III, 2-hydroxy- IV and 2-acyloxyethylsulfonamides V were obtained and their structures confirmed by IR, 1H-NMR, and elemental analysis. The in vitro and in vivo bioactivities against two Botrytis cinerea strains, DL-11 and HLD-15, which differ in their sensitivity to procymidone, were evaluated. The in vitro activity results showed that the EC50 values of compounds V-1 and V-9 were 0.10, 0.01 mg L−1 against the sensitive strain DL-11 and 3.32, 7.72 mg L−1 against the resistant strain HLD-15, respectively. For in vivo activity against B. cinerea, compound V-13 and V-14 showed better control effect than the commercial fungicides procymidone and pyrimethanil. The further in vitro bioassay showed that compounds III, IV and V had broad fungicidal spectra against different phytopathogenic fungi. Most of the title compounds showed high fungicidal activities, which could be used as lead compounds for further developing novel fungicidal compounds against Botrytis cinerea.Sulfonyl-containing compounds, which exhibit a broad spectrum of biological activities, comprise a substantial proportion of and play a vital role, not only in medicines but also in agrochemicals. As a result increasing attention has been paid to the research and development of sulfonyl derivatives. A series of thirty-eight 2-substituted phenyl-2-oxo-  Sulfonyl-containing compounds, which exhibit a broad spectrum of biological activities, comprise a substantial proportion of and play an important role not only in medicines but also in agrochemicals ,2,3,4,5.Phomopsis asparagi, Cladosporium fulvum and Fusarium oxysporum, etc. [Venturia nashicola and Fusarium graminearum [Botrytis cinerea and Corynespora cassiicola.Sulfonamides were the first drugs with a selective effect on bacteria that could be used systemically against bacterial infections . Hence, um, etc. ,24,25,26minearum . Using iminearum  has beenN-substituted-2-oxo-2-phenylethylsulfonamide derivatives were synthesized using combinatorial chemistry, and the fungicidal activity against B. cinerea were evaluated. Therefore, the SAR of substituent groups on the N atom such as monosubstituted anilines, multi-substituted anilines, substituted benzylamines and alkylamines were systematically studied [In our previous work, different scaffolds were introduced onto lead 2-oxocycloalkyl-sulfonamides, and the structure-activity relationships (SAR) were studied ,30,31,32 studied .III, 2-hydroxy- IV and 2-acyloxyethylsulfonamides V  as catalyst.Potassium 2-substituted-phenyl-2-oxoethylsulfonates N-(2-trifluoromethyl-4-chlorophenyl)-2-substituted-phenyl-2-oxoethylsulfonamides III with high fungicidal activity were selected for further reaction with sodium borohydride, giving N-(2-trifluoromethyl-4-chlorophenyl)-2-hydroxy-2-substituted-phenylethyl-sulfonamides IV. Among these compounds N-(2-trifluoromethyl-4-chlorophenyl)-2-hydroxy-2-ethylsulfonamide (IV-5) showed excellent fungicidal activity. Therefore, we further synthesized a series of N-(2-trifluoromethyl-4-chlorophenyl)-2-acyloxy-2-ethyl-sulfonamides V by the reaction of IV-5 with acyl chlorides in the presence of TMEDA and molecular sieves as catalysts. Finally, a total of 38 2-substituted-phenyl-2-oxo-, 2-hydroxy- and 2-acyloxy- ethylsulfonamides were synthesized. Flash chromatography was used for separation and purification and the structures of all the title compounds were confirmed by IR, 1H-NMR, and elemental analysis. The 1H-NMR spectra is available in According to the structure-activity relationships between substituents on the phenyl ring and fungicidal activity, some of these Botrytis cinerea with different sensitivity to procymidone collected from different areas in Liaoning, China were tested by in vitro mycelium growth inhibition assay and in vivo greenhouse pot experiments. Moreover, the fungicidal spectra against seven phytopathogenic fungi  Leonard et Suggs, Pythium aphanidermatum, Phytophthora capsici Leonian, Fusarium graminearum Schw., Corynespora cassiicola and Thanatephorus cucumeris) were also evaluated.The fungicidal activities of all the target compounds against two different strains of N-(2-trifluoromethyl-4-chlorophenyl)-2-substituted-phenyl-2-oxoethylsulfonamides III against two different B. cinerea strains DL-11 (a sensitive strain) and HLD-15 (a resistant strain) were evaluated by an in vitro mycelium growth inhibition assay, and the commercial fungicides procymidone, chlorothalonil and pyrimethanil were used as the positive controls. The synthetic compound (N-(2-trifluoromethyl-4-chlorophenyl)-2-phenyl-2-oxoethyl-sulfonamide (B-1) was also used as the positive control to compare the difference between the pre- and post-structure optimization. The inhibition of mycelial growth was evaluated by measuring colony diameters in the presence and absence of the tested compounds. The results of preliminary bioactivity screening, expressed as the percentage of inhibition, showed that most of the compounds III showed high inhibition rate (more than 70% at 50 mg L−1) and better activity than that of B-1. Based on the experimental results, the EC50 values of the compounds III were next evaluated to confirm their fungicidal effects on the two different kinds of B. cinerea strains DL-11 and HLD-15. The values are listed in The fungicidal activities of 50 values of compounds III against B. cinerea strains DL-11 and HLD-15 were within 1.58–10.81 mg L−1 and 3.48–17.70 mg L−1, respectively. Among them, compounds III-13, with EC50 values of 1.58 mg L−1 and 3.48 mg L−1 against DL-11 and HLD-15, respectively, exhibited the best fungicidal activity against B. cinerea. The commercial fungicide procymidone showed EC50 values of 2.59 mg L−1 and 15.95 mg L−1, chlorothalonil gave EC50 values of 1.66 mg L−1 and 17.52 mg L−1 and pyrimethanil provided EC50 values of 32.73 mg L−1 and 71.25 mg L−1 against the two strains DL-11 and HLD-15, against which B-1, another positive control, provided EC50 values of 14.25 mg L−1 and 22.79 mg L−1. The antifungal activity of compounds III-8, III-14 and III-15 was close to that of the control compound procymidone. With procymidone as a standard fungicide against B. cinerea strain DL-11, the relative activities of compounds III-13 and III-15 were 1.64- and 1.57-fold higher, respectively; with B-1 as a standard fungicide, the relative activities of compounds III-13 and III-15 were 9.02- and 8.69 fold better, respectively. As regards to the control efficiency of B. cinerea strain HLD-15, compounds III-13 and III-15, with 14.42 and 13.89 times higher activity than that of B-1, were 10.09 and 9.72 times better than procymidone, respectively.As shown in N-(2-trifluoromethyl-4-chlorophenyl)-2-hydroxy-2-substituted-phenyl ethylsulfonamides IV and N-(2-trifluoromethyl-4-chlorophenyl)-2-acyloxy-2-ethylsulfonamides V against B. cinerea was also evaluated and the EC50 values are summarized in 50 values of compounds IV against B. cinerea strains DL-11 and HLD-15 were within 0.70–13.33 mg L−1 and 0.61–33.16 mg L−1, respectively, and with compounds V, within 0.01–1279.16 mg L−1 and 3.32–3648.50 mg L−1, respectively. Some of compounds IV and V which were derived from compounds III showed better activity against B. cinerea. Compound V-9 showed the best in vitro fungicidal activity against DL-11, with an EC50 value of 0.01 mg L−1, and IV-5, with an EC50 value of 0.61 mg L−1, exhibited the best activity against HLD-15 in vitro. With procymidone as a standard fungicide against the B. cinerea strain DL-11, the relative activities of compounds V-1 and V-9 were 25.9- and 259-fold higher, respectively. As regards to the control effect of the strain HLD-15, compounds IV-5 and V-1 were 26.15- and 4.8-times more active than procymidone, respectively. Additionally, there is significantly different in the activity level among compounds V. For instance, compound V-1 showed an EC50 values of 0.1 mg L−1 against the B. cinerea strain DL-11, but compound V-6 showed an EC50 value of 131.12 mg L−1.The fungicidal activity of III showed better fungicidal activity against B. cinerea strain DL-11 (sensitive strain) than that against HLD-15 (resistant strain) because of the resistance, but some individual compounds, on the contrary, exhibited better activity against HLD-15. In general, compounds should exhibit better activity against sensitive strains than resistant strains. Moreover, the same compound gave different control effects on the different B. cinerea strains owing to the difference of biological characteristics, pathogenicity and resistant to fungicides, but on the whole, there were consistent trends in the control effects against both DL-11 and HLD-15.Finally, it was important to stress that most compounds III, IV and V against B. cinerea was evaluated and the results listed in III, IV and V showed good inhibition, and some of them exhibited better control than the commercial fungicides procymidone and pyrimethanil. Among them, compounds V-13 demonstrated the best activity, with a control efficiency of 74.6%. Besides, the control efficiencies of V-1, V-6, V-8, V-10, V-13 and V-14 were 70.8%, 71.3%, 70.8%, 71.3%, 74.6% and 73.3%, respectively, which were similar to that of the commercial fungicide procymidone (with a control efficiency of 72.5%).The in vivo fungicidal activity of compounds IV and V was somewhat increased in comparison to compounds III, especially compounds V, all of which showed control efficiencies of more than 60%. In addition, the in vitro fungicidal activity of the tested compounds showed no consistent correlation with that in vivo. Compound V-13, with the high EC50 values of 35.18 mg L−1 in vitro, had the highest control efficiency of 74.6% against the B. cinerea strain DL-11. The reason may be that, there were interactions between reagents and plants in vivo experiments, not just between the reagents and the fungi. Therefore, characteristics of the tested compounds such as polarity, conductibility and degradation in vivo also had influence on the fungicidal activity, which is worthy of further study.The tested results showed that the whole level of the control effects of compounds III, IV and V. We summarize the bioassay results of inhibitory activities of these compounds against seven phytopathogenic fungi at 50 mg L−1. Overall, the title compounds showed different levels of fungicidal activity. Compounds III-2, III-10, III-16, IV-5 and V-10 exhibited wide spectrum antifungal activity and compound V-10 displayed excellent activities against all the tested phytopathogentic fungi. Moreover, some compounds showed highly specific pathogen activity, whereby compounds III-1 and III-4 exhibited activities against Phytophthora capsici Leonian with inhibition rates 90.4% and 91.0%, respectively. It is noteworthy that nearly all compounds III showed excellent fungicidal activity against Phytophthora capsici Leonian and Fusarium graminearum Schw., and most of the inhibition rate was more than 70%. In general, compounds V possessed an improvement of fungicidal activity in comparison to compounds III. However, compounds V were only slightly active against Exserohilum turcicum (Pass.) Leonard et Suggs and Pythium aphanidermatum, being less active than compounds III. Besides, some individual compound showed unexpected antifungal activity against a certain pathogen. For example, Compound V-16 exhibited good activity against P. aphanidermatum with inhibition rate of 72.9%, although the inhibition rates of most compounds V were less than 25%.Different kinds of phytopathogenic fungi were chosen to evaluate the fungicidal spectrum of the title compounds III, IV and V against different phytopathogentic fungi, EC50 values of some highly bioactive compounds were evaluated against four different phytopathogentic fungi . Compound B-1 and chlorothalonil were used as the positive controls. As shown in III-6, III-10, III-13 and V-10 exhibited the best fungicidal activity against the four fungi, with EC50 values of 1.41, 0.99, 7.25, 2.25 mg L−1; 1.05, 0.60, 8.65, 4.47 mg L−1; 0.79, 1.86, 8.62, 2.74 mg L−1 and 2.41, 1.07, 6.96, 3.22 mg L−1, respectively.Based on the inhibition rate of compounds III, IV and V were accomplished in short reaction times and using mild reaction conditions. Oleum was often used as an efficient sulfonating agent, but lots of byproducts are generated during the sulphonation process with that reagent. In our reactions, sulfur trioxide-dioxane adduct was used as the sulfonating agent, prepared by a standard procedure , and high yields of products were achieved. Besides, we also tried to react sulfur trioxide-pyridine adduct with the acetophenones, but the yields of this reaction were very low, and in some cases no product was obtained. A tentative inference from this result was that the relative stability and reactivity of sulfur trioxide-pyridine adduct and sulfur trioxide-dioxane adduct has an influence on the sulfonation of acetophenones. Moreover, oxalyl chloride was used as the chlorinating agent in these experiments to give the corresponding sulfonyl chlorides and acyl chlorides, which were reacted with DMF catalysis. The reaction proceeded smoothly in high yields and the product separation and purification were simple. In most cases, the title compounds III, IV and V were obtained in acceptable yield.High yield syntheses of compounds 1H-NMR spectra of N-(2-trifluoromethyl-4-chlorophenyl)-2-substituted-phenyl-2-oxoethyl-sulfonamides III showed some characteristic peaks, i.e., the 1H chemical shifts of the phenyl rings in the sulfonamides were seen at around 7.50 ppm. The 1H-NMR spectrum of compounds III showed a singlet signal around 4.70 ppm, which was assigned to the methylene protons between the sulfonyl and carbonyl groups. Additionally, a signal appearing at approximately 7.25 ppm was assigned to the NH group in the sulfonamide functionality  and infrared (IR) absorption spectroscopy. The ionality a.