Rhodotorula rubra KCh 82 has been carried out. Nine of the substrates underwent an effective enantioselective reduction to the respective (R)-alcohols according to Prelog’s rule, with the exception of 2-chloro-1,2-diphenylethan-1-one that was not transformed by this strain. The expected reduction proceeded without dehalogenation, leading to the respective (R)-halohydrins in high yields. The use of this biocatalyst yielded (R)-2-bromo-1-phenyl-ethan-1-ol (enantiomeric excess (ee) = 97%) and its derivatives: 4'-Bromo- (ee = 99%); 4'-Chloro- (ee > 99%); 4'-Methoxy- (ee = 96%); 3'-Methoxy- (ee = 93%); 2'-Methoxy- (ee = 98%). There were also obtained and characterized 2,4'-dichloro-, 2,2',4'-trichloro- and 2-chloro-4'-fluoro-phenyetan-1-ol with >99% of enantiomeric excesses.Biotransformation of ten α-haloacetophenones in the growing culture of the strain  R configuration at the carbon with an OH group). Both formoterol and salmeterol are used in treatment of chronic obstructive pulmonary disease, being highly selective β2-adrenergic receptor agonists. They are administered via inhalation . Tn and ee .2e) was obtained in 97%–99% yield and with 90%–92% of ee by chemical synthesis, using triethylamine and chiral ruthenium complexes  in 10% H2SO4. The products were separated by column chromatography using silica gel  and hexane/acetone mixture  as the developing system. Composition of biotransformation mixtures was established by gas chromatography (GC) on Agilent Technologies 7890 A GC instrument , fitted with a flame ionization detector (FID) and a chiral column Chirasil-Dex CB (Agilent) 25 m × 0.25 mm × 0.25 μm film thickness. Temperature of injector: 200 °C; temperature of detector: 250 °C was the same for all compounds. To determinate the composition and enantiomeric excesses of product mixtures the following temperature programs were used (The course of biotransformation was controlled by means of Thin Layer Chromatography (TLC). Analytical TLC was carried out on silica gel G 60 Fere used .3. Optical rotations were measured with an Autopol IV automatic polarimeter . Absolute configurations of the products were determined by comparison of their optical rotation values with literature data.Reference samples of the racemic alcohols were prepared by reducing the ketones with sodium borohydride in methanol. NMR spectra were recorded on a DRX 600 MHz Bruker spectrometer  and measured in CDCl3 (3 × 10 mL). The extracts were dried over MgSO4, concentrated in vacuo and analyzed by GC. All the experiments were repeated three times.Erlenmeyer flasks (300 mL), each containing 100 mL of the medium consisting of 3 g glucose and 1 g aminobac dissolved in water, were inoculated with a suspension of microorganisms and then incubated for 3–7 days at 25 °C on a rotary shaker (190 rpm). After full growth of the culture 20 mg of a substrate dissolved in 1 mL of acetone was added. After 1, 3, 6, 9 days of incubation under the above conditions, portions of 10 mL of the transformation mixture were taken out and extracted with CHCl3 (3 × 300 mL), dried (MgSO4) and concentrated in vacuo. The transformation products were separated by column chromatography and analyzed .The same transformations were performed on the preparative scale in 2000 mL flasks, each containing 500 mL of the cultivation medium. The cultures were incubated under the same conditions and then 200 mg of substrates dissolved in 10 mL of acetone were added to the grown cultures. After incubation the mixtures were extracted with CHClR)-(−)-2-Bromo-1-phenylethan-1-ol (2a): A three-day transformation of substrate 1a (100 mg) in the culture R. rubra KCh 82 yielded 91 mg of compound 2a: c = 1.32 CHCl3) (97% ee), ([1H NMR (CDCl3) δ: 2.68 , 3.59 , 3.68 , 4.96 , and 7.35–7.45 .(7% ee), -(−)-2-Bromo-1-(4'-bromophenyl)-ethan-1-ol (2b): A one-day transformation of substrate 1b (100 mg) in the culture R. rubra KCh 82 yielded 93 mg of compound 2b: c = 14.1 CHCl3) (99% ee), ([1H NMR (CDCl3) δ: 2.72 , 3.52 , 3.62 , 4.91 , 7.28–7.38 , and 7.53–7.59 .(9% ee), -(−)-2-Bromo-1-(4'-chlorophenyl)-ethan-1-ol (2c): A three-day transformation of substrate 1c (100 mg) in the culture R. rubra KCh 82 yielded 79 mg of compound 2c: c = 3.94 CHCl3) (98% ee), ([1H NMR (CDCl3) δ 2.80 , 3.52 , 3.61 , 4.92 , and 7.36–7.48 .(8% ee), -(−)-2-Chloro-1-(4'-chlorophenyl)-ethan-1-ol (2d): A three-day transformation of substrate 1d (100 mg) in the culture R. rubra KCh 82 yielded 93 mg of compound 2d: c = 1.47 CHCl3) (99% ee), 1H NMR (CDCl3) δ 2.71 , 3.63 , 3.73 , 4.90 , and 7.36–7.59 .(R)-(−)-2-Chloro-1--ethan-1-ol (2e): A one-day transformation of substrate 1e (100 mg) in the culture R. rubra KCh 82 yielded 93 mg of compound 2e: c = 2.55 CHCl3) (99% ee), ([1H NMR (CDCl3) δ: 2,84 , 3.55 , 3.90 , 5.29 , 7.34 , 7.40 , and 7.60 .(R)-(−)-2-Chloro-1-(4'-fluorophenyl)-ethan-1-ol (2f): A one-day transformation of substrate 1f (100 mg) in the culture R. rubra KCh 82 yielded 88 mg of compound 2f: c = 1.10 CHCl3), (98% ee) ([1H NMR (CDCl3) δ 2.62 , 3.65 , 3.74  4.74 , 7.09 , and 7.40 .(98% ee) -(−)-2-Bromo-1-(2'-methoxyphenyl)-ethan-1-ol (2g): A one-day transformation of substrate 1g (100 mg) in the culture R. rubra KCh 82 yielded 88 mg of compound 2g: = c = 1.26 CHCl3), (98% ee), 1H NMR (CDCl3) δ: 2.93 , 3.56 , 3.79 , 3.89 , 5.18 , 6.92 , 7.02 , 7.33 , and 7.47 .(R)-(−)-2-Bromo-1-(3'-methoxyphenyl)-ethan-1-ol (2h): A one-day transformation of substrate 1h (100 mg) in the culture R. rubra KCh 82 yielded 73 mg of compound 2h: c = 1.36 CHCl3) (94% ee); 1H NMR (CDCl3) δ: 2.52 , 3.56 , 3.66 , 3.84 , 4.93 , 6.89 , 6.98–7.09 , and 7.32 .(R)-(−)-2-Bromo-1-(4'-methoxyphenyl)-ethan-1-ol (2i): A one-day transformation of substrate 1i (100 mg) in the culture R. rubra KCh 82 yielded 76 mg of compound 2i: c = 1.96 CHCl3), (96% ee); ([S); 1H NMR (CDCl3) δ: 2.61 , 3.56 , 3.63 , 3.84 , 4.91 , 6.92–6.95 , and 7.33–7.36 .(6% ee); ‑halohydrins with high both yields and enantiomeric excesses, which were fully characterized spectroscopically and are potential synthons for synthesis of β-adrenergic receptor agonists. It should be underlined that (R)-halohydrins were single products of these reactions. No side products were observed.The presented results revealed that the strain


Z-α-fluorovinyl Weinreb amide unit with a benzothiazolylsulfanyl substituent at the allylic position was assembled. This was achieved via condensation of two primary building blocks, namely 2-(benzo[d]thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylacetamide (a Julia-Kocienski olefination reagent) and 2-(benzo[d]thiazol-2-ylthio)acetaldehyde . This condensation was highly Z-selective and proceeded in a good 76% yield. Oxidation of benzothiazolylsulfanyl moiety furnished a second-generation Julia-Kocienski olefination reagent, which was used for the introduction of the second olefinic linkage via DBU-mediated condensations with aldehydes, to give -dienamides in 50%–74% yield. Although olefinations were 4Z-selective, -2-fluoro-2,4-dienamides could be readily isomerized to the corresponding 5-substituted -2-fluoro-N-methoxy-N-methylpenta-2,4-dienamides in the presence of catalytic iodine.Modular synthesis of regiospecifically fluorinated 2,4-diene Weinreb amides, with defined stereochemistry at both double bonds, was achieved via two sequential Julia-Kocienski olefinations. In the first step, a  The conjugated diene and polyene amide structural units are found in many naturally occurring compounds that possess biological activity . These cFluorine is an attractive substituent in pharmaceuticals, in agrochemicals, and in materials chemistry ,15,16 du1, d]thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylacetamide (1) with a 2-(heteroarylthio)ethanal -2-fluoro-N-methoxy-N-methylacetamide (1), was synthesized as reported [d]thiazol-2-ylthio)acetaldehyde (2), was initially attempted via the dioxolane derivative of 2. Although the dioxolane derivative of 2 could be readily prepared from 2-(bromomethyl)-1,3-dioxolane and the sodium salt of 2-mercapto-1,3-benzothiazole, attempts at deprotection of 2-[methylthio]benzo[d]thiazole under various conditions proved unsuccessful. Therefore, synthesis via the dimethyl acetal was considered , CBr4 (entry 2), or PTSA (entry 3), no hydrolysis was observed. Reaction of 3 with 4 M HCl at 40 °C resulted in complete consumption of 3, but aldehyde 2 was isolated in a low 20% yield after column chromatography (entry 4). Subsequently, we found that compound 2 is unstable under chromatography conditions, on silica gel and alumina thiazol-2-ylthio)-2-fluoro-N-methoxy-N-methylbut-2-enamide (4) as the only stereoisomer. Comparably, exclusive Z-selectivity has also been observed in NaH-mediated condensations of 1 with aldehydes thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylacetamide (1) was prepared from the known 2-(benzo[d]thiazol-2-ylsulfonyl)-N-methoxy-N-methylacetamide thiazole3. To a solution of the sodium salt of 2-mercapto-1,3-benzothiazole  in DMF (20 mL) was added 2-bromo-1,1-dimethoxyethane , and the mixture was allowed to stir at 40 °C for 4 h. Upon completion of the reaction, as observed by TLC, the reaction mixture was diluted with EtOAc and washed with water. The aqueous layer was extracted with EtOAc (3 × 30 mL). The combined organic layer was washed with saturated NaHCO3 (30 mL), brine, dried over anhydrous Na2SO4, and evaporated. The crude product was purified by column chromatography using 20% EtOAc in hexanes to obtain compound 3  as a colorless viscous liquid. Rf : 0.48. 1H-NMR (CDCl3): δ 7.85 , 7.75 , 7.41 , 7.29 , 4.72 , 3.58 , 3.44 . 13C-NMR (CDCl3): δ 166.4, 153.2, 135.5, 126.1, 124.4, 121.6, 121.1, 103.0, 54.3, 35.5. HRMS (ESI) calcd for C11H14NO2S2 [M+H]+ 256.0460, found 256.0462.2-(Benzo[d]thiazol-2-ylthio)acetaldehyde2. To a stirred solution of 2-benzo[d]thiazole  in acetone (48 mL), was slowly added a mixture of HCl  and water (5.2 mL) at rt. The mixture was stirred for 40 min at 50 °C. Upon completion of the reaction, as observed by TLC, the mixture was cooled to 5 °C and the reaction was quenched by portion-wise addition of solid NaHCO3 up to the neutralization point, and then passed through a bed of anhydrous Na2SO4. The anhydrous Na2SO4 bed was washed with a minimum amount of acetone, the combined eluent was dried over anhydrous Na2SO4, and concentrated under reduced pressure. Crude product 2  was used in the next step without purification. Rf : 0.29. 1H-NMR (CDCl3): δ 9.73 , 7.85 , 7.76 , 7.42 , 7.32 , 4.09 .