Compound 8 is then coupled with aromatic and heteroaromatic diazonium salts to afford the corresponding aryl- and heteroaryl-4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitriles 12a,b and 13a–c. Structural assignments to the dyes were made using NMR spectroscopic methods. A high temperature dyeing method was employed to apply these dyes to polyester fabrics. Most of the dyed fabrics tested displayed very good light fastness levels and good wash fastness. Finally, the biological activity of the prepared dyes against Gram positive bacteria and Gram negative bacteria were evaluated.A series of novel azo-disperse dyes containing alkylhydrazonopyridinone structures were synthesized. 4-Methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitrile ( Arylhydrazonopyridinones are now replacing arylazopyrazolones in the classical dye industry. Moreover, reasonable solubility of these derivatives in lipophilic solvents gives these dyes high potential for utilization in dye diffusion thermal transfer printing . Pyridin8) as a good precursor to novel aryl- and heteroaryl-4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitriles and their application as disperse dyes for the dyeing of polyester fabrics. The antibacterial activities of these dyes were also studied.Moreover, the pyridinone nucleus has been proven to constitute the active part of several biologically active compounds ,10. Keep8 and convert it to the corresponding aryl- or heteroaryl-4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbo-nitriles 12a,b and 13a–c, which were anticipated to be excellent new disperse dyes for the dyeing of polyester fabrics employing a high temperature dyeing method. We introduce a one-pot three component condensation for the synthesis of alkylhydrazonopyridinone 8 under reflux and solvent-free conditions. Thus, the reaction of ethyl acetoacetate, ethyl cyanoacetate, and propyl amineunder reflux without solvent for 6 hours afforded compound 8 in good yield. The product was identified by IR and 1H-NMR spectroscopic data. Compound 8 may be exist in another tautmeric form 9 and in solution there is a very fast equilibrium between them -4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbo-nitrile (12a). Red crystals from toluene (88%), m.p. 252–255 °C, IR (KBr): 3407 (OH), 2222 (CN) and 1657 (CO) cm−1; 1H-NMR : δ = 0.85 , 1.51 , 2.45 , 3.78 , 6.86 , 7.55 , 10.02  14.84 ; 13C-NMR (DMSO-d6): δ = 160.5, 160.0 (2CO), 158.7, 157.5, 133.0, 121.5, 119.2, 116.3, 115.4, 98.2, 29.3 (CH2), 20.4 (CH2). 16.1 (CH3), 11.2 (CH3). MS (EI): m/z (%) = 312 . 295 (8), 270 (52), 121 (24), 108 (43), 93 (24). HRMS: m/z (EI) for C16H16N4O3; calcd. 312.1216; found: 312.1216.4-Methyl-2,6-dioxo-1-propyl-5-(p-tolylhydrazono)-1,2,5,6-tetrahydropyridine-3-carbonitrile (12b). Yellow crystals from ethanol (86%), m.p. 215–218 °C, IR (KBr): 3432 (NH), 2223 (CN) and 1679 (CO) cm−1; 1H-NMR : δ = 0.88 , 1.52 , 2.32 , 2.50 , 3.78 , 7.27 , 7.58 , 14.62 ; MS (EI): m/z (%) = 310 . 293 (9), 268 (26), 177 (12), 106 (32), 91 (56). HRMS: m/z (EI) for C17H18N4O2; calcd. 310.1424; found: 310.1424.4-Methyl-2,6-dioxo-1-propyl-5-(thiazol-2-yl-hydrazono)-1,2,5,6-tetrahydropyridine-3-carbonitrile (13a). Yellow crystals from toluene (65%), m.p. 210 °C, IR (KBr): 3405 (NH), 2224 (CN) and 1684 (CO) cm−1; 1H-NMR : δ = 0.87 , 1.50 , 2.47 , 3.79 , 7.48 , 7.67 , MS (EI): m/z(%) = 303 . 288 (7), 218 (12), 190 (90), 149 (17), 86 (27). HRMS: m/z(EI) for C13H13N5O2S; calcd. 303.0782; found: 303.0782.4-Methyl-5-[(5-methylisoxazol-3-yl)-hydrazono]-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitrile (13b). Yellowish-green crystals from ethanol (80%), m.p. 266–268 °C, IR (KBr): 3429 (NH), 2225 (CN) and 1688 (CO) cm−1; 1H-NMR : δ = 0.88 , 1.51 , 2.44 , 2.45 , 3.79 , 6.67 , 14.24, 14.62 ; MS (EI): m/z (%) = 301 . 286 (8), 216 (52), 190 (72), 149 (42), 68 (42). HRMS: m/z (EI) for C14H15N5O3; calcd. 301.1163; found: 301.1163.4-Methyl-5-[. Yellow crystals from ethanol (82%), m.p. 271–272 °C, IR (KBr): 3431 (NH), 2232 (CN) and 1684 (CO) cm−1; 1H-NMR : δ = 0.88 , 1.51 , 2.47 , 2.50 , 3.78 , 14.20 , 14.60 , MS (EI): m/z (%) = 333 . 291 (12), 256 (12), 205 (32), 149 (12), 97 (36). HRMS: m/z (EI) for C13H15N7O2S; calcd. 333.1002; found: 333.1002.Scoured and bleached polyester 100% was used. The fabric was treated before dyeing with a solution containing non-ionic detergent (5 g/L) and sodium carbonate (2 g/L) in a ratio of 50:1 at 60 °C for 30 min, then thoroughly washed with water and air dried at room temperature.The dye baths were prepared from the dye (1.0% weight of fibre) to a final liquor of 40:1, w/w. The pH value of the bath was adjusted to 4–5 with acetic acid (10%). The polyester fabrics, previously wetted, were placed into the liquor at 25 °C–30 °C. The temperature was raised to 130 °C at the rate of 2 °C/min, and dyeing continued for 60 min. After cooling, the dyed fabrics were then washed and dried .The colorimetric parameters  of the dThe dyed fabrics were tested, employing ISO standard methods . Wash faStaphylococcus auerus and Bacillus subtilus (Gram positive bacteria), Escherichia coli, Klebsiellae pneumoniae and Pseudomonas aeruginosa (Gram negative bacteria) were employed in the test. An aliquot of each bacterial strain (0.1 mL) was inoculated and spread on nutrient agar (NA). The inoculated plates were supplied with 100 µL of each of the tested dyes with concentrations of 0.25, 0.50, 1.00, 2.00 and 3.00 mg·mL−1. The dyes were placed in 4 mm wells produced by sterile cork borer. The NA plates were incubated at 37 °C for 24 h. The zones of inhibition around the wells were determined and the averages based on three replicas were recorded.The antimicrobial activities of four disperse dyes were tested against five different microbial cultures using the agar-well diffusion technique . Pure cu8) was synthesized by one-pot synthesis using ethyl cyanoacetate, propylamine, and ethyl acetoacetate. Compound 8 is then coupled with aromatic and heteroaromatic diazonium salts to afford the corresponding aryl- and heteroaryl-4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitriles. The dyes produced in this manner were then applied to polyester fabrics by using a HT dyeing method. The dyed fabrics, which displayed yellow to light green hues on polyester fabrics, have very good fastness level to light and good fastness to wash. Finally, the biological activity of the synthesized dyes against Gram positive bacteria, Gram negative bacteria was discussed. In summary, 4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitrile (


The structures of the isolated new compounds were elucidated based on extensive spectroscopic analyses. The cytotoxic activities of the 19 sesquiterpenes on three cell lines, human acute promyelocytic leukemia HL-60, human leukemic K562, and human colon cancer HCT-116 cells, were evaluated in vitro. Compound 4 exhibited potent cytotoxic activity against HL-60, K562, and HCT116 cell lines with IC50 values of 3.61 μΜ, 17.13 μΜ and 10.15 μΜ, respectively, and the other compounds displayed moderate activity.Two new β-dihydroagarofuran-type sesquiterpenes ( Celastrus orbiculatus Thunb is a traditional herbal medicine used as a treatment for early tumors, and as a sedative and hypnotic , CinO  .13C-NMR : 73.1 (C-1), 22.5 (C-2), 26.5 (C-3), 33.5 (C-4), 89.7 (C-5), 78.2 (C-6), 48.8 (C-7), 34.7 (C-8), 69.8 (C-9), 53.0 (C-10), 82.5 (C-11), 16.6 (C-12), 65.5 (C-13), 25.9 (C-14), 30.5 (C-15). AcO , CinO ; HR-ESIMS m/z 565.2461 [M + Na]+ .1β,13-Diacetoxy-9α-cinnamoyloxy-β-dihydroagarofuran (2). Colorless crystals; max 2930, 1748, 1721, 1625, 1450, 1364, 1236, 1079, 1024, 721 cm−1; UV (CH2Cl2) λmax 232, 270.1H-NMR : 5.52 , 5.20 , 4.52 , 4.47 , 1.08 , 1.39 , 1.19 , 1.45–2.26 , AcO , CinO . 13C-NMR : 73.4 (C-1), 22.8 (C-2), 26.8 (C-3), 40.1 (C-4), 87.0 (C-5), 36.7 (C-6), 43.6 (C-7), 33.9 (C-8), 69.9 (C-9), 50.6 (C-10), 82.0 (C-11), 17.4 (C-12), 65.0 (C-13), 24.4 (C-14), 30.3 (C-15). AcO , CinO . HR-ESIMS m/z 507.2936 [M + Na]+ .3)  was used as the positive control. All chemical regents used in this research were obtained from Laibo Chemical Company, Ltd., Shenyang, China.1β,6α-diacetoxy-9α-benzoyloxy-β-dihydroagarofuran (3) , 1β-acet3) [4) , 1β-acet [4) [5) , 1β,6α,8 [5) [6) , 1β,8α-d [6) [7) , 1β,2β-d [7) [8) , 1β,6α-d [8) [9) , 1β,6α,1[9) [10) , 1β-acet10) [11) , 1β,2β,111) [12) , 1β,2β,612) [13) , 1β,2β,613) [14) , 1β,2β,614) [15) , 1β-acet15) [16) . 1β,6α-d16) [17) , 1β-cinn17) [18) , 1β,8β-d18) [19)  were iso2 and 95% air. Stock solutions of the compounds for anti-proliferative assay were prepared in DMSO at an initial concentration of 50 or 100 mM.HL-60, K526, and HCT116 cell lines were used in this research. The three cell lines were purchased from American Type Culture Collection . HL-60, K526 and HCT116 were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and incubated at 37 °C in an atmosphere of 5% CO1–19 on cell proliferation was assessed by 3--2,5-diphenyl-tetrazolium bromide (MTT) assay [The effect of Compounds T) assay . Briefly50 values of the compounds inhibiting cell viability over 50% at a concentration of 100 μM were calculated. All cytotoxic activity data were analyzed by SPSS (20.0) and expressed as mean ± S.D.The IC1–2) and seventeen known compounds isolated from the fruit of C. orbiculatus. The structures of the two new compounds were characterized by an extensive analysis of 1D and 2D NMR and HRESIMS data, which is reported for the first time. In addition, Compound 4 exhibited a stronger toxic effect than 5-FU, most of these compounds exhibited moderate effects against HL-60, K562, and HCT116 cells as shown by an MTT assay. This showed that β-dihydroagarofuran sesquiterpenes are an important series of candidate compounds for anti-cancer drug research.In summary, the chemical constituents of the anti-proliferative fraction were investigated and 19 β-dihydroagarofuran sesquiterpenes were obtained, including two new compounds (


