.58 (C4), 162.34 (C(O)NH), 150.14(C2), 137.84 (Ph), 136.5 (C6), 128.85 (Ph), 128.73 (Ph), 126.93 (Ph), 116.79 (CN), 116.35 (CN), 111.33 (C5), 85.76 (C4´), 84.14 (C1´), 67.50 (C3´), 67.08 (C5´), 62.72 (CH2OAc), 50.52(C spiro), 41.96 (CH2NH), 35.19 (CH2Ph), 26.86 (C2´), 20.49 (C(O) CH3), 12.45 (CH3) ppm. 31P-NMR : δ = -2,53; -2,76; -3,17; -3,35 ppm . ESI+-MS: m/z [M + H]+ obsd. 867.8 calcd. 867.3. ESI+-MS: m/z [M + Na]+ obsd. 889.2785, calcd. 889.2780.N-morpholino)ethanesulfonic acid (MES) buffers. The ionic strength of the solutions was adjusted to 0.1 mol L-1 with sodium chloride. The hydronium ion concentration of the buffer solutions was calculated with the aid of the known pKa values of the buffer acid under the experimental conditions. The initial substrate concentration was ca. 0.4 mmol L-1. The reactions were carried out in sealed tubes immersed in a thermostated water bath (37.0 ± 0.1 °C). The oxonium ion concentration of the reaction solution (3.5 mL) was adjusted with glycine, 2-[4-(2-hydroxyethyl)piperazin-1-yl)]ethanesulfonic acid (HEPES) and 2-(-1) in a 2-[4-(2-hydroxyethyl)piperazin-1-yl)]ethanesulfonic acid (HEPES) buffer (0.040/0.024 mol L-1) at pH 7.5 and 37 °C. The samples (200 μL) withdrawn at appropriate intervals were made acidic (pH 2) with 1 mol L-1 aqueous hydrogen chloride to inactivate the enzyme and to quench the hydrolysis, cooled in an ice-bath and filtered with minisart RC 4 filters (0.45 μm). The composition of the samples was analyzed on an ODS Hypersil C18 column , using a mixture of acetic acid/sodium acetate buffer (0.045/0.015 mol L-1) and MeCN, containing ammonium chloride (0.1 mol L-1). A good separation of the product mixtures of 2 was obtained on using a 5 min isocratic elution with the buffer containing 2% MeCN, followed by a linear gradient (23 min) up to 40.0% MeCN. Signals were recorded on a UV-detector at a wavelength of 267 nm. The reaction products were identified by the mass spectra (LC/MS) using a mixture of water and acetonitrile containing a formic acid (0.1%) as an eluent . The first order rate constants for the non-enzymatic hydrolysis of triester 1 were obtained by applying first-order rate-law to the diminution of the concentration of the starting material. The enzymatic deacetylations obeyed first-order kinetics at the high HLE concentrations employed.The enzymatic hydrolysis was carried out with Hog Liver Esterase propyl groups undergoes esterase triggered breakdown to 5´-TMP more readily than its 3-acetyloxy-2,2-bis-(ethoxycarbonyl)propyl counterpart, but considerably less readily than the 3-acetyloxymethoxy-2,2-bis(ethoxycarbonyl)propyl derived triester. The initial enzymatic deacetylation is followed by release of the resulting 3-hydroxy-2-cyano-2-propyl group as 2-cyano-N-(2-phenyl-ethyl)acrylamide (17). When the reaction is carried out in large excess of glutathione, 17 is largely converted to glutathione conjugate 18. In addition, intramolecular cyclization and hydration occur as side reactions. Thymidine 5´-phosphotriester


Phellinus linteus (PLE) (20 mg/Kg) displayed significant protection against hepatic fibrosis. The present investigation characterized eleven new ionone derivatives, phellinulins D–N (4–14), from the P. linteus mycelium extract and the relative stereochemical structures were constructed according to the spectroscopic and spectrometric analytical results. Some purified compounds were examined for their inhibitory effects on activated rat hepatic stellate cells (HSCs) and several isolates did exhibit significant protection. The results indicated that the mycelium of P. linteus could be explored as a hepatoprotective drug or healthy food candidate in the near future.In the dimethylnitrosamine (DMN)-induced hepatic fibrosis Wistar rat model, the mycelium extract of  Phellinus linteus) displayed the hepatoprotective and antihepatotoxic effects +) corresponding to the formula C15H24O4. Comparison of the UV, IR, 1H and 13C-NMR data of 9 with phellinulin E (5) inferred that the terminal methylene group at C-5 in 5 was oxidized to a dihydroxylation product in 9 , 3.29 ; δC 70.1 (C-13), 73.8 (C-5)). It was further evidenced by the 2J, 3J-HMBC crosspeaks from CH2-13 to C-4, -5 and -6. In the NOESY spectral analysis  exhibited similar UV, IR and 1H-NMR spectroscopic characteristics as compared to those of 5, except the disappearance of one set of carbon-carbon double bond. The HR-ESI-MS analysis of 10 exhibited a sodiated ion peak at m/z 259.1676 established the molecular formula of 10 as C15H24O2. The 1H-NMR spectrum of 10 displayed two aliphatic fragment signals, in which the connections were proved by its COSY spectrum, including three mutually coupled methylene groups at δ 1.23 , 1.47 , 1.54 , 2.03 ; and five mutually coupled protons at δ 1.54 , 1.64 , 1.70 , 1.94, 2.13 , respectively. The stereochemical configuration of C-6 was established by NOESY spectral study ) to C-7, -9, -10 and -12 confirmed the location of hydroxyl group at C-8. Comparatively, 12 is a 3-hydroxylated analog of 10 according to the HMBC analysis in which exhibited correlations from H-4 (δ 1.95) to C-3 (δ 68.2) and H-2 (δ 1.42) to C-3. Furthermore, the stereochemical configurations of 11 and 12 were determined by NOESY analysis  was purified as optically active colorless syrup  was assigned the molecular formula C15H26O3 according to its HRESIMS data, in which was one more oxygen atom than that of 13. Comparison of its 1H-NMR spectra with that of 13 suggested that compound 14 was the 2-hydroxyl derivative of 13, confirmed by the HMBC spectroscopic analysis that exhibited 2J, 3J-correlations from H-2 ) to C-1, -4; and from CH3-14 to C-1, -2, -6, respectively. Although the configuration of H-2 was established as β by its coupling constant and NOESY spectrum , nineteen known compounds, including phellilins A–C (15–17) [18) [H-furan-2-one [p-hydroxybenzaldehyde [In addition to phellinulins A–N ( (15–17) , γ-ionyl17) [18) , ergosta17) [18) , (−)-cin17) [18) , ergoste17) [18) , 6β-hydr17) [18) , cinnamo17) [18) , 5-hydroan-2-one , p-hydroaldehyde , lumichraldehyde , phenylaaldehyde , phenylaaldehyde , phenylaaldehyde , prolinealdehyde , phenylaaldehyde , prolinealdehyde  and phenaldehyde  were alsP. linteus (PLE) exhibited protective effect against hepatic fibrosis in experimental animals in the preliminary study. To validate the inhibition of PLE on hepatic fibrosis in vivo, the histological changes in rat liver tissues were investigated and the experimental results evidenced the protective effect . Chemical shifts are shown in δ values (ppm) with tetramethylsilane as an internal standard. The HR-ESI-MS spectra were taken on a Bruker Daltonics APEX II 30e spectrometer (positive-ion mode) . Column chromatography (CC) was performed on silica gel   and pTLC (preparative thin layer chromatography) was executed on precoated Si gel 60 F254 plates , using UV light to visualize the spots. Melting points were determined using Yanagimoto MP-S3 apparatus  without corrections. Optical rotations were measured using a Jasco DIP-370 digital polarimeter . UV spectra were recorded at room temperature on a Hitachi UV-3210 spectrophotometer , respectively. IR spectra were obtained with a Shimadzu FT-IR DR-8011 spectrophotometer . P. linteus was provided and identified from GeneFerm Biotechnology Co. Ltd. in Taiwan in August 2003. A voucher specimen (Wu 2003010001) has been deposited in the Herbarium of National Cheng Kung University, Tainan, Taiwan.The dried powder of fermentation cultivated mycelium of 6-Week-old male Wistar rats were purchased from a commercial animal breeder . The rats were acclimated for 1 week and housed in an environmentally controlled room at 22 ± 2 °C, 55 ± 10% relative humidity and in 12-hr light/dark cycles. The rats were randomly grouped into four groups of three rats each . Saline and DMN was intraperitoneally injected to rats at 10 mg/kg body weight for 3 consecutive days each week for 4 weeks and the rats of the treatment groups were fed with silymarin (100 mg/kg) and PLE (20 mg/kg) for four weeks via oral route from the first week of DMN exposure. The Committee on Research Involving Animal Subjects of Chang Gung University, Taiwan, approved all experiments in compliance with the standards of Chang Gung University's Committee for the Use and Care of Animals.The liver tissue fixed with 10% neutral-buffered formalin was embedded in paraffin and sliced into 5-μm sections that were stained with hematoxylin-eosin and Masson’s trichrome for a histological assessment. The histological images were observed with optical microscopy  in nonconsecutive, randomly chosen ×200 or ×100 histological fields. The digital photomicrographs were then processed with DP-72 (Olympus).P. linteus  was refluxed with ethanol (4 L × 5 × 4 h). The extract was then filtered with Whatman No. 1 filter paper and the ethanol extracts were combined and concentrated at 40 °C under reduced pressure to obtain the ethanol extract . PLE was further dissolved in water and partition with chloroform to give chloroform  and water extracts (280 g). The PLEC extract was subjected to silica gel column chromatography eluted with n-hexane: ethyl acetate (4:1) to afford six fractions (Frs. 1–6).The dried mycelium powder of 1) (7.1 mg), J (10) (11.0 mg), M (13) (1.1 mg) and γ-ionylideneacetic acid (18) (1.1 g), respectively.Fraction F3 was isolated by silica gel CC with gradient mixture of chloroform and methanol (29:1 to 1:1) to afford three sub fractions (F3-1–F3-3). F3-1 and F3-2 were further purified by silica gel CC and preparative TLC to yield phellinulins A (2) (8.0 mg).Fraction F4 was performed silica gel CC with gradient mixture of chloroform and methanol (20:1 to 1:1) to afford five sub fractions (F4-1–F4-5). F4-1 was purified by silica gel CC and preparative TLC to yield ergosta-,22t)-trien-3β-ol (106.0 mg) and cinnamolide (8.0 mg). F4-3 was further purified by preparative TLC with chloroform and methanol (29:1) to yield phellinulin B (15) (7.0 mg), B (16) (1.0 mg) and C (17) (10.0 mg), respectively. F5-2 was purified by silica gel CC and preparative TLC to yield phellinulins C (3) (1.0 mg), D (4) (3.0 mg), E (5) (12.0 mg), F (6) (32.1 mg), G (7) (12.6 mg), H (8) (9.1 mg) and I (9) (6.0 mg), respectively. F5-3 was further isolated by silica gel CC eluted with chloroform and methanol (15:1) and subsequent preparative TLC purification of the resulted minor fractions to afford phellinulins K (11) (62.1 mg), L (12) (1.0 mg) and N (14) (11.0 mg). F5-4 was further purified by silica gel CC and preparative TLC to yield ergosterol peroxide (5.5 mg), 6β-hydroxycinnamolide (1.0 mg), 3β-hydroxycinnamolide (21.0 mg), 5-hydroxy-4- phenyl-5H-furan-2-one (3.0 mg) and p-hydroxybenzaldehyde (2.0 mg), respectively. Recrystallization of the sub fraction F5-5 produced 7,8-dimethylisoalloxazine (3.0 mg).Fraction F5 was subjected to silica gel CC eluted with gradient mixture of chloroform and methanol (19:1 to 1:1) to produce five sub fractions (F5-1–F5-5). F5-1 was further purified by preparative TLC with chloroform and methanol (29:1) to yield obtain phellilins A  to produce three sub fractions (F6-1–F6-3). Subsequent CC on silica gel of the sub fractions 6-1 and 6-2 eluted by the chloroform and methanol mixture resulted in several dipeptide derivatives, including phenylalanine-leucine (1.0 mg), phenylalanine-isoleucine (3.0 mg), phenylalanine (1.9 mg), proline-valine (26.0 mg), phenylalanine-proline (2.0 mg), proline-leucine (4.0 mg) and phenylalanine-alalinine (11.0 mg), respectively.Phellinulin D (4). Colorless powder (CHCl3); mp 92–94 °C; c 0.3, MeOH); UV (MeOH) λmax (log ε) 221.2 (2.99) nm; IR (neat) νmax 3074, 2931, 2870, 1689, 1647, 1438, 1238 cm−1; 1H-NMR  and 13C-NMR , see m/z 273.1466 .Phellinulin E (5). Colorless syrup (CHCl3); c 0.3, MeOH); UV (MeOH) λmax (log ε) 263.8 (4.02) nm; IR (neat) νmax 3399, 3074, 2943, 1686, 1608, 1254 cm−1; 1H-NMR  and 13C-NMR , see m/z 273.1465 .Phellinulin F (6). White needles (CHCl3); mp 138–140 °C; c 0.3, MeOH); UV (MeOH) λmax (log ε) 263.0 (4.29) nm; IR (neat) νmax 2954, 2870, 1685, 1613, 1250, 1176 cm−1; 1H-NMR  and 13C-NMR , see m/z 