Paullinia pinnata L. are used in West Africa as traditional remedies for a variety of diseases including infestations with soil-transmitted helminths. Based on the results of an ethnopharmacological survey in Ghana, an aqueous acetone (70%) extract was investigated for its anthelmintic and phytochemical properties. Partitioning of the crude extract followed by several fractionation steps of the ethyl acetate phase using Sephadex® LH-20, fast centrifugal partition chromatography, RP-18-MPLC and HPLC led to isolation of six oligomeric A-type procyanidins (1 to 6). To determine the anthelmintic activity, the crude extract, fractions and isolated compounds were tested in vitro against the model organism Caenorhabditis elegans. A significantly better activity was observed for the trimeric A-type procyanidin (1) compared to a B-type trimer. However, this effect could not be generalized for the tetrameric procyanidins, for which the type of the interflavan-linkage (4→6 vs. 4→8) had the greatest impact on the bioactivity. Besides the procyanidins, three novel compounds, isofraxidin-7-O-α-l-rhamnopyranosyl-(1″→6′)-β-d-glucopyranoside (17), 4-methoxycatechol-2-O-(5′′-O-vanilloyl-β-apiofuranosyl)-(1′′→2′)-β-glucopyranoside (18) and a 6-(3-methoxy-4-hydroxyphenyl)-hexane-2,4-diol-2-O-hexoside (19) were isolated together with further ten known compounds (7 to 16), mainly coumarins and coumarinolignans. Except for 3-β-d-glucopyranosyloxy-4-methyl-2(5H)-furanone (15), none of the isolated compounds has previously been described for P. pinnata. The anthelmintic activity was attributed to the presence of procyanidins, but not to any of the other compound classes. In summary, the findings rationalize the traditional use of P. pinnata root extracts as anthelmintic remedies.Extracts from the roots of  Paullinia pinnata L. (Sapindaceae) is a woody climber growing in tropical regions worldwide. In West Africa and Tanzania, preparations from different parts of the plant have traditionally been used as remedies for the treatment of various diseases, such as dysenterie, nausea, bacterial and parasitic infections, wounds, infertility, cancer and neurological disorders −), unlike m/z 863.2 as measured for compounds 1 and 2. Characteristic fragments of m/z 285.0 were observed, resulting from a quinone methide cleavage of the doubly linked top unit + ion at m/z 565.1615 corresponding to C26H33O15. A fragment of m/z 169.0506 pointed towards a methoxylated hydroxybenzoic acid (C8H9O4) which had been attached to a pentose moiety (m/z 283.0842). 1H NMR data revealed two AMX spin systems . Two singlets  could be assigned to two methoxy groups attached to C-3′′′ and C-4 of the aromatic rings resp. via HMBC data. Two methylene groups and a TOCSY spectrum showing a spin system of seven protons only for the hexose moiety strongly suggested an apiofuranosyl moiety as the pentose residue which was attached to C-2′ of the hexosyl group and esterified with vanillic acid at position C-5′′  to δc 152.42 (C-2) revealed the linkage position of the hexosyl group to the methoxycatechol. Complete assignments of 1H and 13C NMR signals are given in 18 is thus tentatively identified as 4-methoxycatechol-2-O-(5′′-O-vanilloyl-β-apiofuranosyl)-(1′′→2′)-β-glucopyranoside and represents a novel natural substance.Further signals of the disaccharide were assigned by HMBC, COSY and TOCSY experiments. Coupling constants of 19  fragmenting to m/z 241.1457 [M—hexose] + and m/z 163.0752, seemed to be a glycosylated hydroxyphenolic compound. Again, a three-proton singlet  showing a long-range correlation to an aromatic ring as