(Z)-4-(Benzo[d]thiazol-2-ylthio)-2-fluoro-N-methoxy-N-methylbut-2-enamide4. To a stirred solution of 2-(benzo[d]thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylacetamide  and 2-(benzo[d]thiazol-2-ylthio)acetaldehyde  in dry THF (48.0 mL) at −78 °C (dry ice/iPrOH), was added LHMDS  dropwise under a nitrogen atmosphere. The mixture was allowed to stir at −78 °C (dry ice/iPrOH) for 2 h and checked for the disappearance of 1 by 19F-NMR . Since 19F-NMR showed the presence of 1, more LHMDS  was added and the mixture was allowed to stir at −78 °C for an additional 1 h at which time complete consumption of sulfone 1 was observed by 19F-NMR. The reaction was quenched by the addition of saturated aq. NH4Cl, the solvent was partially removed under reduced pressure, and the mixture was extracted with EtOAc (3×). The combined organic layer was washed with 5% aq. NaOH, followed by water and brine, and then dried over Na2SO4. The organic layer was concentrated under reduced pressure and the crude product was purified by column chromatography using 10%, 15%, and 20% EtOAc in hexanes, to afford compound 4 as a yellow wax . Rf : 0.54. 1H-NMR (CDCl3): δ 7.87 , 7.75 , 7.43–7.40 , 7.32–7.28 , 6.22 , 4.15 , 3.69 , 3.22 . 13C-NMR (CDCl3): δ 165.2, 161.8 , 153.3, 152.4 , 135.7, 126.3, 124.6, 121.9, 121.2, 113.0 , 62.1, 33.9, 26.9 . 19F-NMR (CDCl3): δ −119.9 . HRMS (ESI) calcd for C13H14FN2O2S2 [M+H]+ 313.0475, found 313.0480.(Z)-4-(Benzo[d]thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylbut-2-enamide5. H5IO6  was dissolved in CH3CN (80.0 mL) by vigorous stirring at rt for 20 min. CrO3  was added and the reaction mixture was stirred for an additional 5 min to give an orange-colored solution. A solution of (Z)-4-(benzo[d]thiazol-2-ylthio)-2-fluoro-N-methoxy-N-methybut-2-enamide  in CH3CN (10.0 mL) was added dropwise to this mixture, resulting in an exothermic reaction and the formation of a yellowish precipitate. After complete addition, the mixture was stirred for 3 h, at which time TLC showed complete consumption of amide 4. The mixture was filtered through a Celite pad, the pad was washed with CH3CN, and the filtrate was concentrated under reduced pressure. Water was added to the residue and the mixture was extracted with EtOAc (3 × 30 mL). The combined organic layer was washed with saturated aq. NaHCO3 (5 × 30 mL) and brine (30 mL), and dried over anhydrous Na2SO4. The organic layer was concentrated under reduced pressure and the crude product was purified by column chromatography using 15%, 20%, and 25% EtOAc in hexanes to afford compound 5 as a white solid . Rf : 0.43. 1H-NMR (CDCl3): δ 8.24 , 8.02 , 7.67–7.59 , 5.96 , 4.43 , 3.53 , 3.17 . 13C-NMR (CDCl3): δ 164.8, 160.8 , 155.6 , 152.8, 137.2, 128.4, 127.9, 125.8, 122.6, 102.7 , 62.1, 51.1 , 33.6. 19F-NMR (CDCl3): δ −113.6 . HRMS (ESI) calcd for C13H14FN2O4S2 [M+H]+ 345.0374, found 345.0376.General experimental procedure. To a stirred solution of aldehyde (0.20 mmol) in dry CH2Cl2 (10.0 mL) was added DBU  and the mixture was cooled to 0 °C. A solution of (Z)-4-(benzo[d]thiazol-2-ylsulfonyl)-2-fluoro-N-methoxy-N-methylbut-2-enamide  in dry CH2Cl2 (10.0 mL) was then added slowly, dropwise (over about 2 h). The reaction mixture was allowed to stir overnight at 0 °C. After completion of the reaction, the solvent was evaporated under a stream of nitrogen gas, and the 1H and 19F-NMR spectra of the crude product mixture were recorded for determination of the E/Z ratio. The combined E/Z product mixture was purified by column chromatography. For eluting solvents see the specific compound headings. The mixture of (4E)- and (4Z)-isomers was analyzed and characterized based on the 1H-NMR; the assignment and integration of specific olefinic proton(s) allowed for other signals to be assigned based on the integration, along with a comparison to the pure -isomer . Assignment of the 19F-NMR signals to the (4E)- and (4Z)-isomers was based on the integration.-2-Fluoro-N-methoxy-N-methyl-5-penta-2,4-dienamide6a. Isomer ratio of (4E)-6a:(4Z)-6a = 35:65. Column chromatography using 8%, 10%, and 15% EtOAc in hexanes gave a mixture of -6a as a white solid . Rf : 0.49 for (4E)-6a and 0.58 for (4Z)-6a. 1H-NMR (CDCl3): δ 7.85–7.80 -isomer and 4H, (4Z)-isomer), 7.68 -isomer), 7.50–7.45 -isomer and 3H, (4Z)-isomer), 7.21 -isomer), 7.06 -isomer), 6.97 -isomer), 6.91 -isomer), 6.72 -isomer), 6.66 -isomer), 3.80 -isomer), 3.76 -isomer), 3.29 -isomer), 3.25 isomer). 19F-NMR (CDCl3): δ –121.2 -isomer), −123.4 -isomer). HRMS (ESI) calcd for C17H17FNO2 [M+H]+ 286.1238, found 286.1242.-2-Fluoro-N-methoxy-N-methyl-5-(4-methoxyphenyl)penta-2,4-dienamide6b. Isomer ratio of (4E)-6b:(4Z)-6b = 23:77. Column chromatography using 8%, 15%, and 20% EtOAc in hexanes gave a mixture of -6b as a pale yellow solid . Rf : 0.35. 1H-NMR (CDCl3): δ 7.42 -isomer), 7.29 -isomer), 6.99 -isomer), 6.95 -isomer), 6.91–6.87 -isomer and 2H, (4E)-isomer), 6.75 -isomer), 6.69 -isomer), 6.65 -isomer), 6.47 -isomer), 3.83 -isomer), 3.82 -isomer), 3.77 -isomer), 3.76 -isomer), 3.27 -isomer), 3.25 -isomer). 19F-NMR (CDCl3): δ −122.6 -isomer), −125.2 -isomer). HRMS (ESI) calcd for C14H17FNO3 [M+H]+ 266.1187, found 266.1191.-2-Fluoro-N-methoxy-N-methyl-5-(4-nitrophenyl)penta-2,4-dienamide6c. Isomer ratio of (4E)-6c:(4Z)-6c = 4:6. Column chromatography using 10% and 15% EtOAc in hexanes gave a mixture of -6c as a yellow solid . Rf : 0.47 for (4E)-6c and 0.72 for (4Z)-6c. 1H-NMR (CDCl3): δ 8.23 -isomer), 8.21 -isomer), 7.60 -isomer), 7.48 -isomer), 7.23 -isomer), 6.86–6.61-isomer and 2H, (4E)-isomer), 3.79 -isomer), 3.77 -isomer), 3.28 -isomer), 3.25 -isomer). 19F-NMR (CDCl3): δ −118.4 -isomer), −119.6 -isomer). HRMS (ESI) calcd for C13H14FN2O4 [M+H]+ 281.0932, found 281.0935.-2-Fluoro-N-methoxy-N-methyl-5-(thiophen-2-yl)penta-2,4-dienamide6d. Isomer ratio of (4E)-6d:(4Z)-6d = 1:9. Column chromatography using 8% and 10% EtOAc in hexanes gave a mixture of -6d as a yellow solid . Rf : 0.34 for (4E)-6d and 0.37 for (4Z)-6d. 1H-NMR (CDCl3): δ 7.36 -isomer), 7.27 -isomer), 7.27–7.25 -isomer), 7.12 -isomer), 7.09 -isomer), 7.04 -isomer), 7.00 -isomer), 6.92 -isomer), 6.86 -isomer), 6.76 -isomer), 6.61 -isomer), 6.42 -isomer), 3.78 -isomer), 3.77 -isomer), 3.28 -isomer), 3.26 -isomer). 19F-NMR (CDCl3): δ −120.7 -isomer), −123.7 -isomer). HRMS (ESI) calcd for C11H13FNO2S [M+H]+ 242.0646, found 242.0647.-2-Fluoro-N-methoxy-N-methyl-7-phenylhepta-2,4-dienamide 6e. Isomer ratio of (4E)-6e:(4Z)-6e = 15:85. Column chromatography using 6%, 8%, and 12% EtOAc in hexanes gave a mixture of -6e as a colorless oil . Rf : 0.30. We were unable to resolve the 1H-NMR signals for both isomers, so the signals reported are for the major -isomer. For the minor isomer, only diagnostic vinylic C4-H could be unequivocally assigned . 1H-NMR (CDCl3) of -isomer: δ 7.30–7.27 , 7.20–7.18 , 6.68 , 6.34 , 5.80 , 3.74 , 3.24 , 2.73 , 2.55 . Minor -isomer (CDCl3): δ 6.04 ppm . 19F-NMR (CDCl3): δ −124.2 -isomer), −125.7 -isomer). HRMS (ESI) calcd for C15H19FNO2 [M+H]+ 264.1394, found 264.1407.General experimental procedure. To a stirred solution of the -isomer mixture 6 in dry CHCl3 was added I2 (7–10 mol %) and the mixture was stirred at rt. The reaction was monitored by 19F-NMR and when only one isomer was observed, the reaction mixture was diluted with EtOAc (30 mL). The mixture was washed with water, saturated aq. Na2S2O3 (2 × 10 mL), and dried over Na2SO4. The organic layer was concentrated under reduced pressure to afford the desired -isomer. HMQC data were obtained for -6c and -6d, and where unequivocally assigned, through-bond C–F couplings are reported as 1JCF, 2JCF, etc. Due to the close structural similarity, these couplings are also reported for -6a and -6b, where possible.-2-Fluoro-N-methoxy-N-methyl-5-penta-2,4-dienamide -6a. Isomerization was performed with -6a  in CHCl3 (8.0 mL) using I2 , in a reaction time of 3 h, to yield isomer -6a as a white solid . Rf : 0.49. 1H-NMR (CDCl3): δ 7.83–7.80 , 7.69 , 7.50–7.46 , 7.21 , 6.97 , 6.72 , 3.80 , 3.29 . 13C-NMR (CDCl3): δ 162.8 , 150.2 , 137.9 , 134.1 , 133.7, 128.7, 128.4, 128.0 , 127.9, 126.8, 126.7, 123.6, 119.6 ,118.2 , 62.1 , 34.3. 19F-NMR (CDCl3): δ −123.4 .-2-Fluoro-N-methoxy-N-methyl-5-(4-methoxyphenyl)penta-2,4-dienamide -6b. Isomerization was performed with -6b  in CHCl3 (8.0 mL) using I2 , in a reaction time of 3 h, to yield isomer -6b as a pale yellow solid . Rf : 0.35. 1H-NMR (CDCl3): δ 7.42 , 6.95 , 6.88 , 6.75 , 6.65 , 3.83 , 3.77 , 3.26 . 13C-NMR (CDCl3): δ 162.9 , 160.4, 149.5 , 137.5 , 129.5, 128.7, 118.7 , 117.2, 114.5, 62.1, 55.6, 34.3. 19F-NMR (CDCl3): δ –125.2 .-2-Fluoro-N-methoxy-N-methyl-5-(4-nitrophenyl)penta-2,4-dienamide -6c. Isomerization was performed with -6c  in CHCl3 (1.6 mL) using I2 , in a reaction time of 1.5 h, to yield isomer -6c as a yellow solid . Rf : 0.47. 1H-NMR (CDCl3): δ 8.21 , 7.60 , 7.23 , 6.83 , 6.65 , 3.79 , 3.28 . 13C-NMR (CDCl3): δ 162.3 , 151.8 , 147.6, 142.9, 134.7 , 127.6, 124.4, 123.6 , 116.8 , 62.2 , 34.2. 19F-NMR (CDCl3): δ −119.6 .-2-Fluoro-N-methoxy-N-methyl-5-(thiophen-2-yl)penta-2,4-dienamide -6d. Isomerization was performed with -6d  in CHCl3 (7.5 mL) using I2 , in an overnight reaction, to yield isomer -6d as an off-white solid . Rf : 0.34. 1H-NMR (CDCl3): δ 7.26 , 7.09 , 7.00 , 6.92 , 6.86 , 6.61 , 3.77 , 3.26 . 13C-NMR (CDCl3): δ 162.7 , 150.1 , 142.1 , 130.4 , 128.1 , 128.0, 126.5 , 118.9 , 117.9 , 62.1 , 34.3. 19F-NMR (CDCl3): δ −123.7 .Z,4E)-2-fluoro-2,4-dienamides. This was achieved via two sequential Julia-Kocienski olefinations. In the first olefination, a Z-α-fluorovinyl Weinreb amide unit, with a benzothiazolylsulfanyl substituent at the allylic position, was assembled. For this, two key building blocks, a known fluorinated Julia-Kocienski reagent with a Weinreb amide moiety (1) and 2-(benzo[d]thiazol-2-ylthio)acetaldehyde (2), a precursor to the second Julia-Kocienski reagent, were reacted. Condensation proceeded with high Z-stereoselectivity and in a good 76% yield. Oxidation of the sulfide to the sulfone furnished the “second-generation” Julia-Kocienski olefination reagent, which underwent reactions with aldehydes to furnish the dienamides in 50%–74% yields. The second set of olefinations proceeded under DBU-mediated conditions and with Z-stereoselectivity. Isomeric -mixtures underwent iodine-mediated isomerization to a single -dienamide isomer. Although the method was performed with a Weinreb amide moiety, it is potentially applicable to other amides as well. Moreover, due to versatile chemistry of the Weinreb amide moiety, the -2-fluoro Weinreb dienamides are potentially useful synthetic building blocks, which can undergo a variety of conversions leading to more complex molecules. In this context, the present method offers a straightforward access to synthetically valuable entities that are not otherwise easily prepared by traditional synthetic approaches to Weinreb amides.In conclusion, we have developed a highly modular approach to (2


E-halimadiene-3α,15-diol (1), and 5(10),14-halimadiene-3α,13ξ-diol (2), one labdane, 12-(3-methyl-furan)-labd-8(17)-en-19-oic acid (3), together with sixteen known compounds were isolated from the barks of Dysoxylum densiflorum. All compounds were elucidated by extensive spectroscopic analysis. Three new diterpenoids, including two halimanes, 5(10),13Supplementary material is available for this article at 10.1007/s13659-013-0025-8 and is accessible for authorized users. Supplementary material, approximately 797 KB.