−1 and 1700 cm−1. The NH stretching vibration was around 3250 cm−1 to 3350 cm−1. The spectra of compounds IV, which were obtained in high yield (more than 85%) by reduction of compounds III, showed significant changes compared with those of compounds III. The signal of the methylene group in the 1H-NMR spectra appeared as a multiplet due to the effect of the chiral carbon in the side chain. Therefore, the 1H-NMR chemical shifts of the CH2 combined with the sulfonyl and CH linked with the hydroxyl group were at about 3.40 ppm and 5.30 ppm, respectively. A signal appearing at approximately 3.20 ppm was assigned to the OH group  of methylene were of chemical unequivalent. The 1H-NMR analysis showed that the coupling constant of Ha and Hb was 1J = 7.35 Hz, and that of Ha and Hc, Hb and Hc were 2J = 1.56 Hz, 3J = 4.83 Hz, respectively , it could be seen that the substituents on the benzene ring might exert a greater influence on the activity. As for N-(2-trifluoromethyl-4-chlorophenyl)-2-substituted-phenyl-2-oxoethylsulfonamides III, the biological experiments showed that the introduction of any substituent in the phenyl ring increased the inhibition activity to Botrytis cinerea. The activity was significantly increased when the benzene ring contained fluorine-containing groups (F and CF3) which show significant functions in organisms due to their osmotic and electronic effects, etc. The number of fluorine-containing groups also influenced the antifungal activity. Thus, when the substituent was fluorine, the fungicidal activity increased with the increase of the number of substituents. If the substituent was a trifluoromethyl group, contrarily the fungicidal activity decreased. Moreover, there was no significant difference between different positions of halogen atoms on the benzene ring and their fungicidal activities. However, for trifluoromethyl, it seemed that the compound with a trifluoromethyl group on the para position of the benzene ring showed higher activities than when it was at the meta position, and the compound with a trifluoromethyl group on the para position exhibited the best activity. We also revealed that the strong electron-donating capacity of the substituent on the phenyl ring reduced the fungicidal activity of compounds III. For example, compounds III-1 and III-3  had lower fungicidal activity than the compound III-13 . The structure-activity relationship of N-(2-trifluoromethyl-4-chlorophenyl)-2-hydroxy-2-substituted-phenyl ethyl-sulfonamides IV was similar to that of compounds III, while for N-(2-trifluoromethyl-4-chlorophenyl)-2-acyloxy-2-ethylsulfonamides V, it could be seen that the acyloxy substituent showed higher fungicidal activity when it was a small group. The compounds with an alkylacyloxy groups showed higher activities than that with an arylacyloxy group, and generally substituted benzoyloxy compounds showed lower activity than benzoyloxy compounds. For instance, compounds V-1, V-10 and V-15 provided EC50 values of 0.10, 1.18 and 20.33 mg L−1 against the Botrytis cinerea strain DL-11, respectively. The fungicidal activities showed no significant difference between compounds V with the change of substituent position of the substituted benzoyloxy group.From the results of the fungicidal activity screening against III and IV, most of which had inhibition rates of more than 75.0% against Botrytis cinerea, although for compounds V, there were obvious differences between their individual fungicidal activities. This suggests that these compounds have different modes of action, which need to be further studied. Additionally, the results of in vitro and in vivo fungicidal activities were inconsistent. For example, with procymidone as a standard fungicide against Botrytis cinerea strains DL-11, the relative toxicity of compound V-9 was 259, which represented the best activity in vitro. However, the in vivo activity of V-10 only showed the control efficiency of 65.4%, and the positive control procymidone had a control efficiency of 58.8%. Further investigation are needed to explain these findings.It is worth mentioning that the fungicidal activities showed no significant difference between compounds 3 on Bruker 400 or 600 MHz spectrometers , using tetramethylsilane (TMS) as an internal standard. Elemental analyses were determined on a Vario EL III elemental analyser .The melting points were measured using an X-5 melting-point apparatus . Infrared (IR) spectra were recorded in potassium bromide disks on a Spectrum 65 spectrophotometer . Nuclear magnetic resonance (NMR) spectra were recorded in CDClII, were prepared from readily commercially available substituted acetophenones I by sulfonation with a sulfur trioxide-dioxane adduct and neutralization with potassium carbonate according to the method given in references [The synthetic route of the title compounds is shown in ferences ,29,30. TII (0.03 mol) and N,N-dimethylformamide  in dichloromethane , oxalyl chloride (0.033 mol) was added dropwise at −5 °C. The mixture was stirred at room temperature for 3 h. After cooling in an ice-water bath, it was filtered under reduced pressure. The filtrate was added dropwise to a solution of 2-amino-5-chlorobenzotrifluoride (0.015 mol) and triethylamine  in CH2Cl2 (20 mL) at −5 °C. After stirring at room temperature for 3 h, the reaction mixture was washed successively with 3 mol L−1 hydrochloric acid , saturated sodium bicarbonate solution (15 mL) and saturated salt water (15 mL), and then dried over sodium sulfate. After evaporating the solvent under vacuum, the crude product was further purified by silica column chromatography to give pure compounds III-1 to III-17 using the mixture of ethyl acetate and petroleum ether (10:1) as the eluent.To a slurry of potassium 2-substituted0phenyl-2-oxoethylsulfonate N-(2-Trifluoromethyl-4-chlorophenyl)-2-(2-methylphenyl)-2-oxoethylsulfonamide (III-1): white solid, 88.1% yield: m.p. 59–61 °C; IR (KBr) νmax 3255, 3045, 2917, 1689, 1338, 1130 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.25 , 4.71 , 7.37 , 7.29–7.75 ; Elemental anal. calc. for C16H13ClF3NO3S (%): C, 49.05; H, 3.34; N, 3.58; found: C, 49.33; H, 3.20; N, 3.78.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-methoxylphenyl)-2-oxoethylsulfonamide (III-2): light brown solid, 83.6% yield: m.p. 83–85 °C; IR (KBr) νmax 3350, 3080, 2968, 1670, 1344, 1161 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.90 , 4.71 , 7.37 , 6.97–7.92 ; Elemental anal. calc. for C16H13ClF3NO4S (%): C, 47.13; H, 3.21; N, 3.43; found: C, 47.36; H, 3.48; N, 3.66.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(2-fluorophenyl)-2-oxoethylsulfonamide (III-3): white solid, 78.9% yield: m.p. 122–124 °C; IR (KBr) νmax 3296, 3033, 2941, 1689, 1311, 1116 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.78 , 7.17–7.20 , 7.30–7.97 ; Elemental anal. calc. for C15H10ClF4NO3S (%): C, 45.52; H, 2.55; N, 3.54; found: C, 45.68; H, 2.34; N, 3.74.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(3-fluorophenyl)-2-oxoethylsulfonamide (III-4): light yellow solid, 81.5% yield: m.p. 109–111 °C; IR (KBr) νmax 3342, 3078, 2974, 1687, 1311, 1122 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.72 , 7.25 , 7.35–7.77 ; Elemental anal. calc. for C15H10ClF4NO3S (%): C, 45.52; H, 2.55; N, 3.54; found: C, 45.36; H, 2.37; N, 3.68.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(2-chlorophenyl)-2-oxoethylsulfonamide (III-5): light yellow solid, 76.6% yield: m.p. 85–87 °C; IR (KBr) νmax 3253, 3064, 2927, 1689, 1307, 1168 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.84 , 7.25 , 7.29–7.75 ; Elemental anal. calc. for C15H10Cl2F3NO3S (%): C, 43.71; H, 2.45; N, 3.40; found: C, 43.90; H, 2.59; N, 3.28.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(3-chlorophenyl)-2-oxoethylsulfonamide (III-6): light yellow solid, 68.4% yield: m.p. 109–111 °C; IR (KBr) νmax 3340, 3056, 2974, 1687, 1338, 1116 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.72 , 7.25 , 7.45–7.91 ; Elemental anal. calc. for C15H10Cl2F3NO3S (%): C, 43.71; H, 2.45; N, 3.40; found: C, 44.01; H, 2.19; N, 3.25.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-bromophenyl)-2-oxoethylsulfonamide (III-7): light yellow solid, 78.9% yield: m.p. 89–91 °C; IR (KBr) νmax 3292, 3033, 2926, 1687, 1396, 1120 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.71 , 7.25 , 7.55–7.81 ; Elemental anal. calc. for C15H10BrClF3NO3S (%): C, 39.45; H, 2.21; N, 3.07; found: C, 39.25; H, 2.03; N, 3.24.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(2-trifluoromethylphenyl)-2-oxoethylsulfonamide (III-8): light yellow solid, 74.3% yield: m.p. 79–81 °C; IR (KBr) νmax 3298, 3032, 2943, 1705, 1315, 1114 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.62 , 7.19 , 7.55–7.78 ; Elemental anal. calc. for C16H10ClF6NO3S (%): C, 43.11; H, 2.26; N, 3.14; found: C, 43.35; H, 2.05; N, 3.36.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(3-trifluoromethylphenyl)-2-oxoethylsulfonamide (III-9): white solid, 77.6% yield: m.p. 85–87 °C; IR (KBr) νmax 3348, 3027, 2972, 1689, 1338, 1184 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.77 , 7.24 , 7.55–8.18 ; Elemental anal. calc. for C16H10ClF6NO3S (%): C, 43.11; H, 2.26; N, 3.14; found: C, 43.31; H, 2.04; N, 2.99.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-trifluoromethylphenyl)-2-oxoethylsulfonamide (III-10): light yellow solid, 79.8% yield: m.p. 116–118 °C; IR (KBr) νmax 3311, 3021, 2960, 1680, 1323, 1134 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.75 , 7.22 , 7.57–8.05 ; Elemental anal. calc. for C16H10ClF6NO3S (%): C, 43.11; H, 2.26; N, 3.14; found: C, 42.92; H, 2.50; N, 3.32.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-nitrophenyl)-2-oxoethylsulfonamide (III-11): light yellow solid, 70.5% yield: m.p. 119–121 °C; IR (KBr) νmax 3273, 3010, 2954, 1703, 1396, 1122 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.77 , 7.17 , 7.58–8.39 ; Elemental anal. calc. for C15H10ClF3N2O5S (%): C, 42.62; H, 2.38; N, 6.63; found: C, 42.84; H, 2.18; N, 6.47.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-oxoethylsulfonamide (III-12): white solid, 71.5% yield: m.p. 89–91 °C; IR (KBr) νmax 3356, 3078, 2972, 1687, 1313, 1176 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.69 , 7.21 , 7.30–7.82 ; Elemental anal. calc. for C15H9ClF5NO3S (%): C, 43.55; H, 2.19; N, 3.39; found: C, 43.82; H, 1.98; N, 3.54.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-oxoethylsulfonamide (III-13): light yellow solid, 75.5% yield: m.p. 93–95 °C; IR (KBr) νmax 3334, 3011, 2926, 1680, 1338, 1122 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.67 , 7.19 , 7.17–7.77 ; Elemental anal. calc. for C15H9ClF5NO3S (%): C, 43.55; H, 2.19; N, 3.39; found: C, 43.72; H, 2.35; N, 3.18.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-oxoethylsulfonamide (III-14): light yellow solid, 80.6% yield: m.p. 106–108 °C; IR (KBr) νmax 3290, 3017, 2946, 1687, 1340, 1120 cm−1; 1H-NMR , δ (ppm) 4.80 , 7.20 , 7.38–7.75 ; Elemental anal. calc. for C15H9Cl2F3NO3S (%): C, 40.34; H, 2.03; N, 3.14; found: C, 40.58; H, 2.31; N, 3.32.