273.1465 .Phellinulin G (7). Colorless syrup (CHCl3); c 0.1, MeOH); UV (MeOH) λmax (log ε) 258.8 (4.28) nm; IR (neat) νmax 3360, 2959, 2928, 1690, 1609, 1250 cm−1; 1H-NMR  and 13C-NMR , see m/z 273.1464 .Phellinulin H (8). White powder (CHCl3); mp 125–127 °C; c 0.1, MeOH); UV (MeOH) λmax (log ε) 260.0 (3.89) nm; IR (neat) νmax 3360, 2947, 2866, 1686, 1609, 1254 cm−1; 1H-NMR  and 13C-NMR , see m/z 273.1465 .Phellinulin I (9). Colorless needles (CHCl3); mp 180–183 °C; c 0.2, MeOH); UV (MeOH) λmax (log ε) 264.0 (3.50) nm; IR (neat) νmax 3418, 2947, 2928, 2859, 1686, 1451, 1250 cm−1; 1H-NMR  and 13C-NMR , see m/z 291.1574 .Phellinulin J (10). White powder (CHCl3); mp 59–60 °C; c 0.1, MeOH); UV (MeOH) λmax (log ε) 264.2 (4.57) nm; IR (neat) νmax 2940, 2870, 1690, 1640, 1424, 1253 cm−1; 1H-NMR  and 13C-NMR , see m/z 259.1676 .Phellinulin K (11). Colorless syrup (CHCl3); c 0.8, MeOH); UV (MeOH) λmax (log ε) 219.2 (3.74) nm; IR (neat) νmax 2932, 2866, 1694, 1647, 1246 cm−1; 1H-NMR  and 13C-NMR , see m/z 275.1622 .Phellinulin L (12). Colorless syrup (CHCl3); c 0.2, MeOH); UV (MeOH) λmax (log ε) 219.0 (3.66) nm; IR (neat) νmax 2947, 2874, 1690, 1643, 1427, 1246, 675 cm−1; 1H-NMR  and 13C-NMR , see m/z 275.1621 .Phellinulin M (13). Colorless syrup (CHCl3); c 0.2, MeOH); UV (MeOH) λmax (log ε) 202.4 (3.35); 264.4 nm; IR (neat) νmax 2936, 2870, 1708, 1647, 1447, 1292 cm−1; 1H-NMR  and 13C-NMR , see m/z 261.1828 .Phellinulin N (14). Colorless syrup (CHCl3); c 0.1, MeOH); UV (MeOH) λmax (log ε) 264.6 (3.49), 220.4 (3.69) nm; IR (neat) νmax 3363, 2939, 2870, 1709, 1385, 1256, 1261 cm−1; 1H-NMR  and 13C-NMR , see m/z 277.1781 .2. A total of 1 × 105 cells were seeded in 24-well plates for 24 h and made quiescent by incubating in medium containing 0.2% FBS overnight. After treating with 40 μM of tested compounds for 72 h, isopropanol solution mixed with tetrazolium salt was added to the wells and incubated for additional 4 h at 37 °C. The optical density of the dissolved material was measured spectrophotometrically at 570 nm and assays were performed at least in triplicate [The immortalized rat myofibroblast cell line HSC-T6 was a kind gift of Dr. Scott L. Friedman . The HSC-T6 cells were maintained in Waymouth medium containing 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere of 5% COiplicate .Experimental results were expressed as mean ± standard deviation (SD) of three parallel measurements .


Xylaria sp., the effect of culture conditions on metabolite production was investigated. A shift in the production of five known cytochalasin-type analogues 1–5 to six new α-pyrone derivatives, xylapyrones A–F (compounds 6–11), from a solid to a liquid medium was observed. These compounds were identified by analysis of 1D and 2D NMR and HRMS data. Compounds 1–3 showed moderate cytotoxicity against HepG2 and Caski cancer cell lines with IC50 values ranging from 25 to 63 μM and compounds 4–11 were found to be inactive, with IC50 values >100 μM.Seeking a strategy for triggering the cryptic natural product biosynthesis to yield novel compounds in the plant-associated fungus  This paper describes the isolation, structure elucidation, and cytotoxic activities of the isolated compounds 1–11 , cytochalasin K (2), cytochalasin Z16 (3), rosellichalasin (4) and cytochalasin Z17 (5) were identified by comparison of the corresponding 1H- and 13C-NMR data with published results . The NMR spectroscopic data  of 8 was replaced by an oxygenated methine unit at δH 4.23  and δC 69.5(CH) in 9. Unexpectedly, no HMBC correlation from H1-7 (δH 4.23) to any other carbon signal was observed, so the location of the hydroxyl group was deduced to be C-7 according to the COSY correlations from OH-7 (δH 5.60) via H1-7 (δH 4.23) to H2-8 (δH 1.65 and 1.54) and a strong HMBC correlation from H2-8 (δH 1.54) to C-6 (δC 168.3) (9 was identified as shown in Compound C 168.3) . Consequ10) and xylapyrone F (11) were determined straightforwardly as they were very similar to those of 8. Their NMR data . Cytochalasin-type compounds have been reported as promising lead compounds for anticancer drug discovery that reduce the proliferation of different cancer cell lines such as P388, A549 and KB cells +, 1013 [2M+Na]+.Cytochalasin K (2): White amorphous solid; C28H33NO7; UV (MeOH) λmax (logε) 224 (3.38), 258 (2.42) nm; 13C-NMR (CDCl3): δ 211.5 (C-17), 170.0 (C-1), 149.0 (C-22), 142.5 (C-20), 136.7 (C-1'), 133.6 (C-14), 131.8 (C-6), 129.4 (C-13), 129.3 (C-2' & 6'), 129.0 (C-3' & 5'), 127.2 (C-4'), 125.3 (C-5), 120.5 (C-19), 86.2 (C-9), 77.2 (C-18), 70.1 (C-7), 59.0 (C-3), 50.0 (C-8), 48.4 (C-4), 44.2 (C-10), 41.0 (C-16), 39.0 (C-15), 24.6 (C-25), 20.2 (C-24), 17.7 (C-11), 14.0 (C-12); ESIMS m/z 518 [M+Na]+, 1013 [2M+Na]+.Cytochalasin Z16 (3): White amorphous solid; C28H33NO5; UV (MeOH) λmax (logε) 244 (3.73) nm; 13C-NMR (CDCl3): δ 206.1 (C-17), 171.3 (C-1), 170.0 (C-21), 149.1 (C-6), 143.7 (C-18), 138.1 (C-1'), 137.9 (C-14), 132.9 (C-19), 130.2 (C-2' & 6'), 129.8 (C-3' & 5'), 128.0 (C-4'), 126.6 (C-13), 115.8 (C-12), 84.1 (C-9), 69.9 (C-7), 54.4 (C-3), 50.8 (C-8), 49.9 (C-4), 44.6 (C-10), 40.6 (C-15), 40.3 (C-16), 37.2 (C-20), 32.7 (C-5), 18.5 (C-23), 15.0 (C-11), 13.5 (C-24); ESIMS m/z 486 [M+Na]+, 949 [2M+Na]+.Rosellichalasin (4): White amorphous solid; C28H33NO5; UV (MeOH) λmax (logε) 244 (3.75) nm; 13C-NMR (CDCl3): δ 205.4 (C-17), 171.6 (C-1), 169.0 (C-21), 143.0 (C-18), 136.5 (C-1'), 135.2 (C-14), 131.7 (C-19), 129.5 (C-2' & 6'), 128.8 (C-3' & 5'), 127.1 (C-4'), 125.6 (C-13), 84.5 (C-9), 60.1 (C-3), 57.1 (C-6), 53.8 (C-7), 48.9 (C-4), 46.9 (C-8), 44.1 (C-10), 39.7 (C-5), 39.6 (C-15), 36.4 (C-20), 35.9 (C-16), 19.5 (C-12), 17.3 (C-11), 12.8 (C-23), 12.7 (C-24); ESI-MS m/z 486 [M+Na]+, 949 [2M+Na]+.Cytochalasin Z17 (5): White amorphous solid; C28H33NO5; UV (MeOH) λmax (logε) 242 (3.71) nm; 13C-NMR (CDCl3): δ 205.5 (C-17), 171.6 (C-1), 168.9 (C-21), 143.2 (C-18), 137.1 (C-14), 137.0 (C-1'), 132.9 (C-6), 131.5 (C-19), 129.2 (C-2' & 6'), 128.9 (C-3' & 5'), 127.1 (C-4'), 126.4 (C-13), 125.1 (C-5), 83.7 (C-9), 69.6 (C-7), 59.2 (C-3), 50.0 (C-8), 49.6 (C-4), 