indicated by an AMX spin system  pointed to the presence of an aromatic methoxy group. A similar chemical shift in the 13C spectrum as observed for the methoxylated aromatic carbon (C-3′) and the absence of a signal in the HSQC spectrum suggested an additional aromatic hydroxy group at position C-4′. The HSCQ spectrum also showed the presence of four methylene groups in total, three of which appeared upfield as part of a hexyl chain and one further downfield within the carbohydrate moiety as determined via COSY, HSQC and HMBC data. The absence of a methylene group and a downfield shift of C-4 (δ 68.30) and C-2 (δ 76.09) indicated the presence of a hydroxyl group at these positions. Additionally, an HMBC correlation from the anomeric proton of the hexosyl residue to C-2 supports an O-glycosylation at this site. Due to the small amount of substance available, the identitiy of the hexosyl moiety as well as stereochemical properties of the compound could not be further determined. Based on the data obtained, 19 was tentatively identified as 6-(3-methoxy-4-hydroxyphenyl)-hexane-2,4-diol-2-O-hexoside and also represents a novel compound. Assignments of 1H and 13C NMR signals are given in Finally, 50 1.1 mg/mL) than the crude extract (LC50 1.9 mg/mL) and the aqueous partition (LC50 2.9 mg/mL). The respective dose-response curves are shown in ® LH20 pointed to oligomeric PACs as the main active compounds. A mortality rate of at least 70% was observed for all PAC containing fractions at 1 mg/mL while the activity did not vary significantly among these fractions.Regarding the bioactivity, the EtOAc partition showed much stronger anthelmintic effects (LC1) representing the major A-type trimer was assayed in direct comparison to procyanidin C1 (epicatechin-(4β→8)-epicatechin-(4β→8)-epicatechin), a B-type trimer previously isolated within another study  (Δε): 210 (−37.48), 230 (+18.62), 270 (−2.25). ESITOFMS m/z 863.1813 [M − H]− . 1H and 13C NMR see Cinnamtannin B1 (2), epicatechin--ent-catechin-(4β→8)-epicatechin): off-white amorphous powder. CD (MeOH) λmax [nm] (Δε): 208 (−34.56), 234 (+37.16), 273 (−1.72). ESITOFMS m/z 863.1860 [M − H]− . 1H and 13C NMR see Aesculitannin B (3), epicatechin-(4β→6)-epicatechin--epicatechin-(4β→8)-epi- catechin. Light brown amorphous powder. CD (MeOH) λmax [nm] (Δε): 208 (−114.87), 234 (+37.16), 278 (−8.85). ESITOFMS m/z 1153.2819 [M + H]+ . 1H and 13C NMR after peracetylation (3a) see Pavetannin C1 -epicatechin-(4β→8)-epicatechin (4), off-white amorphous powder. CD (MeOH) λmax [nm] (Δε): 205 (−29.70), 234 (+33.51), 274 (−5.05). ESITOFMS m/z 1151.2468 [M − H]− . 1H and 13C NMR see Epicatechin-(4β→8)-epicatechin-(2β→5), epicatechin--[epicatechin-(4β→6)]-epicatechin- (4β→8)-epicatechin): off-white amorphous powder. CD (MeOH) λmax [nm] (Δε): 212 (−33.14), 235 (+22.77), 279 (−2.06). ESITOFMS m/z 1151.2605 [M − H]− . 1H and 13C NMR see Parameritannin A-1 -epicatechin--epicatechin (6), off-white amorphous powder. ESITOFMS m/z 861.1719 [M − H]− . 1H NMR (methanol-d4) δ: 7.43 ), 7.41 ), 7.17 ), 7.09 ), 7.03 ), 6.98 ), 6.88 , 6.82 ), 6.78 ), 6.16 ), 6.09 ), 6.06 ), 6.02 ), 4.68 ), 4.53 ), 4.26 ), 4.10 ), 3.98 ), 2.96 ), 2.82 ).Epicatechin-(2β→7), brownish yellow amorphous powder. UV  λmax 220, 324. ESITOFMS m/z 163.0400 [M + H]+ . 1H NMR (chloroform-d) δ: 7.63 , 7.36 , 6.80 , 6.78 , 6.26 . Data were identical to those of a reference substance .Umbelliferone (8), yellow amorphous powder. UV  λmax 203, 227, 296s, 344. ESITOFMS m/z 193.0490 [M + H]+ . 1H NMR (chloroform-d) δ: 7.60 , 6.92 ), 6.85 , 6.27 , 6.12 , 3.96 . 