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(2-chloro-3-fluorophenyl)-2-oxoethylsulfonamide (III-15): white solid, 72.8% yield: m.p. 115–117 °C; IR (KBr) νmax 3309, 3014, 2916, 1730, 1338, 1122 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.53 , 7.15, 7.22–7.78 ; Elemental anal. calc. for C15H9Cl2F4NO3S (%): C, 41.88; H, 2.11; N, 3.26; found: C, 42.01; H, 1.97; N, 3.42.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-fluoro-3-trifluoromethylphenyl)-2-oxoethylsulfonamide (III-16): light yellow solid, 84.6% yield: m.p. 99–101 °C; IR (KBr) νmax 3354, 3072, 2964, 1685, 1350, 1174 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.73 , 7.20 , 7.37–8.23 ; Elemental anal. calc. for C16H9ClF7NO3S (%): C, 41.44; H, 1.96; N, 3.02; found: C, 41.66; H, 2.10; N, 3.29.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-oxoethylsulfonamide (III-17): light yellow solid, 69.7% yield: m.p. 119–121 °C; IR (KBr) νmax 3354, 3072, 2964, 1685, 1350, 1174 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.77 , 7.15 , 7.57–8.35 ; Elemental anal. calc. for C17H9ClF9NO3S (%): C, 39.74; H, 1.77; N, 2.73; found: C, 39.55; H, 1.52; N, 2.99.N-(2-trifluoromethyl-4-chlorophenyl)-2-phenyl-2-oxoethylsulfonamide (0.01 mol) in methanol (30 mL) at 0–5 °C, sodium borohydride solution (0.016 mol NaBH4 + 8 mL of 1% NaOH + 8 mL of CH3OH) was added dropwise. The mixture was stirred at 5–25 °C for 1–3 h. After methanol was evaporated in vacuum, the residue was dissolved in ethyl acetate (80 mL), washed with 5% HCl (30 mL) and water (20 mL), and dried over sodium sulfate. After the solvent was evaporated under vacuum, the crude product was recrystallized from acetone/petroleum ether to give pure compounds IV-1 to IV-5.To a solution of N-(2-Trifluoromethyl-4-chlorophenyl)-2-phenyl-2-hydroxyethylsulfonamide (IV-1): white solid, 87.2% yield: m.p. 95–96 °C; IR (KBr) νmax 3242, 3061, 1369, 1315, 1128, 975; 1H-NMR (400 MHz), δ (ppm) 3.03 , 3.37–3.67 , 5.35–5.36 , 7.06–7.79 ; Elemental anal. calc. for C15H13ClF3NO3S (%): C, 47.44; H, 3.45; N, 3.69; found: C, 47.69; H, 3.56; N, 3.49.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(3-fluorophenyl)-2-hydroxyethylsulfonamide (IV-2): white solid, 88.6% yield: m.p. 78–80 °C; IR (KBr) νmax 3241, 3024, 1368, 1316, 1134, 984; 1H-NMR (400 MHz), δ (ppm) 3.12 , 3.36–3.54 , 5.35 , 6.96–7.79 ; Elemental anal. calc. for C15H12ClF4NO3S (%): C, 45.29; H, 3.04; N, 3.52; found: C, 45.06; H, 3.21; N, 3.67.N-(2-Trifluoromethyl-4-chlorophenyl)-2-(4-nitrophenyl)-2-hydroxyethylsulfonamide (IV-3): white solid, 94.1% yield: m.p. 110–111 °C; IR (KBr) νmax 3256, 3054, 1385, 1320, 1122, 956; 1H-NMR (400 MHz), δ (ppm) 3.35–3.47 , 5.45–5.47 , 6.91–8.25 ; Elemental anal. calc. for C15H12ClF3N2O5S (%): C, 42.41; H, 2.85; N, 6.60; found: C, 42.58; H, 3.01; N, 6.48.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-hydroxyethylsulfonamide (IV-4): white solid, 88.7% yield: m.p. 104–105 °C; IR (KBr) νmax 3215, 3042, 1349, 1320, 1156, 964; 1H NMR (400 MHz), δ (ppm) 3.19 , 3.32–3.50 , 5.30–5.32 , 6.92–7.79 ; Elemental anal. calc. for C15H11ClF5NO3S (%): C, 43.33; H, 2.67; N, 3.37; found: C, 43.54; H, 2.39; N, 3.47.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-hydroxyethylsulfonamide (IV-5): white solid, 98.8% yield: m.p. 98–100 °C; IR (KBr) νmax 3273, 3025, 1396, 1311, 1122, 989; 1H-NMR (400 MHz), δ (ppm) 3.25 , 3.34–3.47, 5.31–5.33 , 6.75–7.78 ; Elemental anal. calc. for C15H11ClF5NO3S (%): C, 43.33; H, 2.67; N, 3.37; found: C, 43.16; H, 2.88; N, 3.24.N-(2-trifluoromethyl-4-chlorophenyl)-2--2-hydroxyethyl-sulfonamide (0.01 mol), N,N,N′,N′-tetramethylenediamine  and 3Å molecular sieves (2 g) in dry CH2Cl2 (30 mL), acyl chloride (0.011 mol) was added dropwise at room temperature. The mixture was stirred at room temperature for 2 h. The reaction was quenched with ice water (20 mL × 2), filtered and dried over anhydrous magnesium sulfate. After filtering and evaporating the solvent under vacuum, the crude product was purified by silica gel chromatography using petroleum ether/ethyl acetate  as eluent to obtain pure compounds V-1 to V-16.To the solution of N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-acetoxyethylsulfonamide (V-1): white solid, 65.5% yield: m.p. 96–98 °C; IR (KBr) νmax 3350, 3097, 2954, 1761, 1307, 1220 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.11 , 3.38–3.68 , 6.27–6.29 , 7.55–7.58 , 6.78–7.75 ; Elemental anal. calc. for C17H13ClF5NO4S (%): C, 44.60; H, 2.86; N, 3.06; found: C, 44.42; H, 3.01; N, 3.25.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-benzoyloxyethylsulfonamide (V-2): white solid, 51.8% yield: m.p. 93–95 °C; IR (KBr) νmax 3307, 3032, 2952, 1724, 1338, 1269 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.53–3.88 , 6.77–6.95 , 6.77–6.95 , 7.55–7.76 , 7.44–7.4 , 7.50–8.01 ; Elemental anal. calc. for C22H15ClF5NO4S (%): C, 50.83; H, 2.91; N, 2.69; found: C, 50.67; H, 3.12; N, 2.48.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(2-methylbenzoyloxy)ethylsulfonamide (V-3): white solid, 56.3% yield: m.p. 69–71 °C; IR (KBr) νmax 3325, 3028, 2931, 1747, 1311, 736 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.54 , 3.51–3.82 , 6.44–6.45 , 6.78–8.05 ; Elemental anal. calc. for C23H17ClF5NO4S (%): C, 51.74; H, 3.21; N, 2.62; found: C, 51.92; H, 3.10; N, 2.81.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(3-methylbenzoyloxy)ethylsulfonamide (V-4): white solid, 81.1% yield: m.p. 114–116 °C; IR (KBr) νmax 3253, 3033, 2945, 1747, 1338, 734 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.41 , 3.53–3.89 , 6.45–6.48 , 6.77–6.79 ; Elemental anal. calc. for C23H17ClF5NO4S (%): C, 51.74; H, 3.21; N, 2.62; found: C, 51.59; H, 3.43; N, 2.75.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(4-methylbenzoyloxy)ethylsulfonamide (V-5): white solid, 81.8% yield: m.p. 137–139 °C; IR (KBr) νmax 3307, 3032, 2928, 1716, 1338, 827 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.43 , 3.52–3.87 , 6.45–6.47 , 6.77–7.88 ; Elemental anal. calc. for C23H17ClF5NO4S (%): C, 51.74; H, 3.21; N, 2.62; found: C, 51.88; H, 3.10; N, 2.57.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(4-methoxybenzoyloxy)ethylsulfonamide (V-6): white solid, 60.5% yield: m.p. 118–120 °C; IR (KBr) νmax 3307, 3057, 2943, 1716, 1305, 829 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.52–3.55 , 3.83–3.87 , 3.88 , 6.43–6.45 , 6.75–7.95 ; Elemental anal. calc. for C23H17ClF5NO5S (%): C, 50.24; H, 3.12; N, 2.55; found: C, 50.49; H, 2.94; N, 2.72.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-ethylsulfonamide (V-7): white solid, 71.8% yield: m.p. 120–122 °C; IR (KBr) νmax 3307, 3054, 2927, 1745, 1392, 821 cm−1; 1H-NMR (400 MHz), δ (ppm) 2.37 , 2.51 , 3.51–3.85 , 6.42–6.44 , 6.77–7.83 ; Elemental anal. calc. for C24H19ClF5NO4S (%): C, 52.61; H, 3.50; N, 2.56; found: C, 52.74; H, 3.41; N, 2.68.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(2-chloropropionyloxy)ethylsulfonamide (V-8): light yellow solid, 62.7% yield: m.p. 124–126 °C; IR (KBr) νmax 3502, 3115, 2997, 1691, 1303, 1155 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.30–3.40 , 3.77–5.82 , 6.58–7.85 ; Elemental anal. calc. for C18H14Cl2F5NO4S (%): C, 42.70; H, 2.79; N, 2.77; found: C, 42.56; H, 2.91; N, 2.58.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-dichloroacetoxyethylsulfonamide (V-9): white solid, 51.4% yield: m.p. 150–152 °C; IR (KBr) νmax 3102, 3010, 2956, 1714, 1384, 1170 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.93–4.62 , 6.53 , 5.99–6.02 , 6.35–7.88 ; Elemental anal. calc. for C17H11Cl3F5NO4S (%): C, 38.77; H, 2.11; N, 2.66; found: C, 38.86; H, 2.33; N, 2.48.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-trichloroacetoxyethylsulfonamide (V-10): white solid, 80.9% yield: m.p. 168–170 °C; IR (KBr) νmax 3082, 3010, 2954, 1705, 1379, 1224 cm−1; 1H-NMR (400 MHz), δ (ppm) 4.05–4.71 , 6.32–6.34 , 6.88–7.79 ; Elemental anal. calc. for C17H10Cl4F5NO4S (%): C, 36.39; H, 1.80; N, 2.50; found: C, 36.51; H, 2.02; N, 2.31.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(2-fluorobenzoyloxy)ethylsulfonamide (V-11): white solid, 75.6% yield: m.p. 118–120 °C; IR (KBr) νmax 3307, 3095, 2956, 1743, 756, 686 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.52–3.88 , 6.49–6.51 , 6.78–7.93 ; Elemental anal. calc. for C22H14ClF6NO4S (%): C, 49.13; H, 2.62; N, 2.60; found: C, 49.32; H, 2.51; N, 2.84.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(3-fluorobenzoyloxy)ethylsulfonamide (V-12): white solid, 32.9% yield: m.p. 97–99 °C; IR (KBr) νmax 3290, 3099, 2953, 1741, 752, 684 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.51–3.97 , 6.45–6.47 , 6.77–7.81 ; Elemental anal. calc. for C22H14ClF6NO4S (%): C, 49.13; H, 2.62; N, 2.60; found: C, 48.97; H, 2.54; N, 2.77.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(2-chlorobenzoyloxy)ethylsulfonamide (V-13): white solid, 49.6% yield: m.p. 99–101 °C; IR (KBr) νmax 3327, 3080, 2931, 1741, 754, 692 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.52–3.87 , 6.45–6.48 , 6.87–7.90 ; Elemental anal. calc. for C22H14Cl2F5NO4S (%): C, 47.67; H, 2.55; N, 2.53; found: C, 47.43; H, 2.38; N, 2.69.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(3-chlorobenzoyloxy)ethylsulfonamide (V-14): white solid, 69.5% yield: m.p. 114–116 °C; IR (KBr) νmax 3273, 3048, 2935, 1747, 1338, 742 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.52–3.88 , 6.45–6.48 , 6.78–7.95 ; Elemental anal. calc. for C22H14Cl2F5NO4S (%): C, 47.67; H, 2.55; N, 2.53; found: C, 47.45; H, 2.78; N, 2.39.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(2-trifluoromethylbenzoyloxy) ethylsulfonamide (V-15): white solid, 79.8% yield: m.p. 81–83 °C; IR (KBr) νmax 3308, 3095, 2948, 1747, 1319, 779 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.51–3.84 , 6.45–6.48 , 6.75–7.90 ; Elemental anal. calc. for C23H14ClF8NO4S (%): C, 46.99; H, 2.40; N, 2.38; found: C, 46.78; H, 2.64; N, 2.14.N-(2-Trifluoromethyl-4-chlorophenyl)-2--2-(3-trifluoromethylbenzoyloxy) ethylsulfonamide (V-16): white solid, 43.9% yield: m.p. 119–121 °C; IR (KBr) νmax 3273, 3054, 2931, 1747, 1338, 694 cm−1; 1H-NMR (400 MHz), δ (ppm) 3.52–4.15 , 6.49–6.51 , 6.79–8.27 ; Elemental anal. calc. for C23H14ClF8NO4S (%): C, 46.99; H, 2.40; N, 2.38; found: C, 47.11; H, 2.26; N, 2.51.B. cinerea were tested by a mycelium growth inhibition assay and greenhouse pot experiments, respectively. The Botrytis cinerea strains  were isolated from damaged parts of tomato in a greenhouse in different areas of Liaoning, China, and cultured on potato dextrose agar (PDA) for many generations. The commercial fungicides procymidone (with a purity of 96%), pyrimethanil (with a purity of 95%) and chlorothalonil (with a purity of 96%) provided by the Shenyang Research Institute of the Chemical Industry, National Pesticides Engineering Research Centre, were used as positive controls. Excel 2007  was used to analyze the bioassay data. Analysis of difference significances was performed using SPSS v.18.0 .The in vitro and in vivo fungicidal activities of all the title compounds against III, IV and V was assessed using a radial growth test on potato dextrose agar (PDA). The compounds were dissolved in acetone and mixed with sterile molten PDA to obtain five concentrations of 100, 25, 6.25, 1.56 and 0.39 mg L−1. PDA with different concentrations of the test compounds was poured into 90 mm Petri dishes (15 mL per dish), which were then inoculated with 5 mm plugs of B. cinerea. The plugs were obtained from a PDA culture plate by punching at the edge of the mycelia colony. Three replicates were used per treatment. The commercial fungicides were used as positive controls. After an incubation period of 72 h at 23 °C under a regular 12:12 h light:dark regimen, mycelia growth diameters were measured and the inhibition percentages relative to the control with 1% acetone were calculated. The inhibition rate was determined following a standard method [50 values were calculated using log-probit analysis.The in vitro fungicidal activity of compounds d method . The EC5B. cinerea was evaluated in the greenhouse. B. cinerea was maintained on PDA medium at 4 °C. The culture plates were cultivated at 24 ± 1 °C. Germination was conducted by soaking cucumber seeds in water for 2 h at 50 °C and then keeping the seeds moist for 24 h at 28 °C in an incubator. When the radicles were 0.5 cm, the seeds were grown in plastic pots containing a 1:1 (v/v) mixture of vermiculite and peat. Cucumber plants