43.7 (C-10), 40.1 (C-16), 39.7 (C-15), 37.4 (C-20), 17.5 (C-11), 17.2 (C-23), 14.1 (C-12), 13.0 (C-24); ESI-MS m/z 486 [M+Na]+, 949 [2M+Na]+.Xylapyrone A (6): white amorphous powder; UV (MeOH) λmax (logε) 201 (0.16), 280 (2.94) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 254.1147 [M]+ .Xylapyrone B (7): white amorphous powder; UV (MeOH) λmax (logε) 201 (0.14), 280 (2.93) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 256.1308 [M]+ .Xylapyrone C (8): white amorphous powder; UV (MeOH) λmax (logε) 201 (0.15), 280 (2.93) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 212.1045 [M]+ .Xylapyrone D (9): white amorphous solid; UV (MeOH) λmax (logε) 201 (0.14), 280 (2.98) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 228.0991 [M]+ .Xylapyrone E (10): pale yellow oil; UV (MeOH) λmax (logε) 201 (0.15), 280 (2.99) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 198.0886 [M]+ .Xylapyrone F (11): white amorphous powder; UV (MeOH) λmax (logε) 201 (0.17), 280 (2.99) nm; 1H-NMR (DMSO-d6) and 13C-NMR (DMSO-d6) spectral data see m/z 184.0729 [M]+ .l-glutamine, 100 µg/mL streptomycin, and 100 U/mL penicillin. Cultures were incubated in a humidified atmosphere of 5% CO2 at 37 °C. Cells (1 × 104/well) were seeded in supplemented culture medium (100 μL/well) in a 96-well plate and incubated for 24 h. The medium was then replaced with a test compound-containing medium, and the cells were further incubated for 48 h. All experiments were run in parallel with controls (0.1% DMSO without test compounds) and the cell viabilities were evaluated by MTT assays. The absorbance of formazan formed was measured at 570 nm by a microplate reader. The concentrations resulting in 50% inhibition of cell proliferation/survival (IC50) as measured by this assay are given in The cancer cell lines CaSki and HepG2 were obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Science . All cells were maintained in RPMI-1640 medium supplemented with 10% fetal calf serum, 25 mM HEPES buffer, 2 mmol/L Xylaria genus is an ubiquitous filamentous fungus, often isolated from marine environments and terrestrial sources, that can produce various types of secondary metabolites, including typical cytochalasins [Xylaria sp. Pukachaisirikul et al. described the isolation of one known α-pyrone derivative [1–5 were isolated as the major metabolites from a solid rice medium culture of Xylaria sp. BM9. However, applying liquid culture conditions to Xylaria sp. BM9 resulted in the identification of six new α-pyrone derivatives, xylapyrones A–F (compounds 6–11), with different biogenetic origin, in which its PKS genes for the biosynthesis of complex liquids were actived. Xylapyrones A–F were evaluated for in vitro cytotoxicity against two cancer cell lines (Hep-G2 and Caski) and found to be inactive. Further bioassay evaluation of the compounds against fungi and other targets are ongoing, so it is an issue that we don’t have sufficient amounts of these compounds to determine the absolute stereochemistry of compounds 6, 7 and 9.The halasins , terpenohalasins , benzofuhalasins , xanthonhalasins , and cychalasins . Long-chhalasins ,23. A ferivative . In this


Phytophthora cinnamomi is a phytopathogen that causes extensive damage in different crops, and therefore, produces important economic losses all around the world. Chemical fungicides are a key factor for the control of this disease. However, ecological and environmental considerations, as well as the appearance of strains that are resistant to commercial fungicides, have prompted the quest for new antifungal agents which are of low ecological impact. In this work, a series of new 2-allylphenol derivatives was synthesized, and their structures were confirmed by FT-IR, NMR, and MS. Some of the synthesized compounds, more specifically nitro derivatives, exhibit strong growth inhibition of P. cinnamomi with EC50 as low as 10.0 µg/mL. This level of activity is similar to that exhibited by METALAXYL MZ 58 WP, a commonly-used commercial fungicide; therefore, these compounds might be of agricultural interest due to their potential use as fungicides against P. cinnamomi. The results indicate that this activity depends on the chemical structures of the 2-allylphenol derivatives, and that it is strongly enhanced in molecules where nitro and hydroxyl groups adopt a -para configuration. These effects are discussed in terms of the electronic distribution of the aromatic ring induced by substituent groups. Phytophthora represent some of the most devastating crop diseases worldwide. All known species of Phytophthora are plant pathogens  (100%).This compound was obtained by the acetylation of 2-AP. To a stirred solution of 2-AP  in dichloromethane (10 mL) was added 4-3 (yellow oil) was obtained by CC : IR (cm−1): 3080 (=C-H); 1768 (C=O); 1522 (C=C); 1344 (CH3); 1184 (C-O); 1166 (C-O-C). 1H-NMR: 7.25 ; 7.18 ; 6.93 ; 6.88 ; 6.09–5.99 ; 5.11–5.06 ; 3.85 ; 3.42 . 13C-NMR: 157.2 (C-1); 137.0 (C-2′); 129.7 (C-5); 128.6 (C-2); 127.3 (C-3); 120.5 (C-4); 115.3 (C-3′); 110.3 (C-6); 55.3 (OCH3); 34.2 (C-1′). EI-MS (+) m/z non-detected.This compound was obtained by the methylation of 2-AP  in the presence of dry acetone (120 mL), potassium carbonate  and dimethyl sulfate . Compound 4; reddish oil ; Fraction II: Compound 9; brown oil . Compound 4: IR (cm−1): 3201 (OH); 3085 (=C-H); 2977 (C-H); 1609 (C=C); 1539 (-NO2); 1448 (C=C); 1329 (N=O); 1249 (C-O-C). 1H-NMR: 10.90 ; 7.99 ; 7.46 ; 6.92 ; 6.03–5.93 ; 5.13–5.09 ; 3.48 .13C-NMR: 153.3 (C-6); 137.5 (C-3); 135.1 (C-2′); 131.4 (C-1); 123.1 (C-5); 119.5 (C-4); 112.5 (C-3); 116.8 (C-3′); 33.6 (C-1′). EI-MS (+) m/z 179 [M +] (100%) Compound 9: IR (cm−1): 3218 (OH); 3105 (=C-H); 2921 (C-H); 1610 (C=C); 1331 (CH2); 1552 (-NO2); 1432 (N=O). 1H-NMR: 11.40 ; 8.95 ; 8.34 ; 6.03–5.93 ; 5.27–5.19 ; 3.57 . 