13C NMR (chloroform-d) δ: 161.51 (C-2), 150.32 (C-7), 149.87 (C-9), 144.06 (C-6), 143.25 (C-4), 113.42 (C-3), 111.60 (C-10), 107.44 (C-5), 103.27 (C-8), 56.43 (6-OCH3).Scopoletin (9), light brown amorphous powder. UV  λmax 205, 230, 280, 311. ESITOFMS m/z 139.0378 [M + H]+ . 1H NMR (methanol-d4) δ: 9.67 , 7.30 , 7.29 , 6.89 . 13C NMR (methanol-d4) δ: 193.00 (CHO), 154.54 (C-4), 147.40 (C-3), 130.45 (C-1), 126.63 (C-6), 116.30 (C-5), 115.09 (C-2).Protocatechuic aldehyde (10), pale yellow amorphous powder. UV  λmax 206, 235s, 283s, 328. ESITOFMS m/z 387.1072 [M + H]+ . 1H NMR (acetone-d6) δ: 7.88 , 7.81 , 7.16 , 7.01 , 6.91 , 6.84 , 6.27 , 5.10 , 4.25 , 3.89  3.88 , 3.84 , 3.57 . 13C NMR (acetone-d6) δ: 160.93 (C-2), 148.63 (C-3′), 148.38 (C-4′), 146.97 (C-6), 145.22 (C-4), 139.77 (C-9), 138.72 (C-7), 133.29 (C-8), 128.51 (C-1′), 121.93 (C-6′), 115.96 (C-5′), 114.42 (C-3), 112.53 (C-10), 112.29 (C-2′), 101.79 (C-5), 79.60 (C-8′), 77.64 (C-7′), 61.55 (C-9′), 56.60 (3‘-OCH3), 56.46 (6-OCH3).Cleomiscosin A (11), pale yellow amorphous powder. UV  λmax 210, 235s, 327. ESITOFMS m/z 417.1189 [M + H]+ . 1H NMR (acetone-d6) δ: 7.88 , 7.43 , 6.87 , 6.85 , 6.27 , 5.08 , 4.37 , 4.28 , 3.90 , 3.86 , 3.84 , 3.58 . 13C NMR (acetone-d6) δ: 160.96 (C-2), 149.00 (C-3‘/5‘), 147.01 (C-6), 145.25 (C-4), 139.75 (C-9), 138.76 (C-7), 138.62 (C-8) 137.76 (C-4‘), 133.37 (C-8), 114.49 (C-3), 112.50 (C-10), 127.35 (C-1‘), 106.57 (C-2‘/6‘), 101.84 (C-5), 79.59 (C-8‘), 77.96 (C-7‘), 61.62 (C-9‘), 56.88 (3‘/5‘-OCH3), 56.66 (6-OCH3).Cleomiscosin C (12), pale yellow amorphous powder. UV  λmax 204, 235s, 280s, 328. ESITOFMS m/z 387.1080 [M + H]+ . 1H NMR (acetone-d6) δ: 1H NMR  δ 7.87 , 7.80 , 7.17 , 7.02 , 6.90 , 6.86 , 6.23 , 5.10 , 4.28 , 3.90  3.89 , 3.88 , 3.57 . 13C NMR (acetone-d6) δ: 160.83 (C-2), 148.71 (C-3′), 148.60 (C-4′), 147.09 (C-6), 145.15 (C-4), 138.59 (C-7), 139.91 (C-9), 139.84, 138.76, 137.78, 133.37 (C-8), 128.54 (C-1′), 121.97 (C-6′), 116.00 (C-5′), 114.48 (C-3), 112.45 (C-10), 112.29 (C-2′), 102.03 (C-5), 79.77 (C-8′), 77.28 (C-7′), 61.71 (C-9′), 56.72 (6-OCH3), 56.50 (3′-OCH3).Cleomiscosin B (13), pale yellow amorphous powder. UV  λmax 210, 235s, 331. ESITOFMS m/z 417.1193 [M + H]+ . 1H NMR (acetone-d6) δ: 1H NMR (acetone-d6) δ: 7.87 , 6.87 , 6.87 , 6.24 , 5.08 , 4.30 , 3.89 , 3.88 , 3.86 , 3.57 . 13C NMR (acetone-d6) δ: 159.76 (C-2), 151.18, 149.02 (C-3′/C-5′), 146.59 (C-6), 147.10, 146.84, 145.19 (C-4), 139.88 (C-9), 138.58 (C-7), 137.75 (C-4′), 134.01 (C-8), 127.29 (C-1′), 114.45 (C-3), 111.94 (C-10), 108.36, 106.56 (C-2′/C-5′), 102.05 (C-5), 79.88 (C-8′), 77.55 (C-7′), 61.58 (C-9′), 56.87 (3‘/5‘-OCH3), 56.71 (6-OCH3).Cleomiscosin D (14), white amorphous powder. UV  λmax 220, 278. ESITOFMS m/z 291.0867 [M + H]+ . 1H NMR (methanol-d4) δ 6.97 , 6.80 , 6.76 , 5.94 , 5.91 , 4.82 , 4.18 , 2.86 , 2.74 .Epicatechin (d-Glucopyranosyloxy-4-methyl-2(5H)-furanone (15), white amorphous powder. UV  λmax 224. ESITOFMS m/z 277.0969 [M + H]+ . 1H NMR (deuterium oxide) δ: 5.08 , 4.84 , 3.91 , 3.75 , 3.56 , 3.54–3.46 , 2.09 . 13C NMR (deuterium oxide) δ 172.20 (C-2), 144.42 (C-4), 136.95 (C-3), 101.50 (C-1′), 76.39 (C-5′), 75.47 (C-3′), 73.13 (C-2′), 71.36 (C-5), 69.35 (C-4′), 60.54 (C-6′), 9.98 (4-CH3).3-β-16), yellow amorphous powder. UV  λmax 204, 226, 289, 339. ESITOFMS m/z 355.1065 [M + H]+ . 1H NMR (deuterium oxide) δ: 8.00 , 7.31 , 7.25 , 6.44 , 5.27 , 3.97 , 3.95 , 3.79 , 3.74–3.64 , 3.57 . 13C NMR (deuterium oxide) δ: 165.19 (C-2), 149.28 (C-9), 146.45 (C-6), 146.24 (C-4), 114.33 (C-10), 113.75 (C-3), 110.31 (C-5), 104.02 (C-8), 100.39 (C-1′), 76.72 (C-3′), 75.87 (C-5′), 73.10 (C-2′), 69.63 (C-4′), 60.81 (C-6′), 56.72 (6-OCH3).Scopolin (O-α-l-rhamnopyranosyl-(1”→6′)-β-d-glucopyranoside (17), yellow amorphous powder. UV  λmax 226, 291, 337. ESITOFMS m/z 531.1772 [M + H]+ . 