Mesua beccariana afforded a novel cyclodione coumarin, beccamarin, together with two known xanthones, mesuarianone, mesuasinone, two anthraquinones, 4-methoxy-1,3,5-trihydroxy-anthraquinone and 2,5-dihydroxy-1,3,4-trimethoxyanthraquinone and one coumarin, mammea A/AB. The structures were elucidated by 1D and 2D NMR and MS techniques.Our ongoing investigations on the stem bark of  Mesua is a genus of flowering plants in the family of Clusiaceae, native to tropical southern Asia. Common names for Mesua include ironwood and rose chestnut. They are evergreen shrubs or small trees growing up to 13 m tall, with leaves arranged in opposite pairs. The blossoms are white and give off a nice fragrance. Although Mesua species have been investigated before, reports on phytochemical constituents of Mesua species are few − , which corresponds to a molecular formula of C24H22O6. The UV spectrum supported the coumarin skeleton due to its maxima absorptions at 209 (5.23), 229 (5.27), 281 (5.36) and 348 (5.45). The FTIR spectrum gave absorptions of chelated hydroxyl , carbonyl , saturated C-H stretch (2971 cm−1) and aromatic ring , which reflected similarity to typical IR bands for coumarins.Beccamarin 1,  was isol1H-NMR spectrum indicated a monosubstituted phenyl group, deduced by the presence of three sets of triplets at δ 7.38 , 7.37  and 7.30 . In the 13C-NMR spectrum, the signals at δ 127.2 , 127.7  and 128.2 (C-4”) belong to the phenyl group. Meanwhile, δ 7.30 (H-2” & H-6”) was correlated to δ 156.8 (C-4) via a 3J correlation and δ 7.37 (H-3” & H-5”) was correlated to δ 139.5 (C-1”) (3J correlation). All these observations place the phenyl group at C-4 of the main coumarin skeleton.The 1H, 13C and HMBC spectra revealed a low field hydroxyl group . This OH group correlated to δ 164.8 (C-8) via a 2J correlation in the HMBC spectrum. A 3J correlation to δ 102.6 (C-7) was also observed. Hence, the hydroxyl group was assigned to C-8. The 1H, HMQC and HMBC experiments gave one sharp singlet proton signal at δ 5.93 and this has direct connectivity to δ 112.1 (C-3) and long range couplings to δ 102.6 (C-4a), 139.5 (C-1”) and 159.7 (C-2). Thus, this singlet proton was positioned at C-3.The 1H and COSY spectra suggested the occurrence of a methylbutanol moiety with a set of correlated signals which are δ 1.33 , 1.46 , 1.58  and 4.61  with δ 1.46  coupling to δ 4.61 . The HMQC and HMBC spectrum indicated that δ 4.61 (H-2’) has connectivities with δ 21.0 (C-5’), 25.8 (C-4’), 43.6 (C-1’) and 113.7 (C-5); δ 1.46 (H-3’) correlates with δ 43.6 (C-1’) and 91.9 (C-2’); δ 1.58 (H-4’) has connectivities with δ 21.0 (C-5’), 43.6 (C-1’), 91.9 (C-2’) and 113.7 (C-5); δ 1.33 (H-5’) correlates with δ 25.8 (C-4’), 43.6 (C-1’), 91.9 (C-2’) and 113.7 (C-5). This information justifies a 3-methylbutan-2-ol group to be attached to the coumarin ring at position C-5 (δ 113.7) leaving C-6 and C-7 to form a fused ring. The 1H-NMR experiment demonstrated the presence of a multiplet and doublet signal at the upfield region . The signal at δ 3.77 (H-10) was directly bonded to δ 39.2 (C-10) whereas the signal at δ 1.15 (H-12) was directly bonded to δ 18.8 (C-12) as seen in the HMQC spectrum. The HMBC spectrum showed H-10 to have connectivities with δ 18.8 (C-12), 209.9 (C-11) and 210.1 (C-9); Meanwhile, H-12 has connectivities with δ 39.2 (C-10), 209.9 (C-11) and δ 210.1 (C-9). This implies the presence of 2-methylcyclopentane-1,3-dione. This substituent group has to be placed at C-6 and C-7 since C-5 carries the 3-methylbutan-2-ol group. The left ring of the coumarin is thus substituted at C-7 and C-6 by the above dione substituent group. ---chromen-2-one.Meanwhile, the NOESY experiment shows correlations between H-4’ and H-2’ suggesting that they have a similar configuration. The HMBC and NOESY correlations for 3 as solvent and tetramethylsilane (TMS) as internal standard. Ultraviolet spectra were recorded in EtOH on a Shimadzu UV-160A, UV-Visible Recording Spectrophotometer. Infrared spectra were measured using the universal attenuated total reflection (UATR) technique on a Perkin-Elmer 100 Series FT-IR spectrometer. EIMS were recorded on a Shimadzu GCMS-QP5050A spectrometer. NMR spectra were obtained on a Unity JEOL 500 MHz FT-NMR spectrometer using CDClMesua beccariana was collected from the Sri Aman district in Sarawak, Malaysia. The plant material was identified by Associate Professor Dr Rusea Go, Biology Department, Faculty of Science, Universiti Putra Malaysia.The stem bark of n-hexane and extracted successively with dichloromethane, ethyl acetate and methanol. The extracts were dried under reduced pressure using a rotary evaporator to yield hexane (15.6 g), dichloromethane (21.2 g), ethyl acetate (15.8 g) and methanol (80.5 g) extracts. Each of these extracts was chromatographed over a silica gel column using a stepwise gradient system . The hexane extract was subjected to vacuum column chromatography over silica gel with a stepwise gradient of hexane/dichloromethane (hexane/CH2Cl2) and dichloromethane/ethyl acetate (CH2Cl2/EA). The eluted fraction A  was then subjected to a flash column chromatography using hexane/CH2Cl2 to obtain fraction B  to give beccamarin. Beccamarin was crystallized by repeated crystallizations from hexane. The hexane extract provided beccamarin , mesuarianone , mesuasinone  while the ethyl acetate extract gave two anthraquinones, 4-methoxy-1,3,5-trihydroxyanthraquinone  and 2,5-dihydroxy-1,3,4-trimethoxyanthraquinone  and a coumarin, mammea A/AB .The three kg of milled, air-dried and powdered sample was defatted with Beccamarin (1). Yellow solid. UV (EtOH) λmax nm (log ε): 209 (5.23), 229 (5.27), 281 (5.36) and 348 (5.45). IR νmax (cm-1): 3400, 2971, 1741, 1605 and 1466. MS m/z (rel. int.): 406 [M+] (12), 392 (28), 377 (16), 350 (24), 349 (100), 293 (19), 43 (18). For 1H and 13C-NMR spectra, see Mesuarianone (2). Yellow solid. UV (EtOH) λmax nm (log ε): 208, 280 and 333.8. IR νmax (cm−1): 3392, 2971, 2923, 1639, 1574, 1475. MS m/z (rel. int.): 460 [M+] (39), 445 (100), 377 (89), 361 (10), 347 (11), 323 (12), 203 (8), 181 (61), 91 (6), 77 (5), 69 (27), 55 (9). The 1H- and 13C-NMR (CDCl3) spectral data are consistent with published data  (14), 431 (4), 391 (8), 363 (100), 307 (21), 154 (12), 69 (10). The 1H and 13C NMR (CDCl3) spectral data are consistent with published data (100), 268 (87), 257 (10), 243 (38), 212 (27), 180 (30). The 1H and 13C NMR (CDCl3) spectral data are consistent with published data (100), 315 (60), 312 (5), 297 (20), 287 (22), 272 (24), 227 (20), 58 (23). The 1H and 13C NMR (CDCl3) spectral data are consistent with published data (2), 392 (18), 377 (10), 349 (100), 293 (10). The 1H and 13C NMR (CDCl3) spectral data are consistent with published data[18), 377 , 349 (101), along with two xanthones, two anthraquinones and another coumarin were isolated from the stem bark of Mesua beccariana. A novel cyclodione coumarin, beccamarin (


Dioscorea zingiberensis. Among the three oligosaccharides, the crude oligosaccharide prepared by acid hydrolysis of WEP was found to be the most efficient elicitor to enhance the production of palmarumycins C12 and C13 in liquid culture of endophytic fungus Berkleasmium sp. Dzf12. When OW was applied to the medium at 300 mg/L on day 3 of culture, the maximal yields of palmarumycin C12 (87.96 mg/L) and palmarumycin C13 (422.28 mg/L) were achieved on day 15 of culture, which were 9.83 and 3.24-fold in comparison with those (8.95 and 130.43 mg/L) of control, respectively. The results indicate that addition of the oligosaccharides from the host plant D. zingiberensis should be an effective strategy for enhancing production of palmarumycins C12 and C13 in liquid culture of endophytic fungus Berkleasmium sp. Dzf12.Three crude oligosaccharides were respectively prepared by acid hydrolysis of three polysaccharides, which were water-extracted polysaccharide (WEP), sodium hydroxide-extracted polysaccharide (SEP) and acid-extracted polysaccharide (AEP) from the rhizomes of  OWcrude oligosaccharide prepared by acid hydrolysis of the water-extracted polysaccharideOScrude oligosaccharide prepared by acid hydrolysis of the sodium hydroxide-extracted polysaccharideOAcrude oligosaccharide prepared by acid hydrolysis of the acid-extracted polysaccharideWEPwater-extracted polysaccharideSEPsodium hydroxide-extracted polysaccharideAEPacid-extracted polysaccharideTFAtrifluoroacetic acidBerkleasmium sp. Dzf12, isolated from the healthy rhizomes of the medicinal plant Dioscorea zingiberensis, five spirobisnaphthalenes have been successfully obtained that show antimicrobial activity , 7.49 , 7.01 , 6.96 , 6.81 , 6.76 , 6.46 , 5.31 , 3.64 , 3.61 . 13C-NMR  δ (ppm): 97.2 (C-1), 52.4 (C-2), 54.0 (C-3), 64.4 (C-4), 121.5 (C-4a), 148.6 (C-5), 117.4 (C-6), 117.4 (C-7), 150.0 (C-8), 118.3 (C-8a), 147.4 (C-1'), 108.8 (C-2'), 127.9 (C-3'), 120.3 (C-4'), 133.9 (C-4a'), 120.1 (C-5'), 127.7 (C-6'), 108.6 (C-7'), 147.4 (C-8'), 112.1 (C-8a'). The structure , mp 230 °C. Its molecular formula was determined as C20H14O7  by its HR-ESI-MS. 1H-NMR  δ (ppm): 7.64 , 7.62 , 7.52 , 7.10 , 7.04 , 6.74 , 6.24 , 6.00 , 5.88 , 4.98 , 4.69 , 3.17 . 13C-NMR  δ (ppm):95.1 (C-1), 52.7 (C-2), 55.2 (C-3), 59.7 (C-4), 70.7 (C-4a), 60.6 (C-5), 125.3 (C-6), 144.8 (C-7), 188.8 (C-8), 62.2 (C-8a), 145.2 (C-l'), 109.1 (C-2'), 127.9 (C-3'), 120.7 (C-4'), 133.8 (C-4a'), 120.7 (C-5'), 127.7 (C-6'), 108.7 (C-7'), 145.5 (C-8'), 111.5 (C-8a'). The structure . The data were submitted to analysis of variance (one-way ANOVA) to detect significant differences by PROC ANOVA of SAS version 8.2. The term significant has been used to denote the differences for which D. zingiberensis were successfully applied to the liquid cultures of Berkleasmium sp. Dzf12 for enhancing the production of palmarumycins C12 and C13. There were some reports about the oligosaccharide elicitation of fungal secondary metabolite production, such as the oligosaccharides derived from sodium alginate and locust-bean gum [D. zingiberensis had obvious enhancing effects on palmarumycin biosynthesis in Berkleasmium sp. Dzf12, although the elicitation mechanism was not clear. As the oligosaccharide employed in this study was a mixture, further separation and identification of the bioactive oligosaccharide monomers are necessary. The results indicate that enhancement of palmarumycin accumulation in liquid culture of Berkleasmium sp. Dzf12 by the oligosaccharides from host plant D. zingiberensis could be an effective strategy for large-scale production of palmarumycins C12 and C13 in the future. By comparing with our previous study on the enhancing effects of the polysaccharides from D. zingiberensis on palmarumycin C13 production [i.e., OW) has a better stimulating effect than its corresponding polysaccharide . This indicated that the oligosaccharide fragments in polysaccharide may be the active components to stimulate secondary metabolism that should be further studied.In this work, the oligosaccharides prepared from the polysaccharides isolated from the rhizomes of bean gum ,25. Howeoduction , the oli