13C-NMR: 157.3 (C-1); 139.7 (C-4); 133.8 (C-6); 133.2 (C-2′); 130.9 (C-3); 119.7 (C-5); 118.7 (C-3′); 33.6 (C-1′). EI-MS (+) m/z 224 [M+] (100%).The nitration of 2-AP  was performed in dichloromethane (5 mL) with sulfonitric mixture (2 mL) at 2 °C. Two fractions were obtained by CC. Fraction I: Compound 4. First, 2-AP  was dissolved in dichloromethane (100 mL) and added to a mixture containing potassium hydrogen sulfate , sodium nitrate , and wet silica . The mixture was stirred under reflux for 24 h. The complete disappearance of the starting product was confirmed by TLC . The solid product was filtered through silica and washed with dichloromethane; the solvent was evaporated in a vacuum to yield a reddish oil. The pure product  was obtained by CC .Because of the low reaction yield obtained with the previous reaction, the following alternative synthetic path was used to obtain compound 2 (1.0 g 5.71 mmol) was carried out in dichloromethane (8 mL) using sulfonitric mixture (2 mL) at 2 °C. Two fractions were obtained by CC. Fraction I: Compound 5; yellow oil ; Fraction II: Compound 7; reddish oil . Compound 5: IR (cm−1): 3080 (=C-H); 2930 (C-H); 1768 (C=O); 1522 (N-O); 1344 (N-O); 1184 (C-O); 1165 (C-O-C). 1H-NMR: 7.95 ; 7.54 ; 7.32 ; 5.92–5.82 ; 5.15–5.07 ; 3.38 ; 2.37 .13C-NMR: 168.3 (CO); 142.4 (C-1); 135.9 (C-6); 135.5 (C-3); 134.5 (C-2′); 121.0 (C-2); 117.5 (C-3′); 34.4 (C-1′); 20.7 (CH3CO). EI-MS (+) m/z 43 (CH3CO); 179 (C9H8NO3); 221 [M+] Compound 7: IR (cm−1): 3085 (=C-H); 2915 (C-H); 1767(C=O); 1638 (C=C); 1521 (N-O); 1344 (N-O); 1183 (C-O); 1166 (C-O-C). 1H-NMR: 8.14–8.10 ; 7.23 ; 5.93–5.83 ; 5.16 ; 5.12 ; 3.37 ; 2.34  13C-NMR: 168.2 (CO); 153.6 (C-4); 145.5 (C-1); 134.0 (C-2′); 133.8 (C-2); 125.6 (C-3); 123.3 (C-6); 122.8 (C-5); 117.7 (C-3′); 34.3(C-1′); 20.8 (CH3CO). EI-MS (+) m/z equal Compound 5.Nitration of compound 4  was methylated in dry acetone (45 mL) with potassium carbonate  and dimethylsulfate . Compound 6 (reddish oil) was obtained by CC . Compound 6: IR (cm−1): 3085 (=C-H); 2937 (C-H); 1602 (NO2); 1523 (C=C); 1353 (N=O). 1H-NMR: 7.63 ; 7.43 ; 7.15 ; 6.00–5.90 ; 5.15–5.06 ; 3.88 ; 3.48 . 13C-NMR: 151.4 (C-2); 144.2 (C-3); 136.4 (C-1);135.7 (C-2′); 135.0 (C-6); 123.8 (C-5); 123.4 (C-4); 117.0 (C-3′); 62.6 (OCH3); 33.5 (C-1′). EI-MS (+) m/z 193 [M+] (100%).Compound 7  in methanol (5 mL) and K2CO3  was stirred for 4 h at room temperature; then, HCl (0.1 M) was added to achieve pH = 2. The organic phase was extracted with CH2Cl2 (3 × 15 mL), washed with water, dried over Na2SO4, filtered, and evaporated. Subsequently, the mixture was purified by CC eluting with hexane + ethyl acetate mixtures of increasing polarity. Compound 8; yellow oil : IR (cm−1): 3353 (O-H); 2929 (C-H); 2361 (C=C); 1586 (NO2); 1493 (C=C); 1334 (N=O); 1280 (C-O). 1H-NMR: 8.06 ; 8.05 ; 6.88 ; 6.03–5.99 ; 5.26–5.19 ; 3.46 ; 13C-NMR: 159.7 (C-1); 141.6 (C-4); 134.6 (C-2′); 126.4 (C-2); 126.3 (C-3); 124.3 (C-5); 118.0 (C-3′); 115.8 (C-6); 34.6 (C-1′). EI-MS (+) m/z 179 [M+] (100%).A solution containing 10 was obtained by methylation of 9  with dimethylsulfate  in dry acetone (36 mL) and presence of potassium carbonate . Compound 10 (reddish oil) was obtained by CC . Compound 10: IR (cm−1): 3094 (=C-H); 2924 (C-H); 1593 (C=C); 1530 (-NO2); 1477 (C=C); 1342 (N=O); 1264 (C-O-C). 1H-NMR: 8.57 ; 8.30 ; 6.00–5.90 ; 5.26 ; 5.17 ; 3.98 ; 3.55 . 13C-NMR: 156.3 (C-6); 143.0 (C-3); 142.4 (C-1); 138.4 (C-2′); 133.9 (C-5); 128.8 (C-2); 119.5 (C-4); 63.0 (OCH3); 33.8 (C-3′). EI-MS (+) m/z 238 [M+] (100%).Compound 1–10) against P. cinnamomi were tested according to a procedure previously reported [v/v) solution and added to petri dishes (d = 50 mm) containing a potato dextrose agar medium  at 50 °C. The final concentrations of all the assayed compounds were: 10, 25, 50, 150, and 250 µg/mL. Solvent (1% ethanol) was taken as the negative control (C−), whereas METALAXIL MZ 48 WP  commercial fungicide was used as positive control (C+) at the same concentrations and under the same conditions as those of the tested compounds. A mycelium agar disc (d = 4 mm) of the pathogen fungi was placed upside down in the center of PDA and incubated for six days at 23 °C in the dark. Colony diameters were measured when the colonies on the C− plates covered 100% of the agar surface. The inhibition percentages of mycelial growth for each compound were calculated by using a previously-described method [50) was obtained for each compound by fitting %I and concentration to a dose-response equation  were obtained after molecular mechanic (MM) optimization using ChemDraw 3D 15.0.All 2-AP derivatives synthetized were optimized using Gaussian 09 and the DFT-B3LYP-6-31G  basis set, and verified by frequency calculations (without imaginary frequency) in gas phase calculus. After optimization, the electrostatic potential map of each compound was plotted. The lipophilicity index  changes by attaching electron-acceptor groups to the aromatic ring, or by chemical modification of the hydroxyl group. In addition, this activity is strongly enhanced in those molecules where nitro and hydroxyl groups adopt a -para configuration. These effects are mainly attributed to changes in the electronic distribution on the aromatic ring induced by substituent groups.In this work, a series of phenylpropanoids derived from 2-AP was synthetized and evaluated for its antifungal properties on mycelial growth of P. cinnamomi at such low concentrations, which convert these molecules in potential precursors of useful fungicides for agricultural applications focused to control the infection caused for P. cinnamomi. However, further work is needed to increase the yields of reactions, leading to the most active compounds, and to determine the mechanism of action of these nitro derivatives.It is worth mentioning that few known fungicides exhibit activity against