used for inoculations were at the stage of two seed leaves. The tested compounds were confected to 2.5% EC formulations, which were diluted to concentration of 500 μg/mL with water to obtain the solutions. Tested compounds and commercial fungicides were sprayed with a hand spray on the surface of the seed leaves. Water sprayed seed leaves were set as the CK. After drying, the upper sides of the leaves were inoculated with 5 mm plugs of B. cinerea, which was maintained on PDA. The plants were maintained at 24 ± 1 °C and above 80% relative humidity in the greenhouse. The fungicidal activity was then evaluated.The in vivo fungicidal activity of the title compounds against III, IV and V against phytopathogentic fungi  Leonard et Suggs, Pythium aphanidermatum, Phytophthora capsici Leonian, Fusarium graminearum Schw., Corynespora cassiicola and Thanatephorus cucumeris) was assessed using the mycelium growth test on PDA. The compounds were dissolved in acetone and mixed PDA to obtain final concentration of 50 mg L−1. Chlorothalonil was used as the positive control. Other test conditions were the same with the method given in the mycelial growth.The fungicidal activity of compounds 1H-NMR, IR and elemental analysis. In vitro and in vivo fungicidal activities against 2 different B. cinerea strains were tested. The results showed that some title compounds exhibited better fungicidal activities than that of the lead compound B-1. Among them, compound V-13 showed better in vivo fungicidal activity than the commercial fungicides procymidone and pyrimethanil. In addition, these new compounds had a broad spectrum of fungicidal activity. Bioassays showed that some target molecules might be used as new lead compounds for further development of novel fungicides against B. cinerea. Finally, as we all know, sulfonamides are more used as herbicides, while its application as fungicides is less common in agrochemicals. In our preliminary research, we found that Botrytis cinerea control using sulfonamides may involve a new mode of action mode [In summary, 38 2-substituted-phenyl-2-oxo-, 2-hydroxy- and 2-acyloxyethylsulfonamides were designed and synthesized. Their structures were characterized by ion mode ,37, whic


Dendroidone (1) and dendroidiol (2) were isolated from samples collected at Biscaia Inlet, Angra dos Reis, Rio de Janeiro and at Manguinhos Beach, Serra, Espírito Santo, respectively. Debromoelatol (3), obtusane (4) and -2,3,5,9-tetramethyltricyclo[6.3.0.01.5]undecan-2-ol (5) were obtained from specimens collected at Vermelha Beach, Parati, Rio de Janeiro. The structures of new compounds were elucidated by extensive NMR  and high resolution mass spectrometry analysis. Additionally, the absolute configuration of compound 2 was assigned by X-ray analysis. Full spectroscopic data is described for the first time for compound 3. Anti-inflammatory and antimycobacterial activities of compounds 2–5 were evaluated. Compounds 3–5 inhibited the release of inflammatory mediator NO while TNF-α levels were only affected by 3. All compounds tested displayed moderate antimycobacterial action.Two new chamigrane sesquiterpenes  Laurencia, family Rhodomelaceae, order Ceramiales undecan-2-ol (5) from Vermelha Beach, Parati, Rio de Janeiro (population C). Compounds 2–5 were tested for anti-inflammatory and antimycobacterial activities.The organic extracts were subjected to chromatographic separations yielding dendroidone . The molecular formula was established as C15H20BrClO2 on the basis of HR-APCI-MS data , implying five degrees of unsaturation. The infrared (IR) spectrum exhibited absorptions of hydroxyl  and carbonyl  groups. The 1H-NMR spectrum displayed signals corresponding to two methyl groups at δH 1.09 (H3-12) and 1.11 (H3-13), two methines geminal to heteroatoms at δH 4.67 (H-10) and δH 4.21 (H-9) and three olefinic hydrogens at δH 5.16 (H-14a), δH 4.77 (H-14b) and one highly deshielded at δH 7.24 (H-15), suggesting it was part of a conjugated system. 13C and HSQC experiments revealed the presence of two methyls, four aliphatic methylenes, two deshielded methines, two quaternary sp3 carbons and five sp2 carbon resonances that were assigned to a ketone [δC 197.6 (C-4)] and two olefins . The positions of the bromine and the hydroxyl groups were deduced from the carbon chemical shifts of carbons at δC 69.6 (C-10) and 72.0 (C-9), respectively  and to δC 48.7 (C-6), 69.6 (C-10) and 43.0 (C-11).Compound H 2.81 (H-2a) and δH 1.76 (H-1b) on the COSY spectrum along with HMBC correlations from δH 2.46 (H-5b) to δC 197.6 (C-4) and from δH 1.76 (H-1b) to δC 48.7 (C-6). Based on the 13C-NMR data, a chlorine atom was assigned to C-15 undec-3(15)-ene-4-one which represents a new chemical entity, which was trivially named dendroidone.Ring B was proposed based on correlations between methylenes protons at δ to C-15 . The Z c2 was isolated as colorless crystals,  = −19.2° , with the molecular formula C15H24BrClO2 deduced by HR-ESI-MS from the pseudomolecular ion peak [M+Na]+ at m/z 375.0417 , requiring three degrees of unsaturation. The IR spectrum exhibited absorptions of a hydroxyl group at 3,463 cm−1. The 1H-NMR spectrum (H 5.25 (H-14a) and 4.97 (H-14b), three tertiary methyl groups at δH 1.03 (H3-13), 1.06 (H3-12) and 1.21 (H3-15) and three hydrogens of heterosubstituted carbons at δH 4.07 (H-9), 4.23 (H-4) and 4.52 (H-10). The 13C-NMR spectrum and DEPT-135 experiment revealed the presence of fifteen carbon atoms corresponding to three methyls, five methylenes, three methines and four quaternary carbons, including two olefinic carbons and four carbons attached to heteroatoms. 1H-1H COSY and HMBC correlations indicated that the first ring was similar to compound 1 while the second was proposed based on 1H-1H COSY correlations between δH 4.23 (H-4) and 2.10  and correlations from δH 1.21 (H3-15) to δC 33.8 (C-2), 70.3 (C-3) and 67.4 (C-4) in HMBC spectrum. Chemical shifts of C-3 and H-4/C-4 indicated the presence of a tertiary alcohol and chlorine substitution, respectively undec-3,9-diol, for which the trivial name dendroidiol was proposed. Further X-ray crystallographic data of 2 confirmed the suggested structure and defined the absolute configuration as 3R, 4S, 6S, 9R, 10S as depicted (2 = 0.060(7)Å; ϕ2 = 352(7)° and q2 = 0.042(5) Å; ϕ2 = 233(7)°, respectively undecan-2-ol (5) were isolated and identified by comparison of their spectroscopic and physical data to those reported in the literature  and 13 [n-hexane–CH2Cl2 50:50], respectively. Fraction 22 (336.0 mg) was submitted to gel filtration column on Sephadex LH-20 with a mixture of n-hexane–CH2Cl2–MeOH . This process resulted on the separation of five sub-fractions (22.1–22.5). Fraction 22.3 (111.0 mg) was further chromatographed in silica gel column chromatography (7.0 g) using a gradient of CH2Cl2–EtOAc  resulting in five sub-fractions (22.3.1–22.3.5). Compound 3 (23.0 mg) was identified from fraction 22.3.1, eluted in CH2Cl2.Vermelha Beach crude extract (1.9 g) was purified by silica gel column chromatography (52.0 g) using a mixture of Dendroidone (1). Colorless oil;  = +6,0° ; IR (film) νmax 3457, 2927, 1675 cm−1; 1H-NMR and 13C-NMR data, see m/z 320 (2), 318 (2), 313 (3), 311 (2), 308 (3), 306 (15), 304 (12), 269 (3), 267 (9), 249 (26), 221 (19), 207 (75), 183 (33), 175 (22), 171 (24), 157 (23), 147 (38), 143 (59), 133 (27), 131 (28), 129 (35), 119 (66), 117 (32), 115 (34), 107 (50), 105 (63), 93 (32), 91 (96), 85 (95), 83 (41), 79 (58), 77 (61), 41 (100); HR-APCI-MS [M+H]+m/z 349.0381 .Dendroidiol (2). Colorless crystal; m.p. 116 °C;  = −19.2° ; IR (film) νmax 3463, 2971, 1918, 1640, 1453, 757, 622 cm−1; 1H-NMR and 13C-NMR data, see m/z 334 (2), 316 (5), 314 (6), 299 (7), 297 (5), 253 (4), 237 (15), 235 (46), 217 (10), 199 (25), 173 (5), 157 (16), 133 (22), 119 (42), 107(73), 105 (74), 85 (91), 69 (47), 55 (53), 43 (100). HR-ESI-MS [M+Na]+m/z 375.0417 .Debromoelatol (3). Colorless oil;  = +9.0° ; IR (film) νmax 3391, 2918, 2849, 1702, 1464, 1296, 939, 758, 720 cm−1; 1H-NMR and 13C-NMR data, see m/z 254 [M+] (1), 239 (15), 237 (6), 236 (34), 223 (10), 221 (29), 219 (6), 210 (8), 209 (4), 208 (25), 201,15 (27), 195 (11), 173 (31), 119 (88), 105 (74), 91 (91), 85 (100).2 reported in this paper have been deposited at the Cambridge Crystallographic Data Center, under the reference number CCDC 950138. Copies of the data can be obtained, free of charge, on application to the Director, CCDC, 12 Union Road, Cambridge CB2 1EZ, UK, fax: +44 1223 336033 or data_request@ccdc.cam.ac.uk.X-ray diffraction data was carried out in Nonius Kappa CCD diffractometer at room temperature with radiation MoKα. The collect was performed utilizing Collect software  and the 2 was crystallized from n-hexane to give colorless crystals. Crystal data: C15H24O2ClBr, M = 351.7, colorless block, size 0.30 × 0.26 × 0.16 mm3, T = 293(2) K, Orthorhombic, space group P212121, a = 9.2197(8) Å, b = 11.2080(8) Å, c = 15.6121(10) Å, V = 1613.3(2) Å3, Z = 4, Dc = 1.448 g/cm3, µ = 2.71 mm−1, F(000) = 728, 14001 reflections measured in the range 3.14° ≤ θ ≤ 25.65°, completeness θmax = 99.7%, 3044 independent, with Rint = 0.045; 178 parameters; Final agreement factors: R1 = 0.037 [F2 > 2σ(F2)], wR2 = 0.079 and GOOF = 1.09; largest difference peak and hole = 0.73, −0.62 eÅ−3. Flack parameter value x = −0.024(13)  method [The murine peritoneal macrophage cell line RAW 264.7 was obtained from the American Type Culture Collection  and grown at 37 °C and 5% COess test . As a po] method . The cytLDH assay was used to evaluate the toxicity of the studied samples towards macrophage cultures. The release of LDH (cytoplasmic enzyme lactate dehydrogenase) from RAW 264.7 cells treated with samples was determined using 50 µL of cell culture supernatant collected 24 h after the treatment, as described in the previous section . The LDHL. dendroidea. Compound 1 represents a new compound with a chloroenone group while compound 2 displayed a chlorohydrin function. Full spectroscopic data is described for the first time for compound 3. Moreover, compound 4 significantly suppressed NO production in LPS-stimulated RAW 264.7 macrophages and also displayed antimycobacterial action against M. bovis BCG. Thus, present data showed that L. dendroidea is a promising source of immunomodulatory and antimycobacterial drugs.In conclusion, the present study resulted in the isolation of five sesquiterpenes from Brazilian specimens of


The structures of the new compounds were established by extensive spectroscopic analyses, including 1D and 2D nuclear magnetic resonance (NMR) spectroscopy and data comparison with related structures. Compounds 8 and 14 were isolated from a natural source for the first time. The isolated metabolites were shown to be not cytotoxic against a limited panel of cancer cells. Compound 9 showed anti-inflammatory activity by reducing the expression of proinflammatory cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) proteins in lipopolysaccharide (LPS)-stimulated mouse leukaemic monocyte macrophage (RAW 264.7) cells.Six new polyoxygenated cembrane-based diterpenoids, stellatumolides A–C ( Sarcophyton have been well recognized to be a rich source of structurally unique and bioactive diterpenes, in particular cembranoids c 0.1, CHCl3), was isolated as a colorless oil and exhibited a sodium adduct ion peak at m/z 371.1832 [M + Na]+ from high-resolution electrospray ionization mass spectrometry (HRESIMS). Thus, the molecular formula C20H28O5 was established. The infrared (IR) absorption band at νmax 3445 cm−1 and the ion peak appearing in the ESIMS at m/z 353 [M − H2O + Na]+ indicated the presence of one hydroxy group in the molecule. The 20 carbon signals in the 13C nuclear magnetic resonance (NMR) spectrum . The α,β-unsaturated γ-lactone moiety was deduced from NMR signals at δC 171.1, 157.9, and 128.8 (each C), and IR absorption at νmax 1760 and 1680 cm−1. Moreover, one trisubstituted double bond was found by NMR signals at δC 130.6 (C), 129.2 (CH), and δH 5.26  , C-1 , and C-15 ; H3-18 to C-3, C-4, and C-5; H3-19 to C-7, C-8, and C-9; and H3-20 to C-11, C-12, and C-13 established the carbon skeleton of 1. The unresolved 2H signal (δH 4.27) present in the 1H NMR spectrum, measured in CDCl3, could be well resolved by measuring 1 in C6D6 , resulting in two signals at δH 4.19  and 3.93 , which were further assigned to oxymethines H-3 and H-7, respectively. Therefore, the HMBC correlation from H-3 to C-8 clearly indicated