N-1- and N-2-methyl derivatives of OM. Both compounds, of which N-2 isomer of olmesartan dimedoxomil is a novel impurity of OM, were synthesized and fully characterized by differential scanning calorimetry (DSC), infrared spectroscopy (IR), nuclear magnetic resonance spectroscopy (NMR) and high-resolution mass spectrometry/electrospray ionization (HRMS/ESI). Their 1H, 13C and 15N nuclear magnetic resonance signals were fully assigned. The molecular structures of N-triphenylmethylolmesartan ethyl (N-tritylolmesartan ethyl) and N-tritylolmesartan medoxomil, the key intermediates in OM synthesis, were solved and refined using single-crystal X-ray diffraction (SCXRD). The SCXRD study revealed that N-tritylated intermediates of OM exist exclusively as one of the two possible regioisomers. In molecular structures of these regioisomers, the trityl substituent is attached to the N-2 nitrogen atom of the tetrazole ring, and not to the N-1 nitrogen, as has been widely reported up to the present. This finding indicates that the reported structural formula of N-tritylolmesartan ethyl and N-tritylolmesartan medoxomil, as well as their systematic chemical names, must be revised. The careful analysis of literature spectroscopic data for other sartan intermediates and their analogs with 5-(biphenyl-2-yl)tetrazole moiety showed that they also exist exclusively as N-2-trityl regioisomers.During the process development for multigram-scale synthesis of olmesartan medoxomil (OM), two principal regioisomeric process-related impurities were observed along with the final active pharmaceutical ingredient (API). The impurities were identified as  Potential advantages of this drug include once-daily dosing, an absence of significant adverse reactions, a well-tolerated side effect profile, and a cost-effective average wholesale price.Elevated blood pressure is one of the most important causes of death and disability worldwide, accounting for 7.6 million premature deaths and 92 million disability-adjusted life years annually . Controli.e., losartan, valsartan, irbesartan and candesartan, are 5-(biphenyl-2-yl)tetrazole derivatives. [2′-(N-Triphenylmethyltetrazol-5-yl)biphenyl-4-yl]methyl bromide methyl substituent methyl methyl , 60:40 fl]methyl  and 47.3l methyl bromide (2), the commercially available in kg scale intermediate to the syntheses of sartans, should be tritylated at the 2-position of tetrazole. The crystal structure of [2′-(N-triphenylmethyltetrazol-5-yl)biphenyl-4-yl]methyl bromide obtained according to the procedure given by Aldrich was solved using a single crystal grown from butan-2-one methyl bromide (2) and bromides 2 described in the literature in many cases shows differences (see N-1-position of 5-(biphenyl-2-yl)tetrazole was found. Thus, the starting bromide 2 for syntheses of sartans is [2′-(2-triphenylmethyl-2H-tetrazol-5-yl)biphenyl-4-yl]methyl bromide.Alkylation of 5-phenyltetrazole with trityl chloride under alkaline conditions as well as with triphenylmethanol under acidic conditions should lead exclusively to the 5-phenyl-2-trityltetrazole ,21. Theran-2-one . Althoug 2 later . The com1 with the bromide 2 in the presence of K2CO3 and KI in DMF (3. The SCXRD measurement of this product showed that the molecule contains tetrazole ring tritylated at N-2 position (3 is ethyl 4-(1-hydroxy-1-methylethyl)-2-propyl-1-[2′-(2-triphenylmethyl-2H-tetrazol-5-yl)biphenyl-4-yl]methyl-1H-imidazole-5-carboxylate. The next intermediate in the synthesis of OM, N-tritylolmesartan medoxomil (6), is obtained in two steps from 3 methyl 4-(1-hydroxy-1-methylethyl)-2-propyl-1-[2′-(2-triphenylmethyl-2H-tetrazol-5-yl)biphenyl-4-yl]-methyl-1H-imidazole-5-carboxylate methanol with the bromide 2 in the presence of K2CO3 in DMF, reported in chemical databases as both N-1- (CAS 124751-00-4) and N-2-trityl (CAS 133909-99-6), is tritylated at N-2 of tetrazole, which was unambiguously proved by SCXRD a et al.  as N-1-ma et al.  is none oisomers . Withoutterature ,34, syntterature . However9 and 10 could be determined only on the basis of the detailed analysis of the multinuclear magnetic resonance results since in their case the attempts of obtaining crystals suitable for SCXRD measurements failed. For these compounds careful analysis of the 1H-, 13C-, 15N-NMR chemical shifts and 1H/13C (1H/15N) gradient selected HSQC and HMBC (g-HSQC and g-HMBC) correlations were done. Differentiation of 9 and 10 was possible using cross-peaks coming from 1H/13C g-HMBC experiment of methylene group of medoxomil substituent at the tetrazole ring. In the case of 9 methylene protons at 5.46 ppm (in CDCl3) correlate with carbons at 9.3, 130.7 and 140.5 ppm (10 (3) correlate with medoxomil carbons at 8.9, 130.3 and 140.0 ppm. However, other additional correlation is observed in 2D g-HMBC experiment. The fourth cross-peak at 4.67 ppm/154.6 ppm identifies tetrazole carbon atom and allows 10 to assign the structure of 1,5-disubstituted isomer methyl bromide  marketed as N-(triphenylmethyl)-5-[4′-(bromomethylbiphenyl)-2-yl]tetrazole, i.e., without specified trityl position, was purchased from Huanggang Luban Pharmaceutical Co. Ltd. . Potassium dihydrogen phosphate (V) , phosphoric acid (V) (85 wt % in H2O), acetonitrile and tetrahydrofuran of HPLC grade were purchased from Sigma-Aldrich  and POCh S.A.  chemical companies. Deionized water was prepared using MilliQ plus purification system . Potassium bromide (FT-IR grade), deuterated chloroform and deuterated dimethylsulfoxide were purchased from Merck KGaA . The course of all reactions and the purity of products were checked by thin-layer chromatography (TLC). Analytical TLC was performed on silica gel DC-Alufolien Kieselgel 60 F254 , with mixtures of hexanes, ethyl acetate, dichloromethane and methanol, in various ratios as developing systems. Compounds were visualized by UV light (λ = 254 nm). Column chromatography was carried out on silica gel  with mixtures of ethyl acetate, methanol and dichloromethane in varying ratios as eluents.Ethyl 4-(1-hydroxy-1-methylethyl)-2-propylimidazole-5-carboxylate .FT-IR spectra were taken for KBr pellets on a Nicolet Impact 410 FT-IR spectrophotometer .HRMS spectra were recorded on an AMD 604 Inectra Gmbh  and a Mariner PE Biosystem ESI-TOF  spectrometers.H-tetrazol-5-yl)biphenyl-4-yl]methyl-1H-imidazole-5-carboxylate (3) and methyl 4-(1-hydroxy-1-methylethyl)-2-propyl-1-[2′-(2-triphenylmethyl-2H-tetrazol-5-yl)biphenyl-4-yl]methyl-1H-imidazole-5-carboxylate (6) were collected on an Oxford Diffraction X-calibur  with Ruby detector . Monocrystals of 3 and 6, suitable for the XRD experiment, were obtained from acetone-methanol and acetone solution, respectively. The data were collected at 100 K using an Oxford Cryosystem device  by rotation of 180° around the C11–N71 bond (CH2-imidazole N1 bond). In 6, hydrogen atoms bonded to methyl groups  are disordered over two positions. The ratio was refined assuming that the sum of both components of the disorder is equal to 1. Non-hydrogen atoms were refined with anisotropic displacement parameters. SIMU, SADI and ISOR restraints were applied for the disordered atom. All H atoms were found in Δρ maps or placed at calculated positions. Before the last cycle of refinement, all H atoms were fixed and were allowed to ride on their parent atoms. For clarity, the minor component of the disordered imidazole units in 3, and the acetone molecule in 6 solvate, are omitted in 3 and 6 were visualized with Mercury CDS 3.3 software  and an eluent B . UV detection was performed at 250 nm and the injection volume was set as 10 μL. Individual stock solutions of standards and samples were prepared in acetonitrile at the concentration of 1.0 mg/mL.Determination of dimedoxomil impurities nographs ,18. The nographs ,43. A ty7) and olmesartan (8) from commercially available substrates 1, 2 and 5 is shown in 3 and 6–8 as well as their physical and spectroscopic data are provided in the Supplementary Materials.The synthesis of OM , KI  and medoxomil chloride (5)  were added to a suspension of olmesartan  in DMF (60 mL). After being stirred at room temperature for 22 h, TLC analysis (30% MeOH/AcOEt) indicated the disappearance of the starting material 8 and formation of two reaction products characterized by similar, but different from OM (7), polarity. The reaction mixture was diluted with H2O (50 mL) and the products were extracted with AcOEt (2 × 50 mL). The combined organic phases were dried over anhydrous MgSO4, filtered and concentrated under reduced pressure to give an oily residue. The crude mixture of products  was purified by column chromatography over silica gel with 50%–100% AcOEt/hexanes gradient elution to afford N-2-medoxomil derivative 9 , a mixture of N-2- 9 and N-1-medoxomil 10 derivatives (2.09 g) and N-1-medoxomil derivative 10 . Total reaction yield was 95%.K9: FT-IR (KBr) ν: 3438, 2968, 2933, 2874, 1828, 1738, 1708, 1675, 1529, 1463, 1433, 1392, 1308, 1227, 1131, 1033, 1066, 765 cm−1.1H-NMR  δ: 7.84 , 7.56 , 7.49 , 7.44 , 7.11 , 6.82 , 5.46 , 5.44 , 4.90 , 2.72 , 2.19 , 2.08 , 1.74 , 1.62 (CH3)2), 0.98 .13C-NMR  δ: 165.9 (tetrazole C-5), 160.8 (>C=O), 160.0 (imidazole C-4), 152.3 (imidazole C-2), 151.8 (medoxomil >C=O of ester group), 151.3 (medoxomil >C=O at N-2 of tetrazole), 141.3 (biphenyl C-1′), 140.6 (medoxomil C-4 or C-5 of ester group), 140.5 (medoxomil C-4 or C-5 at N-2 of tetrazole), 140.0 (biphenyl C-1), 135.7 (biphenyl C-4), 132.8 (medoxomil C-4 or C-5 of ester group), 130.8 (biphenyl C-6′), 130.7 (medoxomil C-4 or C-5 at N-2), 130.4 (biphenyl C-5′), 129.6 , 127.8 (biphenyl C-4′), 125.5 (biphenyl C-2′), 125.0 , 116.0 (imidazole C-5), 70.4 (-C(OH)(CH3)2), 54.0 (-CH2O-), 49.2 (imidazole >N-CH2-), 45.1 (tetrazole -CH2-N<), 29.2 (CH3)2), 29.1 (-CH2CH2CH3), 21.4 (-CH2CH2CH3), 13.8 (-CH2CH2CH3), 9.3 (medoxomil CH3-5 at N-2 of tetrazole), 9.3 (medoxomil CH3-5 of ester group).1H-NMR  δ: 7.77 , 7.63 , 7.55 , 7.47 , 7.05 , 6.84 , 5.90 , 5.42 , 5.21 , 5.07 , 2.65 , 2.17 , 2.08 , 1.63 , 1.48 (CH3)2), 0.90 .13C-NMR  δ: 164.7 (tetrazole C-5), 160.7 (>C=O), 157.6 (imidazole C-4), 151.7 (medoxomil >C=O of ester group), 151.5 (medoxomil >C=O at N-2 of tetrazole), 151.0 (imidazole C-2), 140.9 (biphenyl C-1′), 140.4 (medoxomil C-4 or C-5 of ester group), 140.4 (medoxomil C-4 or C-5 at N-2 of tetrazole), 139.0 (biphenyl C-1), 136.2 (biphenyl C-4), 132.8 (medoxomil C-4 or C-5 of ester group), 131.2 (medoxomil C-4 or C-5 at N-2 of tetrazole), 130.8 (biphenyl C-6′), 130.5 (biphenyl C-5′), 130.3 (biphenyl C-3′), 129.1 , 127.8 (biphenyl C-4′), 125.4 (biphenyl C-2′), 125.2 , 116.1 (imidazole C-5), 69.6 (-C(OH)(CH3)2), 54.1 (-CH2O-), 48.0 (imidazole >N-CH2-), 44.9 (tetrazole -CH2-N<), 29.7 (CH3)2), 28.3 (-CH2CH2CH3), 20.6 (-CH2CH2CH3), 13.6 (-CH2CH2CH3), 8.7 (medoxomil CH3-5 of ester group), 8.6 (medoxomil CH3-5 at N-2 of tetrazole).15N-NMR  δ: 0.0 (tetrazole N-3), −76.7 (tetrazole N-1), −99.2 (tetrazole N-2), −120.0 (imidazole N-3), −211.6 (imidazole N-1).m/z 671.2454 .HRMS (ESI) 10: mp 143.84–151.62 °C (AcOEt/hexanes), peak 147.57 °C, heating rate 10.00 °C/min .Lit. mp 126–130 °C (acetone) .FT-IR  ν: 3383, 3028, 2967, 2935, 2875, 1836, 1821, 1808, 1743, 1676, 1533, 1467, 1397, 1371, 1313, 1230, 1192, 1151, 1085, 1003, 957, 769.Lit. IR (KBr) ν: 3385, 2931, 2875, 1816, 1742, 1531 .1H-NMR  δ: 7.72 , 7.63 , 7.58 , 7.53 , 7.12 , 6.87 , 5.53 , 5.40 , 4.91 , 4.67 , 2.63 , 2.12 , 1.89 , 1.61 (CH3)2), 0.95 .13C-NMR  δ: 160.7 (>C=O), 160.6 (imidazole C-4), 154.6 (tetrazole C-5), 152.4 (imidazole C-2), 151.7 (medoxomil >C=O of ester group), 151.0 (medoxomil >C=O at N-1 of tetrazole), 141.0 (biphenyl C-1′), 140.6 (medoxomil C-4 or C-5 of ester group), 140.0 (medoxomil C-4 or C-5 at N-1 of tetrazole), 137.7 (biphenyl C-1), 137.1 (biphenyl C-4), 132.8 (medoxomil C-4 or C-5 of ester group), 132.2 (biphenyl C-5′), 131.1 (biphenyl C-3′), 130.6 (biphenyl C-6′), 130.3 (medoxomil C-4 or C-5 at N-1 of tetrazole), 129.0 , 128.4 (biphenyl C-4′), 126.2 , 121.6 (biphenyl C-2′), 115.9 (imidazole C-5), 70.4 (-C(OH)(CH3)2), 53.9 (-CH2O-), 48.9 (imidazole >N-CH2-), 39.8 (tetrazole -CH2-N<), 29.2 (-CH2CH2CH3), 29.1 (CH3)2), 21.2 (-CH2CH2CH3), 13.8 (-CH2CH2CH3), 9.3 (medoxomil CH3-5 of ester group), 8.9 (medoxomil CH3-5 