Fruits. The structures of these compounds were established by 1D and 2D NMR spectra and HR-ESI-MS. To the best of our knowledge, this is the first scientific report of diterpenes from Arenga genus.Three new  Arenga pinnata (Wurmb.) Merr. are tall evergreen trees belonging to the genus Arenga of the family Palmae. They are widely distributed in Southeast Asian countries, including China. A. pinnata fruits are the fruits of the A. pinnata. +. The acid hydrolysis of 1 liberated d-glucose, which was identified by HPLC analysis using an optical rotation detector. 1H-NMR spectrum , δH 0.86 ; two olefinic proton signals at δH 5.0 , δH 5.39 ; an anomeric proton signal at δH 4.40 . The 13C-NMR and DEPT spectrum  and C-3, C-4, C-5, C-19 (COOH); H-20 (CH3) and C-1, C-5, C-9, C-10; H-15 and C-7, C-8, C-9, C-16; H-14 and C-12, C-13, C-16; H-17 and C-16. Thus, structure 1 was confirmed as shown in Compound spectrum  of 1 shospectrum  of 1 shoskeleton . The comand 62.9 . The 1H- B and C . The conJ5,6 = 6.5 Hz and J6,7 = 6.5 Hz) and H-7  observed in the 1H-NMR spectrum were the same as those of H-6  and H-7  in 7β, 16α, 17-trihydroxy-ent-kauran-6α, 19-olide +. The acid hydrolysis of 2 liberated d-glucose, which was identified by HPLC analysis using an optical rotation detector. 1H- and 13C-NMR spectra indicated that the structure of 3 was similar to that of 2, except for a 11β-hydroxy group  and 17-hydroxy group , as well as the presence of a 11-methylene group , and 17-(β-glucopyranosyl oxy) group . The connectivity of these partial structures and the functional groups were investigated by analysis of the HMBC of 3. As shown in 3) and C-3, C-4, C-5, C-19 (COOH); H-20 (CH3) and C-1, C-5, C-9, C-10; H-15 and C-7, C-8, C-9, C-16; H-7 and C-8, C-14, C-15; H-17 and C-13, C-16, C-1′. Thus, the structure of 3 was confirmed as shown in Compound 3 was assigned by analysis of the NOESY spectrum. The 1H- and 13C-NMR spectra showed similar data for 3 and 2. Likewise, it was determined that H-6 was in the β-orientation and that H-7 was in the α-orientation. The NOESY of 3 showed the cross-peaks between H-6 and H-18, H-5, H-9; H-7 and H-20. The results showed that the stereochemistry of 3 is similar to the stereochemistry of 2. Therefore, the structure of 3 was determined to be as shown . The other compounds were characterized as pseudaminic acid (4), 12α-(β-d-glucopyranosyl)-7β-hydroxy-kaurenolide (5), paniculoside (6), agittarioside b (7), orychoside B (8) by comparing their NMR spectroscopic data with the literature values . UV spectra were recorded on a PerkinElmer Lambda UV-365 instrument . IR spectra were recorded on a PerkinElmer Spectrum Two spectrometer  with KBr disks. Preparative HPLC  was performed on 5C18 MS-II . NH2 column , silica gel , Amberlite IRA-400  and ODS  were used for column chromatography. Detectors  and  were used in the HPLC. Precoated silica GF 254 plates  were used for TLC analysis. All the solvents were of analytical grade .NMR spectra were measured on a Bruker AV-400 spectrometer  with TMS as an internal standard. High-resolution ESI-MS mass spectra were carried out on an AB SCEIX Triple-TOFTM 5600A. pinnata fruits were collected from Guangxi in China during September 2017, and authenticated by Prof. Weiming Wang of the Heilongjiang Research institute of Chinese Medicine. The fruitage had been deposited at the Heilongjiang Research institute of Chinese Medicine.The A. pinnata fresh fruits (30.0 kg) were extracted with 70% EtOH (200 L × 3 h × 3 times). The combined extract was concentrated under vacuum yielding a residue (3.0 kg) which was dissolved in H2O (12 L) and extracted sequentially with petroleum, chloroform, ethyl acetate and n-butanol (12 L × 3 h × 5 times). The eluate was separately concentrated in vacuo to give a petroleum syrup (109.0 g), chloroform syrup (123.0 g), ethyl acetate syrup (205.0 g), and an n-butanol syrup (380.0 g). In this study, we only separated the n-butanol layer. The n-butanol (380.0 g) extract was subject to column chromatography on silica gel (4460.0 g) and eluted with CH2Cl2/MeOH (20:1 (80.0 L), 10:1 (110.0 L), 5:1 (120.0 L), 3:1 (100.0 L), 2:1 (80.0 L) and 1:1 (60.0 L), v/v) to afford six fractions (fractions A (36.0 g), B (96.7 g), C (90.2 g), D (40.1 g), E (21.2 g), F (20.1 g). The TLC and HPLC were used to observe each of the fractions, and similar fractions were combined to afford A1-A6, B1-B5, C1-C6, D1-D6, E1-E10, F1-F6. Fraction B4 (21.7 g) was eluted by Rp-18 (600.0 g) (MeOH/H2O 2:8 (1.4 L)→3:7 (2.0 L)→4:6 (2.7 L)→5:5 (2.0 L)→6:4 (2.0 L)→7:3 (1.4 L)→8:2 (1.4 L)→9:1 (0.8 L)→1:0 (1.0 L), v/v) to afford nine subfractions (subfractions B4-1–B4-9). Subfraction B4–6 further purified by a preparative RP-HPLC  to give 1 . Subfractions B4–5 were further purified by a preparative RP-HPLC  to give 2 . Fraction C1 (8.0 g) was eluted by Rp-18 (600.0 g) (MeOH/H2O 2:8 (1.4 L)→3:7 (2.0 L)→4:6 (2.7 L)→5:5 (2.7 L)→6:4 (2.0 L)→7:3 (1.4 L)→8:2 (1.4 L)→9:1 (0.8 L)→1:0 (1.0 L), v/v) to afford nine subfractions (subfractions C1-1–C1-9). Subfractions C1–3 were further purified by a preparative RP-HPLC  to give 4 . Subfraction C1–5 were further purified by a preparative RP-HPLC  to give 5 . Fraction D3 (18.0 g) was eluted by Rp-18 (600.0 g) (MeOH/H2O 2:8 (1.4 L)→3:7 (2.0 L)→4:6 (2.7 L)→5:5 (2.7 L)→6:4 (2.7 L)→7:3 (1.4 L)→8:2 (1.4 L)→9:1 (0.8 L)→1:0 (1.0 L), v/v) to afford nine subfractions (subfractions D3-1–D3-9). Subfractions D3–6 were further purified by a preparative RP-HPLC  to give 3 . Subfraction D3–6 were further purified by a preparative RP-HPLC  to give 8 . Subfractions D3–4 were further purified by a preparative RP-HPLC  to give 6 . Subfractions D3–5 were further purified by a preparative RP-HPLC  to give 7 .The 1). Yellow amorphous powder. Gave −1; 1H- and 13C-NMR  data, see m/z 513.2651 [M + H]+  (Arenterpenoids A (13.2655) .2). Yellow amorphous powder. Gave −1; 1H- and 13C-NMR  data, see m/z 385.2186 [M + H]+  (Arenterpenoids B (85.2182) .3). White amorphous powder. Gave −1; 1H- and 13C-NMR  data, see m/z 531.2769 [M + H]+  (Arenterpenoids C (31.2761) .1,3) (2.0 mg) were in 1.0 mL HCl and were each heated under reflux for 3 h. After cooling, the two mixtures were separately filtered with Amberlite IRA-400 to give a solution. Assigned with AcOEt to get two layers. The aqueous layer was evaporated to dryness under vacuum, and then subjected to HPLC analysis using an NH2 column and an optical-rotation detector. d-glucose was confirmed by comparison of the tR with that of an authentic sample (mobile phase: MeCN/H2O 85: 15 (v/v); flow rate: 0.8 mL/min; tR = 12.8 min )The isolated compounds (A. pinnata fruits were mainly used to treat rheumatism and bone pain. This study obtained eight diterpene compounds from A. pinnata fruits, including three new diterpenes and five known ones. This also reveals some structural characteristics of the chemical constituents in the A. pinnata fruit, which provides some clues for further clarifying the composition of the components and correlations of the relative plant species. In this study we have made this contribution to discover active ingredients and leading compounds and additionally provided an experimental and scientific basis of drug design and drug discovery of the A. pinnata fruits.As a traditional Chinese medicine,