1H and 13C NMR see Isofraxidin-7-O-(5′′-O-vanilloyl-β-apiofuranosyl)-(1′′→2′)-β-glucopyranoside (18), white amorphous powder. UV  λmax 220, 265, 290. ESITOFMS m/z 585.1850 [M + H]+ . 1H and 13C NMR see 4-Methoxycatechol-2-O-hexoside (19), UV  λmax 225, 280. ESITOFMS m/z 403.1993 [M + H]+ . 1H and 13C NMR see 6-(3-Methoxy-4-hydroxyphenyl)-hexane-2,4-diol-2-C. elegans (N2 Bristol strain) were maintained as described by Stiernagle [Escherichia coli OP50 strain as a food source [Cultures of wildtype iernagle . Monoxend source . The in d source . Crude eTo assess the stability of the tested procyanidin trimers, test solutions were collected after the incubation period, worms were allowed to sink and the supernatants were centrifuged at 10000 rpm 5 min prior to LC-MS analysis.® Ver. 3 . Mean values of mortality rates were compared by a one-way ANOVA test followed by a Tukey’s Test for multiple comparison. A p-value < 0.05 compared to the negative control was considered to be significant. Nonlinear regression  was performed to obtain best-fit values for the LC50.Statistical analysis of the data obtained from the in vitro assay was performed using GraphPad PrismP. pinnata as anthelmintic remedies. A-type procyanidins are not only the major class of secondary metabolites, but are also responsible for the anthelmintic activity of the extract. While A-type PACs are not generally superior compared to B-types, the interflavan linkage connecting the flavanol units seems to have a great impact on the bioactivity of the molecule.Im summary, the current findings rationalize the traditional use of root extracts from 17, 18 and 19, mainly belonged to the class of coumarins and phenolic glycosides. Despite its popular medicinal use and frequent phytochemical investigations of the drug, with exception of 15, none of the isolated compounds has previously been described for P. pinnata.Further substances identified, including the three novel compounds


E)-1,3-dimethyl-2-[(4-nitrophenyl)triaz-2-enylidene]-2,3-dihydro-1H-imidazole was synthesized by coupling 1,3-dimethylimidazolium iodide with 1-azido-4-nitro benzene. The title compound has monoclinic (C2/c) symmetry and an E conformation about the –N= N– bond.The triazene derivative, ( E)-1,3-dimethyl-2-[3-(4-nitrophenyl)triaz-2-enylidene]-2,3-dihydro-1H-imidazole, C11H12N6O2, has monoclinic (C2/c) symmetry at 100 K. This triazene derivative was synthesized by the coupling reaction of 1,3-dimethylimidazolium iodide with 1-azido-4-nitro benzene in the presence of sodium hydride  and characterized by 1H NMR, 13C NMR, IR, mass spectrometry, and single-crystal X-ray diffraction. The molecule consists of six-membered and five-membered rings, which are connected by a triazene moiety (–N=N—N–). In the solid-state, the molecule is found to be planar due to conjugation throughout the molecule. The extended structure shows two layers of molecules, which present weak intermolecular interactions that facilitate the stacked arrangement of the molecules forming the extended structure. Furthermore, there are several weak pseudo-cyclical interactions between the nitro oxygen atoms and symmetry-adjacent H atoms, which help to arrange the molecules.The title compound ( Crystals suitable for X-ray analysis were grown from the slow evaporation of a THF/hexane mixture yielding air-stable, red-colored crystals.1-Azido-4-nitro benzene : δ 8.12 , 7.43 , 7.13 , 3.62 . 