an ether linkage between C-3 and C-8  and the γ-lactone ring (C-2/C-16) of 1. From the above findings, detailed HMBC spectrum (1 was established.Stellatumolide A (spectrum  were att11.0 Hz) . The cor systems . The het and C-8 . In consspectrum , and 2D spectrum , the pla1 were proposed from analysis of nuclear Overhauser effect spectroscopy (NOESY)  with H-7 , measured in C6D6  established the β-orientation of the C-1 residue in the oxetane, and hence the R* configuration of C-2. Furthermore, one of the methylene protons at C-6  exhibited a NOESY correlation with H-7, and was therefore characterized as H-6β, while the other  was assigned as H-6α. H-6α showed NOE interactions with both H3-18 and H3-19; therefore, H3-18 and H3-19 were also positioned on the α-face. Due to the high flexibility of the macrocycle ring structure of 1, a conformational search at the molecular mechanics level was carried out using CONFLEX® -3′-one moiety in the molecule.The relative configurations at C-2, C-3, C-4, C-7, and C-8 of  in C6D6 , indicatCONFLEX® ,30 softwormula 1 . Compoun2), [α]c 0.3, CHCl3), possessed the molecular formula C20H28O4 as indicated by HREIMS at m/z 332.1985, implying seven degrees of unsaturation. Similarly to compound 1, the strong IR absorptions at νmax 3420, 1752, and 1666 cm–1 indicated the presence of hydroxyl, ester carbonyl, and olefinic groups, respectively. The carbonyl group was identified as an α,β-unsaturated ester from the 13C NMR signals of three nonprotonated carbons at δC 170.3, 152.2, and 123.3. Moreover, two trisubstituted double bonds were deduced from NMR signals at δC/δH 147.8 (C), 130.9 (C), 117.4 (CH)/5.16 , and 129.1(CH)/ 4.97   and heteronuclear multiple quantum coherence (HMQC) experiments, showed 20 carbon signals, assigned to four methyls, six methylenes, one 3sp oxymethine, two olefinic methines, and seven quaternary carbons . As for compound 1, three partial structures of consecutive spin systems were distinguished by analysis of COSY correlations  to C-3  and H-3  to C-2  and C-1 , the C-2/C-3 location of the oxytrisubstituted double bond could be assigned. Furthermore, the HMBC correlations from H3-17  to C-1, C-15 , and the carbonyl C-16 , confirmed the C-1/C-2 location of the γ-lactone ring. In considering the degrees of unsaturation and molecular formula, an ether linkage was placed between C-4 and C-8.Stellatumolide B  . The 13Celations . Examina and C-8 . From th2 as 4,8-epoxy-7β-hydroxy-cembra-1(15),2,11-trien-16,2-olide. Careful investigation of NOESY correlations in combination with measuring distances between relevant protons in the MM2 energy-minimized model enabled resolution of the relative configuration of compound 2 , H2-10 with H3-19, and H3-19 with H-7  revealed that the methyl groups at C-4 and C-8 were on the same face as H-7, and assuming that H3-18 was α-oriented, both C-7 and C-8 were therefore of the S* configuration. Moreover, the NOE correlations displayed by H3-17/H-14, H3-18/H-3, and H3-20/H-10 assigned the geometry of the double bonds at C-1/C-15, C-2/C-3, and C-11/C-12 as Z, Z, and E, respectively. On the basis of the above findings, the structure of stellatumolide B (2) was elucidated. Further detailed examination of 2D NMR correlations  establismpound 2 . The NOE3) was found to have the same molecular formula (C20H28O4) as compound 2 and the same hydroxyl and unsaturated γ-lactone functionalities on the basis of its HREIMS and IR spectra. The NMR data of 3  in comparison with that of 2, and the variational J values of 9.5 and 5.0 Hz at H-7 of 3 relative to that of 2 (7.0 and 3.0 Hz). After interpretation of the 2D NMR spectra, in particular the NOE correlations including that of H3-18/H3-19, compound 3 was established as the 7-epimer of compound 2.Stellatumolide C (ata of 3  were fou4) was isolated as a colorless oil  and possessed the molecular formula C20H30O3, as determined by HRESIMS . The NMR spectroscopic features  spectroscopic correlations , and a downfield shift at C-13 (ΔδC + 3.8 ppm), in 4 relative to 17 were observed. Also, H-14 in 4 exhibited a significant downfield shift (ΔδH − 0.46 ppm) relative to that of 17. The E geometry, instead of Z geometry as in 17, for the C-14/C-1 double bond was thus suggested for 4, and was consequently confirmed by the NOE interaction of H-14 with H3-17. Furthermore, detailed NOE correlation analysis of 4  welations , and waspound 17  31] was was4) wasis of 4  confirmeectively .m/z 348.2301, M+) and NMR data (5) to be C21H32O4, with six degrees of unsaturation. The 13C NMR of 5 displayed 21 carbon signals ascribable to five methyls (including that of a methoxy group), seven methylenes, three methines, and six quaternary carbons. In addition, a trisubstituted epoxide (δC/δH 62.0/2.84 and 60.7), a tetrasubstituted epoxide , two trisubstituted double bonds , a quaternary dioxycarbon , and an oxymethylene  were disclosed. The COSY and HMBC correlations, as depicted in 5 (δC 107.4), containing a methoxy group , as supported by the HMBC correlation from protons of methoxy group to C-2. The gross structure of 5 was thus confirmed. The upfield chemical shifts (δC < 20 ppm) of C-18 and C-20 and NOE correlations . The NOE interactions of H-16β with 2-OCH3 protons, and H3-17 with H2-16, H-13, (δH 2.10) and H-14 (δH 1.82) reflected the β-orientations of the methoxy at C-2 and the oxygen of the epoxide at C-1/C-15. The significant NOE response of the olefinic H-3 with H-7 and other NOE correlations  c 0.1, CHCl3); IR (neat) νmax 3445, 2925, 2857, 1760, 1680, 1455, 1379, 1259, 1227, 1092, 1037 and 762 cm–1; UV (MeOH) λmax 204 (log ε = 3.5); 13C  and 1H NMR  data, see 13C  δ 170.0 , 157.8 , 130.8 , 129.9 , 129.5 , 114.0 , 92.1 , 83.8 , 72.2 , 71.2 , 41.2 , 40.1 , 39.1 , 32.1 , 24.7 , 22.6 , 23.2 , 20.8 , 16.5 , 8.7 ; 1H  δ 5.15 , 4.19 , 3.93 , 2.29 , 2.12 , 2.01 ,1.89 , 1.80 , 1.78 , 1.73 , 1.68 , 1.56 , 1.50 , 1.49 , 1.30 , 1.22 , 1.08 ; ESIMS m/z 371 [M + Na]+, 353 [M − H2O + Na]+; HRESIMS m/z 371.1832 [M + Na]+ .Stellatumolide A (2): colorless oil; [α]c 0.3, CHCl3); IR (neat) νmax 3420, 2928, 2859, 1752, 1666, 1457, 1376, 1063, and 1020 cm–1; UV (MeOH) λmax 275 (log ε = 4.3); 13C  and 1H NMR  data, see m/z 332.1985 .Stellatumolide B (3): colorless oil; [α]c 0.3, CHCl3); IR (neat) νmax 3419, 2965, 2927, 2856, 1749, 1666, 1457, 1377, 1063, and 1019 cm–1; UV (MeOH) λmax 275 (log ε = 4.3); 13C  and 1H NMR  data, see m/z 332.1989 : colorless oil; [α]c 0.3, CHCl3); IR (neat) νmax 3421, 2927, 2863, 1729, 1453, 1380, 1247, 1136, 1043 and 767 cm–1; 13C  and 1H NMR  data, see m/z 341.2091, [M + Na]+ .Stellatumonin A (5): colorless oil; [α]c 0.2, CHCl3); IR (neat) νmax 2959, 2927, 2857, 1734, 1455, 1385, and 1049 cm–1; 13C  and 1H NMR  data, see m/z 348.2301 .Stellatumonin B (6): colorless oil; [α]c 0.1, CHCl3); IR (neat) νmax 2925, 2859, 1720, 1673, 1619, 1447, 1298, 1253 and 1093 cm–1; UV (MeOH) λmax 203 (log ε = 4.4); 13C  and 1H NMR  data, see m/z 346.2143 . Cytotoxicity assays of the isolated metabolites were performed using 3--2,5-diphenyltetrazolium bromide (MTT) colorimetric method ,48. Comp2. RAW 264.7 cells were seeded onto a 6-well plate with 2 × 106 cell per well and cultured for 24 h. The cells were pretreated with the compound 9 for 1 h and treated with lipopolysaccharide  from Escherichia coli 055:B5  in the presence or absence of the compound 9 . After 24 h LPS treatment, the cell lysates were prepared using Cell Lysis Buffer . The expression of COX-2 and iNOS proteins was measured using Western blotting analysis. RAW264.7 cells were purchased from the Food Industry Research and Development Institute . Cell were cultured in DMEM, supplemented with RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin G, and 100 μg/mL streptomycin , at 37 °C in an incubator with 5% COProtein concentrations were measured using a bicinchoninic acid (BCA) protein assay kit . Protein extracts were boiled, loaded into sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and electrotransferred to polyvinylidene fluoride (PVDF) membranes. After blocking in 5% nonfat milk in TBST buffer  for 1 h, the membranes were incubated with antibodies for COX-2 (#4842), iNOS (#13120), and β-actin (#3700)  at 4 °C with gentle agitation overnight. The membranes were then washed and incubated with horseradish peroxidase-labelled rabbit and mouse secondary antibodies  for 2 h at room temperature. After successive washes, the membranes were developed with an enhanced chemoluminescence (ECL) kit , and blots were visualized using a LAS3000 system . Densitometric analysis was performed with ImageJ software .p values < 0.05 were considered statistically significant.The data are expressed as the mean ± SD. One-way ANOVA followed by Tukey’s post-hoc test  was used to compare multiple groups according to the experiments. 1–6, together with ten known related compounds (7–16), were isolated from the Formosan soft coral Sarcophyton stellatum. Compound 1 possessed an unusual spiroketal unit between the oxetane (C-2/C-4) and the γ-lactone ring (C-2/C-16) in the cembranoid, suggesting the potential to discover more new molecular structures from marine organisms and warrant the further discovery of new medicines from the ocean. Compound 9 could effectively inhibit the accumulation of the proinflammatory COX-2 and iNOS proteins in LPS-stimulated RAW264.7 macrophage cells in a dose-response manner. Thus, compound 9 might be useful for discovery of an effective anti-inflammatory agent. Six new polyoxygenated cembrane-based diterpenoids,


E-triene-3β-ol (1), was isolated along with cholesterol from a Nephthea sp. Bornean soft coral. The structure of compound 1 was elucidated on the basis of spectroscopic analysis and comparison of the data with those of the related compounds.A new 4α-methyl sterol, 4α-methyl-ergosta-6,8(14),22 Lemnalia, Paralemnalia, Capnella, Lithophyton, Dendronephthya, Scleronephthya, Stereonephthya and Nephthea have received considerable attention from organic chemists , six olefinic carbons  and one OH-bearing carbon . The NMR and HRMS data could thus account for three of the seven degrees of unsaturation, suggesting the tetracyclic nature of 1.Compound 1 were detected in the HSQC experiment. The 1H–1H COSY spectrum exhibited partial structures a, b, c and d . HMBC correlations between H3-18 and C-12/C-13/C-17 revealed partial structure b could be connected to c through a quaternary carbon (C-13). Furthermore, HMBC correlations between H3-26/C-24, H3-27/C-24 and H3-28/C25 established the connection of partial structure c with d. Partial structures a and c were connected through fully substituted double bond between C-8 and C-14 by HMBC cross-peaks between H-7/C-8, H2-15/C-8, H2-15/C-14 and H3-18/C-14. Based on available spectroscopic data obtained for this compound, there were no other available connection option then of C-8 to C-9. HMBC correlations from H3-29 to C-3, C-4 and C-5 confirmed the existence of a 4α-methyl group. Based on these findings, the planar structure of 1 was concluded as shown in All C–H correlations of  c and d . Confirm1 was deduced from the NOESY experiment as well as the coupling constants in the 1H-NMR spectrum. The coupling constant between H-22 and H-23 (J = 15.1 Hz) suggested the double bond to have E configuration. Furthermore, as shown in 3-19, H-3/H-5, H-4/H3-19, H-5/H-9, H-5/H3-29, H-9/H-12α, H-11β/H3-18, H-11β/H3-19, H-12β/H3-21 and H3-18/H-20 revealed the relative configurations for each ring junction and chiral center. The configuration at C-24 was proposed by comparison of its NMR data with those of model compounds ,22E-triene-3β-ol.The relative stereochemistry of compound ompounds  21,22].,22.1 was1H-NMR (600 MHz) and 13C-NMR (150 MHz) spectra were recorded with a JEOL ECA 600, with TMS as internal standard. HR-ESI-TOFMS spectrum was obtained with LCMS-IT-TOF (Shimadzu) in ESI mode. HPLC was conducted on a Waters 600 using UV detector, Luna 5μ Phenyl-hexyl (10.0 × 250 mm) and Luna 5μ C18(2) 100A (10.0 × 250 mm). Preparative TLC was performed with silica gel plates . Silica gel  was used for column chromatography. Analytical TLC was performed on Merck Kieselgel 60 F254. Spots were visualized by UV light or by spraying with a 5% phosphomolybdic acid-ethanol solution.Optical rotations were measured on an AUTOPOL IV automatic polarimeter . Nephthea sp. was collected from Sepanggar Island, Sabah , on January 24, 2008. The voucher specimen was deposited in the herbarium of Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah (BORNEENSIS).A specimen of 2O. The EtOAc fraction was further partitioned with hexane and 90% MeOH. The hexane fraction (1.0 g) was fractionated by Si gel column chromatography with a step gradient of hexane and EtOAc in the ratio of 9:1, 8:2, 7:3, 1:1 and EtOAc. The fraction (237 mg) eluted with hexane/EtOAc (8:2) was further separated by a combination of preparative TLC with CHCl3 and HPLC (Luna 5μ Phenyl-hexyl) with 80% MeCN to afford compound 1 (1.8 mg). The fraction (20 mg) eluted with hexane/EtOAc (7:3) was separated by repeated preparative TLC with CHCl3 and hexane/EtOAc (3:1) to give compound 2 (3.2 mg).The fresh soft coral (400 g wet wt) was extracted with MeOH at room temperature for 7 days. The crude extract was evaporated under reduced pressure and the residue was partitioned between EtOAc and H25D -36.9 ; HR-TOFMS m/z 411.3593 [M+H]+ ; 1H-NMR and 13C-NMR spectral data: see White amorphous solid; [α]