R,3R)-2,3-BD, meso-2,3-BD and -2,3-BD. Microbial fermentative processes have been reported for -2,3-BD and meso-2,3-BD production.Butane-2,3-diol  is a fueland platform biochemical with various industrial applications. 2,3-BD exists in three stereoisomeric forms: -2,3-BD from glucose was acquired by whole cells of recombinant Escherichia coli coexpressing the α-acetolactate synthase and meso-butane-2,3-diol dehydrogenase of Enterobacter cloacae subsp. dissolvens strain SDM. An optimal biocatalyst for -2,3-BD production, E. coli BL21 (pETDuet–PT7–budB–PT7–budC), was constructed and the bioconversion conditions were optimized. With the addition of 10 mM FeCl3 in the bioconversion system, -2,3-BD at a concentration of 2.2 g/L was obtained with a stereoisomeric purity of 95.0 % using the metabolically engineered strain from glucose.The production of -2,3-BD from glucose. The results demonstrated that the method developed here would be a promising process for efficient -2,3-BD production.The engineered The online version of this article (doi:10.1186/s13068-015-0324-x) contains supplementary material, which is available to authorized users. R,3R)-2,3-BD, meso-2,3-BD and -2,3-BD. Optically pure 2,3-BD can act as an excellent building block in asymmetric synthesis of valuable chiral chemicals )/([M] + [S] + [R])) × 100 %, where [S], [M] and [R] represent the concentrations of -2,3-BD, meso-2,3-BD and -2,3-BD, respectively.Samples were withdrawn periodically and centrifuged at 12,000×eviously . The GC


H-benzo[d]imidazoles, possessing trimethoxyphenyl pharmacophore, were synthesized to evaluate their biological activities as tubulin inhibitors. Cytotoxic activity of the synthesized compounds 7a-f was assessed against several human cancer cell lines, including MCF-7 (breast cancer cell), HEPG2 (liver hepatocellular cells), A549 , T47D  and fibroblast. According to our results, HEPG2 seems to be the most sensitive, while MCF7 was the most resistant cell line to the compounds. All the compounds expect 7b, possessed satisfactory activity against HEPG2 with mean IC50 values ranging from 15.60 to 43.81 µM. A new series of 1,2-diaryl-4,5,6,7-tetrahydro-1 Twisting around a hypothetical axis, usually 13 protofilaments constitute a hallow microtubule cylinder with H-donor groups on positions 3 or 4 are essential for optimum activity  from the African willow tree, mcaffrum . CA-4  values are calculated in hertz (Hz) and spin multiplicities are given as s (singlet), d (double), t (triplet) and m (multi). The mass spectral measurements were performed on a 6410Agilent triple quadrupole mass spectrometer (LCMS) with an electrospray ionization (ESI) interface.All chemicals, reagents and solvents used in this study were purchased from Merck AG and Aldrich Chemical Company and were used without further purifications. Melting points were determined with a Thomas–Hoover capillary apparatus. Infrared spectra were acquired using a Perkin Elmer Model 1420 spectrometer. A Brucker FT-500 MHz instrument  was used to acquire 1HNMR spectra with TMS as internal standard. Chloroform-D and DMSO-DChemistryGeneral procedure for the preparation of compounds 7a-fAmixture of 3,4,5-trimethoxyaldehyde , 1,2-cyclohexadione , an aniline derivatives  and ammonium acetate in15 mL glacial acetic acid was placed in a 20 mL microwave reactor and stirred under microwave irradiation (150 W) at approximately 120 oC for 6 minutes. The completion of the reaction was monitored by TLC. The reaction mixture was then concentrated under reduced pressure in a water bath. The dark sticky residue was subjected to a column chromatography using a 30 cm silica gel column as stationary phase and Ethyl actate/Hexan (5:1) as mobile phase to obtain the purified compounds 7a-f.1-(4-Methoxyphenyl)-2--4,5,6,7-tetrahydro-1H-benzoimidazole (7a)m/z: 395.1HNMR : δ 1.85-1.91 , 2.38-2.40, 2.75-2.78 ,3.66 , 3.84 , 3.86 , 6.65 , 6.97-6.98 , 7.16-7.17 .Yield,14%;brown powder; mp 110-111 oC; IR (KBr disk): γ (cm-1) 1578 (C=N), 1600-1400 (aromatic), 1130 (C-N);LC-MS (ESI) 1-(3-Methoxyphenyl)-2--4,5,6,7-tetrahydro-1H-benzoimidazole (7b)m/z: 395.1HNMR : δ1.82-1.87 , 2.39 and 2.72 ,3.61 , 3.75 , 3.80 , 6.62 , 6.73 , 6.80-6.82 , 6.92-6.94 , 7.32-7.35 .Yield,10%;red liquid; IR (KBr disk): γ (cm-1) 1596 (C=N), 1600-1400 (aromatic), 1141 (C-N); LC-MS (ESI) 4--phenol (7c)m/z: 381.1HNMR : δ 1.75 , 2.28 and 2.55 , 3.53 , 3.61 , 6.58 , 6.85-6.87 , 7.08-7.09 ,8.3 .Yield: 15.3%, off-white powder; mp: 259-260 oC; IR (KBr disk): γ (cm-1) 3383 (OH), 1600-1400 (aromatic), 1223 (C-N); LC-MS (ESI)1-(4-Fluorophenyl)-2--4,5,6,7-tetrahydro-1H-benzoimidazole (7d)m/z: 383.1H NMR : δ 1.81-1.83 , 2.33 and 2.69 ,3.60 , 3.78 , 6.54 , 7.15-7.16 , 7.09-7.13 . Yield,23.3%;pink powder; mp 152-154 oC; IR (KBr disk): γ (cm-1) 1685 (C=N), 1600-1400 (aromatic); LC-MS (ESI) 1-(4-Chlorophenyl)-2--4,5,6,7-tetrahydro-1H-benzoimidazole (7e)m/z: 399,401 . 1HNMR : δ 1.86-1.90 , 2.41 and 2.76 ,3.66 , 3.85 , 6.59 , 7.17-7.19 , 7.44-7.46 .Yield, 24%;brown powder; mp 157-158 oC; IR (KBr disk): γ (cm-1) 1584 (C=N), 1600-1400 (aromatic), 1135 (C-N); LC-MS (ESI) 1-(3-Chlorophenyl)-2--4,5,6,7-tetrahydro-1H-benzoimidazole (7f)m/z: 399,401 . 1H NMR : δ 1.87-1.92 , 2.43 and 2.76 ,3.67 , 3.87, 6.61 , 7.10-7.14 , 7.41 -7.44 .Yield, 16%; yellow powder; mp289-290 oC; IR (KBr disk): γ (cm-1) 1586 (C=N), 1600-1400 (aromatic), 1130 (C-N); LC-MS (ESI) Anticancer activity2 supplemented with 10% heat inactivated fetal bovine serum (FBS), 100 U/mL penicillin and 100 µg/mL streptomycin. Cell viability was evaluated using MTT method which is based on the reduction of 3--2,5-diphenyltetrazolium bromide (MTT) dye to purple formazan crystals by mitochondrial succinate dehydrogenase enzyme in living cells. The cells were seeded into 96-well plates at a concentration of 6000 cells/well and allowed to incubate for 24 h. The medium was then discarded and different concentrations of the test compounds dissolved in complete medium, were added to each well. After further incubation for 24 h at 37 °C, the medium was aspirated and 100 µL MTT (2 mg/mL) was added to the wells and incubated for 3 h at 37 °C. The produced formazan crystals were dissolved in 100 µL of DMSO. Plates were incubated for 20 min at 37 °C and the optical densities were read at 570 nm with a reference wavelength of 630 nm as background using a spectrophotometer plate reader . Colchicine was used as positive control and DMSO as the solvent of the test compounds and its final concentration was less than 0.2%. IC50 was calculated by calibration curve using Prism software. All the tests were performed in triplicates.Fibroblast, MCF7 (human breast cancer), HEPG2 (human liver carcinoma), A549 (human lung cancer) and T47D (human breast cancer) cell lines were purchased from Pasteur Institute, Tehran, Iran. The cells were grown in RPMI1640 medium at 37 °C under 5% COMolecular modeling (docking) studiesAll the synthesized compounds were subjected to molecular modeling study and their interactions with the colchicine binding site of β-tubulin were simulated. The tubulin structure was downloaded from the PDB data bank (http://www.rcsb.org/ - PDB code: 1SA0) , 32. AllH-benzo[d]imidazole derivatives were synthesized based on the synthetic procedure described previously by Zarghi et al. , HEPG2 (liver hepatocellular cells), A549 , T47D  and fibroblast following a previously described procedure . The res50) of each compound against each cancerous cell with that of the normal human Fibroblast cells (Table 1). Results were expressed as selectivity index (SI). Compounds 7c, 7d and 7e appear to be the most selective against HEPG2. The highest selectivity was, however, observed for compound 7c against HEPG2 cell line (SI~ 4.1).Compound 7e exhibited the most potent activities against HEPG2. Selectivity of the cytotoxic activity of the six synthesized compounds was assessed by comparing the cytotoxic activity (ICMolecular modeling studies comparing colchicine and the synthesized compound 7a were carried out. The structures were superimposed by a least square fit of the 3,4,5-trimethoxyphenyl moieties. The results are shown in


Their structures were identified by extensive spectroscopic analysis. Compounds 8 and 16 showed significant selective cytotoxicity against human histiocytic lymphoma U937 cell line with IC50 values of 4.9 and 8.8 μM, respectively. In addition, this is the first time to report that 8, 15 and 16 had mild antibacterial activity against Escherichia coli and Bacillus subtilis, and 15 showed potent antilarval activity against barnacle Balanus amphitrite larval settlement.Eight new chromones, engyodontiumones A–H ( Engyodontium album, isolated from a marine sediment sample collected in the South China Sea , attracted our attention. Preliminary experiments demonstrated that the extract of the culture medium of the strain showed toxicity towards brine shrimp and antibacterial activity. Further investigation on the chemical constituents of the extract resulted in the isolation of eight new chromones (1–8) and three new phenol derivatives (9–11) together with eight known compounds: sydowinin A (12) +). Its NMR data (C 56.8) and the absence of a hydroxyl proton signal in 1, which indicated that 1 was a chromone skeleton. In the HMBC spectrum  with C-2 suggested a methoxy group located at C-2, and correlations of δH 4.60  with C-5/C-6/C-7 suggested a hydroxymethene group attached at C-6. In addition, HMBC correlations of δH 3.87  with C-14 combined with the chemical shift of C-1 , suggested a methoxycarbonyl group located at C-1. Therefore, the structure of 1 was determined as shown.Engyodontiumone A  had a molecular formula of C16H11ClO6 as inferred from its HRESIMS (m/z 335.0309 [M + H]+) showing peaks at m/z 335.0309 and 337.0264 with the height ratio of 3:1. Its NMR data (C 148.9 in sydowinin B to δC 125.5 in 2) suggesting the loss of the hydroxyl functionality. Combined with the molecular formula and HMBC spectral data  was deduced from its HRESIMS (m/z 321.0972 [M + H]+). Its NMR data (7 with the only difference is the presence of one extra methine (δH 3.84 and δC 43.5) and the absence of one oxygenated quaternary carbon (δC 74.3 (C-1) in aspergillusone B), which suggested that 3 had the same skeleton as aspergillusone B with the only difference being the substitution at C-1. This was proved by the HMBC spectrum (H 3.84) with C-3/C-9/C-11. The coupling constant values of JH-1,H-2 (6.0 Hz) and JH-2,H-3  indicated that H-1 and H-2 were equatorial–axial relationship and required the placement of H-1 and H-2 in pseudo equatorial and axial orientations +). Its NMR data  and one oxygenated methine  and absence of two methylenes. The HMBC spectrum . In addition, correlations of δH 2.37 with C-5/C-6/C-7 suggested that a methyl group CH3-16 was attached at C-6. The coupling constant values of JH-1,H-2 (5.5 Hz), JH-2,H-3 (5.5 Hz) and JH-3,H-4  indicated the equatorial–axial relationship for H-1/H-2 and H-2/H-3 placing H-1, H-2 and H-3 in pseudo equatorial, axial and equatorial orientations +). Its NMR data  and JH-2,H-3  indicated the equatorial–equatorial relationship for H-1 and H-2 placing of H-1 and H-2 in pseudo equatorial orientation  had a molecular formula of C16H16O8 as deduced from its HRESIMS (m/z 337.0922 [M + H]+). Its NMR data (4 with the only difference of one methyl group (CH3-16) oxygenated to be methylene . This was supported by the HMBC spectrum (see H 5.50) with C-5/C-6/C-7. The coupling constant values of JH-1, H-2 (4.5 Hz), JH-2,H-3 (4.5 Hz) and JH-3,H-4  indicated the axial–equatorial relationship for H-1/H-2 and H-2/H-3 placing H-1, H-2 and H-3 in pseudo equatorial, axial and equatorial orientations, respectively  had the same molecular formula of C16H16O8 as 6, which was deduced from its HRESIMS (m/z 337.0922 [M + H]+). Its NMR data , JH-2,H-3 (1.5 Hz), and JH-3,H-4  indicated that axial–equatorial relationship for H-1 and H-2 and equatorial–equatorial relationship for H-2 and H-3 placing H-1, H-2 and H-3 in pseudo axial, equatorial and equatorial orientations, respectively  had the same molecular formula of C16H14O7 as AGI-B4 [m/z 321.0496 [M + H]+). Its NMR data (H 4.60 in AGI-B4 to δH 5.01 in 8) and the coupling constant value JH-1,H-2 (from 3.5 Hz in AGI-B4 to 10.0 Hz in 8), suggesting 8 is an epimer of AGI-B4 at C-2. Further detailed analysis of HMBC spectrum  indicated that H-1 and H-2 were in a cis-configuration. We also failed to determine the absolute structure of 8 by Mosher ester methods as compound 3.Engyodontiumone H  had the NMR data  also shotrum see  proved t9 was established as C15H16O4 by its HRESIMS (m/z 261.1122 [M + H]+). Its NMR data showed close similarity to those of cordyol C +). Its 1H NMR spectrum exhibited characteristic signals for a trisubstituted benzene ring , 7.33 , 7.09 ) and a trisubstituted double-bond olefinic proton ). The 13C NMR and DEPT spectral data revealed 15 carbon signals including six quaternary carbons, four methines, two methylenes, and three methyls. These data showed similarity to those of hydroxysydonic acid +). Its NMR data were very similar to those of 10 except for the chemical shift changes of H-9, C-3 and C-14. Further detailed analysis of the HMBC spectrum suggested that 11 and 10 had the same planar structure. The chemical shift of C-14 (δC 23.8) indicated that the double bond between C-7 and C-8 was in a Z-configuration + .Engyodontiumone B (2): Yellow needles; UV (MeOH) λmax (log ε) 372 (2.75), 287 (3.05), 259 (3.52), 234 (3.55) nm; FT-IR: 3394, 1674 cm−1; 1H and 13C NMR data, see m/ z 335.0309 [M + H]+ .Engyodontiumone C (3): Brown needles; c 0.5, MeOH); UV (MeOH) λmax (log ε) 328 (3.38), 239 (4.13), 203 (4.06) nm; FT-IR: 3417, 1728, 1697, 1651 cm−1; 1H and 13C NMR data, see m/z 321.0972 [M + H]+ .Engyodontiumone D (4): Brown needles; c 0.3, MeOH); UV (MeOH) λmax (log ε) 325 (3.38), 322 (3.38), 238 (4.07), 230 (4.05), 202 (4.07) nm; FT-IR: 3456,1728,1658, 1627 cm−1; 1H and 13C NMR data, see m/z 321.0974 [M + H]+ .Engyodontiumone E (5): Brown needles; c 0.4, MeOH); UV (MeOH) λmax (log ε) 329 (3.41), 258 (3.97), 238 (4.10), 203 (4.07) nm; FT-IR: 3336,1724,1654, 1620 cm−1; 1H and 13C NMR data, see m/z 321.0968 [M + H]+ .Engyodontiumone F (6): Brown needles; c 0.3, MeOH); UV (MeOH) λmax (log ε) 328 (3.42), 238 (4.17), 203 (4.06) nm; FT-IR: 3334, 1732, 1651 cm−1; 1H and 13C NMR data, see m/z 337.0922 [M + H]+ .Engyodontiumone G (7): Brown needles; c 0.2, MeOH); UV (MeOH) λmax (log ε) 328 (3.14), 238 (3.94), 202 (3.94) nm; FT-IR: 3336, 1728, 1651 cm−1; 1H and13C NMR data, see m/z 337.0922 [M + H]+ .Engyodontiumone H (8): Brown needles; c 1, MeOH); UV (MeOH) λmax (log ε) 344 (3.31), 272 (3.98), 213 (3.90), 203 (3.97) nm; FT-IR: 3420, 1735, 1658 cm−1; 1H and13C NMR data, see m/z 319.0922 [M + H]+ .2-methoxyl cordyol C (9): Yellow amorphous solid; UV (MeOH) λmax (log ε) 360 (2.64), 356 (2.64), 274 (3.20) nm; FT-IR: 3271, 1678 cm−1; 1H NMR  δH: 9.35 , 6.49 , 6.26 , 6.24 , 6.17 , 6.06  , 3.60 , 2.16  and 2.14 ; 13C NMR  δC: 158.7 , 158.2 , 151.0  , 148.3 , 139.7 , 137.3 , 133.1 , 113.2 , 112.4 , 110.1 , 108.0 , 100.9 , 60.1 , 21.1  and 20.6 . HRESIMS m/z 261.1122 [M + H]+ .Engyodontiumone I (10): Yellow gum; UV (MeOH) λmax (log ε) 302 (3.40), 255 (3.64), 211 (4.16) nm; FT-IR: 3402, 3132, 1689, 1651, 979, 948 cm−1; 1H NMR  δH: 7.40 , 7.33 , 7.09 , 5.49 , 2.18 , 1.93 , 1.48 , 1.11 ; 13C NMR  δC: 167.2 , 154.3 , 136.7 , 133.4 , 130.8 , 129.9 , 129.2 , 120.1 , 116.1 , 68.8 , 43.1 , 29.2 , 23.1 , 16.4 ; HRESIMS m/z 265.1426 [M + H]+ .Engyodontiumone J (11): Pale yellow gum; UV (MeOH) λmax (log ε) 297 (3.26), 244 (3.57), 207 (4.10) nm; FT-IR: 3344, 1689, 1612 cm−1; 1H NMR  δH: 7.42 , 7.35 , 7.03 , 5.44 , 2.18 , 1.93 , 1.81 , 1.37 , 0.95 ; 13C NMR  δC: 167.2 , 154.2 , 133.3 , 132.5 , 130.1 , 129.6 , 128.7 , 119.7 , 116.0 , 68.5 , 43.4 , 29.0 , 24.0 , 23.8 ; HRESIMS m/z 265.1428 [M + H]+ .2–8, 12–14, 16) were tested for cytotoxicity against Hela, MCF-7, HepG2 and Huh7 human carcinoma cell lines using a MTT -2,5-diphenyl-2-H-tetrazolium bromide) method, and against U937 human carcinoma cell line using a CCK-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5--2H-tetrazolium, monosodium salt) method. The media for U937 and the other four cell lines were RPMI1640 and DMEM, respectively. Cells in the logarithmic phase were seeded into 96-well plates in a 100 μL volume of medium, maintained at 37 °C in a 5% CO2 incubator overnight, then treated with the corresponding compounds or vehicle control at the indicated concentration. After 48 h of incubation, 5 μL of MTT or CCK-8 was added into the 96-well plates and incubated for additional 2–3 h, then the absorbance was measured in a microplate reader at 450 nm and 650 nm. Cell toxicity was evaluated as the ratio of the absorbance of the sample to that of the control. The IC50 were further calculated using Graphpad Prism 5 .Chromones (1−19 were tested against common bacteria Bacillus subtilis and Escherichia coli using a standard disc diffusion assay [8 CFU/mL) was mixed with 20 mL of corresponding nutrient agar and poured in 100 mm sterile petri plates. Sterile 6 mm diameter circular discs of filter paper were loaded with 25 μg of tested compound dissolved in methanol, evaporated to dryness, and then placed onto the seeded bacterial plates. An additional set of discs with 10 μg penicillin was used as the positive controls. The agar plates were incubated for 18 h at 37 °C. The inhibition zones were measured. Antibacterial activities of on assay . Briefly5 CFU/mL) in the starch culture-medium. Tested samples and penicillin (antibiotic control) were dissolved in DMSO and dispensed into 96-well plates to create a dilution series  in a 100 μL volume of starch culture-medium. Following incubation at 37 °C for 12–24 h, the tubes were examined for visible bacterial growth as evidenced by turbidity. The lowest concentration of test compound that prevented 100% growth represented the minimal inhibitory concentration (MIC). The experiment was run in 3 replicates.The minimum inhibitory concentration (MIC) of active compounds were determined by a microbroth dilution method . Briefly−1 for preliminary bioassay. To define the EC50 values of antilarval compounds found in the preliminary bioassay, different dilutions of the tested compounds were further prepared to the concentrations ranging from 1 to 200 μg mL−1 in autoclaved FSW (filtered seawater). About 20 competent larvae were added to each well in 1 mL of the test solution. Wells containing only FSW with DMSO served as the controls. The plates were incubated at 27 °C for 24 h. The percentage of larval settlement was determined by counting the settled, live individuals under a dissecting microscope and expressing the result as a proportion of the total number of larvae in the well [Larval settlement bioassays were performed using sterile 24-well polystyrene plates. Tested samples were dissolved in DMSO to a concentration of 25 μg mLthe well .1–8) and three new phenol derivatives (9–11) together with eight known polyketides were isolated from the culture medium of the deep-sea-derived fungus Engyodontium album DFFSCS021. Among then, 8 and 16 showed significant selective cytotoxicity against U937 cell line with IC50 < 10 μM. This is the first time to report that 8, 15 and 16 had mild antibacterial activity against E. coli and B. subtilis with MIC ≤ 64.0 μg/mL, and 15 showed potent antilarval activity against barnacle B. amphitrite larval settlement with EC50 value of 19.1 μg/mL and low toxicity.In conclusion, eight new chromones (


A green, efficient synthesis of 5-arylidene rhodanine and 2,4-thiazolidinedione derivatives without using any external catalyst in polyethylene glycol (PEG) at 80°C has been described. Reaction procedure is very simple, short, and obtained yields are very high. Rhodanines and thiazolidinediones both are aprivileged class of molecule, and they show large number of biological activities. The most significant position of these molecules seems to be as they are asubset of commercially employed noninsulin-dependent diabetes mellitus (NIDDM), insulin sensitizing agents  such as N-acetylmuramate/L-alanine ligase inhibitor [β-lactamase [Furthermore, rhodanine derivatives possess anticonvulsant, antibacterial, antiviral, and antidiabetic activities –3 Some onhibitor , aldose nhibitor , β-lactaactamase , and JNKactamase , while sactamase . Therefoactamase –12.Unlike rhodanine, 2,4-thiazolidinedione derivatives also have remarkable biological activities like antidiabetic , antibacNitrones (imine oxides) are reputed as 1,3-dipoles and are extensively explored for the synthesis of five membered heterocycles by combining them with several types of multiple bonds –27. Apar1a and rhodanine (10 mmol) 2a was stirred at 80°C in polyethylene glycol (PEG) for 20 min to afford corresponding arylidene rhodanine 3a  via addition-elimination process.First of all, a series of nitrones was prepared using a variety of aldehyde and hydroxyl amine as per already reported method . A mixtu2O, THF, PEG, DMF, and so forth and in absence of solvent. Conclusively, in case of PEG best results were obtained with high yields in minimum reduced time. Keeping optimized reaction conditions, a variety of aldehydes with rhodanine/2,4-thiazolidinedione were reacted to afford 5-arylidenerhodanines 3a–g and 5-arylidene-2,4-thiazolidinediones 3h–n with excellent yields  1H NMR spectra were recorded on a BRUKER AVANCE II 400 NMR Spectrometer using tetramethylsilane (TMS) as internal standard. The progress of the reaction was monitored by thin layer chromatography (TLC) using silica gel G (Merck). Reagent-grade chemicals were purchased from a commercial source and used without further purification. Yields refer to the yield of the isolated products. Melting points were determined in open capillaries in paraffin bath and are uncorrected. Infrared (IR) spectra were recorded in KBr discs on a Perkin-Elmer 240C analyzer. A mixture-nitrone (10 mmol) 1, rhodanine or 2,4-thiazolidinedione (10 mmol) 2, and polyethylene glycol (5 mL) was stirred at 80°C temperature for appropriate time see . Reactio3a): IR (KBr): 3390, 1709, 1669, 1600, 1429, 1200 cm−1. 1H NMR  δH: 13.57 , 7.68 , 7.48–7.79 . (5Z)-5-Benzylidene-2-thioxo-1,3-thiazolidine-4-one (3f): 1H NMR  δH: 13.71 , 7.61 , 7.52 , 7.08 , 3.09 .(5Z)-5-(4-Methoxybenzylidene)-2-thioxo-1,3-thiazolidine-4-one ((3h): IR (KBr) cm−1:  3155 (NH), 3049, 879 , 2868 , 1739, 1691 (C=O). 1H NMR  δH: 8.27 , 7.86 , 7.26 . MS m/z (%): 206 (M + 1).(5Z)-5-Benzylidene-1,3-thiazolidine-2,4-dione 3m): IR (KBr) 3211, 1725, 1689 cm−1. 1H NMR  δH: 12.50 , 7.69 , 7.50 , 7.11 , 3.49 . (5Z)-5-(4-Methylbenzylidene)-1,3-thiazolidine-2,4-dione (


C-glycosyl-type D-mannoside analogues, namely 1-propargylamino- and 1-phenylamino-1-deoxy-α-D-manno-heptopyranose. They were investigated as ligands of type 1-fimbriated E. coli bacteria by means of molecular docking and bacterial adhesion studies. It turns out that Amadori rearrangement products have a limited activity as inhibitors of bacterial adhesion because the β-C-glycosidically linked aglycone considerably hampers complexation within the carbohydrate binding site of the type 1-fimbrial lectin FimH.The Amadori rearrangement was employed for the synthesis of  C-glycosyl-type pentose analogues) without the need of hydroxy group protection  can be obtained in exclusively one anomeric form as well as in excellent yields  is the reaction in which aldohexoses react with suitable amines under acidic catalysis to 1-amino-1-deoxyketohexoses (otection . For a lt yields  2–3]. T–3. TC-gl1) but not β-mannosides. Mannosides with an aromatic aglycone, such as p-nitrophenyl α-D-mannoside (pNPMan) and 4-methylumbelliferyl α-D-mannoside (3) show an improved affinity to FimH due to π–π-stacking interactions of the aromatic moiety with the so-called tyrosine gate at the entrance of the carbohydrate binding site, formed by Y48 and Y137. Additional interactions exerted by extended aglycone portions can further improve ligand affinity for FimH; for example ortho-chloro substitution of the phenyl ring (compounds 2 and 5), a squaric acid partial structure (compound 4) or heterocyclic substituents such as in indolinylphenyl mannoside 5 as recently introduced .–17.mannol groups –19 follo8 with an amine of choice allows an efficient and versatile approach towards D-manno-configured C-glycosyl-type glycoconjugates. In our study, we have employed two different amines in the Amadori rearrangement with 8, propargylamine and aniline. Under typical conditions for this reaction D +13.2 ; 1H NMR  δ 3.84 , 3.82 , 3.80 , 3.74 , 3.72–3.69 , 3.62 , 3.58 , 3.10 , 2.96 , 2.76 ; 13C NMR  δ 97.4 (C-2), 80.3 (C-9), 74.8 , 74.5 (C-10), 72.9 (C-4), 68.2 (C-5), 62.8 (C-7), 55.4 (C-1), 38.6 (C-8); ESIMS (m/z): calcd for [C10H17NO6 + H]+, 248.1134; found, 248.113 [M + H]+.1-(N-Phenyl)amino-1-deoxy-α-D-manno-hept-2-ulose (10): To a solution of D-glycero-D-galacto/D-talo-heptopyranoses 8a and 8b  in a mixture of EtOH (1 mL), 1,4-dioxane (0.2 mL) and water (2 drops), aniline  and acetic acid  were added and the reaction mixture was stirred at 70 °C for 48 h. Complete consumption of the starting material was indicated by TLC . The solvents were removed under reduced pressure and subsequent column chromatography  gave 1-phenylamino ketose 10  in a yield of 24%. [α]D +21.5 ; 1H NMR  δ 7.11 , 6.75 , 6.65 , 3.90 , 3.85 , 3.87–3.83 , 3.78–3.76 , 3.75 , 3.63 , 3.43 , 3.27 ; 13C NMR  δ 150.2, 130.0, 118.7, 114.8 , 98.9 (C-2), 74.9 (C-6), 73.3 (C-3), 72.9 (C-4), 68.7 (C-5), 63.0 (C-7), 51.4 (C-1). ESIMS (m/z): calcd for [C13H19NO6 + H]+, 286.1291; found, 286.129 [M + H]+.File 1NMR spectra, boiassays and molecular docking.