13C NMR : δ 159.1, 150.7, 143.2, 125.5, 120.8, 118.9, 35.7. UV/Vis : λ (ɛ) = 450 nm. HRMS : m/z calculated for C11H12N6O2Na [M + Na]+ 283.0914, found 283.0918.IR (neat) ν 3435, 1572, 1382, 1360, 1233 cmUiso(H) = 1.2Ueq(C) or 1.5Ueq(C-methyl).Crystal data, data collection and structure refinement details are summarized in Table 110.1107/S2056989021000426/jy2004sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021000426/jy2004Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021000426/jy2004Isup4.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021000426/jy2004Isup4.cmlSupporting information file. DOI: 2055595CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The structures and the relative configurations of the isolated natural products have been determined based on extensive analysis of their NMR and MS data. The cytotoxic, anti-inflammatory, anti-angiogenic, and neuroprotective activity of compounds 3–7, 9–12, 14–20, and 22–26, as well as their effect on androgen receptor-regulated transcription was evaluated in vitro in human tumor and non-cancerous cells. Steroids 22 and 23 showed significant cytotoxicity in the low micromolar range against the HeLa and MCF7 cancer cell lines, while migration of endothelial cells was inhibited by compounds 11, 12, 22, and 23 at 20 µM. The results of the androgen receptor (AR) reporter assay showed that compound 11 exhibited the strongest inhibition of AR at 10 µM, while it is noteworthy that steroids 10, 16, and 20 displayed increased inhibition of AR with decreasing concentrations. Additionally, compounds 11 and 23 showed neuroprotective activity on neuron-like SH-SY5Y cells.Six new ( The Red Sea, one of the warmest and most saline marine habitats, is an extension of the Indian Ocean, located between the Arabian Peninsula and Africa. The entire coastal reef complex extends along a 2000-km shoreline and is characterized by a high degree of chemodiversity, including more than 200 soft and 300 hard coral species . DespiteSinularia have been extensively studied as sources of new bioactive compounds, most often leading to the isolation of diterpenes and steroids with noteworthy levels of bioactivity +. Compound 2 shared quite similar spectroscopic features with 1. In particular, all signals attributed to the steroidal nucleus, with the most prominent being the signals of the two angular methyls H3-18 and H3-19 , the methyl at C-4 (δH 1.04), the oxymethine H-3 (δH 3.10), and the two olefinic protons H-9 and H-11 , were also evident in the 1H NMR spectrum of compound 2  in the side chain of 2. The correlations observed in the COSY spectrum of 2 identified the relevant spin systems; however, metabolite 2 was proven unstable and degraded prior to the acquisition of heteronuclear NMR spectra. Nevertheless, the high structural similarity of 2 with compound 1 renders safe the proposed identification of 2 as (9E)-3β-hydroxy-4α,24-dimethyl-8,9-seco-5α-cholesta-9(11),24(28)-dien-8-one.Compound mpound 2 . The mos6, isolated as a white amorphous solid, possessed the molecular formula C29H50O2, as suggested by its HR-APCIMS and NMR data. The spectroscopic data of 6 were quite similar to those of the previously reported metabolites 5 and 7  and two olefinic methines (δH/C 5.09/135.7 and 5.16/131.8) that was assigned on the basis of the COSY and HMBC correlations. The E geometry of the Δ22 double bond was supported by the large coupling constant of H-22/H-23 (J = 15.3 Hz). The enhancements of H-3/H-5, H-3/H3-29, H-4/H3-19, H-5/H-9, H-5/H3-29, H-9/H-12α, H-9/H-14, H-12β/H3-18, and H3-18/H-20 observed in the NOESY spectrum verified the trans fusion of rings A/B, B/C, and