Garcinia mangostana resulted in the successful isolation of one new prenylated xanthone, mangaxanthone B (1), one new benzophenone, mangaphenone (2), and two known xanthones, mangostanin (3) and mangostenol (4). The structures of these compounds were elucidated through analysis of their spectroscopic data obtained using 1D and 2D NMR and MS techniques.A detailed chemical study on the ethyl acetate and methanol extracts of the stem bark of  Garcinia, Mesua and Cratoxylum  were observed in compound 1 after further examination of the DEPT spectrum.Meanwhile, the DEPT experiment indicated that this compound is composed of three methine (3J correlations between the chelated hydroxyl group with C-9a (δC 102.9) and C-2 (δC 110.3) and a 2J correlation with C-1 (δC 160.8) were observed. This allows the assignment of the hydroxyl group to C-1 [δH 13.74 ]. The two prenyl moiety methyl groups resonating at δH 1.76 and 1.62 are correlated to the neighbouring C-3' (δC 130.6) and C-2' (δC 122.6) in the HMBC experiment. Moreover, the linkages between H-1' [δH 3.32 ] and C-2' (δC 122.6) as well as C-3' (δC 130.6) are seen. The linkages between H-2' [δH 5.25 ] and C-4' (δC 17.1) and C-5' (δC 25.1) are also clearly seen in the experiment ] with C-2 (δC 110.3) and the 3J correlations of H-1' [δH 3.32 ] with C-3 (δC 162.2) indicated that the prenyl moiety is positioned at C-2. In the COSY analysis, the coupling of H-1' and H-2' was very strong.The HMBC long range periment . These cδH 1.28 ] gave clear correlations with C-2'' (δC 44.9) and C-3'' (δC 69.7) indicating the methyls to be attached to C-3''. H-2'' [δH 1.70 ] gave cross peaks to C-4''(δC 28.5), C-5'' (δC 28.5), C-3'' (δC 69.7) and C-1'' (δC 22.2). Meanwhile H-1'' [δH 3.42 ] showed a 2J correlation with C-2'' (δC 44.9) and a 3J correlation with C-3'' (δC 69.7). These long range correlation signals allow us to conclude a 3-hydroxy-3-methylbutyl moiety ] to C-7 (δC 144.1), C-8 (δC 138.4), C-8a (δC 111.4) and the cross-peak of H-2'' [δH 1.70 ] to C-8 (δC 138.4) in the HMBC analysis were suggestive of the 3-hydroxy-3-methylbutyl moiety location at C-8. In addition, the two aromatic methoxyl groups [6-OCH3 (δH 4.00) and 7-OCH3 (δH 3.79)] are assigned at C-6 and C-7 because of their 3J correlations with δC 158.6 (C-6) and δC 144.1 (C-7). The two remaining aromatic methine protons of the xanthone skeleton were determined to be at the two remaining available carbons, C-4 and C-5. The long range correlations between H-4 (δH 6.38) and C-2 (δC 110.3), C-3 (δC 162.2), C-4a (δC 154.9), C-9 (δC 182.0) and C-9a (δC102.9) places the δH 6.38 (H-4) singlet at C-4. The signal at δH 6.90 (H-5) was assigned to C-5 based on its HMBC correlations with C-5a (δC 155.4), C-6 (δC 158.6), C-7 (δC 144.1), C-8 (δC 138.4), C-8a (δC 111.4) and C-9 (δC 182.0). Compound 1 differs from a closely related compound, garcinone D ) analysis. The FTIR spectrum exhibited a strong absorption at 1728 cm−1, which suggested the presence of a carbonyl group, and another strong absorption at 2924 cm−1 representing the aromatic C-H stretching band. An aromatic benzene chromophore which is of a benzophenone skeleton, was confirmed by the maximum absorption peaks of 211, 214 and 309 nm in the UV-Visible spectrum.Compound 1H-NMR spectrum of compound 2 showed five aromatic proton signals which resonated at δH 5.96 , δH 6.52  and δH 6.39 . After a detailed inspection on the 13C NMR and DEPT spectra, it was found that the signals in the 13C-NMR spectrum indicated 14 carbons which consisted of one methoxyl (δC 54.6), five methine  and eight quaternary carbons . The benzophenone skeleton of compound 2 was demonstrated by signals resonating at δC 198.4 for the carbonyl group, as well as signals at δC 158.0, 161.5 and 163.3 for the hydroxylated aromatic carbons ] showed three-bond connectivities with, C-4' (δC 105.7), and C-7 (δC 198.4). Protons H-2' and H-6' also gave two-bond connectivities with C-3' and C-5' (δC 158.0 × 2) respectively. Meanwhile H-4' has 2J correlations with C-3' and C-5' (δC 158.0 × 2) and 3J correlations with C-2' and C-6' (δC 106.5 × 2). The aromatic proton of H-3 [δH 5.96 ] exhibited three-bond (3J) and two-bond (2J) connectivities with C-1 (δC 106.4) (3J), C-5 (δC 95.3) (3J) and C-4 (δC 163.3) (2J) respectively. H-5 [δH 5.96 ] also showed correlations with C-1 (δC 106.4) (3J), C-3 (δC 91.1) (3J), C-4 and C-6 (δC 163.3 × 2) (2J) in the HMBC spectrum. Moreover, the methoxyl group was assigned to C-2 because of the 3J HMBC correlation of the methoxyl proton [δH 3.54 ] with C-2 (δC 161.5). The observation of a 2J correlation between H-3 and C-4 implies position 3 to be occupied by H-3. Moreover, we also observed a 2J correlation between H-5 and C-6 as well as between H-5 and C-4. Therefore, another proton is situated at C-5. 4-OH and 6-OH carbons have the same chemical shift values, thus justifying that the molecule is symmetrical. Hence, the two OH groups are at C-6 and C-4 and not at C-2. Hence, the structure of this compound was elucidated as -methanone and it was given the trivial name mangaphenone.The structure of compound trum See . The aroGarcinia mangostana was collected from Melaka, Malaysia. A herbarium specimen (RG221) was deposited at the Herbarium in the Biology Department of UPM.The stem bark of 1H, 13C, DEPT) and 2D  NMR spectra were recorded on a Unity INOVA 500 MHz NMR instrument using tetramethylsilane (TMS) as the internal standard. EIMS spectra were obtained using a Shimadzu GCMS model QP2010 Plus spectrophotometer. The ultraviolet spectra were recorded on a Shimadzu UV-160A UV-Visible Recording Spectrophotometer. Infrared spectra were obtained using the universal attenuated total reflection (UATR) technique on a Perkin-Elmer 100 Series FT-IR spectrometer. Melting points were measured through Leica Galen III microscope which was equipped with Testo 720 temperature recorder.The 1D (Garcinia mangostana (2.0 kg) was first de-fatted using hexane followed by extraction with ethyl acetate  for 72 h at room temperature then with 70% methanol  for another 72 h. The three extracts were concentrated to give 14.22 g of dark brown residue of EtOAc extract and 278.96 g of dark brown residue of MeOH extract. The EtOAc extract was subjected to vacuum column chromatography by eluting with a stepwise gradient system of hexane, chloroform (CHCl3), ethyl acetate and methanol to afford 6 fractions. The fourth fraction was further fractionated through column chromatography using hexane–CHCl3 and CHCl3–MeOH to give 8 fractions. The last fraction was subjected to repeated chromatography by eluting with hexane–EtOAc (7:3) and CHCl3–MeOH (9.8:0.2) to furnish compounds 1, 2 and 3. Meanwhile, the dry MeOH extract (278.96 g) was suspended in H2O and then partitioned with n-butanol . The n-BuOH soluble portion (2.56 g) was chromatographed in a polarity gradient manner  and afforded eight fractions. Fraction 2 was further purified through column chromatography by using CHCl3–MeOH (9:1) and compound 4 was thus obtained.The air-dried powdered stem bark of Mangaxanthone B (1). Yellow crystals; m.p. 194–195 °C; UV (EtOH) λmax (log ε): 209 (4.40), 245 (4.44), 262 (4.42), 316 (4.29) and 353 (3.79) nm; IR ν (cm−1): 3447, 3249, 2938, 1601, 1457, 1277 and 826; 1H-NMR  and 13C-NMR , see m/z (rel. int.): 442(32), 424(28), 381(74), 369(43), 368(33), 354(23), 353(100), 327(53) and 325(33).Mangaphenone (2). Brownish-yellow crystals; m.p. 245–246 °C; UV (EtOH) λmax (log ε): 211 (4.11), 214 (4.12) and 309 (3.77) nm; IR ν (cm−1): 3599, 2924, 1728, 1261, 804 and 730; 1H-NMR  and 13C-NMR , see m/z (rel. int.): 276(67), 260(50), 168(70), 167(100), 153(22) and 69(44).Mangostanin (3). Yellow amorphous powder. Spectral data are in agreement with the literature [terature .Mangostenol (4). Yellow solid; m.p. 159–160 °C. Spectral data are in agreement with the literature [terature .Garcinia mangostana. Biological evaluation of these compounds is under way.A new prenylated xanthone, mangaxanthone B and a new benzophenone, mangaphenone, were isolated along with two known xanthones, mangostanin and mangostenol, from the stem bark of


Tamarix ramosissima Ledeb, a traditional herbal medicine used for rheumatoid arthritis (RA) treatment in northwest China, led to the discovery of a new phenolic aromatic rings substituted lactam, tamaractam (1), together with the previously reported compounds cis-N-feruloyl-3-O-methyldopamine (2) and trans-N-feruloyl-3-O-methyldopamine (3). The structures of the compounds were determined by high resolution electrospray ionization mass spectroscopy (HRESIMS) and 1D and 2D-NMR experiments, as well as comparison with the literature data. The effects of the three compounds on the viability of RA fibroblast-like synoviocytes (RA-FLS) were assessed by 3--2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Pro-apoptosis effect of compound 1 in RA-FLS was further investigated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, activated caspase-3/7 level assessment using luminescence assay, and sub-G1 fraction measurement using flow cytometry. It was found that these three compounds displayed variable proliferation inhibitory activity in RA-FLS, and compound 1 exhibited the strongest effect. Compound 1 could remarkably induce cellular apoptosis of RA-FLS, increase activated caspase-3/7 levels, and significantly increase sub-G1 fraction in the cell cycle. The results suggested that compound 1 may inhibit the proliferation of RA-FLS through apoptosis-inducing effect, and these compounds may contribute to the anti-RA effect of T. ramosissima.Chemical investigation of  Tamarix ramosissima, commonly known as tamarisk or rose willow, is a shrub or dungarunga belonging to the family Tamaricaceae. The Tamarix species have strong adaptability to the arid desert environment and saline and alkaline soil. They are not only excellent sand fixing plants, but also a good species for soil and water conservation and afforestation in saline-alkali soil + and 1H- and 13C-NMR spectral analyses, requiring eleven degrees of unsaturation. The 1H-NMR spectrum of compound 1 (H 3.51 and 3.67), one methylene (δH 3.78 and 3.03) and an adjacent methine (δH 4.53), six aromatic proton signals which belong to two ABX coupling systems  +; 1H-NMR : δH 6.74 , 6.67 , 6.58 , 2.63 , 3.29 , 7.70 , 6.71 , 7.09 , 6.48 , 5.76 , 3.71 , 3.73 , 8.09 ; 13C-NMR : δC 130.2 (C-1), 112.7 (C-2), 147.4 (C-3), 144.8 (C-4), 115.3 (C-5), 120.7 (C-6), 34.7 (C-7), 40.5 (C-8), 126.7 (C-1′), 114.2 (C-2′), 146.8 (C-3′), 147.4 (C-4′), 114.9 (C-5′), 124.4 (C-6′), 136.8 (C-7′), 120.7 (C-8′), 166.2 (C-9′), 55.5 (2 × -OCH3).Trans-N-feruloyl-3-O-methyldopamine (3): White amorphous powder; ESIMS m/z: 344 [M + H]+; 1H-NMR : δH 6.78 , 6.69 , 6.60 , 2.66 , 3.37 , 7.11 , 6.80 , 6.98 , 7.33 , 6.45 , 3.74 , 3.79 , 7.99 ; 13C-NMR : δC 130.3 (C-1), 112.8 (C-2), 147.5 (C-3), 144.9 (C-4), 115.4 (C-5), 120.8 (C-6), 34.8 (C-7), 40.6 (C-8), 126.4 (C-1′), 110.8 (C-2′), 147.9 (C-3′), 148.4 (C-4′), 115.7 (C-5′), 121.6 (C-6′), 138.9 (C-7′), 119.0 (C-8′), 165.4 (C-9′), 55.5 (2 × -OCH3).Human fibroblast-like synoviocytes from RA patients (RA-FLS) were purchased from Cell Applications, Inc.,  and cultured with a synoviocyte growth medium (Cell Applications). RA-FLS obtained from the 3rd to 5th passages were used for experiments. The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Ethics Committee of Ningxia Medical University (Project identification code: 2014-036).1–3 was dissolved in dimethyl sulfoxide . RA-FLS were seeded in 48-well plates