These results indicated that both potency and selectivity of COX-2 inhibitory activity were affected by the type and size of amine on C-5 of imidazothiazole ring. Our data identified N,N-dimethyl-1-(6-(4-(methylsulfonyl)phenyl)imidazothiazol-5-yl)methanamine (6a) as a potent and selective COX-2 inhibitor . Our results indicated that both potency and selectivity of COX-2 inhibitory activity were affected by the type and size of amine on C-5 of imidazothiazole ring.A new series of imidazothiazole analogs containing a methyl sulfonyl COX-2 pharmacophore was synthesized and evaluated for their COX-2 inhibitory activity. According to  Also, we have recently reported a group of imidazopyridine derivatives with remarkable potency and selectivity. In this series, the most potency and selectivity of COX-2 inhibition belongs to 2-(4-(methylsulfonyl)phenyl)-3-(morpholinomethyl)H-imidazopyridine   can relieve inflammation, fever, and pain by inhibiting cyclooxygenase. Cyclooxygenase (COX) is the key enzyme in the biosynthesis of inflammation mediators, prostaglandins . There aelecoxib , Rofecoxelecoxib  and Valdelecoxib . Increaselecoxib . Accordirimidine  exhibite> 10 µM) . We repoivatives . In thisH-indole  was the = 217.1) . In prese on C-5 .Materials and Methods1HNMR spectra with TMS as internal standard were acquired with Bruker FT-500 MHz instrument  values are estimated in hertz (Hz) and spin multiples are given as s (singlet), d (double), t (triplet), q (quartet), and m (multiplet). The mass spectral measurements were performed on a 6410 Agilent LCMS triple quadrupole mass spectrometer (LCMS) with an electrospray ionization (ESI) interface. Microanalyses, determined for C and H, were within ±0.4% of theoretical values. All reagents used in this study were purchased from Merck AG and Aldrich Chemical companies without further purifications. Melting points were determined with a Thomas–Hoover capillary apparatus. Perkin Elmer Model 1420 spectrometer was used to acquire Infrared spectra. Preparation of 4-(methylthio)acetophenone (2)3 (87 mmol) was suspended in 200 mL CHCl3 and stirred until disperse uniformly. After cooling to 10 ºC, 6.2 mL (87 mmol) acetyl chloride was added, followed by dropwise addition of 7.5 mL (80 mmol) thioanisole when the temperature was under 4 ºC and allowed the mixture to stir at room temperature for 2 h. The mixture was poured into crushed ice, organic layer was separated with CHCl3 and washed with saturated NaHCO3 solution and finally dried over anhydrous Na2SO4. The solvent was evaporated under reduced pressure. The precipitate was filtered and washed with n-hexane. Yield 87%; white crystalline powder; mp: 81-82 ºC; IR (KBr disk): υ (cm-1) 1690 (C=O); LC-MS (ESI) m/z: 167 .11.5 g AlCl4-(Methylsulfonyl)acetophenone (3)3 (3 × 25 mL). The organic layer was washed twice with saturated NaHCO3 solution and dried over anhydrous Na2SO4. The solvent was evaporated and white precipitate was recrystallized in ethanol. Yield 95%; white crystalline powder; mp: 128-130 ºC; IR (KBr disk): υ (cm-1) 1148, 1309 (SO2), 1681 (C=O); LC-MS (ESI) m/z: 198.9 .To a solution of 4 g of 1 (24 mmol) in 20 mL THF, mixture of 20 g oxone in THF and water (1:1) was added and stirred at room temperature for 2 h. THF was removed under reduced pressure and extracted with CHClα-Bromo-4-(methylsulfonyl)acetophenone (4)3. The bromine was added drop wise. After the reaction was completed (monitored by TLC), the solvent was evaporated under reduced pressure and the precipitate was recrystallized in ethanol. Yield 83%; white crystalline powder; mp: 125-127 ºC; IR (KBr disk): υ (cm-1) 1165, 1308 (SO2) 1710 (C=O); LC-MS (ESI) m/z: 276.7 .Dissolve 2 g (10.1 mmol) of 2 in 20 mL CHCl6-(4-(Methylsulfonyl)phenyl)imidazothiazole (5)2CO3 (7.25 mmol) and 0.36 g 2-aminothiazole (3.4 mmol) were added and refluxed for 24 hours. The precipitate was filtered off and washed with water. Yield 70.5%; brown powder; mp: 145.5-147 ºC; IR (KBr disk): νcm-1 1149, 1298 (SO2); 1HNMR (CDCl3): δ ppm 3.08 , 6.94 , 7.51 , 7.89-8.01 ; LC-MS (ESI) m/z: 279.0 . Anal. Calcd. for C12H10N2O2S2: C, 51.78; H, 3.62; N, 10.06. Found: C, 51.52; H, 3.85; N, 10.25.To 1 g of 3 (3.62 mmol) in ethanol, 0.38 g Na6-(4-(Methylsulfonyl)phenyl)imidazothiazole derivatives (6a-g)4  was added and the reaction mixture stirred at 50 ºC for reaction completion. The reaction mixture was cooled under 10 ºC and pH was adjusted to ~8-9 with 20% sodium hydroxide solution. The solid was filtered and washed with water. The crude was purified by chromatography to give compounds (6a-g) (Yield 50-70%).At 0 ºC, acetic acid, ~36% formalin solution (1.37 mmol) and appropriate aliphatic amine 1.37 mmol) were added in the flask. Then, N,N-Dimethyl-1-(6-(4-(methylsulfonyl)phenyl)imidazothiazol-5-yl)methanamine (6a)-1) 1156, 1316 (SO2); 1HNMR (CDCl3): δ ppm 2.28 , 3.08 , 3.79 , 6.85 , 7.68 , 7.98 ; LC-MS (ESI) m/z : 336.0 . Anal. Calcd. for C15H17N3O2S2: C, 53.71; H, 5.11; N, 12.53. Found: C, 53.51; H, 5.35; N, 12.84.Yield 61%; Yellow powder; mp: 123-124 ºC; IR (KBr): 1 ν (cmN-Ethyl-N-((6-(4-(methylsulfonyl)phenyl)imidazothiazol-5-yl)methyl)ethanamine (6b)-1) 1152,1304 (SO2); 1HNMR (CDCl3): δ ppm 1.04-1.07 , 2.54-2.58 , 3.12 , 3.97 , 6.86 , 7.81 ,8.01 ; LC-MS (ESI) m/z : 364.0 . Anal. Calcd. for C17H21N3O2S2: C, 56.17; H, 5.82; N, 11.56. Found: 