C/D and suggested the axial orientation of the hydroxy group at C-8. The configuration at C-24 was proposed as R due to the fact that the difference in the chemical shifts of C-26 and C-27 was 0.3 ppm and that C-28 resonated at 18.0 ppm −. The high degree of similarity of the spectroscopic data of metabolite 8 , one oxygenated methine (δH/C 4.72/79.9), one oxygenated methylene , one olefinic methine (δH/C 5.36/119.4), and one non-protonated olefinic carbon (δC 142.0). The COSY cross-peaks of H-20/H3-21, H-20/H-22, H-22/H-23, H-25/H3-26, and H-25/H3-27, in combination with the HMBC correlations of H3-21 with C-17, C-20, and C-22, of H-23, H-25, H3-26, and H3-27 with C-24 and of H2-28 with C-23, C-24, and C-25 verified the side chain. In accordance with the literature, H-22 was assigned to be on the opposite side of H3-21, as also suggested by the chemical shift of C-23 which resonated at 119.4 ppm. Instead, when H-22 and H3-21 are co-planar, C-23 is shielded, resonating at 115–116 ppm −, dictating the molecular formula C29H48O5. The spectroscopic data of metabolite 13 related to the steroidal nucleus -22α,28-epidioxy-4α,24-dimethyl-5α-cholest-23-en-3β,8β,11β-triol.Compound  nucleus  displaye spectra c. Additi20, isolated as a white amorphous solid, had the molecular formula C28H44O2, as indicated by its HR-ESIMS and NMR data. In the 1H NMR spectrum of metabolite 20 evident were only one methyl on a non-protonated carbon (δH 0.75), three methyls on tertiary carbons , one hydroxymethylene (δH 3.59 and 3.81), one oxygenated methine (δH 3.56), three olefinic methines  and an exomethylene group (δH 4.79 and 4.83). The spectroscopic data of 20 . On the basis of the above, metabolite 20 was identified as (22E)-24-methyl-cholesta-5,22,24(28)-trien-3β,19-diol.Compound ta of 20  closely  spectra d verifie3, 5, 7, 9–12, 14–19, and 21–26 were identified by comparison of their spectroscopic and physical characteristics with those reported in the literature as 4α,24-dimethyl-5α-cholest-24(28)-en-3β-οl (3) + .(9E)-3β-Hydroxy-4α,24-dimethyl-8,9-seco-5α-cholesta-9(11),24(28)-dien-8-one (2): White amorphous solid; 1H NMR data, see m/z 429.3725 [M + H]+ .-4α,24-Dimethyl-5α-cholest-22-en-3β,8β-diol (6): White amorphous solid; + .(22E)-22α,28-Epidioxy-4α,24-dimethyl-5α-cholest-23-en-3β,8β-diol (8): White amorphous solid; − .(23E)-22α,28-Epidioxy-4α,24-dimethyl-5α-cholest-23-en-3β,8β,11β-triol (13): White amorphous solid; − .(23E)-24-Methyl-cholesta-5,22,24(28)-trien-3β,19-diol (20): White amorphous solid; − . was cultivated in DMEM and Ham’s F12 Nutrient Mixture , as previously described [2 incubator at 37 °C using the standard trypsinization procedure twice or three times a week.The tested compounds were dissolved in DMSO to afford 10 mM stock solutions. Human cervical carcinoma (HeLa) and human breast adenocarcinoma (MCF7) cell lines were purchased from European Collection of Authenticated Cell Cultures  and cultivated in Dulbecco’s Modified Eagle Medium (DMEM) , as previously reported . Human uescribed . Cells wescribed ,30, to r50 values were calculated as previously reported [In cytotoxicity assays, cancer cells were treated with six different concentrations of each tested compound for 72 h. Cells were stained with resazurin and ICreported . Triplicreported .An enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of the cell adhesion molecule ELAM in HUVEC cells after 30 min of incubation with the tested compounds and 4 h of stimulation with TNFα, as previously described .The scratch test was performed with HUVEC cells and evaluated after 20 h of treatment, as previously