at N-1 of tetrazole).1H-NMR  δ: 7.76 , 7.64 , 7.04 , 6.85 , 5.40 , 5.21 , 5.19 , 5.04 , 2.58 , 2.08 , 1.78 , 1.56 , 1.47 (CH3)2), 0.86 .13C-NMR  δ: 160.7 (>C=O), 157.6 (imidazole C-4), 154.0 (tetrazole C-5), 151.6 (medoxomil >C=O of medoxomil ester), 151.2 (medoxomil >C=O at N-1 of tetrazole), 151.0 (imidazole C-2), 141.1 (biphenyl C-1′), 140.4 (medoxomil C-4 or C-5 of ester group), 139.7 (medoxomil C-4 or C-5 at N-1 of tetrazole), 137.2 (biphenyl C-1), 137.1 (biphenyl C-4), 132.8 (medoxomil C-4 or C-5 of ester group), 131.8 (biphenyl C-5′), 131.0 , 130.7 (medoxomil C-4 or C-5 at N-1 of tetrazole), 128.5 , 128.0 (biphenyl C-4′), 125.6 , 121.6 (biphenyl C-2′), 116.2 (imidazole C-5), 69.6 (-C(OH)(CH3)2), 54.1 (-CH2O-), 47.9 (imidazole >N-CH2-), 39.8 (tetrazole -CH2-N<), 29.6 (CH3)2), 28.2 (-CH2CH2CH3), 20.5 (-CH2CH2CH3), 13.5 (-CH2CH2CH3), 8.7 (medoxomil CH3-5 of ester group), 8.2 (medoxomil CH3-5 at N-1 of tetrazole).15N-NMR  δ: −11.1 (tetrazole N-2), −119.6 (imidazole N-3), −149.4 (tetrazole N-1), −212.3 (imidazole N-1).m/z 671.2463 .HRMS (ESI) 2CO3 , n-Bu4NBr (cat.) and medoxomil chloride 5  were added to a solution of olmesartan  in Me2CO (40 mL). After heating at reflux for 12 h, TLC analysis (30% MeOH/AcOEt) indicated the disappearance of the starting material 8 and formation of two reaction products characterized by similar, but different from OM (7) polarity. The reaction mixture was cooled to room temperature and filtered. The resulting solid was washed with Me2CO (10 mL). The combined filtrate and washings were concentrated under reduced pressure, and the residue was portioned between AcOEt (50 mL) and H2O (50 mL). The acetate layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a dark-brown oily residue . An unsuccessful attempt to purify the crude mixture of N-2- (9) and N-1- (10) medoxomil derivatives and acetone condensation products by column chromatography on silica gel with 10% MeOH/CH2Cl2 elution was made. A dark-brown oily starting material was recovered. After the addition of Me2CO, no solid of dimedoxomil derivatives precipitated out of solution.K7) formed from the olmesartan acid (8) were described. The impurities were identified as isomeric N-2- (9) and N-1- (10) methyl derivatives of OM. The N-2-medoxomil impurity 9 is a new compound that has never been synthesized, identified and characterized. The structure of regioisomeric N-1-medoxomil impurity 10 was disclosed in the literature. However, either there were no synthetic procedures and physicochemical data, or presented data do not prove the revealed structure. Thus, both compounds 9 and 10 were synthesized, separated and fully characterized. The structures of the impurities, the substrate 2 and the intermediates 3 and 6 in the synthesis of OM were determined by crystallographic and/or spectroscopic methods. The careful analysis of ours and reported analytical data  allowed disproving the structural dualism of the tetrazole tritylated compounds 2, 3 and 6, existing in the literature data and chemical databases, and undoubtedly established their exact structure as N-2-tritylated tetrazole regioisomers. The analysis also evidently shows that the tritylated intermediates of other sartans with 2-(tetrazol-5-yl)biphenyl moiety possess triphenylmethyl substituent at tetrazole N-2 position.Two principal process-related impurities of olmesartan medoxomil API (OM,


On the basis of spectroscopic analysis and chemical methods, their structures were identified as 20-O-acetyl-8,14-seco-penupogenin-8-one 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopyranosyl-(1→4)-β-D-cymaropyranoside (1), 2′,3′-Z-gagaminine 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaro-pyranosyl-(1→4)-α-L-diginopyranosyl-(1→4)-β-D-cymaropyranoside (2), 17-O-acetyl-kidjoranin 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-cymaro-pyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→4)-β-D-digitoxopyranoside (3), gagaminine 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-digino-pyranosyl-(1→4)-β-D-cymaropyranoside (4) and wilfoside D1N (5).Three new steroidal glycosides, cyanoauriculosides F, G and H ( Cynanchum auriculatum is a famous traditional medicine widely used in south China for the prevention of hair graying, strengthening sinews and bones, and enhancing immunity + arising from the aglycone moiety. From 1H-NMR, 13C-NMR, HSQC and HMBC data, one acetyl group was also identified by the observation of a proton signal at δH 1.96  and two carbon signals at δC 170.5, 21.6. Comparison of the 13C-NMR data of the aglycone portion of 1 with that of penupogenin + arising from the aglycone moiety. From 1H-NMR and 13C-NMR, one nicotinoyl group  was also found in the aglycone moiety. The 1H-NMR data of the aglycone portion of 2 was compared with the data of gagaminine  in the structure of 3. Thus, the aglycone of compound 3 was proposed to be 17-O-acetylkidjoranin, which was supported by NOE correlations between H-21  and H-11′ , between H-16  and H-11′ . The 17-α configuration was confirmed by the observation that the carbonyl carbon of the α-linked methyl ketone at C-17 appears at δ 210.2, compared with δ 216 ppm for the β configuration D20 = + 19.2 ; For 1H-NMR and 13C-NMR spectroscopic data (in C5D5N), see -m/z 1,165.5694 . 2′,3′-Z-gagaminine 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopyranos-yl-(1→4)-β-D-cymaropyranoside (2). White amorphous powder; UV λmax (MeOH) nm (logε): 203 (4.32); [α]D20 = + 21.6 ; For 1H-NMR and 13C-NMR spectroscopic data (in CD3OD), see +m/z 1,216.6033 .17-O-acetyl-kidjoranin 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-cymaro- pyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→4)-β-D-digitoxopyranoside (3). White amorphous powder; UV λmax (MeOH) nm (logε): 278 (4.37); [α]D20 = - 60.2 ; For 1H-NMR and 13C-NMR spectroscopic data (in C5D5N), see + m/z 1,267.6268 .C.auriculatum Royle ex Wight, including thirteen C-21 steroidal glycosides. The anti-tumour activity of these C-21 steroidal glycosides compounds has been studied. Further research on isolation and identification of more bioactive compounds will be helpful to understand this traditional medicine.Twenty compounds were isolated from the dry roots


The synthesized compounds were characterized by various spectroscopic techniques viz IR, HR-MS, 1H NMR, and 13C NMR. The time required for completion of reaction in MM varied from 1 to 5 min as compared to CM which required 10–40 h. All the synthesized compounds were screened for antifungal activity against Sclerotium rolfsii and Alternaria solani. In vitro fungicidal activity revealed that compound 3o (ED50 = 23.24 mg L−1) was found to be most active against S. rolfsii. But in case of A. solani, compound 3c (ED50 = 29.9 mg L−1) showed highest activity. The nematicidal activity revealed that the compound 3b was more potent with LC50 values of 10.67, 7.30, and 4.55 ppm at 24, 48, and 72 h, respectively. 2D-Quantitative Structural Activity Relationship (2D-QSAR) analysis of these ferrocenyl chalcones was carried out by developing three different models namely Partial Least Squares , Multiple Linear Regression  and Principal Component Regression . Statistical significance and predictive ability of these models were assessed by internal and external validation and also verified by leave one-out cross-validation. QSAR study revealed that MLR for S. rolfsii , PLS for A. solani  and PCR for M. incognita  were the best model. The physico-chemical parameters were calculated using VLife MDS 4.6 software. QSAR study could be employed for structure optimization to achieve better activity.A new microwave method (MM) has been developed for the synthesis of a series of 16 substituted ferrocenyl chalcones using acetylferrocene ( Chalcones and their corresponding heterocyclic analogs were reported to exhibit several biological activities such as antibacterial, antifungal, insecticidal, nematicidal, anti-oxidant, antiplasmodial, antitumor, and anthelmintic due to occurrence of highly reactive unsaturated carbonyl moiety in skeleton  are the most destructive pathogens in terms of yield loss due to their broad host range i.e., vegetables, cereals, pulses, etc. . They have been reported to cause extensive damage to the tune of 20–32%  with PDA detector using isocratic solvent system (methanol-water: 98:2) was used for determining the purity of synthesized ferrocenyl chalcones. 1H-NMR and 13C-NMR studies were carried out on JEOL 400 MHz Spectrospin spectrometer instrument, data were processed using Delta software and tetramethylsilane (TMS) was used as internal standard.Acetylferrocene and different benzaldehydes were procured from Aldrich. All other solvents and chemicals were of analytical grade and were used as received unless otherwise noted. Reactions were observed by thin layer chromatography (TLC) on pre-coated Merck silica gel 60F+ equipped with TurboIonSpray (TIS), SCIEX ExionLC, and PDA detector. C-18 column  was used for separation of compounds. The column was eluted with methanol and water  with 0.1% formic acid at a flow rate of 0.3 mL/min. The column oven temperature was set at 40°C. Infrared (IR) spectra were recorded on Bruker alpha FT-IR spectrophotometer, values were expressed as υmax cm−1. Melting point apparatus was used for determination of melting points. ED50 and LC50 values were calculated by SPSS statistical package. All computational activities were performed by using Vlife MDS QSAR plus 4.6 software in ASUS VivoBook (windows 10 OS and Intel Core i5).High Resolution Mass Spectrometry (HRMS) was performed by AB SCIEX Triple TOF™ 5,600The synthesis of the ferrocenyl chalcones series was done by the base-catalyzed Claisen-Schmidt reaction according to the method reported in literature. The general method of synthesis is reproduced below , 4-hydroxybenzaldehyde (2p)] was synthesized as follows: Acetylferrocene  and KOH (0.2 g) were dissolved in ethanol (5 mL) in a round bottom flask and stirred at room temperature for 10 min, followed by drop wise addition of ethanolic solution of equimolar amounts of different benzaldehydes (2a-2o),  with continuous stirring. The stirring ranged between 1 and 40 h for different reactions at room temperature  , 2H-3, 4-dihydropyran (8 mmol), and pyridinium p-toluenesulfonate (0.2 mmol) were taken in DCM (10 mL) and kept for stirring at room temperature for 4 h , followed by removal of solvent under vacuum. These protected benzaldehydes (2i and 2p) were then treated with acetylferrocene as per the method described above to get the corresponding ferrocenyl chalcones.3i and  for 4 h . After cEquimolar amounts of acetylferrocene  in 5% ethanolic KOH (5 mL) with minimum ethanolic solution of the different benzaldehydes were added in microwave vial (10 mL). The vial was placed in microwave synthesis reactor  with the conditions; Temp. 100°C, rpm-600, and time 1–5 min. Monitoring of reaction was carried out by TLC in ethyl acetate: hexane (1:4 solvent systems). Reaction mixture was worked up as described above.f: 0.50 . IR (cm−1): 1,645 (CO), 1,593 (C = C). 