at a density of 3 × 104 cells/well, and were treated with 0.01, 0.1, 1, 5, and 10 μM of compounds 1–3 or DMSO vehicle only in a serum free medium. After 24 and 48 h of incubation, 2.5 mg/mL of 3--2,5-diphenyl-tetrazolium bromide (MTT) solution (Sigma-Aldrich) was added to the wells, and the cells were then incubated for 2 h. The absorbance of each well was measured by a Bio-Rad 680 microplate reader  at 570 nm. Based on MTT assay results, compound 1 was employed in the subsequent apoptosis-inducing experiments.Compounds 1 for 48 h and then fixed with 4% paraformaldehyde for 30 min at room temperature. After washing with phosphate buffer saline (PBS), permeabilization solution  was added for a 2 min reaction and cells were incubated with terminal deoxynucleotidyl transferase (TdT) and biotin-11-dUTP for 1 h at 37 °C. The nuclear morphology was observed by fluorescence microscopy IX71 . The apoptosis index was calculated based on the percentage of TUNEL-positive cells in 1000 RA-FLS.Apoptosis in RA-FLS cells was measured by TUNEL assay , according to the manufacturer′s protocol. Briefly, RA-FLS were incubated with 0.1 and 1 μM of compound 4 cells/well in 96-well plates and treated with 0.1 and 1 μM of compound 1 for 48 h. After incubation, cells were treated for 2 h with reconstituted Caspase 3/7-Glo reagent. Then the luminescence signal generated after cleavage of DEVD-aminoluciferin substrate by caspase 3/7 was detected using a luminometer plate reader .The levels of activated caspase-3/7 in RA-FLS were further assessed with a Caspase-Glo kit  , accordi5 cells/well), in triplicate, and then treated with 0.1 and 1 μM of compound 1 for 48 h. Cells were harvested, washed twice with PBS, and the cellular DNA stained with 200 μL PI . After incubation at 4 °C for 20 min, the cells were analyzed by flow cytometry  and sub-G1 fractions were computed using ModFit LT version 4.0 software .The induction of apoptosis was confirmed by a flow cytometric approach using propidium iodide (PI) as fluorescent stain . Brieflyp < 0.05.All statistical tests were performed using SPSS11.0 statistical software . The level of statistical significance was determined by using one-way analysis of variance (ANOVA). Results were expressed as the mean ± S.E.M. of three independent experiments performed in triplicate. Differences were considered significant when 2O extract of the traditional herbal medicine T. ramosissima yielded the new compound tamaractam (1), along with the previously reported cis-N-feruloyl-3-O-methyldopamine (2) and trans-N-feruloyl-3-O-methyldopamine (3). The structures of the isolated compounds were determined by HRESIMS and 1D and 2D-NMR data, as well as by comparison with the available data in the literature. These compounds displayed variable proliferation inhibitory activity, and compound 1 showed remarkable apoptosis-inducing effect on RA-FLS.The investigation of the 70% EtOH/H


S)-2’,7-dihydroxy-5,8-dimethoxyflavanone (1) and (S)-2-(4-hydroxyphenyl)-6-methyl-2,3-dihydro-4H-pyran-4-one (2), were isolated from the ethanol extract of Scutellaria barbata. Their structures were elucidated on the basis of spectroscopic analysis.Two new phenols, 2( Scutellaria barbata (family Labiatae) and some other species from the same genus have been widely used in China, India, Nepal and other Asian countries for a long time as a traditional Chinese medicine or a folk remedy for the treatment of different diseases  –19.1 . UV (MeOH): λmax (log ε) 207 (3.76), 285 (3.02) nm. IR (KBr): νmax 3266, 1614, 1583, 1510, 1458, 1416, 1367, 1270, 1099, 992, 748 cm-1. 1H-NMR (DMSO) and 13C NMR (DMSO) data, see m/z 317.1024 .2(S)-2-(4-Hydroxyphenyl)-6-methyl-2,3-dihydro-4H-pyran-4-one (2), powder; [α] –21.7 . 1H NMR (DMSO) and 13C NMR (DMSO) data, see m/z 227.0683 .(S)- 2’,7-dihydroxy-5,8-dimethoxyflavanone (1) and (S)-2-(4-hydroxyphenyl)-6-methyl-2,3-dihydro-4H-pyran-4-one (2), were isolated from the ethanol extract of Scutellaria barbata. The discovery of compounds 1-2 is a further addition to the diverse plant phenolic compounds. Their biological activities are evaluating in progress. In conclusion, two new phenols, 2(


Configuration and optical purity of 1a (%ee) was found to vary depending on the geographic distribution of ascidian colonies. An improved method for preparing N,O-diarenoyl derivatives of 1a was optimized. The EC ECD method was found to be complementary to the MPA-NMR method at different ranges of %ee.Sample configurations of distaminolyne A ( Cyclodehydration of AA benzamides is known to occur with inversion of configuration at C-2 under certain conditions that activate the secondary OH group  for intramolecular nucleophilic SN2 displacement by the carboxamide oxygen D for sample and standard have long been used for determinations of %ee, but they become flawed when the magnitudes of the specific rotations are low, or when sample solutions of different concentrations induce non-linearity. The complementarity of the two methods used in this work—ECD and CDA derivatization with MPA—for establishing %ee and AC of 1-AAs, such as 1a, is apparent from a simple semi-quantitative analysis of the sources of error and observation of the discrepancies between the two methods (εsample/∆εstandard = 0). The sign and magnitude of non-zero split-CE signal (~4%ee and higher) indicate configuration and non-racemic mixtures, respectively, with errors in the latter associated only with absolute A values and the signal-to-noise of the measurement.The value of %ee, defined by a ratio (Equation (1)) of the sum and difference of concentrations,  methods . The ECDN,O-di-arenoyl derivatives has two advantages over [α]D for determination of %ee: Δε is a molar quantity that conforms to the Beer–Lambert law and the absolute magnitude of the Δε signal (or A) is typically larger. Consequently, EC ECD is preferred for AC determination and quantitation of %ee, but there are limitations. Errors in %ee will be larger for samples near 100%ee where the ratio is obtained from Δεsample and Δεstandard values that differ little in magnitude. MPA-derivatization—although requiring the preparation of separate S- and R- derivatives—is equally reliable for assigning AC of 1a. For optical purity, however, the MPA method is better suited for measurements within a different range of %ee values (closer to 50%ee) where cumulative bias in integral measurement (C (+) or C (−)) due to instrumental factors  is lowest. EC ECD of vicinal 1a is unusual, and suggestive of a biosynthesis from lipid and amino acid precursors that differs from conventional long-chain base biosynthesis at one or more later steps. The proposed biosyntheses of 1-AA natural products  that undergoes stereospecific reduction of the keto group by NADPH and subsequent oxidative desaturation -sphingosine. The finding of heterogeneous enantiomeric compositions of different samples of products a is modeproducts b 1]: Co: Co1a isd-configuration. By comparison, the proposed biosynthesis of 1a would appear to proceed by activation of a hypothetical long-chain diyne-ene C17 fatty acid CoA ester 16, followed by decarboxylative condensation with Gly instead of Ser, and reduction of the resultant ketone 17 by NADPH. Unlike 15, the proposed intermediate aminoketone 17 undergoes reduction with lower stereofidelity than reduction of 15. In contrast, all natural 2-AAs appear to be biosynthesized with hallmark high stereofidelity even when the relative and absolute configurations differ from species to species D.The foregoing results should have broader applications to both stereochemical analysis of other aminoalkanols from Nature and synthesis. While reliable, prior analyses of 2-AAs exploiting ECD of gosines; ) suffere3 (stabilized with amylenes) or CF3CH2OH (TFE), purchased from Acros Organics . Optical rotations were recorded on a JASCO P2000 digital polarimeter  at the D-line of Na emission on solutions in quartz cells (0.100 dm pathlength). UV-visible spectra were recorded on a JASCO V630 dual beam spectrometer  in quartz cuvettes (1.00 cm or 0.100 cm pathlength). Electronic circular dichroism spectra were recorded on either a Jasco J810 spectropolarimeter  or an Applied PhotoPhysics Chirascan with solutions in quartz cells (0.100 or 0.500 cm). HRMS were acquired on two instruments: a Bruker micrOTOF Q II mass spectrometer  using external calibration of sodium formate clusters, and an Agilent 6230 TOFMS  with a Jetstream electrospray ionization source, calibrated by using Agilent ESL-L Low Concentration Tuning Mix . 1H-NMR spectra were recorded at 500.16 MHz on a Jeol ECA 500 spectrometer, at 399.91 MHz on a Mercury 400, at 400.13 MHz on a Bruker Avance AVIII 400 MHz or at 500.19 MHz on a Bruker Avance AVIII-HD 500 MHz spectrometer . 13C-NMR spectra were recorded at 125.69 MHz on a Varian VX 500 spectrometer  equipped with an Xsens 13C{1H} cryoprobe. Spectra were referenced to solvent . Dry pyridine was distilled from CaH2 under an atmosphere of N2, and dry CH2Cl2, CH3CN were obtained by passage through commercial basic alumina cartridges under an atmosphere of Ar. Jacobsen’s catalysts +, .hed data ; (+)-HREDistaminolyne A : 27 mg, 0.2% dry wt; c 0.73, MeOH); The 1H NMR  spectrum was identical with the published data [m/z 262.2161 [M + H]+, .Distaminolyne A : 7 mg, 0.1% dry wt; c 0.70, MeOH); The 1H NMR  spectrum was identical with the published data [m/z 262.2159 [M + H]+, .Distaminolyne A : 8.5 mg, 0.04% dry wt; c 0.275, MeOH); 1H NMR  δ 5.89–5.83 , 5.12–5.08 , 5.06–5.03 , 3.76–3.73 , 3.03 , 2.77 , 2.35 , 2.28–2.24 , 1.56–1.29 ; 13C NMR  δ 137.8, 116.3, 78.0, 77.1, 68.7, 66.7, 66.3, 46.1, 36.0, 33.7, 30.5, 30.1, 29.8, 29.5, 26.3, 19.6; (+)-HRESIMS m/z 262.2157 [M + H]+, .N,O-Dibenzoyl derivative1b: Distaminolyne A 1a (NP-12-8-1)  was dibenzoylated with benzoic acid according to Method B to obtain, after RP C18 chromatography (1:9 water/methanol) dibenzoyl derivative 1b, 2.6 mg (40%). c 0.26, MeOH); UV (MeOH) λmax (log ε) 203 (4.28), 228 (4.19), 267 (3.55), 282 (3.44); The 1H NMR  spectrum was identical with the published data [m/z 492.2500 [M + Na]+, .N,O-Dibenzoyl derivative1b: Distaminolyne A 1a (NP-12-9-1)  was dibenzoylated with benzoic acid according to Method B to obtain, after RP C18 chromatography (1:9 water/methanol) dibenzoyl derivative 1b, 1.8 mg (40%). c 0.18, MeOH); UV (MeOH) λmax (log ε) 204 (4.05), 228 (3.98), 266 (3.31), 281 (3.17); The 1H NMR  spectrum was identical with the published data [m/z 492.2504 [M + Na]+, ., 281 3.1; The 1H N,O-Dibenzoyl derivative1b: Distaminolyne A 1a (NP-12-10-1)  was dibenzoylated with benzoic acid according to Method B to obtain, after RP C18 chromatography (1:9 water/methanol) dibenzoyl derivative 1b, 2.6 mg (41%). c 0.26, MeOH); UV (MeOH) λmax (log ε) 205 (4.34), 226 (4.23), 267 (3.70), 282 (3.59); The 1H NMR  spectrum was identical with the published data [m/z 492.2504 [M + Na]+, .S)-MPA derivative1c: Distaminolyne A 1a (NP-9-161-3)  was treated with (S)-MPA according to Method B to yield 3.4 mg (80%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.41 .(R)-MPA derivative1d: Distaminolyne A 1a (NP-9-161-3)  was treated with (R)-MPA according to Method B to yield 2.4 mg (56%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.41 .(S)-MPA derivative1c: Distaminolyne A 1a (NP-12-8-1)  was treated with (S)-MPA according to Method B to yield 3.6 mg (84%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.41 .(R)-MPA derivative1d: Distaminolyne A 1a (NP-12-8-1)  was treated with (R)-MPA according to Method B to yield 3.8 mg (89%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.41 .(S)-MPA derivative1c: Distaminolyne A 1a (NP-12-9-1)  was treated with (S)-MPA according to Method B to yield 2.3 mg (54%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.40 .(R)-MPA derivative1d: Distaminolyne A 1a (NP-12-9-1)  was treated with (R)-MPA according to Method B to yield 3.5 mg (82%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.59 , 4.41 .(S)-MPA derivative1c: Distaminolyne A 1a (NP-12-10-1)  was treated with (S)-MPA according to Method B to yield 2.4 mg (75%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.60 , 4.40 .(R)-MPA derivative1d: Distaminolyne A 1a (NP-12-10-1)  was treated with (R)-MPA according to Method B to yield 2.8 mg (88%) of crude product. 1H NMR  diagnostic CαH resonances of bis-MPA: δ 4.79 , 4.71 , 4.60 , 4.40 .