reported .AR-transcriptional reporter assays were performed on 22Rv1-ARE14 cells after 24 h of incubation, as previously described [v/v) were used as healthy controls.Cell viability of neuron-like SH-SY5Y cells growing in 96-well plates (7000 cells/well) 24 h after treatment was evaluated using the Calcein AM assay , with mip-values or ANOVA followed by Tukey’s multiple comparison test  being new natural products. Among them, 1 and 2 display the rare 8,9-seco-cholestane steroidal nucleus. To the best of our knowledge, dictyoneolone, isolated from a sponge of the genus Dictyonella, is the only previously reported 4-methyl-8,9-seco-cholestane derivative [22 and 23, while compounds 11, 12, 22, and 23 inhibited the migration of endothelial cells at 20 µM. Most of the compounds with activity against cancer cells also showed cytotoxicity toward normal cells (BJ), except for compounds 11, 12, 14, 15, and 18. Compared to cisplatin, these compounds have therefore a broad therapeutic window due to low or zero cytotoxicity on normal human fibroblasts. Moreover, metabolite 22 was more active against HeLa cells than cisplatin used as positive control. Furthermore, the effect of the isolated metabolites on AR-regulated transcription was evaluated in vitro in human tumor and non-cancerous cells, with compound 11 exhibiting the strongest inhibition of AR at 10 µM. It is worth-noting that metabolites 10, 16, and 20 displayed increased inhibition of AR with decreasing concentrations. In addition, compounds 11 and 23 showed neuroprotective activity on neuron-like SH-SY5Y cells.The chemical analysis of the organic extract of the soft coral rivative . Evaluat


Smilax glabra flavonoids extract (SGF) using preparative HPLC, MS, 1H NMR and 13C NMR, determine the contents of these flavonoids using UPLC, then compare their pharmacological activities in vitro. We obtained six flavonoids from SGF: astilbin (18.10%), neoastilbin (11.04%), isoastilbin (5.03%), neoisoastilbin (4.09%), engeletin (2.58%) and (−)-epicatechin (1.77%). The antioxidant activity of six flavonoids were evaluated by determining the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2′-Azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS+) radical scavenging activity and ferric reducing antioxidant power (FRAP). In addition, the anti-inflammatory activity of six flavonoids were evaluated by determining the production of cytokines , nitric oxide (NO) using enzyme linked immunosorbent assay and the NF-κB p65 expression using Western blotting in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The results showed that (−)-epicatechin, astilbin, neoastilbin, isoastilbin and neoisoastilbin had strong antioxidant activities, not only in DPPH and ABTS+ radicals scavenging capacities, but in FRAP system. Furthermore, all the six flavonoids could significantly inhibit the secretion of IL-1β, IL-6, NO (p < 0.01) and the protein expression of NF-κB p-p65 (p < 0.01) in LPS-stimulated RAW264.7 cells. This study preliminarily verified the antioxidant and anti-inflammatory activities of six flavonoids in S. glabra.This study aimed to isolate, prepare and identify the main flavonoids from a standardized  Smilax glabra Roxb. is a dried rhizome of Liliaceae plant. Hundreds of years ago, it had been used as food and folk medicine in some Asian countries, such as China, Japan, Korea and Vietnam. S. glabra was widely used in traditional Chinese medicine (TCM) for the treatment of syphilis, hypertonia, nephritis, heavy metal poisoning and other diseases in China [S. glabra has a variety of biological activities, such as