1H-NMR : δ 4.22 , 4.63 , 4.94 , 7.29 , 7.45 , 7.67 , 7.76 .13C- NMR : δ 69.65 (C-2″ & C-5″), 69.78 (C-6″), 72.74 (C-3″ & C-4″), 80.66 (C-1″), 123.64 (C-α), 128.61 (C-3′ & C-5′), 128.89 (C-2′ & C-6′), 130.11 (C-4′), 134.90 (C-1′), 139.75 (C-β), 191.96 (CO). HR-MS for C19H16FeO [M+H]+m/z: Calcd 317.0623; Observed 317.0639.It was obtained as a red colored solid in 81% yield; m.p.:140–142°C, Rf: 0.42 . IR (cm−1): 1,649 (CO), 1,591 (C = C). 1H-NMR : δ 4.22 , 4.64 , 4.93 , 7.28 , 7.61 , 7.63 , 7.68 . 13C-NMR : δ 69.70 (C-2″ & C-5″), 69.80 (C-6″), 72.83 (C-3″ & C-4″), 80.59 (C-1″), 123.45 (C-α), 124.43(C-4′), 130.55 (C-2′ & C-6′), 131.82 (C-3′ & C-5′), 134.24 (C-1′), 138.41 (C-β), 191.88 (CO). HR-MS for C19H15BrFeO [M+H]+m/z: Calcd 394.9728; Observed 394.9739.It was obtained as an orange colored solid in 87% yield; m.p.:160–162°C, Rf: 0.43 . IR (cm−1): 1,649 (CO), 1,589 (C = C). 1H-NMR : δ 4.22 , 4.63 , 4.93 , 7.20 , 7.22 , 7.65 , 7.81 .13C-NMR : δ 69.28 (C-2″ & C-5″), 69.99 (C-6″), 72.95 (C-3″ & C-4″), 80.87 (C-1″), 115.62 (C-3′ & C-5′), 123.79 (C-α), 130.60 (C-2′ & C-6′), 131.07 (C-1′), 138.34 (C-β), 163.64 (C-4′),192.45 (CO). HR-MS for C19H15FFeO [M+H]+m/z: Calcd 335.0529; Observed 317.0544.It was obtained as a red colored solid in 79% yield; m.p.:138–140°C, Rf: 0.60 . IR (cm−1): 1,653 (CO), 1,591 (C = C). 1H-NMR : δ 4.19 , 4.52 , 4.62 , 6.64 , 7.24 , 7.42, 7.47, 7.51, 7.53 .13C-NMR : δ 68.24 (C-2″ & C-5″), 69.12 (C-6″), 72.99 (C-3″ & C-4″), 80.35 (C-1″), 124.22 (C-α), 125.89 (C-5′), 127.20 (C-6′), 128.56 (C-3′), 130.6 (C-4′), 133.73 (C-2′), 136.61 (C-1′), 139.85 (C-β), 192.87 (CO). HR-MS for C19H15ClFeO [M+H]+m/z: Calcd 351.0233; Observed 351.0231.It was obtained as a chocolate colored solid in 80% yield; m.p.:125–126°C, Rf: 0.50 . IR (cm−1): 1,653 (CO), 1,595 (C = C). 1H-NMR : δ 4.24 , 4.66 , 4.94 , 7.24 , 7.31 , 7.44 , 7.69 , 7.96 , 7.97 .13C-NMR : δ 69.71 (C-2″ & C-5″), 69.86 (C-6″), 72.99 (C-3″ & C-4″), 80.35 (C-1″), 125.09 (C-2′), 126.51 (C-α), 128.20 (C-5′), 128.56 (C-4′), 131.66 (C-6′), 133.23 (C-3′), 134.21 (C-1′)137.14 (C-β), 191.70 (CO). HR-MS for C19H15BrFeO [M+H]+m/z: Calcd 394.9728; Observed 394.9724.It was obtained as a red colored solid in 83% yield; m.p.:120–122°C, Rf: 0.28 . IR (cm−1): 1,645 (CO), 1,585 (C = C). 1H-NMR : δ 3.84 , 4.21 , 4.60 , 4.92 , 6.99 , 7.17 , 7.63, 7.72 .13C-NMR : δ 55.34 , 69.54 (C-2″ & C-5″), 69.70 (C-6″), 72.48 (C-3″ & C-4″), 80.87 (C-1″), 114.35 (C-3′ & C-5′), 121.26 (C-α), 127.56 (C-1′), 130.37 (C-2′ & C-6′), 139.65 (C-β), 160.92 (C-4′), 191.90 (CO). HR-MS for C20H18FeO2 [M+H]+m/z: Calcd 347.0729; Observed 347.0749.It was obtained as an orange colored solid in 82% yield; m.p.:150–152°C, Rf: 0.40 . IR (cm−1): 1,639 (CO), 1,562 (C = C). 1H-NMR : δ 3.01 2), 4.19 , 4.57 , 4.90 , 6.76 , 7.08 , 7.58 , 7.61 .13C-NMR : δ 40.65 2), 69. 68 (C-2″ & C-5″), 69.97 (C-6″), 72.28 (C-3″ & C-4″), 81.05 (C-1″), 111.79 (C-3′ & C-5′), 117.80 (C-α), 122.62 (C-1′), 130.17 (C-2′ & C-6′), 141.24 (C-β), 151.72 (C-4′), 192.92 (CO). HR-MS for C21H21FeNO [M+H]+m/z: Calcd 360.1045; Observed 360.1049.It was obtained as a chocolate colored solid in 78% yield; m.p.:140–142°C, Rf: 0.30 . IR (cm−1): 1,643 (CO), 1,579 (C = C). 1H-NMR : δ 3.77 , 3.90 , 4.22 , 4.63 , 4.97 , 7.05 , 7.21 , 7.60 . 13C-NMR : δ 55.31 , 60.10 , 69.60(C-2″ & C-5″), 69.76(C-6″), 72.59 (C-3″ & C-4″), 80.95 (C-1″), 106.17 (C-2′ & C-6′), 122.82 (C-α), 129.56 (C-1′), 130.37 (C-4′), 140.21 (C-β), 153.07 (C-3′ & C-5′), 191.79 (CO). HR-MS for C22H22FeO4 [M+H]+m/z:Calcd 407.0940; Observed 407.0931.It was obtained as a chocolate colored solid in 80% yield; m.p.:145–146°C, Rf: 0.37 . IR (cm−1): 1,647 (CO), 1,585 (C = C). 1H-NMR : δ 4.22 , 4.63 , 4.94 , 5.53 , 7.29 , 7.38 , 7.63 .13C-NMR : δ 69.51 (C-2″ & C-5″), 69.77 (C-6″), 72.74 (C-3″ & C-4″), 80.28 (C-1″), 116.21 (C-2′), 118.05 (C-4′), 122.21 (C-6′), 123.91 (C-α), 129.86 (C-5′), 136.15(C-1′), 139.62 (C-β), 157.17 (C-3′), 192.80 (CO). HR-MS for C19H16FeO2 [M+H]+m/z: Calcd 333.0572; Observed 333.0588.It was obtained as a dark red colored solid in 81% yield; m.p.:120–122°C, Rf: 0.34 . IR (cm−1): 1,645 (CO), 1,579 (C = C). 1H-NMR : δ 4.21 , 4.60 , 4.92 , 5.16 , 7.06 , 7.16 , 7.41 , 7.46 , 7.66 .13C-NMR : δ 69.53 (C-2″ & C-5″), 69.69 (C-6″), 72.48 (C-3″ & C-4″), 77.45 , 80.85 (C-1″), 115.19 (C-3′ & C-5′), 121.37 (C-α), 127.73 , 127.90 , 128.44 , 130.36 , 136.59 , 139.57 (C-β), 159.89 (C-4′), 191.94 (CO). HR-MS for C26H22FeO2 [M+H]+m/z: Calcd 423.1042; Observed 423.1045.It was obtained as an orange colored solid in 82% yield; m.p.:140–142°C, Rf: 0.25 . IR (cm−1): 1,648 (CO), 1,594 (C = C). 1H-NMR : δ 4.24 , 4.55 , 4.94 , 7.24 , 8.01 , 7.77 , 8.04 . 13C-NMR : δ 69.64 (C-2″ & C-5″), 69.95 (C-6″), 72.91 (C-3″ & C-4″), 80.88 (C-1″), 125.68 (C-α), 126.81 (C-3′), 128.62 (C-6′), 129.98 (C-5′), 131.79 (C-1′), 134.12 (C-4′), 137.21 (C-β), 148.98 (C-2′), 192.25 (CO). HR-MS for C19H15FeNO3 [M+H]+m/z: Calcd 362.0474; Observed 362.0476.It was obtained as dark viscous liquid in 71% yield; Rf: 0.31 . IR (cm−1): 1,651 (CO), 1,595 (C = C). 1H-NMR : δ 4.23 , 4.66 , 4.97 , 7.43 , 7.68–7.74 , 8.13 . 8.24 , 8.60 . 13C-NMR : δ 69.53 (C-2″ & C-5″), 69.82 (C-6″), 72.97 (C-3″ & C-4″), 80.41 (C-1″), 122.81 (C-2′), 124.21 (C-4′), 126.28 (C-α), 130.28 (C-5′), 134.79 (C-6′), 136.88 (C-1′), 137.38 (C-β), 148.40 (C-3′), 191.81 (CO). HR-MS for C19H15FeNO3 [M+H]+m/z: Calcd 362.0474; Observed 362.0480.It was obtained as a red colored solid in 80% yield; m.p.:175–176°C, Rf: 0.33 . IR (cm−1): 1,653 (CO), 1,597 (C = C). 1H-NMR : δ 4.24 , 4.68 , 4.96 , 7.39 , 7.72 , 7.97 , 8.27 . 13C-NMR : δ 69.48 (C-2″ & C-5″), 69.85 (C-6″), 72.98 (C-3″ & C-4″), 80.67 (C-1″), 123.77 (C-3′ & C-5′), 125.72 (C-α), 129.20 (C-2′ & C-6′), 136.91 (C-β), 141.21 (C-1′), 148.67 (C-4′),192.41 (CO). HR-MS for C19H15FeNO3 [M+H]+m/z: Calcd 362.0474; Observed 362.0478.It was obtained as a violet colored solid in 82% yield; m.p.:180–182°C, Rf: 0.60 . IR (cm−1): 1,651 (CO), 1,584 (C = C). 1H-NMR : δ 4.24 , 4.66 , 4.88 , 7.29 , 7.31 , 7.33 , 7.49 , 7.66 .13C-NMR : δ 69.46 (C-2″ & C-5″), 69.85 (C-6″), 73.23 (C-3″ & C-4″), 79.83 (C-1″), 123.28 (C-α), 128.75 (C-5′), 130.62 (C-6′), 132.05 (C-3′), 132.34 (C-2′), 132.86 (C-1′), 133.78 (C-4′), 139.36 (C-β), 191.17 (CO). HR-MS for C19H14Cl2FeO [M+H]+m/z: Calcd 384.9843; Observed 384.9813.It was obtained as dark viscous liquid in 76% yield; Rf: 0.60 . IR (cm−1): 1,654 (CO), 1,586 (C = C). 1H-NMR : δ 4.23 , 4.66 , 4.93 , 7.45 , 7.21–7.35 , 7.66 ,7.94–7.98 .13C-NMR : δ 69.72 (C-2″ & C-5″), 70.00 (C-6″), 73.14 (C-3″ & C-4″), 80.35 (C-1″), 124.24 (C-α), 128.07 (C-3′& C-5′), 129.03 (C-4′), 132.51 (C-2′ & C-6′), 135.22 (C-1′), 139.48 (C-β), 192.35 (CO). HR-MS for C19H14Cl2FeO [M+H]+m/z: Calcd 384.9843; Observed 384.9856.It was obtained as dark viscous liquid in 83% yield; Rf: 0.45 . IR (cm−1): 1,645 (CO), 1,571 (C = C). 1H-NMR : δ 4.21 , 4.60 , 4.92 , 5.52 , 7.08 , 7.17 , 7.63 , 7.70 .13C-NMR : δ 69.59 (C-2″ & C-5″), 69.89 (C-6″), 72.28 (C-3″ & C-4″), 80.37 (C-1″), 116.35 (C-3′ & C-5′), 120.78 (C-α), 127.06 (C-1′), 127.37 (C-2′ & C-6′), 139.80 (C-β), 156.92 (C-4′), 192.37 (CO). HR-MS for C19H16FeO2 [M+H]+m/z: Calcd 333.0572; Observed 333.0579.It was obtained as an orange colored solid in 80% yield; m.p.:115–120°C, RA. solani ITCC 4632 and S. rolfsii ITCC 6474 were procured from Indian Type Culture Collection (ITCC) center, Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India and kept at 27°C for at least 4–7 days on Potato Dextrose Agar (PDA) slant. Sub-culturing was done in petriplates and before doing the antifungal activity.Two fungi, In vitro antifungal activity of the all synthesized compounds was carried out on PDA medium by using poisoned food technique  and Mancozeb 75% WP (A. solani) were taken as positive control. Percentage inhibition was calculated by Abott's formula  was calculated by given formulaWhere, I = Percentage inhibition, CF = (90-C)/C × 100, 90 is the diameter (mm) of the petriplate and C is the growth of the fungus (mm) in control.50 (mg L−1) values (Effective Dose for 50% inhibition) were calculated using SPSS statistical Package (v16.0).EDIn vitro nematicidal activity of the all synthesized compounds was carried by water screening method. The root galls were obtained from M. incognita infected plants collected from the glass house of Division of Nematology, ICAR-IARI, New Delhi and were incubated for 2–3 days at 25–30°C for hatching of eggs. After hatching, the number of J2s per 1 mL of aliquot were calculated and diluted to 100 J2s per mL. Synthesized ferrocenyl chalcones (12 mg) were weighed separately and dissolved in 0.5 mL of DMSO and 24.5 mL distilled water with Tween 80 (3.0%) to get a stock solution of 500 ppm. Five test concentrations viz. 125, 62.5, 31.25 15.62, and 7.81 mg L−1 were prepared from stock solution by serial dilution. 1 mL of aliquot (100 J2s/mL) was taken in 24 well culture plate to which 1 mL of test concentrations was added .LC50 value (ppm) was calculated for each compound with the help SPSS statistical Package (v16.0) using ASUS VivoBook laptop.50 (ppm) [pLC50 = –log (LC50)] as dependent variable and 2D descriptors  and test set (5 compounds) with the help of sphere exclusion method  and established the condensation of carbonyl moiety of acetylferrocene with carbonyl moiety of different benzaldehydes. In 13C-NMR, the peaks at δ 117.80–126.28 (C-α), 136.91–141.24 (C-β) for HC = CH and at δ 191.17–192.92 for CO were prominent for all the compounds. In all compounds, the higher chemical shifts values of H-α and C-α as compared to H-β and C-β was due to carbonyl moiety which polarizes the C = C double bond  as compared to conventional method (71–87%) . In MM, 3o i.e., (2E)-1-ferrocenyl-3--prop-2-en-1-one was found to be most active against S. rolfsii, having ED50 value 23.24 mg L−1 as compared to commercial fungicide Hexaconazole 5% SC (ED50 = 8.5 mg L−1) -1-ferrocenyl-3-(4-fluorophenyl)-prop-2-en-1-one, having value ED50 = 29.9 mg L−1 showed highest activity  . But in activity . The corM. incognita ((2E)-1-ferrocenyl- 3-(4-bromophenyl)-prop-2-en-1-one (3b) was more potent with LC50 values of 10.67 ppm as compared to commercial nematicide Carbofuran 3G (LC50 = 4.78 ppm) and Velum Prime 500 SC (LC50 = 2.40 ppm). It was found that mortality was directly proportional to time of exposure and concentration.Although all the synthesized compounds possessed nematicidal activity against ncognita , but theviz. Model-1 (PLS), Model-2 (MLR), and Model-3 (PCR) were developed in 2D-QSAR analysis of fungicidal activity against S. rolfsii and A. solani and nematicidal activity against M. incognita. Fitness plot of each model was used for validation of statistical significance of model 2 = 0.9997, q2 = 0.9962, F_test = 3036.53, r2_se = 0.0122, q2_se = 0.0464, pred_r2 = 0.6201, pred_r2se = 0.3985Where n = 11, DF = 8, rA. solaniModel-1 (PLS)2 = 0.9348, q2 = 0.7499, F_test = 50.2052, r2_se = 0.0727, q2_se = 0.2917, pred_r2 = 0.4856, pred_r2se = 0.2826Where n = 11, DF = 9, rM. incognitaModel-3(PCR)2 = 0.8784, q2 = 0.7728, F_test = 78.9078, r2_se = 0.0528, q2_se = 0.0722, pred_r2 = 0.4818, pred_r2se = 0.1703.Where n = 11, DF = 8, r2) with 