((R)-1-Amino-2-hexanol,5b: 1-Amino-2-alkanols were prepared from the corresponding optically enriched 1,2-epoxides - or -catalyst) by ammoniolysis using a variation of the published procedure for 4a,b + calcd. for C17H21NO2Na+ 294.1465.Oxazoline8: c 0.50, CHCl3); 1H NMR  δ 8.43 , 8.03 , 7.92 , 7.87–7.86 , 7.57-7.7.50 ; ESI TOF HRMS m/z 254.1539 [M+H]+, calcd. for C17H20NO+ 254.1539.N,O-di-(2-Naphthoyl)-1-amino-2-hexanol (R)-9b: Colorless oil, c 0.8, TFE); ECD (TFE) δ 228 (∆ε +53), 236 (0), 244 (−66); 1H NMR  δ 8.65 , 8.27 , 8.09 , 7.97 , 7.91 , 7.79 , 7.62-7.50 , 6.92 , 5.42 , 3.91 , 3.85 , 1.92 , 1.84 , 1.5 , 1.42 , 0.94 ; ESI TOF HRMS m/z 426.2061 [M + H]+ calcd. for C28H28NO3+ 426.2064.Tribenzoyl-1-amino-butanol (S)-10a: A solution of 1-AA (S)-6a  in dry pyridine (2.0 mg) and DMAP  was treated with freshly distilled benzoyl chloride  and the mixture heated at 60 °C under at atmosphere of N2 with stirring for 48 h. The volatiles were removed under high vacuum and the semi-crystalline residue (1.28 g) taken up in CH2Cl2 and filtered through a plug of basic alumina. The eluate was concentrated and the residue separated on a ‘pencil column’  to give a non-polar, clear oil (67 mg) that was further purified by HPLC  to give tribenzoyl derivative (S)-10b . c 2.92, MeOH); 1H NMR  δ 7.81 , 7.39 , 7.33 , 7.23 , 7.12 , 7.02 , 5.65 , 4.53 , 4.25 , 1.87 , 1.06 . 13C NMR (CDCl3) δ 2), 166.4 ), 136.3 , 132.8 , 131.9 , 130.0 , 129.6 , 129.0 , 128.16 , 128.15 , 75.0 , 49.9 , 25.8 , 9.6 ; HRMS m/z 424.1520 [M + Na]+, calcd. for C25H23NO4Na+ 424.1519.Naphthimide (R)-11b: A solution of the 1-AA (R)-5b  in dry pyridine (0.175 mL) was added to a stirred solution of freshly sublimed 2,3-naphthalenedicarboxylic acid anhydride  in dry pyridine (0.325 mL) and the mixture heated to 110 °C under N2 for 48 h. The volatiles were removed under a stream of N2. The residue was divided in half, and each applied to duplicate preparative TLC plates . The plates were developed, twice (2:3 EtOAc/n-hexane) and the silica corresponding to the major fluorescent bands  were scrapped from the plates and extracted with EtOAc. The extracts were combined, filtered and the solvent removed from the filtrate to deliver (R)-11b  as a colorless powder. c 0.64, CHCl3); 1H NMR  δ 8.35 , 8.05 , 7.71 , 3.94 , 3.88 , 3.82 , 2.46 , 1.58–1.48 , 1.42–1.33 , 0.92 ; ESI TOF HRMS m/z 320.1253 [M + Na]+, calcd. for C18H19NO3Na+ 320.1257.O-Naphthoyl-naphthimide (R)-12b: An ice-cold solution of 2-naphthoic acid , DMAP  and EDC∙HCl  in CH2Cl2 (0.5 mL) was stirred for 30 min under an atmosphere of N2, then treated with a solution of naphthimide (R)-11b  in CH2Cl2 (0.3 mL). The solution was allowed to warm to 23 °C and stirred for 64 h, then diluted with additional CH2Cl2 (15 mL) and washed sequentially with 0.2M HCl, NaHCO3 (satd.) and water. After drying the solution (Na2SO4), the volatiles were removed and the residue (4.5 mg) purified by ‘pencil column’ chromatography  to give pure (R)-12b as a colorless oil . c 0.44, TFE); ECD (TFE) δ 216 (∆ε +24.7), 240 (+25.8), 245 (0), 259 (−31.8); 1H NMR  δ 8.55 , 8.29 , 8.00 , 7.98 , 7.94 , 7.85 , 7.83 , 7.85 , 7.83 , 7.66 , 7.57 , 7.52 , 5.52 , 4.16 , 4.02 , 1.85 , 1.49 , 1.40 , 0.92 ; ESI TOF HRMS m/z 452.1847 [M + H]+, calcd. for C29H26NO4+ 452.1856.N,O-di-(2-Naphthoyl)-1-amino-2-decanol (S)-13a: A sample of the 1-AA (S)-3a  was acylated with 2-naphthoic acid (5 equiv.) using Method B to give, after standard workup and purification by pipette column , 13a . c, 0.16, CHCl3)), which displayed 1H and 13C NMR spectra identical with those of (R)-13b [m/z 504.2501 [M + Na]+, calcd. for C32H35NaNO3+ 504.2509. (R)-13b . ESI TOF1H-NMR—revealed complementary advantages. An optimized N,O-dinaphthoyl 1-AA derivative lends clarity of interpretation of ECD spectra with sufficient sensitivity for ‘sub-nanomole scale’ stereoassignments. Chiroptical investigations of multiple samples of the 1-AA, distaminolyne A (1a) extracted from Pseudodistoma spp., showed that configuration and enantiomeric purity varied with collections, a behavior that is unlike 2-AA natural products that, so far, have been found to be stereochemically homogenous. A promiscuous biosynthesis for 1a is proposed to account for the difference.An optimized approach to assignment of absolute configuration of 1-amino-2-alkanols (1-AAs), based on ECD, is presented which avoids problems with prior methods, and streamlines analyses of multiple samples. Comparison and contrast of two methods for chiroptical analysis of 1-AAs—ECD and CDA derivatization-


After this time the resin was drained off, washed with DCM (5 x 1 min) and the deprotection procedure was repeated once more, allowing the mixture to stand for 30 min. Finally, the resin was successively washed with DCM/dioxane-H2O (9:1)/DMF/DCM (5 × 1 min).To the pre-swollen resin, under Ar atmosphere, dried DCM (1 mL) and phenylsilane (25 equiv.) were added. After 5 min, a solution of Pd.To the previously swollen resins 5 and 6 (0.11 mmol), a solution of HBTU (0.275 mmol) or HBPyU (0.33 mmol) and DIEA  in anhydrous DMF (1 mL) was added, and the reaction allowed to stand at r.t. overnight. The resins were drained and washed [DMF /DCM/DMF/DCM (5 × 0.5 min each)].After removing the Fmoc and Alloc group from resins 12-16, the removal of the Fmoc protecting group before cleavage was required. Resins 7-11 and 12-16 were treated with a mixture of TFA-H2O, 19:1 (1 mL for 100 mg) at r.t. for 2 h. After washings with DCM (5 × 1mL), the filtrates were evaporated and the residue solved in water and washed with ethyl acetate. The aqueous phase was lyophilized to give derivatives 17-26, isolated as trifluoroacetate salts. When required, the products were purified through reverse-phase cartridges using mixtures of H2O/ACN as solvent.For resins iBu-Trp(Nps)-Lys-NH2 (17): Yield: 4%. HPLC: Rt = 2.76 min. 1H-NMR (D2O):, δ 0.77 , 0.98 , 1.3-1.4 , 1.81 , 2.5-2.8 , 3.2-3.3 , 3.87-3.94 , 6.47 , 7.01 , 7.1-7.2 , 7.29 , 7.42 , 8.1 . Calc. MW = 540.68, MS ESI+ [M+H]+ m/z: 541.2Bn-Trp(Nps)-Lys-NH (18): Yield: 24%. HPLC: Rt = 3.42 min. 1H-NMR (D2O):, δ 1.03 , 1.3-1.5 , 2.69 , 3.20 , 3.94 , 4.02 , 4.60 , 6.44 , 7.02 , 7.1-7.3 , 7.44 , 8.1 . Calc. MW = 574.69, MS ESI+ [M+H]+ m/z: 575.2N-C(=NH)]-Trp(Nps)-Lys-NH2+ m/z: 527.3)2N]2-C=]-Trp(Nps)-Lys-NH2+ m/z: 583.4H8)2N]2-C=]-Trp(Nps)-Lys-NH2+ m/z: 635.iBu)-NH2H-Trp(Nps)-Lys( (22a): Yield: 8%. 1H-NMR (D2O): δ 0.74 , 0.77 , 1.06 , 1.3-1.4 , 1.85 , 2.6-2.9 , 3.18-3.21 , 3.94 , 4.01 , 6.45 , 7.03 , 7.1-7.2 , 7.31 , 7.45 , 8.1 . Calc. MW = 540.68, MS ESI+ [M+H]+ m/z: 541.2H-Trp(Nps)-Lys(Bn)-NH (23a): Yield: 14%.1H-NMR (D2O): δ 1.04 , 1.3-1.4 , 2.70 , 3.17 , 3.9-4.0 , 4.60 , 6.45 , 7.03 , 7.1-7.3 , 7.45 , 8.1 . Calc. MW = 574.69, MS ESI+ [M+H]+ m/z: 575.2N-C(=NH)]-Trp(Nps)-Lys[C(=NH)-NH2]-NH2+ m/z: 569.3)2N]2-C=]-Trp(Nps)-Lys[=C-[N(CH3)2]2]-NH2+ m/z: 681 (62%), 583 (20%).4H8)2N]2-C=]-Trp(Nps)-Lys[=C-[(C4H8)2N]2]-NH2+ m/z: 786 (36%), 635 (12%). .Recombinant rat TRPV1 and human NMDAR channel expression in Xenopus oocytes and channel blockade: All the procedures have been described in detail elsewhere [-1) was microinjected (V = 50 nL) into defolliculated oocytes (Stage V and VI) as described. Oocytes were functionally assayed 2–4 days after cRNA injection. Whole-cell currents from rat TRPV1-injected oocytes were recorded in standard Ringer solution  with a two-microelectrode voltage-clamp amplifier at 20 ºC. TRPV1 channels were activated by application of 10 µM capsaicin in the absence or presence of compounds (10 µM) at a holding potential (Vh) of −60 mV. Whole-cell currents from oocytes injected with NR1/NR2A  subunits were recorded in standard Ringer solution upon activation with L-glutamate (100 µM) supplemented with glycine (10 µM) at Vh = −60 mV. Responses were normalized with respect to that evoked without channel blockers. Experiments were performed in triplicate; each bar in n (number of oocytes) = 12.lsewhere ,14. BrieN-terminus or on the Lys side-chain. Main steps of these procedures are the AcOH-catalyzed incorporation of the Nps moiety at the indole ring, compatible with TFA labile resins, the reductive amination to N-alkyl derivatives, and the guanidylation reactions with thioureas or uronium salts. Diguanidylated by-products were obtained during the incorporation of guanidino groups on the Lys side-chain, probably due to Fmoc-removal by the free Lys ε–NH2 group under the basic media. From the biological point of view, the presence of substituted guanidino groups at N-terminus led to the best TRPV1 activity/NMDA selectivity ratio. Based on the significant activation of NMDA receptor by compound 20 (~30%), derivative 21 could be considered as a suitable molecule for further modifications in the search for new TRPV1 channel blockers.Convenient solid-phase procedures have been developed for the modification of Trp(Nps)-Lys dipeptides at the


Ligularia duciformis, L. kongkalingensis, and L. nelumbifolia (the d/k/n complex) was studied using eight samples collected in central and northern Sichuan Province of China. Cacalol (14) and epicacalone (15), rearranged eremophilanes, were isolated from the complex for the first time. Two new phenylpropanoids were also obtained. Seven of the eight samples produced phenylpropanoids and the other produced lupeol alone. Two of the seven samples also produced furanoeremophilanes or their derivatives and one produced oplopanes. The geographical distribution of the sesquiterpene-producing populations suggests that the production of sesquiterpenes evolved independently in separate regions. L. limprichtii collected in northern Sichuan was also analyzed and its chemical composition and the sequence of internal transcribed spacers (ITSs) in the ribosomal RNA gene cluster were found to be similar to that in the d/k/n complex and L. yunnanensis, which are morphologically similar.The diversity in root chemicals and evolutionally neutral DNA regions in the complex of  Ligularia are highly diversified in the Hengduan Mountains area of China and their evolution is considered to be continuing , an oxygenated methylene , an E-alkene [δH 6.14  and 6.58 ], and a 1,2,4-trisubstituted aromatic ring  (See H 0.90) and a doublet (δH 0.95) in addition to those of two methylenes and one methine (δH 1.91), the latter of which was observed in octet, suggesting that the acid part was 3-methylpentanoate. The structure was established from COSY observed for 2′-H/3′-H/4′-H/5′-H and 3′-H/6′-H and from HMBC correlations shown in The structure of the new compound Hz)] See . Signals8 was obtained as an inseparable mixture of E- (8a) and Z-isomers (8b), the latter of which was new. Although a full characterization of 8b was not feasible due to signal overlap with 8a, its structure was deduced to be the Z isomer from its 1H-NMR spectrum, as the J-value between 2-H (δH 6.15) and 3-H (δH 7.54) was 11.4 Hz .Compound 4 Hz See . AlthougtR = 10 to 30 min  and 6β-ethoxyfuranoeremophilan-10β-ol (13) were detected at tR = 15.3 and 17.5 min, respectively, in the chromatogram of sample 7  at tR = 15.4 min, which was isolated as the major component -5.8S-ITS2 region of the ribosomal RNA gene. The results are shown in 14) and epicacalone (15) have a rearranged carbon skeleton generated from furanoeremophilane 1H-NMR (CDCl3) δ 1.59 , 1.66 , 1.67 , 1.99-2.11 , 3.87 , 3.90 , 4.59 , 4.61 , 5.03–5.09 , 5.52–5.58 , 6.15 , 6.64 , 6.60 , 6.78 , 7.40 , 7.54 , 9.68 , 10.03 ; 13C-NMR (CDCl3) assigned for 8a: δ 16.4 (C-10′), 17.6 (C-9′), 25.7 (C-8′), 26.4 (C-5′), 39.6 (C-4′), 56.1 (OMe), 69.6 (C-1′), 105.6 , 119.8 (C-2′), 123.9 (C-6′), 127.8 (C-2), 129.4 (C-4), 131.7 (C-7′), 139.8 (C-7), 142.0 (C-3′), 152.9 (C-3), 154.0 , 193.5 (C-1); assigned for 8b: δ 16.4 (C-10′), 17.6 (C-9′), 25.7 (C-8′), 26.4 (C-5′), 39.6 (C-4′), 56.2 (OMe), 69.6 (C-1′), 107.2 , 119.9 (C-2′), 123.9 (C-6′), 129.6 (C-4), 129.9 (C-2), 131.7 (C-7′), 138.5 (C-7), 142.0 (C-3′), 148.7 (C-3), 153.7 , 192.6 (C-1).14) and its derivative, epicacalone (15), were found in the d/k/n complex for the first time, showing that the diversity in the complex is higher than previously known. Two new phenylpropanoids  were also obtained. The basal chemicals in this complex appear to be phenylpropanoids and lupeol, to which sesquiterpenoids are added in some populations. The geographical separation of sesquiterpene-producing populations suggests that the production of this class of compounds evolved independently, perhaps through introgression. L. limprichtii was found to produce a phenylpropanoid, showing similarity to the d/k/n complex, which is consistent with DNA data.Cacalol (