Dorema species are used in Persian traditional medicine. In the present study the total extract from the roots of Dorema hyrcanum Koso-Pol. was investigated for its in-vitro (pLDH assay) and in-vivo (Peters’ 4-days suppressive test) antiplasmodial effects and assessed for cytotoxicity against the normal cell line MDBK (MTT test). The IC50 values for a chloroquine- sensitive (3D7) and a chloroquine- resistant (K1) strain of Plasmodium falciparum were 28.64 and 9.79 µg/mL, respectively. The inhibition percentage of the rodent parasite, Plasmodium berghei, on day 4 in mice was 77.9% and IC50 value on Madin–Darby bovine kidney cells (MDBK cells) was 59.84 µg/mL.Some in-vivo model which resulted in the isolation of an acetophenon (compound 1), one new sesquiterpenoid;The total extract was subjected to a bioassay-guided fractionation protocol based on the in-vivo antiplasmodial effect and the results revealed that naghibione showed good suppression activity, inhibiting 68.1 % of the parasite growth.naghibione (compound 2) and two known sesquiterpenoid derivatives . Their structures were elucidated by spectroscopic analysis, including 1D and 2D NMR experiments and ESI-MS. All compounds were evaluated for  Malaria is a global public health problem whose main victims are children under five years of age in Africa. According to the latest WHO estimates, there were about 219 million cases of malaria in 2010. and an estimated 660000 deaths are included in the flora of Iran. Semi preparative HPLC was performed on a Shimadzu instrument with PRC- ODS C18 (20 mm × 25 cm) column. Silica gel 60  was used for column chromatography. All solvents for semi preparative HPLC were of technical grade and purified by distillation. Plant material D. hyrcanum (Apiaceae), collected in June 2008 from Almed Valley in Golestan District, Iran, were identified by Mr. H. Moazeni from Traditional Medicine and Materia Medica Center (TMRC), Shahid Beheshti University of Medical Sciences, Iran. The voucher specimen 2495 (TMRC) of the plant has been deposited in the herbarium of the TMRC.The roots of ExtractionD. hyrcanum was macerated in methanol (MeOH), for 24 hours with constant shaking, at room temperature. The filtrates of total extract were evaporated to dryness and investigated for its in-vitro and in-vivo antiplasmodial and cytotoxic effects.The powdered dried root of Biological assaysIn-vitro antiplasmodial activityPlasmodium falciparum that were continuously cultured according to the methods described by Trager and Jensen (in-vitro in human blood using parasite lactate dehydrogenase method (pLDH) with slight modifications and chloroquine-sensitive (3D7) strains of d Jensen . Plant efications, 15. The analysis.  In-vivo antimalarial assay Plasmodium berghei infection in mice was employed(17) for evaluating of the samples of D.hyrcanum antiplasmodial activities. Adult male albino mice from the Pasteur Institute of Iran were housed under standard environmental conditions and fed with standard pellets and water. All the procedure was accepted by Shahid Beheshti University of Medical Sciences Ethics Committee and in accordance with the principles for laboratory animal use and care in the European Community guidelines. Peters’ 4-day suppressive test against NICD strain of7 infected erythrocytes. The mice were randomly divided into groups of five per cage and treated during consecutive days with 10 mg/mL of the sample by I.P injection for 4 days. Two control groups were used in this experiment, one treated with chloroquine at dose of 20 mg/Kg as a positive control while the other group was kept untreated as a negative group. On day 5 of the test, thin blood smears were prepared and blood films were fixed with methanol. The blood films were stained with Giemsa, and then microscopically examined. Percentage of parasitaemia was counted based on infected erythrocytes calculated per 1000 erythrocytes.Briefly, the parasites (blood contained parasites) were maintained by serial passage of blood from mouse to mouse. Adult male albino mice weighing 20–25 g were inoculated by intra-peritoneal (I.P) injection with 1×107 infected erythrocytes. The mice were treated during consecutive days with 10 mg/Kg of the sample by I.P injection for 4 days. Two control groups were used in this experiment, one treated with chloroquine at dose of 20 mg/Kg as a positive control while the another group was kept untreated as a negative group. Percentage of parasitaemia was counted based on infected erythrocytes calculated per 1000 erythrocytes and the inhibition percentage of each group expressed in relation to the untreated group.The mice were inoculated by intra-peritoneal (I.P) injection with 10In-vitro cytotoxicity assay50 values were calculated from the drug concentration–response curves. Cytotoxicity of samples was determined using MDBK cells, 18 by tBioassay guided fractionation and isolationD. hyrcanum was subjected to a bioassay-guided fractionation protocol based on the in-vivo model. For this purpose the roots of D. hyrcanum (750 g) were macerated in ethyl acetate (EtOAc) for 24 hours at room temperature with constant shaking (2: 1) for 3 times eluting with hexane, a gradient of hexane- chloroform , a gradient of chloroform- EtOAc , EtOAc- MeOH (5:5) and MeOH to give 14 fractions. Compound 2 (200 mg) was isolated from fraction 2- 6 (237 mg) which was eluted with hexane- chloroform (1:9) to 100% chloroform.t = 23.5min (76 mg). It was purified on a silica gel column  wi'th a gradient of petroleum benzene- DCM , a gradi'ent of dichloromethane- EtOAc , EtOAc- MeOH (5:5), MeOH to yield compounds 3 (18 mg) and 4 (20 mg). All pure compounds were evaluated for in-vivo antiplasmodial effects. Fraction 3 was separated by silica gel column chromatography  eluting with a gradient of petroleum benzene- DCM , a gradient of DCM- EtOAc , EtOAc- MeOH (5:5) and MeOH to give 8 fractions. Fraction 3-2 (2 g), which was eluted with DCM- EtOAc (9:1), was separated by silica gel column chromatography  eluting with a gradient of petroleum benzene- DCM , gradients of dichloromethane- EtOAc , EtOAc- MeOH (5:5) and MeOH. There were obtained 7 sub fractions. Fraction 3-2-3 (230 mg) was separated by semi preparative HPLC  and then isolated the subfraction with R1H-NMR : δ ppm= 6.14 , 6.25 , 5.19 , 13C-NMR: δ =206.4 (C-7), 166.8 (C-4), 164.5 (C-2), 160.2 (C-6), 106.9 (C-1), 103.2 (C-1''), 100.0 (C-1'), 96.0 (C-5), 95.0 (C-3), 76.4 (C-3'), 76.1 (C-5''), 76.0 , 74.0 (C-2''), 73.2 (C-2'), 70.1 (C-4''), 69.6 (C-4'), 68.8 (C-6'), 61.2 (C-6''), 56.3 (C-OMe), 33 (C-8); HR‑ESI‑TOF‑MS (positive): m/z = 506.1631 [M + Na]+ .4-methoxy-6-hydroxyacetophenone-2-O-β-D-gentiobioside (1): Yellow crystal, (E), 10 dodecadiene- 1- dione (2): Yellow oil; 1H-  and 13C‑NMR (125 MHz) data: see + . 1(2-hydroxy-4-methoxy)- 3,7,11- trimethyl-3-vinyl-6S*,3R*-dimethyl-2--furocoumarin (3): yellow oil, 1H-NMR : δ ppm = 7.52 , 6.85 , 6.83, 5.08 , 5.05 , 3.86 , 3.27 , 2.09 , 2.01 , 1.94 , 1.67 , 1.59 , 1.77 , 1.44 , 13C-NMR: δ = 165.4 (C- 9b), 165.2 (C4), 163.2 (C-7), 156.9 (C- 5a), 131.9 (C-8'), 136.0 (C-4'), 124.2 (C-7'), 122.8 (C-3'), 123.7 (C-9), 112.1 (C- 8), 106.3 (C-9a), 103.7 (C-3a), 100.7 (C-6), 96.6 (C-2), 55.2 (C-OMe), 39.0 (C-5'), 42.0 ( C-3), 41.5 (C-1'), 25.7 (C-9'), 22.1 (C-2 '), 20.2 (C-2Me), 17.7 (C-8'Me), 16.0 (C-4'Me), 13.7; HR‑ESI‑TOF‑MS (positive): m/z = 419.2221 [M + Na]+ .2,3-dihydro-7-methoxy-2R*,3R*dimethyl-2--furocoumarin (4): yellow oil, 1H-NMR : δ ppm= 7.52 , 6.85 , 6.83 , 5.19 , 5.05 , 3.86 , 3.22 , 2.24 , 2.01 , 1.94 ,1.67 , 1.63 , 1.60 , 1.90 , 1.48, 13C-NMR: δ = 165.4 (C- 9b), 165.2 (C4), 163.2 (C-7), 156.9 (C- 5a), 131.9 (C-8'), 136.0 (C-4'), 124.2 (C-7'), 122.8 (C-3'), 123.7 (C-9), 112.1 (C- 8), 106.3 (C-9a), 103.7 (C-3a), 100.7 (C-6), 96.6 (C-2), 55.2 (C-OMe), 39.0 (C-5'), 44.4 ( C-3), 35.4 (C-1'), 25.7 (C-9'), 23.1 (C-2'), 25.7 (C-2Me), 17.7 (C-8'Me), 16.0 (C-4'Me), 13.7; HR‑ESI‑TOF‑MS (positive): m/z = 419.2221 [M + Na]+ .2,3-dihydro-7-methoxy-2D. hyrcanum was evaluated for its antiplasmodial activity. In a screening program, its methanolic extract was tested against a chloroquine- sensitive strain of Plasmodium falciparum (3D7) as well as a chloroquine- resistant strain (K1) and showed IC50 values of 28.64 and 9.79 µg/mL, respectively. Furthermore it was evaluated by Peters’ 4-day suppressive test and revealed 77.9% inhibition of P. berghei on day 4 in mice at a concentration of 10 mg/Kg. In a cytotoxicity assay against MDBK cells, it showed an IC50 value of 59.84 µg/mL. In this study for the first time in-vivo model (Peters’ test) was assessed in order to isolate the active compounds. Therefore, the ethyl acetate extract, some of its selected fractions and pure compounds were evaluated for antiplasmodial activity by the in-vivo test and for cytotoxicity , 4.95 , 5.00 , 5.88 . Three aromatic protons at δ 6.40 (br s), 6.42  and 7.65  suggested the presence of a 1, 2, 4-trisubstituted benzene ring, which was supported by the 13C NMR spectrum. The side chain of this ring was also assigned through 13C NMR, HMQC, 1H-1H COSY, and HMBC experiments. In the HMBC spectrum of 2, the correlations of H-2 (δH 2.87) with C-1 (δ C 204.2), C-3 (40.1), and C-13 (145.8); H-8 (δH 1.95) with C-7 (δc 135.5) and C-9 (δc 26.5); H-13 (δH 5.88) and H-14 (δH 4.97) with C-3 (40.1); OH (δH 13.1) with C-3' (δC 100.5), C-1' (δC 115.0), and C-2' (δC 166.2); and H-6' (δH 7.65) with C-1 (δC 204.2) and C-4' (δC 165.85) confirmed the structure of compound 2 (The bio-assay guided fractionation led to the isolation of compound 2sonances  of compompound 2 . E), 10- dodecadiene-1- dione and named naghibione.Therefore, the structure of compound 2 was elucidated as 1-(2-hydroxy-4-methoxyphenyl)-3, 7, 11-trimethyl- 3- vinyl- 6, sesquiterpenes should be considered as lead compounds for further investigation,7-nonadienyl]-furocoumarin and 2,3-dihydro-7-methoxy-2R*,3R*dimethyl-2--furocoumarin, respectively, by comparison with literature data ; a new sesquiterpenoid with antiplasmodial activity has been isolated. Furthermore, two known sesquiterpenoids 3 and 4 were for the first time isolated from this genus. In this study for the first time, Further studies in order to evaluate the mode of action of the pure compounds are under way.