100. The predictive power (q2) of models was determined by LOO (Left Out One) method. F denotes ratio of variance of models and that of error in regression. Models with higher F and lower SE of estimation viz. r2se and q2se were found statistically significant. External validation with pred_r2 > 0.3 confirmed the predictive power of QSAR model. The best model was determined by r2, q2, higher values of pred_r2F_test and , 3(atom with double bond), C (Carbon), N (Nitrogen), O (Oxygen), S (Sulfur), H (Hydrogen), F (Flourine), Cl (Chlorine), and Br (Bromine) with distance ranging from 0 to 7.Alignment Independent (AI) descriptors were calculated as explained in Baumann's paper based on type of atom, bond and molecular topology Balaban, . MaximumModel-2 (MLR) was found to be the best model for QSAR study molecules against S. rolfsii. The developed model indicated that descriptors, Nitrogens Count, Most+ve&–vePotential Distance, DeltaPsiA, and AI descriptor, T_T_C_3 were inversely related to the fungicidal activity (activity . The majModel-1 (PLS) was found to be the best model for QSAR study molecules against A. solani.The developed model indicated that descriptors, T_N_O_6, chiV2, SKMostHydrophobic HydrophilicDistance were inversely related to the fungicidal activity (activity . The majModel-3 (PCR) was found to be the best model for QSAR study molecules against M. incognita. The developed model indicated that descriptors, DeltaAlphaA positively influences the nematicidal activity of test compounds and SaaCHcount & SsOHE-index were negatively influences the activity (activity . These t3i) is new molecule which was characterized by various spectroscopic techniques. The compound 3o was found to be most active against S. rolfsii and the compound 3c showed highest activity against A. solani. The compounds 3b, 3m, 3o, 3f, and 3h were most active against root-knot nematode M. incognita. QSAR studies was performed to find quantitative relationship between fungicidal and nematicidal activity and physicochemical/structural properties of the synthesized chalcones. The developed models were evaluated and validated for their external predictive power and statistical significance. Among the three 2D-QSAR models , MLR, PLS, and PCR analysis showed significant predictive power and reliability for S. rolfsii, A. solani, and M. incognita, respectively. Information and understanding of descriptors affecting fungicidal and nematicidal activity of these ferrocenyl chalcones be used for structure optimization to attain higher activity.This is the first report of microwave assisted synthesis and bioefficacy  evaluations of ferrocenyl chalcones. Out of 16 synthesized compounds (2E)-1-ferrocenyl-3-(3-hydroxyphenyl)-prop-2-en-1-one (All datasets generated for this study are included in the article/NS: conceptualization, draft editing, and supervision. DY: methodology, data creation, analysis, and original draft writing. PK: methodology and analysis. P: methodology and nematicidal analysis. VR: draft editing and supervision. DKa, methodology and fungicidal analysis. DKh: analysis and funding.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Albizia adianthifolia are highly sought after in tropical Africa as herbal medicines. Therefore, the aim of this study was to review the botany, medicinal uses, phytochemistry, and pharmacological properties of A. adianthifolia so as to provide baseline data required for evaluating the therapeutic potential of the species. Information on the botanical profile, medicinal uses, phytochemistry, and pharmacological properties of A. adianthifolia was undertaken using databases such as ScienceDirect, SciFinder, Pubmed, Google Scholar, Medline, SCOPUS, EThOS, ProQuest, OATD, and Open-thesis. Preelectronic literature search of conference papers, scientific articles, books, book chapters, dissertations, and theses was carried out at the University library. Literature search revealed that A. adianthifolia is used as purgative and herbal medicine for diabetes, eye problems, gastrointestinal problems, haemorrhoids, headache, neurodegenerative disorders, reproductive problems in women, respiratory problems, wounds and pain, skin diseases, sexually transmitted infections, and ethnoveterinary medicine. Phytochemical compounds identified from the species include apocarotenoids, chalcone, dipeptide, elliptosides, essential oils, fatty acids, flavonoids, histamine, imidazolyl carboxylic acid, prosapogenins, steroids, triterpene saponins, and triterpenoids. Pharmacological studies revealed that A. adianthifolia extracts and compounds have acetylcholinesterase enzyme inhibitory, anthelmintic, antiamoebic, antibacterial, antimycobacterial, anti-sexually transmitted infections, antifungal, anti-inflammatory, antioxidant, anxiolytic, and antidepressant, cognitive-enhancing, haemolytic, hypoglycemic and antihyperglycemic, immunomodulatory, and cytotoxicity activities. Detailed studies on the pharmacokinetics, in vivo, and clinical research involving compounds isolated from A. adianthifolia and extracts of the species are required.The bark, leaves, and roots of Albizia adianthifolia (Schumach.) W. Wight is a medium to large tree  -3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, 3-O-{O-α-L-arabinopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-O-[β-d-glucopyranosyl-(1→2)]-β-d-glucopyranosyl}-21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 79, 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80, 21-O-{-6-{{3-O--β-d-quinovopyranosyl}oxy}-2,6-dimethylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 81, and 3-O-{O-α-L-arabinopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-O-[β-d-glucopyranosyl-(1→2)]-β-d-glucopyranosyl}-21-O-{-2,6-dimethyl-6-(β-d-quinovopyranosyloxy)octa-2,7-dienoyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 82 which have been shown to be cytotoxic against a large panel of cancer cells -3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, 3-O-{O-α-L-arabinopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-O-[β-d-glucopyranosyl-(1→2)]-β-d-glucopyranosyl}-21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 79, 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80, 21-O-{-6-{{3-O--β-d-quinovopyranosyl}oxy}-2,6-dimethylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 81 and 3-O-{O-α-L-arabinopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-O-[β-d-glucopyranosyl-(1→2)]-β-d-glucopyranosyl}-21-O-{-2,6-dimethyl-6-(β-d-quinovopyranosyloxy)octa-2,7-dienoyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 82, acacic acid 3-O-beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)-2-acetylamino-2-deoxy-beta-D-glucopyranoside 84, acacic acid 3-O-(beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)- [beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl)-21-O-(6(S)-2-hydroxymethyl-6-methyl-6-O-(beta-D-quinovopyranosyl)-2,7-octadienoyl) ester 85, and lupeol 86 which exhibited pharmacological properties -3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77 and 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78 exhibited activities with HD50 values of 17.5 μg/mL and 48 μg/mL, respectively [α-hydroxy-21β-[(2-hydroxybenzoyl)oxy]-3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, and 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80 isolated from the roots of A. adianthifolia using the haemolysis assay against sheep erythrocytes. The compounds 16α-hydroxy-21β-[(2-hydroxybenzoyl)oxy]-3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, and 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80 exhibited haemolytic activities with HC50 values ranging from 12.5 μg/mL to 36.6 μg/mL -beta-D-glucopyranosyl)-21-O-(6(S)-2- hydroxymethyl-6-methyl-6-O-(beta-D-quinovopyranosyl)-2,7-octadienoyl) ester 85, and 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78 isolated from the roots of A. adianthifolia using an in vitro lymphocyte proliferation assay. The cellular proliferation was measured by 3H-thymidine incorporation in Jurkat tumor cell lines (human T cell leukemia). The compounds acacic acid 3-O-beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)- 2-acetylamino-2-deoxy-beta-D-glucopyranoside 84 and acacic acid 3-O-(beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)- [beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl)-21-O-(6(S)-2- hydroxymethyl-6-methyl-6-O-(beta-D-quinovopyranosyl)-2,7-octadienoyl) ester 85 exhibited a dose-dependent immunomodulatory effect in the concentration range of 0.01 μM to 10 μM, whereas compound 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78 showed a lymphoproliferative activity in the concentration range of 0.01 μM to 10 μM -2,5-diphenyl2H-tetrazolium bromide (MTT) cellular viability assay on the human embryonic kidney epithelial  cell line. The cell viability assay indicated that the extracts were nontoxic at 100 mg/ml against the human kidney epithelial cell line, but 110% and 112% cell growth exhibited by aqueous and organic extracts, respectively, appeared to increase cellular activity, which would be effective in wound healing -beta-D-glucopyranosyl)-21-O-(6(S)-2- hydroxymethyl-6-methyl-6-O-(beta-D-quinovopyranosyl)-2,7-octadienoyl) ester 85, 16α-hydroxy-21β-[(2-hydroxybenzoyl)oxy]-3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, and 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80 isolated from the roots of A. adianthifolia on human leukemia T-cells (Jurkat cells) and on splenocytes. The compounds 16α-hydroxy-21β-[(2-hydroxybenzoyl)oxy]-3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77, 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78, and 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80 exhibited cytotoxic activities on Jurkat cells, while the compounds acacic acid 3-O-beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)-2-acetylamino-2-deoxy-beta-D-glucopyranoside 84 and acacic acid 3-O-(beta-D-xylopyranosyl-(1-->2)-beta-D-fucopyranosyl-(1-->6)- [beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl)-21-O-(6(S)-2- hydroxymethyl-6-methyl-6-O-(beta-D-quinovopyranosyl)-2,7-octadienoyl) ester 85 exhibited lymphoproliferative activities on this cell type. Cytotoxic activity on Jurkat cells was observed at 10-1 μM and 1 μM for compound 21-O-{-6-{{4-O--β-d-quinovopyranosyl}oxy}-2-(hydroxymethyl)-6-methylocta-2,7-dienoyl}-3-O-{O-β-D-xylopyranosyl-(1→2)-O-β-d-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-d-glucopyranosyl}acacic acid 28-{O-α-L-arabinofuranosyl-(1→4)-O-[β-d-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl} ester 80 and at 1 μM for compounds 16α-hydroxy-21β-[(2-hydroxybenzoyl)oxy]-3β-[(O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)-2-(acetylamino)-2-deoxy-β-D-glucopyranosyl)oxy]olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)-O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl ester 77 and 16α-hydroxy-21β-[(2-hydroxybenzoyl)-oxy]-3β-[(O-β-D-glucopyranosyl-(1→2)-O-[O-β-D-xylopyranosyl-(1→2)-O-β-D-fucopyranosyl-(1→6)]-β-D-glucopyranosyl)oxy]-olean-12-en-28-oic acid 28-O-α-L-arabinofuranosyl-(1→4)O-[β-D-glucopyranosyl-(1→3)]-O-α-L-rhamnopyranosyl-(1→2)β-D-glucopyranosyl ester 78 [Gengan et al. (2013)  evaluatephocytes . Govendephocytes  evaluatec/DIABLO . The sild PARP-1 . Haddad d PARP-1  evaluateester 78 .Albizia adianthifolia has been used as herbal medicine in tropical Africa for several centuries and significant breakthrough has been made in the last 40 years elucidating the phytochemical and pharmacological properties of the species. However, there are still some research gaps regarding correlating the ethnomedicinal applications of the species with the chemical compounds and pharmacological properties of the compounds and extracts of the species. Detailed studies on the pharmacokinetics, in vivo, and clinical research involving compounds isolated from A. adianthifolia and extracts of the species are required. The bark of A. adianthifolia is known to be toxic [ A. adianthifolia is used as a hunting poison and in the Central African Republic, the bark and leaves of the species are used as fish poison [ A. adianthifolia to establish the toxicity and/or any side effects that can arise when the species and its products are used as herbal medicines.be toxic  and rootbe toxic . Similarh poison . These r