56.35; H, 6.02; N, 11.71.Yield 55.1%; Yellow powder; mp: 132-135 ºC; IR (KBr): ν (cmN-((6-(4-(Methylsulfonyl)phenyl)imidazothiazol-5-yl)methyl)-N-propylpropan-1-amine (6c)-1) 1155,1307 (SO2); 1HNMR (CDCl3): δ ppm 0.84-0.87 , 1.46-1.53 , 2.41-2.43 , 3.12 , 3.96 , 6.86 , 7.76 , 8.01  ; LC-MS (ESI) m/z : 392.1 . Anal. Calcd. for C19H25N3O2S2: C, 58.28; H, 6.44; N, 10.73. Found: C, 58.33; H, 6.57; N, 10.98.Yield 54.9%; Yellow powder; mp: 165-167 ºC; IR (KBr): ν (cm6-(4-(Methylsulfonyl)phenyl)-5-(pyrrolidin-1-ylmethyl)imidazothiazole (6d)-1) 1147,1307 (SO2); 1HNMR (CDCl3): δ ppm 1.82 , 2.57 , 3.11 , 4.03 , 6.87 , 7.78 , 8.00 ; LC-MS (ESI) m/z : 362.00 . Anal. Calcd. for C17H19N3O2S2: C, 56.49; H, 5.30; N, 11.62. Found: C, 56.13; H, 5.51; N, 11.88.Yield 51%; Yellow powder; mp: 144-146 ºC; IR (KBr): ν (cm6-(4-(Methylsulfonyl)phenyl)-5-(piperidin-1-ylmethyl)imidazothiazole (6e)-1) 1151, 1309 (SO2); 1HNMR (CDCl3): δ ppm 1.50 ,1.58-1.63 , 2.46 , 3.12 , 3.84 , 6.87 , 7.78 , 8.02  ; LC-MS (ESI) m/z : 376 . Anal. Calcd. for C18H21N3O2S2: C, 57.58; H, 5.64; N, 11.19. Found: C, 57.78; H, 5.91; N, 11.26. Yield 60%; Yellow powder; mp: 142.6-144 ºC; IR (KBr): ν (cm4-((6-(4-(Methylsulfonyl)phenyl)imidazothiazol-5-yl)methyl)morpholine (6f)-1) 1173, 1331 (SO2); 1HNMR (CDCl3): δ ppm 2.49 , 3.08 , 3.69-3.71 , 3.86 , 6.87 , 7.70 , 7.98 ; LC-MS (ESI) m/z : 378.1 . Anal. Calcd. for C17H19N3O3S2: C, 54.09; H, 5.07; N, 11.13. Found: C, 54.23; H, 5.29; N, 11.33.Yield 69.5%; Yellow powder; mp: 203-206 ºC; IR (KBr): ν phenyl)imidazothiazol-5-yl)methyl)azanediyl)bis(ethan-1-ol) (6g)-1) 1170, 1331 (SO2), 3417 (OH) ; 1HNMR (CDCl3): δ ppm 2.13-2.14 , 2.74-2.76 , 3.13 , 3.66-3.68 , 4.18 , 6.91 , 7.91 , 7.96-7.98 ; 8.02 ; LC-MS (ESI) m/z: 396.1 . Anal. Calcd. for C17H21N3O4S2: C, 51.63; H, 5.35; N, 10.62. Found: C, 51.77; H, 5.65; N, 10.82.Yield 65%; white powder; mp: 168.8-170 ºC; IR (KBr): ν  and hydrogens were added. All the ligand molecules were built by the Builder module and were energy minimized for 1000 iterations reaching a convergence of 0.01 kcal/mol Å. The energy minimized ligands were superimposed on SC-558 in the PDB file of 6COX after which SC-558 was deleted. Searching for the desired binding configuration between the small flexible ligands and the rigid protein is the purpose of docking study. For efficiency, protein residues with atoms greater than 6.0 Å from the docking box were removed. The quality of the docked structures was evaluated by measuring the intermolecular energy of the ligand-enzyme assembly , 33.In-vitro cyclooxygenase (COX) inhibition assayin-vitro and whole cell methods.Enzyme chemiluminescent kit  was used to evaluate inhibition activities of synthesized compounds based on our previous reported procedure . The CayChemistry1), acetyl chloride and aluminium chloride through a Friedel-crafts reaction, 4-(methylthio)acetophenone was obtained (2). (Methylsulfonyl)acetophenone (3) was prepared using oxone in THF/water. The bromination of compound (3) via bromine in CHCl3 at room temperature afforded α-bromo-4-(methylsulfonyl)acetophenone (4). Compound (4) was treated with 2-aminothiazole to yield 6-(4-(methylsulfonyl)phenyl)imidazothiazole (5). The target molecules (6a-g) were synthesized in presence of appropriate aliphatic amines, formalin in acetic acid. The purity of all products was determined by thin layer chromatography using different polarity solvent systems. The structures of the synthesized compounds were confirmed by IR, 1HNMR, and ESI-MS.The synthesis of 6-(4-(methylsulfonyl)phenyl)imidazothiazole derivatives was carried out according to Docking study6f) within the COX-2 binding site were evaluated by a docking experiment. Results obtained from docking procedure of 6a as selective and most potent COX-2 inhibitor among the synthesized compounds into the COX-2 binding site . One of O-atoms of para-SO2Me forms a hydrogen bonding interaction with the amino group of Arg513 (distance = 3.27 Å). Additional docking study showed that 6a and SC558 were superimposed tightly in COX active site  were selective inhibitors of COX-2 isoenzyme with IC50 values in the highly potent 0.08-0.16 µM range. These results indicated that both potency and selectivity of COX-2 inhibitory activity were affected by the type and size of amine on C-5 of imidazothiazole ring. Based on these results N,N-dimethyl-1-(6-(4-(methylsulfonyl)phenyl)imidazothiazol-5-yl)methanamine (6a) was proved to be the most potent (IC50 = 0.08 µM) COX-2 inhibitor among the synthesized compounds. These results showed that replacement of dimethylamino group with larger dialkylamino substituents such as diethylamino (6b) or dipropylamino (6c) decreased both potency and selectivity for COX-2 inhibitory activity. Our results also indicated that compounds having cyclic amino groups such as pyrrolidine (6d), piperidine (6e) or morpholine (6f) showed moderate to high COX-2 selectivity. However, compound 6f having morpholine group had higher potency and selectivity for COX-2 inhibitory activity compared with compound 6e possessing piperidine substituent which might be explained by the ability of morpholine for hydrogen binding with COX-2 active site. Similarly, compound 6g containing diethanolamine with hydrogen binding potential showed high COX-2 inhibitory potency and selectivity. These findings confirm that synthesized compounds should inhibit the biosynthesis of prostaglandins through the cyclooxygenase pathway at sites of inflammation. The ability of 6-(4-(methylsulfonyl)phenyl)imidazothiazole derivatives to inhibit COX-1 and COX-2 was measured using chemiluminescent enzyme assay to investigate the effect of different substituents on COX-2 selectivity and potency . Accordi6a) exhibited highly COX-2 inhibitory potency.The results of this investigation indicated that (i) 6-(4-(methylsulfonyl)phenyl)imidazothiazole is a suitable scaffold to design COX-2 inhibitors, (ii) COX-1/-2 inhibition is sensitive to the type of substituent at C-5 of imidazothiazole ring, (iii) N,N-dimethyl-1-(6-(4-(methylsulfonyl)phenyl)imidazothiazol-5-yl)methanamine (