O-mono-lauryl β-cyclodextrin, the self-organized supramolecular architecture with nano-vesicular morphology in water was investigated by fluorescence spectroscopy, dynamic light scattering and transmission electron microscopy. The cavity-type nano-assembled vesicle and the novel synthetic methods for the preparation of mono-acylated cyclodextrin should be of great interest with regard to drug/gene delivery systems, functional surfactants, and carbohydrate derivatization methods.Amphiphilic cyclodextrins have been synthesized with self-acylating reaction using vinyl esters in dimethylformamide. In the present study no base, catalyst, or enzyme was used, and the structural analyses using thin layer chromatography, nuclear magnetic resonance spectroscopy and mass spectrometry show that the cyclodextrin is substituted preferentially by one acyl moiety at the C2 position of the glucose unit, suggesting that cyclodextrin functions as a regioselective catalytic carbohydrate in organic solvent. In the self-acylation, the most acidic OH group at the 2-position and the inclusion complexing ability of cyclodextrin were considered to be significant. The substrate preference was also observed in favor of the long-chain acyl group, which could be attributed to the inclusion ability of cyclodextrin cavity. Furthermore, using the model amphiphilic building block, 2- Cyclodextrins (CDs) are macrocyclic oligosaccharides consisting of six (in α-), seven (in β-), eight (in γ-) or more α-D-1 → 4) linked glucoses. The structures are shown as a hollow truncated cone shape with the C6 primary hydroxyl groups are located in the narrow rim while the more open rim is crowned by the secondary hydroxyl groups at positions C2 and C3. Based on the torus shape with the hydrophilic exterior and hydrophobic interior, they can incorporate hydrophobic guests in their cavies → 4 linkThe modification of CDs has widened the application fields for binding receptors36812131416Grafting of hydrophobic moieties onto CDs produces amphiphilic CDs which are self-assembled to nanoarchitectures such as nanoparticles, micelles, and vesicles in aqueous media1819O-mono-lauryl β-CDs to self-assemble was evaluated using fluorescence spectroscopy, dynamic light scattering and transmission electron spectroscopy.The present work describes the novel ladle-type CDs based on the self-monolaurylated β-CD at the C2 position and the self-assembled nanovesicles. For the self-acylation, the effects of vinyl laurate concentration, temperature, and solvent are presented and discussed. Other vinyl esters  and carbohydrates  are also investigated. The ability of 2-O-lauryl β-CD whose chemical structure is shown in 2O, mono-2-O-lauryl β-CD was obtained in 17.1% yield and the structure was analyzed by MALDI TOF mass spectrometry and NMR spectroscopy. The pseudo-molecular ion peak at m/z = 1339.9013 as [mono-2-O-lauryl β-CD+Na]+ is shown in its MALDI-TOF mass spectrum  of β-CD cavity are correlated with the methylene protons (H10-H17) of the lauryl chain. In the previous study, lollipop-style mono-6-substituted β-CD has also shown the intramolecular complexes by auto-inclusionFor the self-laurylation in the basic and polar aprotic solvent, DMF, it is assumed that intramolecular hydrogen bonds between OH3′ and OH2 stabilize the oxyanion at the C2 position of β-CD and the alkyl chain of vinyl laurate is inserted into the cavity . Indeed,28The mono-laurylation is regarded to be due to the inclusion property of β-CD. Although the ratio of vinyl laurate to β-CD may be increased, the conversion to mono-laurate in 24 h shows no significant difference . From th31O-lauryl β-CD was investigated at different temperatures including 25, 50, and 75 °C. The mono-acylation was found to be increased and then saturated with increasing temperature, and significant conversions  were achieved in 20 h , α- and γ-CD were compared in the same way for self-laurylation. Glucose and maltodextrin show no product, and only the CD makes the acyl product, indicating the significance of cyclic nature for the self-laurylation. Cyclization may induce the characteristic molecular environment that hydrogen bonding networks inside of the ring make the 2OH of glucose more acidic. The order of produced lauryl CD amount is γ-CD > β-CD > α-CD at every time period . The oxyO-lauryl β-CD is assumed to look like a ladle from the conformational search work in the Maestro software  is regarded as a polarity indicator of the microenvironment around the pyrene. Actually, the I1/I3 value is 0.58 in hexane and 1.87 in water1/I3 value of mono-2-O-lauryl β-CD declined with increasing the concentration, and the inflection point of the curve corresponds to the critical aggregation concentration (cac) (O-lauryl β-CD can be used as effective nanocarrier systems for target hydrophobic substances.The mono-2-software . This laon (cac) . Since tO-lauryl β-CD . Skirt-type β-CD with 2–7 decanoyl substituents at the C2 position makes the self-assembled nanoparticle 219 nm in mean diameterSelf-assembling properties by the amphiphilic β-CD in water were analyzed using the nanoprecipitation method, and investigated by DLS and TEM. The DLS result shows the mean hydrodynamic diameter of the self-assembled architecture as 136 nm with a polydispersity index of 0.07 . The DLSryl β-CD . From thryl β-CD , an end-O-lauryl β-CD was successfully synthesized by the self-acylating method using vinyl laurate without a base, catalyst or enzyme. The structure was analyzed with TLC, NMR, and MALDI TOF mass spectrometry. Throughout the investigations on the effects of substrate variations, temperature, solvent choice, and molecular docking, the main factor for self-acylation is considered to be the most acidic 2OH and the inclusion ability of β-CD. Using the amphiphilic hybrid building block, the supramolecular nanovesicles are formed via self-assembly in water. The self-assembling behavior was assessed using fluorescence spectroscopy, DLS and TEM. The present self-acylating and self-assembling β-CD will be a promising material in food, pharmaceutical, and biomedical applications for its nanovesicular architecture, and the self-acylating concept can be applied to the design of novel catalytic cyclic carbohydrates as well as the modification of other carbohydrates using the enhanced reactivity of characteristic hydroxyl groups induced by the presence of cyclic constraint.In the present study, the ladle-type β-CD, mono-2-N, N-dimethylformamide  was obtained from Alfa Aesar, a Johnson Matthey Company. Organic solvents such as methanol and ethanol were of analytical reagent grade, and the water used was triply distilled. Thin-layer chromatography (TLC) was carried out on Merck Kieselgel 60 F254 analytical plates with the specified solvent system. The detection and visualization are followed by spraying 5% sulfuric acid-ethanol and heating for 2 min at 170 °C.α-D-glucose, maltodextrin, α-CD, vinyl pivalate, vinyl benzoate, pyrene, and dimethylsulfoxide (DMSO) were purchased from Sigma Aldrich. β-CD, γ-CD, vinyl laurate, and vinyl stearate were obtained from Tokyo Chemical Industry Co., Ltd. 2O as eluentβ-CD  was dissolved in 1 mL of DMF, and then vinyl laurate  was added. For other self-acylations of β-CD , vinyl pivalate , vinyl butyrate , vinyl benzoate , and vinyl stearate  were used. For self-laurylation with other carbohydrates, α-CD , γ-CD , glucose  and maltodextrin  were used. After stirring at 50 °C for 24 h, the product was isolated by chromatography on silica gel with 8:5:2 or 8:5:1 (v/v) EtOAc-MeOH-H1H NMR : δ 5.92-5.52 H, O(3)H), 5.03 , 4.90-4.71 , 4.63-4.41 H), 3.89 , 3.66-3.22 , 3.18 , 2.44-2.29 , 1.58-1.51 , 0.90 ; 13C NMR : δ 173.06 (C7), 101.98 (C1), 98.49 (C1′), 81.56 (C4), 80.84 (C4′), 78.23 (C2′), 73.12 (C3), 72.46 (C2), 72.09 (C5), 69.38 (C3′), 59.96 (C6), 35.13 (C8), 17.68 (C9), 13.55 (C10); MALDI-TOF MS: 1227.6432 [mono-2-O-butyryl β-cyclodextrin+Na]+.1H NMR : δ 8.10-7.96 , 7.64-7.58 , 7.52-7.46 , 6.03-5.70 H, O(3)H), 5.20 , 4.94-4.77 , 4.68-4.45 H), 4.12  3.70-3.22 , 3.17 ; 13C NMR : δ 165.97 (C7), 133.08 (C11), 131.29 (C8), 129.81, 129.50 , 128.44, 128.15 , 102.00 (C1), 98.20 (C1′), 81.62 (C4), 80.92 (C4′), 78.11 (C2′), 73.03 (C3), 72.54 (C2), 72.07 (C5), 69.67 (C3′), 59.97 (C6); MALDI-TOF MS: 1261.6154 [mono-2-O-benzoyl β-cyclodextrin+Na]+.1H NMR : δ 6.60-6.10 H, O(3)H), 5.01 , 4.88-4.69 , 4.64-4.38 H), 3.87 , 3.63-3.20 , 2.44-2.26 , 1.49 , 1.23 , 0.85 ; 13C NMR : δ 166.23 (C7), 102.23 (C1), 98.53 (C1′), 81.69 (C4), 80.96 (C4′), 78.38 (C2′), 73.12 (C3), 72.46 (C2), 72.09 (C5), 69.37 (C3′), 59.96 (C6), 33.25 (C8), 31.36 (C22), 29.12-28.53 (C10-21), 24.74 (C9), 22.16 (C23), 14.04 (C24); MALDI-TOF MS: 1423.9385 [mono-2-O-stearyl β-cyclodextrin+Na]+.1H NMR : δ 5.84-5.41 H, O(3)H), 4.97 , 4.85-4.67 , 4.65-4.44 H), 4.00 , 3.64-3.54 , 3.43-3.27 , 2.42-2.29 , 1.50 , 1.25 , 0.86 ; 13C NMR : δ 162.99 (C7), 102.06 (C1), 98.80 (C1′), 82.30 (C4), 81.92 (C4′), 79.40 (C2′), 73.34 (C3), 72.58 (C2), 72.15 (C5), 69.62 (C3′), 60.06 (C6), 35.84 (C8), 31.42 (C16), 29.09-28.68 (C10-15), 24.37 (C9), 22.22 (C17), 14.06 (C18); MALDI-TOF MS: 1177.6322 [mono-2-O-lauryl α-cyclodextrin+Na]+.1H NMR : δ 6.82-6.18 H, O(3)H), 5.16 , 5.11-4.77 , 4.75-4.45 H), 3.80 , 3.69-3.16  2.46-2.28 , 1.50 , 1.25 , 0.85 ; 13C NMR : δ 166.29 (C7), 101.56 (C1), 97.67 (C1′), 81.35 (C4), 80.75 (C4′), 78.94 (C2′), 73.22 (C3), 72.84 (C2), 72.18 (C5), 69.85 (C3′), 59.89 (C6), 33.29 (C8), 31.34 (C16), 29.09-28.57 (C10-15), 24.32 (C9), 22.14 (C17), 14.04 (C18); MALDI-TOF MS: 1501.8729 [mono-2-O- lauryl γ-cyclodextrin+Na]+.1H-NMR, 13C-NMR, and DEPT spectra. The NMR analyses were performed in DMSO-d6 at room temperature. ROESY data was recorded with 256/2048 data points using Bruker Avance 500 MHz spectrometer in DMF-d7 at room temperature.A Bruker Avance 600 MHz spectrometer was used to record the TM STR BioSpectrometry, PerSeptive Biosystems, Framingham, MA, USA) using the positive-ion mode. 2, 5-Dihydroxybenzoic acid (DHB) was used as the matrix.The mass spectrum was obtained using a MALDI-TOF mass spectrometer  and vinyl laurate was built using Maestro software . Molecular docking simulations were performed with the vinyl laurate onto the β-CD under the ligand-flexible mode using the Glide docking moduleO-lauryl β-CD or unmodified β-CD was prepared with concentrations of 0.05 to 12.5 mM in water. A small amount of a hydrophobic probe, pyrene solution in acetone, was added into each sample giving a final concentration of 1 μM in solution. Acetone was not removed, and its final content was 0.1%, v/vFor the fluorescence measurement, mono-2-O-lauryl β-CD (1.25 mM) dissolved in ethanol is dropped slowly through a syringe into water (1 mL) and subjected to magnetic stirring. The nanospheres were formed immediately, and the colloidal suspension obtained was submitted to evaporation under reduced pressure to remove ethanol. The resulting colloidal suspension was stored in closed vials at room temperature.Using the nanoprecipitation techniqueO-lauryl β-CD.The resulting colloid was measured using a Wyatt Technology DynaPro Plate Reader for the assessing the nanoarchitecture size distribution of the mono-2-O-lauryl β-CD was loaded onto a Formvar-coated copper grid (200 mesh) and air-dried. For negative staining, 2% uranyl acetate solution was used. Then, the self-assembled nanoarchitecture was examined using energy-filtering TEM .The colloidal suspension by mono-2-How to cite this article: Cho, E. et al. Regioselective self-acylating cyclodextrins in organic solvent. Sci. Rep.6, 23740; doi: 10.1038/srep23740 (2016).