Compounds 5–8 were isolated for the first time in the genus Rauia. These compounds were characterized on the basis of their spectral data, mainly one and two-dimensional NMR, and mass spectra, also involving comparison with the literature data. Theoretical studies at the DFT level reveal structural parameters for the 1,2,3-trioxole bridge compatible with known structures containing a similar group.A new natural product, a 3,9--type steroid, named rauianodoxy ( Pilocarpus genus that synthesizes large concentrations of the alkaloid pilocarpine used to treat glaucoma , seven methylene  and six methyl [(CH3) × 6] carbon atoms, allowing to deduce the presence of C28H44O2, HRESIMS (positive mode) revealed [M+H]+ at m/z 429.3384  and [M+Na]+ at m/z 451.3235  compatible with the molecular formulae C28H44O3 ]/CH-12 [δC 130.8/δH 6.53 ] and CH-22 [δC 135.2/δH 5.17 ]/CH-23 [δC 132.2/δH 5.23 ] in an stigmasterol skeleton  with 3H-18 (δH 0.84), CH-22 (δC 135.2) with 3H-21 (δH 1.02) and CH-23 (δC 132.2) with 3H-28 (δH 0.92)] confirming the presence of two double bonds in the steroidal-skeleton 6, as shown in Rauianodoxy  in the HMBC spectrum] and 82.2  and the molecular formulae C28H44O3 (two double bonds and pentacyclic skeleton) induced us inevitably to postulate the presence of the O-O-O (O3) group and to classify the new compound as a 1,2,3-trioxocine-type steroid 6  was determined from the coupling constants of relevant hydrogen atoms and from the observed 1H-1H-NOESY, which showed cross-peaks assigned to dipolar interaction , [M+Na]+ at m/z 451.3235 (C28H44NaO3), and the adduct [2M+Na]+ at m/z 879.6479 , was also useful in confirming the new steroidal 3,9--type structure mainly due to fragment with m/z 395.3353 , formed by elimination of a molecule of hydrogen peroxide (H2O2). This fragment was the precursor for the formation of fragment with m/z 377.3225 , a result of the loss of a water molecule (H2O)  . AlternaA few natural ozonides  as precursors in ozonolysis of olefins, have been discovered in higher plant species ,17,18. H6 was mainly characterized by one and two-dimensional NMR and mass spectra, but theoretical studies at the DFT level have also been used. A model was performed to access structural details of the suggested structure and to verify the stability of the 1,2,3-trioxocine bridge connecting two atoms, C3 and C9, spatially not very close to each other in compound 6. The modeling results revealed that the proposed bridge is in fact stable, with bonding distances comparable to those of known compounds (models) containing similar bridges. In the optimized LSDA/pBP86/DN* compound 6 structure, the 1,2,3-trioxocine bridge is part of two adjacent eight-membered rings which adopt two different conformations: a distorted chair-like conformation and a distorted boat-like conformation  and O(3) atoms are attached, C(3) and C(9), which are part of the rigid steroid ring system, and also to 1,3-diaxial steric interactions between the O(1) and O(3) atoms with H atoms. Accordingly, in a simplified model of compound 6, containing only rings A and B -O(2)-O(3) angle is 112.57°, a value compatible with a sp3 character for the oxygen atoms. Thus, the structure of the new 3,9--type steroid isolated from Rauia nodosa was here established for the first time as a novel natural product and named rauianodoxy (6).The 1,2,3-trioxocine bridge is not symmetrical in the steroid derivative, with the O(1)-O(2) distance is equal to 1.507 Å and the O(2)-O(3) distance is equal to 1.471 Å. The corresponding bond distances were almost identical in the model compound 1H- and 13C-NMR spectra were measured on a Brüker Ultrashield 500 Plus spectrometer , operating at 500 (1H) and 125 (13C) MHz. CDCl3 was used as solvent with TMS as internal reference. Chemical shifts are given in the δ scale (ppm) and coupling constants (J) in Hz. One dimensional (1D) 1H- and 13C-NMR spectra were acquired under standard conditions by using a direct detection 5 mm 1H/13C dual probe. Standard pulse sequences were used for two dimensional spectra by using a multinuclear inverse detection 5 mm probe with field gradient.ESI-MS (high resolution) mass spectra were obtained by using a ESI-TOF-MS Shimadzu mass spectrometer , using the positive ion mode of analysis. Chromatographic purifications were carried out by using silica gel 60 (0.063–0.200 mm). i.e., non-hydrogen) atoms, which adds some flexibility within the basis set.A theoretical study was implemented to verify the viability of the molecular structure containing the 1,2,3-trioxocine bridge. The Spartan’10  program was used in all steps of the theoretical study. The structure was initially submitted to a Monte Carlo conformational search with the MMFF94 molecular mechanics method . This prRauia nodosa (Rutaceae) were collected in May 2011 at Vale Cia Reserve, Linhares, Espírito Santo, Brazil. The voucher specimen of R. nodosa was deposited at Vale Cia herbarium, under the code CRVD-3301.Stems of Rauia nodosa (Rutaceae), immediately after collection, were dried at room temperature until a constant weight (about one week) and ground in hammer mills. The dried and powdered material (4.8 kg) was extracted with hexane. The obtained solutions (48 L) were distilled under reduced pressure in a rotary evaporator furnishing 4.47 g of crude hexane extract.Stems of 1+2 (5.1 mg). Fraction 9.5 (260.9 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate furnishing seven fractions and 3+4 (11.5 mg) and the fraction 9.5.6 (55.6 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate yielding compound 7 (29.8 mg).The hexane extract was chromatographed over silica gel column with a gradient of hexane/ethyl acetate to afford twenty one fractions. Fraction 9 (835.6 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate furnishing six fractions. Fraction 9.2 (62.8 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate affording the compounds 5 (5.3 mg). Fraction 10 (292.6 mg) was chromatographed over a silica gel column with a gradient of hexane/ethyl acetate furnishing eight fractions. Fraction 10.8 (94.6 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate yielding compound 6 (4.7 mg). Fraction 19 (126.0 mg) was chromatographed over a silica gel column with a gradient of hexane/ethyl acetate affording eight fractions. The fraction 19.5 was chromatographed over a silica gel column with a gradient of hexane/ethyl acetate the provide compounds 8+9 (8.8 mg) and 10 (3.7 mg).Fraction 16 (227.2 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate affording eleven fractions and the fraction 16.7 (53.2 mg) was rechromatographed over a silica gel column with a gradient of hexane/ethyl acetate yielding compound Rauianodoxy (6).  = −10.2,  and obtained as a yellow oil; HRESIMS (positive mode) m/z 429.3384  and m/z 451.3235 ; 1H- and 13C-NMR data, see Ergosterol 5,8-peroxide (5). 1H-NMR δH (ppm): 1.97–2.0, 1.71–1.73 , 1.86–1.90, 1.50–1.52 , 4.01 , 2.15–2.17, 1.90–1.97 , 6.53 , 6.27 , 1.5 , 1.63–1.67, 1.42–1.45 , 1.98–2.2, 1.23–1.27 , 1.58 , 1.3–1.35 , 1.76–1.80 , 1.23–1.24 , 0.84 , 0.91 , 2.05–2.15 , 1.02 , 5.16 , 5.24 , 1.87–1.90 , 1.47–1.49 , 0.85 , 0.83 , 0.93 ; 13C-NMR δC (ppm): 34.7 (CH2-1), 30.1 (CH2-2), 66.4 (CH-3), 36.9 (CH2-4), 81.0 (C-5), 130.8 (CH-6), 135.4 (CH-7), 51.1 (CH-9), 36.6 (C-10), 20.6 (CH2-11), 39.8 (CH2-12), 44.6 (C-13), 51.7 (CH-14), 20.6 (CH2-15), 29.3 (CH2-16), 56.2 (CH-17), 12.9 (CH3-18), 17.6 (CH3-19), 39.8 (CH-20), 20.9 (CH3-21), 135.2 (CH-22), 132.3 (CH-23), 42.8 (CH-24), 33.1 (CH-25), 20.0 (CH3-26), 19.7 (CH3-27), 18.2 (CH3-28).O-geranylosthenol (7). 1H-NMR δH (ppm): 6.24 , 7.62 , 7.29 , 6.84 , 4.66 , 5.51 , 1.73 , 2.11-2.14 , 2.14–2.19 , 5.10 , 1.59 , 1.69 , 3.57 , 5.27 , 1.90  ,1.67 ; 13C-NMR δC (ppm): 161.5 (C-2), 112.9 (CH-3), 143.8 (CH-4), 125.0 (CH-5), 108.6 (CH-6), 159.63 (C-7), 118.3 (C-8), 152.9 (C-9), 113.3 (C-10), 65.7 (CH2-1'), 119.2 (CH-2'), 141.5 (C-3'), 39.5 (CH2-4'), 26.3 (CH2-5'), 123.7 (CH-6'), 132.4 (C-7'), 17.7 (CH3-8'), 25.8 (CH3-9'), 22.1 (CH2-1''), 121.3 (CH-2''), 131.9 (C-3''),18.0 (CH3-4''), 25.7 (CH3-5''), 16.7 (CH3-3'a).N-methylflindersine (8). 1H-NMR δH (ppm): 8.00 , 7.26 , 7.58 , 7.35 , 6.79 , 5.57 , 1.57 , 3.73 ; 13C-NMR δC (ppm): 122.9 (CH-5), 121.7 (CH-6), 130.8 (CH-7), 113.7 (CH-8), 117.5 (CH-1'), 126.3 (CH-2'), 28.0 (CH3-4'/5'), 29.2 (CH3-N).Zantobungeanine (9). 1H-NMR δH (ppm): 7.63 , 7.17 , 7.08 , 6.78 , 5.56 , 1.53 , 3.93 , 3.96 ; 13C-NMR δC (ppm): 115.6 (CH-5), 122.3 (CH-6), 114.1 (CH-7), 117.8 (CH-1'), 126.4 (CH-2'), 78.5 (C-3'), 28.0 (CH3-4'/5'), 56.3 (CH3O-8), 35.0 (CH3-N).Veprissine (10). 1H-NMR δH (ppm): 7.73 , 6.90 , 6.75 , 5.51 , 1.52 , 3.99 , 3.80 , 3.96 ; 13C-NMR δC (ppm): 162.6 (C-2), 104.0 (C-3), 155.1 (C-4), 119.1 (CH-5), 107.2 (CH-6), 155.6 (C-7), 136.0 (C-8), 134.6 (C-9), 112.1 (C-10), 118.0 (CH-1'), 125.6 (CH-2'), 78.7 (C-3'), 28.0 (CH3-4'/5'), 56.3 (CH3O-7), 61.7 (CH3O-8), 33.6 (CH3-N).Rauia nodosa provided six steroids 1–6, one coumarin 7 and three alkaloids 8–10 isolated in a previous phytochemical investigation. The compounds 5–8 were isolated for first time in the genus Rauia. There are no published records yet of other steroids possessing 1,2,3-trioxocine group yet. Then, the 3,9--type steroid, named rauianodoxy (6) was established as novel natural product and consequently described for the first time on in the literature.The hexane extract from the stems of