Dillenia suffruticosa, which is locally known as Simpoh air, has been traditionally used to treat cancerous growth. The ethyl acetate extract of D. suffruticosa (EADs) has been shown to induce apoptosis in MCF-7 breast cancer cells in our previous study. The present study aimed to elucidate the molecular mechanisms involved in EADs-induced apoptosis and to identify the major compounds in the extract. EADs was found to promote oxidative stress in MCF-7 cells that led to cell death because the pre-treatment with antioxidants α-tocopherol and ascorbic acid significantly reduced the cytotoxicity of the extract (P<0.05). DCFH-DA assay revealed that treatment with EADs attenuated the generation of intracellular ROS. Apoptosis induced by EADs was not inhibited by the use of caspase-inhibitor Z-VAD-FMK, suggesting that the cell death is caspase-independent. The use of JC-1 dye reflected that EADs caused disruption in the mitochondrial membrane potential. The related molecular pathways involved in EADs-induced apoptosis were determined by GeXP multiplex system and Western blot analysis. EADs is postulated to induce cell cycle arrest that is p53- and p21-dependent based on the upregulated expression of p53 and p21 (P<0.05). The expression of Bax was upregulated with downregulation of Bcl-2 following treatment with EADs. The elevated Bax/Bcl-2 ratio and the depolarization of mitochondrial membrane potential suggest that EADs-induced apoptosis is mitochondria-dependent. The expression of oxidative stress-related AKT, p-AKT, ERK, and p-ERK was downregulated with upregulation of JNK and p-JNK. The data indicate that induction of oxidative-stress related apoptosis by EADs was mediated by inhibition of AKT and ERK, and activation of JNK. The isolation of compounds in EADs was carried out using column chromatography and elucidated using the nuclear resonance magnetic analysis producing a total of six compounds including 3-epimaslinic acid, kaempferol, kaempferide, protocatechuic acid, gallic acid and β-sitosterol-3-O-β-D-glucopyranoside. The cytotoxicity of the isolated compounds was determined using MTT assay. Gallic acid was found to be most cytotoxic against MCF-7 cell line compared to others, with IC50 of 36 ± 1.7 μg/mL (P<0.05). In summary, EADs generated oxidative stress, induced cell cycle arrest and apoptosis in MCF-7 cells by regulating numerous genes and proteins that are involved in the apoptotic signal transduction pathway. Therefore, EADs has the potential to be developed as an anti-cancer agent against breast cancer. There is a shift of attention in chemotherapy from the use of a single drug to multi-drugs for the management of various kinds of cancer, which aims to regulate diverse signaling processes responsible for the survival of tumor via suppression or activation of multiple targets simultaneously . The samFailure to induce apoptosis is a crucial factor that leads to the formation of cancer . Hence, Mitochondria-mediated or intrinsic pathway is controlled by the family of Bcl-2 proteins. The roles of pro-apoptotic and anti-apoptotic Bcl-2 family of proteins are significant in maintaining the permeability of mitochondrial membrane, thus governing the activation or abortion of apoptosis . CaspaseAnother signaling pathway that plays an important role in apoptosis is Jun N-terminal kinase (JNK) pathway. It can promote apoptosis either by the regulation of pro-apoptotic genes through distinct transcription factor transactivation in nuclear signaling or by the manipulation of pro- and anti-apoptotic proteins in mitochondria . In contSOD1, SOD2, catalase, Akt1, NF-κB, p53, and p38 MAPK .1H NMR. . δ (ppm): 6.18 , 6.39 , 8.08 , 6.90 , 6.90 , 8.08 . 13C NMR  δ (ppm): 148.1 (C-2), 137.2 (C-3), 177.4 (C-4), 162.6 (C-5), 99.4 (C-6), 165.7 (C-7), 94.6 (C-8), 160.7 (C-10), 123.8 (C-1’), 130.8 (C-2’), 116.4 (C-3’), 158.3 (C-4’), 116.4 (C-5’), 130.8 (C-6’) .1H NMR. . δ (ppm): 6.91 , 6.91 . 13C-NMR . δ (ppm): 121.0 (C-1), 109.0 (C-2), 145.9 (C-3), 138.3 (C-4), 145.9 (C-5), 109.0 (C-6), 168.0 (-COOH) [Identical with the reference [Gallic acid . White crystal. eference ].1H NMR. . δ (ppm): 3.77 , 3.15 , 5.18 , 1.06 , 0.89 , 0.79 , 0.73 , 1.12 , 0.91 , 0.90 . 13C-NMR . δ (ppm): 41.7 (C-1), 65.2 (C-2), 78.4 (C-3), 38.5 (C-4), 48.1 (C-5), 18.1 (C-6), 32.1 (C-7), 38.6 (C-8), 47.4 (C-9), 38.2 (C-10), 23.6 (C-11), 122.4 (C-12), 144.9 (C-13), 42.2 (C-14), 28.7 (C-15), 26.2(C-16), 43.7 (C-17), 40.2 (C-18), 42.9 (C-19), 29.4 (C-20), 35.0 (C-21), 31.1 (C-22), 28.7 (C-23), 22.4 (C-24), 16.8 (C-25), 17.1 (C-26), 26.9 (C-27), 178.5 (C-28), 32.7 (C-29), 26.3 (C-30) phenanthren-3-ol). White powder. 4 was the most cytotoxic towards both MCF-7 and MDA-MB-231 cells, with IC50 36 ± 1.7 μg/mL and 35 ± 1.3 μg/mL, respectively, compared to others. Compound 5 was less cytotoxic than Compound 4, with IC50 of 83 ± 2.1 μg/mL, towards MDA-MB-231 cells. Compound 1, 2, 3 and 6 were least cytotoxic towards MCF-7 and MDA-MB-231 cells at 72 hours (IC50>100 μg/mL).The cytotoxic effects of the isolated compounds on breast cancer cells were determined using MTT assay. As displayed in There are assortment of signals and cellular metabolic events required to induce apoptosis. One of the cellular events capable of inducing apoptosis is the generation of oxidative stress . Sies 3 defined In this study, EADs was found to be less cytotoxic towards MCF-7 cells after pre-treatment with α-tocopherol and ascorbic acid, suggesting that the extract induces oxidative stress as being hypothesized previously . However2O2, lipid peroxides, singlet O2- and O2 , 631), kic acids , 65. Antic acids . The antic acids . Gallic ic acids . Nonetheic acids . As obsecompound .Kip1 and p21Cip1 decreased the proliferation of MCF-7 via G2/M phase cell cycle arrest [in vivo model [Apart from that, flavonoids and phenolic acids are ubiquitous in plants, and they have shown promising anticancer and chemopreventive activities through various mechanisms such as cell cycle arrest, induction of apoptosis, suppression of angiogenesis, antioxidant activity and overcoming multidrug resistance –72. Kaeme arrest . Anothervo model . Based oIn summary, EADs induced oxidative stress in MCF-7 cells that has led to apoptosis via mitochondria-dependent pathway with the loss of mitochondria membrane permeability. Moreover, EADs induced caspase-independent apoptosis and cell cycle arrest that was facilitated by p21 and p53. Additionally, activation of JNK and inhibition of AKT and ERK pathway play an unequivocal role in EADs-induced apoptosis. This unveils the potential of EADs as an anti-breast cancer agent by modulating multiple pathways involved in the induction of apoptosis.


Neosartorya pseudofischeri, which was isolated from the inner tissue of starfish Acanthaster planci, glycerol-peptone-yeast extract (GlyPY) and glucose-peptone-yeast extract (GluPY) media were used to culture this fungus. When cultured in GlyPY medium, this fungus produced two novel diketopiperazines, neosartins A and B (1 and 2), together with six biogenetically-related known diketopiperazines,1,2,3,4-tetrahydro-2,3-dimethyl-1,4-dioxopyrazinoindole (3), 1,2,3,4-tetrahydro-2-methyl-3-methylene-1,4-dioxopyrazinoindole (4), 1,2,3,4-tetrahydro-2-methyl-1,3,4-trioxopyrazino indole (5), 6-acetylbis(methylthio)gliotoxin (10), bisdethiobis(methylthio)gliotoxin (11), didehydrobisdethiobis(methylthio)gliotoxin (12) and N-methyl-1H-indole-2-carboxamide (6). However, a novel tetracyclic-fused alkaloid, neosartin C (14), a meroterpenoid, pyripyropene A (15), gliotoxin (7) and five known gliotoxin analogues, acetylgliotoxin (8), reduced gliotoxin (9), 6-acetylbis(methylthio)gliotoxin (10), bisdethiobis(methylthio) gliotoxin (11) and bis-N-norgliovictin (13), were obtained when grown in glucose-containing medium (GluPY medium). This is the first report of compounds 3, 4, 6, 9, 10 and 12 as naturally occurring. Their structures were determined mainly by MS, 1D and 2D NMR data. The possible biosynthetic pathways of gliotoxin-related analogues and neosartin C were proposed. The antibacterial activity of compounds 2–14 and the cytotoxic activity of compounds 4, 5 and 7–13 were evaluated. Their structure-activity relationships are also preliminarily discussed.The production of fungal metabolites can be remarkably influenced by various cultivation parameters. To explore the biosynthetic potentials of the marine fungus,  Chondrostereum sp., which was isolated from soft coral Sarcophyton tortuosum, grows in different culture media and can produce various novel bioactive hirsutane-type sesquiterpenoids indole (3), 1,2,3,4-tetrahydro-2-methyl-3-methylene-1,4-dioxopyrazinoindole (4), 1,2,3,4-tetrahydro-2-methyl-1,3,4-trioxopyrazinoindole (5), 6-acetylbis(methylthio) gliotoxin (10), bisdethiobis (methylthio)gliotoxin (11), didehydrobisdethiobis(methy1thio)gliotoxin (12) and N-methyl-1H-indole-2-carboxamide (6). Isolation of the extract of GluPY medium gave a new alkaloid, neosartin C (14), known compounds, α-pyrone meroterpenoid pyripyropene A (15), gliotoxin (7) and gliotoxin analogues, acetylgliotoxin (8), reduced gliotoxin (9), 6-acetylbis(methylthio)gliotoxin (10), bisdethiobis(methylthio)gliotoxin (11) and bis-N-norgliovictin (13)  . In this1 was obtained as a yellowish solid. The molecular formula was determined as C14H14N2O3 from the HREIMS peak at m/z 258.0998 [M]+  (−1) and a benzene ring . UV maxima at 211, 243 and 295 nm displayed the conjugated system containing a benzene ring. The 13C NMR and DEPT spectra displayed three methyls, five methines and six quaternary carbons ; δC 125.8, δH 7.43 ; δC 128.2, δH 7.55 ; and δC 122.7, δH 7.72 ) established the partial structure -CH-CH-CH-CH- based on the 1H-1H COSY correlations of H-6/H-7, H-7/H-8 and H-8/H-9  and a quaternary carbon C-10a at δC 127.8 constructed an additional trisubstituted double bond. The HMBC correlations of H-9/C-10, H-10/C-9a, H-10/C-10a and H-10/C-1 revealed the trisubstituted double bond connected to a benzene ring and an amide carbonyl. Methyl group C-11 at δH 3.13 (δC 26.6) showed HMBC correlations with amide carbonyl C-1 and quaternary carbon C-3, so it was connected to amide nitrogen N-2. Methyl group C-12 at δH 1.81 (δC 25.3) was connected to C-3 based on the HMBC correlation with the amide carbon C-4 and quaternary carbon C-3. The methoxyl group was also located at C-3, because it showed an HMBC correlation with C-3  , implyin carbons . Two qua H-8/H-9 . Furtherwith C-3 . Finallywith C-3 . Therefo2 was isolated as a yellowish solid. The molecular formula was established as C14H14N2O4 based on the HREIMS peak at m/z 274.0947 [M]+ and 13C NMR data  in 1 was replaced by an oxymethylene  in 2. Therefore, the structure of 2 was elucidated as 1,2,3,4-tetrahydro-3-methoxyl-3-hydroxylmethyl-2-methyl-1,4-dioxopyrazinoindole, commonly named neosartin B.Compound NMR data . The 13Cmpound 1 . By comp1 and 2 did not show optical activity in circular dichroism (CD) spectra; thus, they existed as a racemic mixture of 3R and 3S.Compounds 3 was isolated as a white solid. The molecular formula was established as C13H12N2O2 based on the LREIMS molecular ion at m/z 228 and the NMR data . Its structure was elucidated as 1,2,3,4-tetrahydro-2-methyl-3-methylene-1,4-dioxopyrazinoindole by analysis of its spectral data (4 was previously obtained as the conversion product of gliotoxin (7) by passing through a column of alkaline alumina at 20 °C indole by careful analysis of the MS and NMR data  and NMR data   . It is u7 was identified as gliotoxin by comparing the data with the literature values + , implying 16 degrees of unsaturation  with the coupling constants being about 7.6, suggesting at least two phenyl groups in the molecule. By analysis of the HMQC spectrum, the 1H and 13C NMR data of each carbon were definitely assigned. The 1H-1H COSY correlations of H-15/H-16, H-11/H-12, H-12/H-13 and H-12/H-14 deduced the presence of two partial structures, -CH2CH- and -CHCH(CH3)2, respectively (C 171.4 (C-10), 163.6 (C-17) and 161.7 (C-19) are amide carbonyl groups. HMBC correlations of H-11/C-10, H-16/C-17, H-21/C-19 revealed that three amide carbonyl groups were connected to -CHCH(CH3)2, -CH2CH- and a disubstituted benzene ring, respectively. The hydroxyl group at δH 4.09 (s) was attached to C-3 (δC 74.8), and C-3 was connected to the methine C-2 (δC 84.8) and C-15 (δC 36.5) based on the HMBC correlations of H-2/C-3 and H-15/C-3. The planar structure of 14 was finally established by the HMBC correlations of H-2/C-10, H-5/C-3, H-15/C-3, H-16/C-19, H-16/C-27 and H-27/C-25 , previously isolated from Aspergillus fumigatus FO-1289, showed very potent inhibition of cholesterol acyltransferase (ACAT).The NMR data of compound ferences . By compropene A . Pyripyrl-phe and l-ser to form dipeptidyl l-phe-l-ser, and under the action of the enzymes, the latter is converted to the corresponding diketopiperazines + calcd. for C14H14O3N2: 258.0999; found 258.0998.Neosartin A (2): Yellowish solid. UV (MeOH) λmax (ε) 212 , 243 , 295  nm. IR: υmax 3301, 3126, 2951, 1706, 1635, 1591, 1577, 1435, 1392, 1360, 1348, 1224, 1151, 1116, 1081, 981, 746, 733 cm−1. 1H and 13C NMR: m/z 274, 260, 243, 229, 215, 202, 188, 172, 156, 143, 130, 115, 103, 89, 83, 72, 57. HREIMS: m/z [M]+ calcd. for C14H14O4N2: 274.0948; found 274.0947.Neosartin B (3): White solid. 1H and 13C NMR: m/z 228, 213, 200, 185, 172, 143, 131, 115, 100, 89, 71, 62, 56.1,2,3,4-Tetrahydro-2,3-dimethyl-1,4-dioxopyrazinoindole (4): White solid. 1H and 13C NMR: m/z 226, 199, 185, 169, 157, 143, 129, 115, 99, 88, 75, 62, 55.1,2,3,4-Tetrahydro-2-methyl-3-methylene-1,4-dioxopyrazinoindole (5): White solid. 1H and 13C NMR: m/z 228, 200, 159, 143, 131, 115, 100, 88, 71, 62, 50.1,2,3,4-Tetrahydro-2-methyl-1,3,4-trioxopyrazinoindole (N-methyl-1H-indole-2-carboxamide (6): White solid. 1H and 13C NMR: m/z 174, 156, 143, 115, 89, 77, 63, 58.7): Pale yellowish solid. 1H NMR : 5.98 , 5.92 , 5.84 , 5.75 , 4.80 , 4.43 , 4.23 , 4.06 , 3.73 , 3.19 , 2.94 ; 13C : 166.0 (C-1), 165.1 (C-4), 130.8 (C-9a), 129.8 (C-7), 123.3 (C-8), 120.1 (C-9), 77.3 (C-3), 75.8 (C-10a), 73.1 (C-6), 69.7 (C-5a), 60.4 (C-12), 36.5 (C-10), 27.5 (C-11). LREIMS: m/z 326, 308, 277, 262, 244, 233, 217, 199, 188, 160, 144, 132, 107, 89, 77, 73, 64, 55, 42.Gliotoxin (8): Yellowish solid. 1H and 13C NMR: Acetylgliotoxin (9): Yellowish solid. 1H and 13C NMR: Reduced gliotoxin (10): Yellowish oil. 1H NMR : 6.16 , 5.90 , 5.75 , 5.01 , 4.23 , 3.76 , 3.60 , 3.09 , 3.04 , 2.90 , 2.29 , 2.12 , 2.05 ; 13C : 170.7 (COCH3-14), 166.4 (C-1), 164.4 (C-4), 133.8 (C-9a), 127.8 (C-7), 123.3 (C-8), 119.8 (C-9), 72.7 (C-3), 72.3 (C-10a), 75.2 (C-6), 65.4 (C-5a), 63.6 (C-12), 40.1 (C-10), 28.7 (C-11), 21.3 (CO3CH-14), 15.0 (C-13), 13.0 (CH3-15).6-Acetylbis(methylthio)gliotoxin (11): Yellowish oil. 1H NMR : 5.85 , 5.65 , 4.86 , 4.30 , 3.82 , 3.09 , 3.01 , 2.92 , 2.22 , 2.19 ; 13C : 166.7 (C-1), 165.9 (C-4), 131.7 (C-9a), 129.5 (C-7), 123.2 (C-8), 119.8 (C-9), 74.2 (C-3), 72.1 (C-10a), 71.4 (C-6), 69.4 (C-5a), 63.4 (C-12), 38.7 (C-10), 28.5 (CH3-11), 15.0 (CH3-13), 13.0 (CH3-15) .Bisdethiobis(methylthio)gliotoxin (12): Yellowish oil. 1H and 13C NMR: Didehydrobisdethiobis(methylthio)gliotoxin (N-norgliovictin (13): White solid. 1H and 13C NMR: Bis-14): Yellowish solid. c =0.0822, MeOH). UV (MeOH) λmax (ε) 314 (2910), 301 (3653), 226 , 212  nm. IR: υmax 3348, 3068, 2970, 2934, 2875, 1724, 1667, 1608, 1476, 1465, 1387, 1326, 1292, 1267, 1248, 1172, 1112, 1076, 1007, 976, 915, 891, 844, 764, 742, 700, 593, 560 cm−1. 1H and 13C NMR: m/z 430, 412, 341, 329, 314, 298, 286, 266, 238, 224, 184, 147, 130, 102, 83, 76, 55. HREIMS: m/z [M]+ calcd. for C24H22O4N4: 430.1636; found 430.1635.Neosartin C (15): Yellowish solid. UV (MeOH) λmax (ε) 320 , 231 , 209  nm. IR: υmax 3409, 2975, 2947, 2883, 1723, 1644, 1580, 1481, 1436, 1394, 1370, 1234, 1159, 1109, 1090, 1074, 1040, 1026, 1009, 984, 960, 925, 873, 809, 763, 704, 687, 650, 603, 583 cm−1. 1H and 13C NMR: m/z 583, 565, 550, 523, 505, 494, 463, 445, 430, 403, 385, 358, 334, 304, 283, 240, 218, 202, 190, 171, 157, 148, 133, 119, 106, 95, 79, 69, 55.Pyripyropene A (6 CFU/mL) in the first well of flat-bottomed 96-well tissue culture plates. The solution was then double diluted. The bacterial culture solution containing the appropriate compound (50 µL) was discarded from the last well in order to ensure a 100-µL volume of bacterial culture in every well. A set of tubes containing only inoculated broth and solvent were kept as controls. The plate was incubated at 37 °C overnight in an electro-heating standing-temperature cultivator before the measurement of the absorbance value. The optical density values at 600 nm were measured using a multifunction microplate reader . Vancomycin and ampicillin sodium were used as positive controls.The MIC values were determined using a broth dilution method (Mueller–Hinton broth) based on the National Committee for Clinical Laboratory Standards (NCCLS) standard. The starting concentrations of the tested compounds were 256 µg/mL (from 256 to 0.25). The solution of compound in DMSO (10 µL) was added to 90 µL of bacterial culture (1 × 10H-tetrazolium)) assay. 5-Fluorouracil was used as the positive control, and DMSO was used as the negative control. 293, HCT116 and RKO cells were suspended in fresh RPMI-1640 medium containing 10% fetal bovine serum and 100 μg/mL penicillin and streptomycin at a cell density of 1 × 105 cells/mL and seeded into 96-well plates each 100 μL/well. The suspension cells, 293, HCT116 and RKO cells, were incubated at 37 °C for 12 h. Then, compounds were added to the cultures at different concentrations; then, the cells were cultured at 37 °C for 72 h. Twenty microliters of MTS/PMS (phenazine methosulfate) were added into each well, incubated at 37 °C for 4 h in a humidified, 5% CO2 atmosphere. The absorbance at 490 nm was recorded using a Thermo Scientific Varioskan Flash Multimode Reader . The data were analyzed with the GraphPad Prism 6 software package [Compounds in DMSO at 50 mM were used in the MTS -5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 package .Neosartorya is a rich source of nitrogen-containing natural products. Interestingly, the dominant metabolites from the GluPY medium were the diketopiperazines with disulfide bonds; however, the main compounds from the GlyPY medium were the diketopiperazines without a disulfide bond. The biosynthetic pathways of the unique alkaloids are complex and diverse. Hopefully, further investigation on the secondary metabolites of Neosartorya pseudofischeri in varied culture conditions supplied with amino acid precursors may find more novel alkaloids and improve their production. Most of the metabolites showed significant antibacterial and cytotoxic activities. Based on the structure-activity relationship analysis, the disulfide bridge, the α-methylene ketone group, the hydroxyl group at C-6 and the thiol groups were considered as the pharmacophores.Alternating the cultivation parameters systematically, such as the components of the media, to enhance the diversity of secondary metabolites produced by the fungus is a well-known, simple and efficient technique. Using this strategy, we isolated and identified fifteen compounds, including twelve diketopiperazine derivatives, a meroterpenoid, an alkaloid with a unique tetracyclic-fused skeleton and an imidazole analogue from the two cultures. The discovery of these compounds provided further evidence that the genus of


Linum scabrellum Planchon (Linaceae) were analyzed. Using NMR spectra and mass spectrometry analyses of the extract we identified eight main constituents: oleic acid (1), octadecenoic acid (2), stigmasterol (3), α-amyrin (4), pinoresinol (5), 6 methoxypodophyllotoxin (6), coniferin (7), and 6-methoxypodophyllotoxin-7-O-β-D-glucopyranoside (8). By using the sulforhodamine B assay, an important cytotoxic activity against four human cancer cell lines, HF6 colon (IC50 = 0.57 μg/mL), MCF7 breast (IC50 = 0.56 μg/mL), PC3 prostate (IC50 = 1.60 μg/mL), and SiHa cervical (IC50 = 1.54 μg/mL), as well as toward the normal fibroblasts line HFS-30 IC50 = 1.02 μg/mL was demonstrated. Compound 6 (6-methoxypodophyllotoxin) was responsible for the cytotoxic activity exhibiting an IC50 value range of 0.0632 to 2.7433 µg/mL against the tested cell lines. Cell cycle studies with compound 6 exhibited a cell arrest in G2/M of the prostate PC3 cancer cell line. Microtubule disruption studies demonstrated that compound 6 inhibited the polymerization of tubulin through its binding to the colchicine site (binding constant Kb = 7.6 × 106 M−1). A dose-response apoptotic effect was also observed. This work constitutes the first investigation reporting the chemical composition of L. scabrellum and the first study determining the mechanism of action of compound 6.The cytotoxic activity and the chemical composition of the dichloromethane/methanol root extract of  Linum of the Linaceae family includes more than 200 globally distributed species. The phytochemical profiles that have been reported for the species of Linum suggest the presence of some lignan-type secondary metabolites 24D  −82.3° , and had the formula C20H22O6 derived from its positive FABMS [M+H]+ ion at m/z 359. IR : 3210, 2870, 1600, 1465 1050. UV λmax (nm): 232, and 280. 1H NMR  δH: 6.89 , 6.88 , 6.81 , 4.73 , 3.09 , 4.24 , 3.87 , 3.90 . 13C NMR  δC: 132.90 , 108.57 , 146.68 , 145.22 , 114.24 , 118.96 , 85.86 , 54.16 , 71.66 . On the basis of the spectral data, compound 5 was identified as (-)-pinoresinol + ion peak at m/z 445 with molecular formula C23H24O9. IR : 3220, 1768, 1600. UV λmax (nm): 272, and 282. 1H NMR  δH: 6.30 , 5.03 , 2.88 , 4.06 , 4.64 , 6.44 , 4.53 , 2.75 , 3.78 , 3.81 , 3.78 , 5.95 . 13C NMR  δC: 124.93 (C-1), 137.04 (C-2), 104.36 (C-3), 149.45 (C-4), 132.84 (C-5), 141.57 (C-6), 70.52 (C-7), 39.01 (C-8), 71.92 (C-9), 134.71 (C-1′), 108.09 (C-2′), 152.61 (C-3′), 134.93 (C-4′), 152.61 C-5′), 108.09 (C-6′), 45.11 (C-7′), 44.53 (C-8′), 174.44 (C-9′), 56.18 (C3′-OMe), 60.77 (C4′-OMe), 56.18 (C5′-OMe), 59.93 (C6-OMe), 101.37 (-O-CH2-O-). On the basis of the spectral data, compound 6 was identified as 6-methoxypodophyllotoxin . Positive FABMS m/z 342 [M + H]+, with molecular formula C16H22O8. IR : 3020, 1060, 940. UV λmax (nm): 256 and 292. [α]24D  −66.3 . 1H NMR  δH: 7.07 , 6.89 , 7.09 , 6.52 , 6.29 , 4.19 , 4.87 , 4.23 , 4.22 , 4.21 , 4.05 , 3.54 , 3.70 , 3.79 . 13C NMR  δC: 150.01 (C-1), 146.54 (C-2), 110.24 (C-3), 132.36 (C-4), 117.11 (C-5), 119.23 (C-6), 128.88 (C-1′), 128.87 (C-2′), 62.36 (C-3′), 55.53 (C2-OMe), 101.69 (C-1′′), 73.92 (C-2′′), 77.07 (C-3′′), 70.69 (C-4′′), 76.93 (C-5′′), 61.82 (C-6′′). On the basis of spectral data, compound 7 was identified as phenylpropane glycoside coniferin (7) . The positive FABMS spectrum showed the [M + H]+ ion peak at m/z 607, with molecular formula C29H34O14. IR : 3020, 1746, 1620, 1050, 920. UV λmax (nm): 276 and 284. 1H NMR  δH: 6.28 , 5.48 , 2.90 , 4.23 , 4.71 , 6.31 , 4.51 , 3.25 , 3.72 , 3.99 , 3.68 , 6.02 , 4.54 , 3.28 , 3.35 , 3.41 , 3.43 , 3.76 , 3.84 . 13C NMR  δC: 124.26 (C-1), 136.28 (C-2), 105.74 (C-3), 149.68 (C-4), 135.89 (C-5), 140.26 (C-6), 72.78 (C-7), 39.87 (C-8), 72.39 (C-9), 136.73 (C-1′), 108.58 (C-2′), 152.89 (C-3′), 136.54 (C-4′), 152.89 (C-5′), 108.58 (C-6′), 45.86 (C-7′), 46.23 (C-8′), 176.67 (C-9′), 56.24 , 60.76 (C4′-OMe), 60.76 (C6-OMe), 102.34 (-O-CH2-O-), 100.46 (C-1′′), 74.87 (C-2′′), 77.85 (C-3′′), 71.67 (C-4′′), 78.13 (C-5′′), 63.08 (C-6′′). On the basis of spectral data, compound 8 was identified as 6-methoxypodophyllotoxin 7-O-β-D-glucopyranoside [From the fraction F-6, compoundhttp://dx.doi.org/10.1155/2015/298463).The obtained data were compared with the literature –43 , while compound 6 (6MPTOX) showed an important cytotoxic activity with IC50 values ranging from 0.0632 to 2.7433 μg/mL against the cancer tested cell lines  and PTOX (0.0024 μM) were used as negative and positive controls, respectively. μM induced a similar effect on G2/M cell arrest with 39.8% and 41.1% of G2/M cell population, respectively. These results suggest that 6MPTOX efficiently arrest cells at G2/M phase.The effect of 6MPTOX in the cell cycle progression of PC3 cells was determined at four concentrations: 0.011 Histone H3 phosphorylated at serine 10 has long been used to identify mitotic cells nuclei in culture cell lines . The proα/β tubulin binding.Our results showed that 6MPTOX induced a mitotic cell arrest by the observed high mitotic index and the G2/M cell arrest, suggesting a similar effect as that displayed by PTOX or Taxol. In order to determine the mechanism of action of 6MPTOX, we analyzed the effect of microtubule polymerization by 6MPTOX through disruption of α-tubulin was observed in PC3 cells by confocal microscopy. Microtubules are in dynamic equilibrium with tubulin dimers as tubulin is polymerized into microtubules and depolymerized as free tubulin. This dynamic equilibrium is targeted by microtubule disrupting agents , which iThese results suggest that 6MPTOX as well as PTOX act as microtubule-destabilizing agents, while cells treated with DMSO (0.02%) demonstrated a normal and intact tubulin organization .α/β-tubulin as does PTOX, 6MPTOX was assayed for its ability to displace MTC from its binding at the colchicine site of α/β-tubulin [Kb = 7.26 × 106 M−1 by displacement of MTC in the colchicine binding site. MTC has Kb = 4.7 × 105 M−1 [To corroborate whether 6MPTOX binds to -tubulin , 55.  Fi 105 M−1 , which iμM did not show any effect on cell death. When the concentration was increased to 0.002 μM some apoptotic bodies were observed , and necrosis . Apoptosobserved ; with 0.necrosis . Three cnecrosis ; H2O2 foc bodies , and a nc bodies . The eff L. scabrellum was established. Eight compounds including lignans, terpenes, and fatty acids were isolated and identified. This study demonstrated the cytotoxic activity of 6MPTOX toward four selected human carcinomas and against a normal fibroblast cell line. Biochemical and biological experiments showed that 6MPTOX specifically arrested PC3 cells in their G2/M phase as well as a higher mitotic index. Moreover, at its lowest concentration it induces apoptosis as shown by an increase in its subG1 cell population and in stained apoptotic cells. Necrotic cell death was observed at higher concentrations. Data presented in this study show that 6MPTOX binds at the colchicine binding site of tubulin with a Kb = 7.26 × 106 M−1, causing disruption of tubulin polymerization. These results showed that 6MPTOX inhibit tubulin polymerization and arrest cells in G2/M as a mechanism of cytotoxic activity in PC3 cells. This is the first report describing the mechanism of action of 6MPTOX.For the first time, the metabolic content of the Mexican speciesSupplementary Material provides all the spectroscopic data of isolated compounds (NMR and MS spectra).