The chemical structures were determined by spectroscopic methods, including HR-EIMS, 1D NMR , 2D NMR , and IR spectroscopy. All isolated compounds were tested for the ability to inhibit LPS-induced nitric oxide production in RAW264.7 macrophages.A new lignan, named acanthosessilin A ( Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The bark and twigs of Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory, and anti-diabetic drugs and are recognized to have ginseng-like activities , confirming 1 to be a lignan. The multiplicity of each carbon was determined using a DEPT experiment. In the aromatic region, six olefin methine carbon signals , two carbonated quaternary carbon signals [δC 135.7 (C-1) and 133.5 (C-1')] and four oxygenated quaternary carbon signals  due to the 1,3,5-tri- and 1,2,4-trisubstituted benzene moieties were observed. The oxygenated methine carbon signal at δC 84.0 (C-7) shifted downfield due to attached to heteroatom (–OH). Also, two oxygenated methylene carbon signals [δC 73.4 (C-9') and 60.4 (C-9)] and two methoxy carbon signals [δC 56.3 ] were observed. In the high magnetic field, two methine carbon signals [δC 54.0 (C-8) and 43.8 (C-8')] and a methylene carbon signal [δC 33.6 (C-7')] were observed. With further analysis of the HSQC and DEPT 135 spectra of 1, the proton and carbon NMR signals could be assigned (1H−1H COSY spectrum indicated key connectives of H-8 (δH 2.34) with H-7 (δH 4.74), H-8' (δH 2.72), H-9a (δH 3.97), and H-9b (δH 3.71) and H-8' (δH 2.72) with H-7'b (δH 2.48), H-9'a (δH 3.82), and H-9'b (δH 3.62) (H 4.74) and C-2 (δC 116.1), C-6 (δC 115.9), and C-9 (δC 60.4) and between H-7'  and C-1' (δC 133.5), C-2' (δC 113.3), and C-6' (δC 122.1)  and the oxygenated quaternary carbon signals of C-3, 5 (δc 149.0) were also identified. The relative stereochemistry of H-8 and H-8' 1 was identified as trans from the lack of NOE effect between H-8 and H-8'. The coupling constant of 6.8 Hz between H-7 and H-8, as well as the optical rotation of 1 ([α]25D = −43.5°) suggested an S configuration at C-7 -3-(4-hydoxy-3-methoxybenzyl)tetrahydrofuran ([α]20D = −49°), supported the above, as reported in the literature [13C-NMR spectra  and NOESY experiment of 1 was very similar to (+) tripterygiol except for the optical rotation ([α]25D = +48.3°) and were comparable to the epi-THF lignan tetrahydrofuran, and named acanthosessilin A. Comparisons of NMR and MS data for the known compounds 2–9 with reported values led to their identification as (−)-sesamin (2)  was subjected to the silica gel CC  to give compound 2 . Subfraction E1-17  was separated by CC  to give compound 3 . Fraction E3  was subjected to the silica gel CC  to give five subfractions (E3-1 to E3-5). CC  of subfraction E3-3  gave 19 subfractions (E3-3-1 to E3-3-19). Subfraction E3-3-13  was separated by CC  to give compound 1 . Fraction E8  was fractionated using silica gel CC  and yielded nine subfractions (E8-1 to E8-9). Subfraction E8-4  was purified using CC  to give compound 4 . Subfraction E8-5  was fractionated using a Sephadex LH 20 CC  and yielded five subfractions (E8-5-1 to E8-5-5). Purification of subfraction E8-5-4  using CC  yielded compound 8 . Subfraction E8-5-5  was separated by CC  to give compound 9 . Fraction E9  was fractionated using silica gel CC  and yielded four subfractions (E9-1 to E9-4). Subfraction E9-4  was chromatographed over RP-18 (5 × 5.5 cm) and eluted with MeOH−H2O  to give twenty subfractions (E9-4-1 to E-9-4-20). Subfraction E9-4-1  was purified over silica gel CC (4 × 12 cm) and eluted with CHCl3−MeOH−H2O  to give compound 10 . Subfraction E9-4-6  was separated by CC  to give compound 6  and compound 7 .The air-dried fruits of Acanthosessilin A (1). Colorless crystals, m.p.: 123−125 °C; [α]25D −43.5° ; CD  λmax nm (∆ε): −0.42 (217), −0.49 (236); UV λmax (MeOH) nm: 280; IR (CaF2 window) cm−1: 3430, 1648, 1512, 1245; EIMS m/z: 360 [M]+; HR-EIMS m/z: 360.1552 [M]+ ; 1H- and 13C-NMR data, see 4 cells/well) for measurement of NO production. The plates were pretreated with various concentrations of samples for 30 min and incubated with LPS (1 μg/mL) for 24 h. The amount of NO was determined by the nitrite concentration in cultured RAW264.7 macrophage supernatants using the Griess reagent. The cell viability was evaluated by MTT reduction.Assays for NO production and cell viability were carried out as previously described . BrieflyS),7-hydroxy-3,5-dimethoxybenzyl]tetrahydrofuran, named acanthosessilin A (1), was isolated from Acanthopanax sessiliflorus, together with eight known lignans. According to previous investigations on Acanthopanax species, we have evaluated the inhibitory activities of all compounds against LPS-induced NO production in RAW264.7 macrophages. All compounds moderately inhibited NO production with IC50 values in the range of 10.34 to 65.07 μM. The results provide a potential explanation for the use of this plant as a herbal medicine in the treatment of inflammatory diseases, and they may be potentially useful in developing new anti-inflammatory agents.The new compound 3--4-[(7


The compounds were effective against the Trypanosomatidae Leishmania amazonensis and Leishmania chagasi and Trypanosoma cruzi, etiologic agents of leishmaniasis and Chagas’ disease, respectively. The quinonemethide triterpenes 1 and 2 exhibited a marked in vitro leishmanicidal activity against promastigotes and amastigotes with 50% inhibitory concentration (IC50) values of less than 0.88 nM. Both compounds showed IC50 lower than 0.3 nM against Trypanosoma cruzi epimastigotes. The selectivity indexes (SI) based on BALB/c macrophages for L. amazonensis and L. chagasi were 243.65 and 46.61 for (1) and 193.63 and 23.85 for (2) indicating that both compounds presented high selectivity for Leishmania sp. The data here presented suggests that these compounds should be considered in the development of new and more potent drugs for the treatment of leishmaniasis and Chagas’ disease.The present study describes the leishmanicidal and trypanocidal activities of two quinonemethide triterpenes, maytenin ( Trypanosoma cruzi (the parasite that causes Chagas’ disease), mostly in Latin America + 421.30 , 1H-NMR : δ = 0.92 , 0.95 , 1.28 , 0.94 , 2.84 , 1.45 ,1.79 , 2.16 , 6.31 , 6.48 , 6.97 . 13C-NMR : δ = 10.24 (C-23), 15.06 (C-30), 19.70 (C-27), 21.54 (C-26), 32.05 (C-19), 28.50 (C-15), 29.93 (C-12), 38.17 (C-17), 40.62 (C-20), 33.78 (C-11), 32.54 (C-28), 35.50 (C-16), 39.03 (C-25), 40.62 (C-13), 42.70 (C-9), 43.53 (C-18), 52.52 (C-22), 44.64 (C-14), 213.54 (C-21), 117.15 (C-4), 118.12 (C-7), 119.77 (C-1), 127.74 (C-5), 133.57 (C-6), 146.08 (C-3), 164.69 (C-10), 168.64 (C-8), 178.40 (C-2).Pristimerin (2). Orange-red amorphous solid; EIMS m/z [M+1]+ 465.30 , 1H-NMR : δ = 0.47 , 1.11 , 1.19 , 1.03 , 1.38 , 2.14 , 6.28 , 6.47 , 6.94 , 3.48 . 13C-NMR : δ = 10.22 (C-23), 32.36 (C-30), 18.32 (C-27), 21.62 (C-26), 30.90 (C-19), 28.65 (C-15), 29.66 (C-12), 30.53 (C-17), 40.41 (C-20), 33.57 (C-11), 31.58 (C-28), 36.39 (C-16), 38.25 (C-25), 39.42 (C-13), 42.93 (C-9), 44.32 (C-18), 34.80 (C-22), 45.04 (C-14), 29.88 (C-21), 117.10 (C-4), 118.12 (C-7), 119.56 (C-1), 127.43 (C-5), 134.02 (C-6), 146.02 (C-3), 164.76 (C-10), 170.03 (C-8), 178.34 (C-2), 51.53 (CH3O).L. amazonensis MPRO/BR/1972/M1841-LV-79, L. chagasi MHOM/BR/1974/PP75 strains and epimastigotes of T. cruzi Y strain were maintained at 28 °C in liver-infusion Tryptose (LIT) supplemented with 10% fetal calf serum (FCS).Promastigotes of 6 parasites/mL in 96 well flat-bottom plates (Costar®). Maytenin and pristimerin were dissolved in DMSO , added to parasite suspension to final concentrations between 1.6 µg/mL and 100 µg/mL and incubated at 28 °C for 72 h. The assays were carried out in triplicate. Pentamidine isethionate purchased from Sigma-Aldrich  was used as reference drug (a 16.7 mg/mL stock solution was prepared in sterile deionized water) and added to parasite suspension to final concentrations between 1.6 and 100 µg/mL. Leishmanicidal effects were assessed by 3--2,5-diphenyltetrasodium bromide (MTT) method with modifications [50).Cultured promastigotes at the end of the exponential growth phase were seeded at 8 × 10ications . Absorba®) at a density of 1 × 105 cells/well (100 µL/well) in RPMI 1640 medium supplemented with 10% heat inactivated fetal calf serum, 25 mM HEPES and 2 mM L-glutamine and incubated for 24 h at 37 °C in a 5% CO2-air mixture. The medium was then removed and the cells were treated with different concentrations of quinonemethides triterpenes. Positive (with pentamidine) and negative (without drugs) controls were included in each experiment. The plates were incubated under the same conditions for 24 h. Subsequently, the MTT colorimetric assay was carried out as described above. Absorbance was read in a 96-well plate reader (Robonik®) at 595 nm. The 50% lethal concentration (LC50) was determined by logarithm regression analysis of the obtained data. The cytotoxicity for macrophages and for promastigotes was compared using the selectivity index (SI), which was determined as the ratio between LC50 for macrophages and IC50 for promastigotes.The cytotoxicity was evaluated on mouse peritoneal macrophages. Peritoneal macrophages were collected from BALB/c mice and were seeded in 96 well flat-bottom plates (Costar5 cells/well on coverslips (13 mm diameter) previously arranged in a 24-well plate in RPMI 1640 medium supplemented with 10% inactivated FCS, and allowed to adhere for 4 h at 37 °C in 5% CO2. Adherent macrophages were infected with promastigotes in the stationary growth phase using a ratio of 5:1 (parasites:cell) at 37 °C in 5% CO2 for 4 h. After that time, the non-internalized parasites were removed by washing, and infected cultures were incubated in RPMI medium or treated with different concentrations of quinonemethide triterpenes for 24 h. The cells were then fixed in a methanol solution and stained with Giemsa. The infection index was determined by multiplying the percentage of infected macrophages by the mean number of amastigotes per infected cell. The concentration that caused a 50% decrease in the infection index compared to the control was determined by regression analysis of the log-transformed data. All experiments involving mice were performed as indicated by the ethical committee of Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara (protocol number CEUA/FCF/Car n°24/2012).Peritoneal macrophages from BALB/c mice were plated at 3 × 10T. cruzi Y strain epimastigote forms. The parasites were grown axenically at 28 °C in LIT medium supplemented with 10% fetal calf serum and harvested during the exponential growth phase (7-day-old culture forms). It was added 1 × 107 parasites/mL to each well of a 96-well microplate and the same volume of LIT medium was used as control. Compounds were dissolved in dimethyl sulfoxide (DMSO) and further added to the microplate for final concentrations from 1 μg/mL to 100 μg/mL. The plates were maintained at 28 °C for 72 h and MTT was performed as described above. MTT assay was prepared as described elsewhere [50 and was determined from sigmoidal regression of the concentration-response curves after 72 h of incubation (Benznidazole was employed as positive control). For the statistical analysis Probit´s method was employed [All experiments were performed with lsewhere . Absorbaemployed .1 and 2 should be further considered in the development of new and more potent drugs for the treatment of leishmaniasis and Chagas’ disease.The results herein presented suggest that the quinonemethide triterpenes