antioxidant, anti-inflammatory, antiviral, antibacterial, hypouricemic, anti-gout, hepatoprotection and cardiovascular protection [in China ,2. In adotection ,4,5,6.S. glabra was rich in flavonoids. In previous investigations, several flavonoids compounds such as astilbin  and taxifolin have been isolated and identified from S. glabra [Flavonoids are the main active components in some medicinal plants and foods, which have various pharmacological functions ,8. S. gl. glabra ,10. More. glabra . Among t+ radical) in vitro [Free radicals are highly reactive intermediates with unpaired electrons. Free radical-mediated oxidative stress was involved in the pathogenesis of a variety of diseases, including cancer, inflammation and neurodegenerative disorders . Some flin vitro ,14.Inflammation is an important contributor to the pathology of various acute and chronic diseases ,16. LipoS. glabra flavonoids extract (SGF) was prepared from the rhizomes of S. glabra [According to TCM theory, the treatment of diseases by traditional Chinese materia medica based on the interaction of multiple components. Therefore, in our previous studies, a standardized . glabra , the fla. glabra . In in v. glabra ,20, whicIn order to clarify the pharmacodynamic material basis and the potential pharmacological mechanisms of SGF, it is necessary to perform further isolation, preparation and identification of the main active compounds in SGF. Furthermore, as mentioned above, a wide variety of diseases are associated with oxidative stress caused by free radicals, as well as inflammation. Thus, this study also evaluated the antioxidant and anti-inflammatory activities of the main active compounds (six flavonoids) in SGF.After optimized the isolation conditions of the six main compounds in SGF by ultra-high-pressure liquid chromatography (UPLC), the PHPLC method was used to isolate and purify these six compounds .1H-NMR and 13C-NMR , 2.46 , 2.67 , 4.00 , 4.70 , 4.73 , 5.71 , 5.89 , 6.65 , 6.89 , 8.71 , 8.80 , 8.89 , 9.10 ; 13C NMR , 27.86 (C-4), 66.09 (C-3), 78.49 (C-2), 94.50 (C-8), 95.01 (C-6), 98.69 (C-10), 113.93 (C-2′), 114.50 (C-5′), 118.01 (C-6′), 130.90 (C-1′), 144.39 (C-3′), 144.55 (C-4′), 155.98 (C-5), 156.28 (C-7), 156.61 (C-9). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [cis-3,3′,4′,5,7-pentahydroxyflavane)), the molecular formula was C15H14O6 , 0.79 , 2.26 , 3.03 , 3.15 , 3.76 , 4.45 , 4.70 , 4.75 , 4.93 , 5.10 , 5.85 , 5.90 , 6.70 , 6.90 , 8.98 , 9.01 , 10.90 , 11.72 ; 13C NMR , 16.49 (C-6″), 68.90 (C-5″), 70.53 (C-2″), 72.04 (C-3″), 75.51 (C-4″), 82.28 (C-3), 94.90 (C-2), 96.06 (C-8), 100.65 (C-6), 101.44 (C-10), 114.09 (C-1″), 114.88 (C-5′), 119.57 (C-2′), 128.58 (C-6′), 145.23 (C-1′), 146.05 (C-3′), 162.91 (C-4′), 164.10 (C-9), 167.43 (C-5), 167.48 (C-7), 196.23 (C-4). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [S,3S-taxifolin 3-O-α-l-rhamnopyranoside), the molecular formula was C21H22O11 , 1.04 , 3.12 , 3.37 , 3.38 , 3.90 , 4.03 , 4.51 , 4.52 , 4.65 , 4.71 , 5.24 , 5.88 , 5.90 , 6.73 , 6.88 , 9.54 , 9.97 , 10.88 , 11.80 ; 13C NMR , 16.45 (C-6″), 69.12 (C-5″), 70.38 (C-2″), 70.77 (C-3″), 72.41 (C-4″), 77.18 (C-3), 82.56 (C-2), 94.86 (C-8), 95.97 (C-6), 100.75 (C-1″), 101.10 (C-10), 114.09 (C-5′), 114.93 (C-2′), 119.09 (C-6′), 127.79 (C-1′), 145.15 (C-3′), 145.98 (C-4′), 162.71 (C-9), 164.12 (C-5), 167.19 (C-7), 194.60 (C-4). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [R,3R-taxifolin 3-O-α-l-rhamnopyranoside), the molecular formula was C21H22O11 , 1.00 , 2.73 , 2.89 , 3.09 , 3.46 , 4.10 , 4.50 , 4.59 , 4.69 , 5.47 , 5.92 , 5.94 , 6.74 , 6.91 , 8.99 , 9.00 , 10.91 , 11.81 ; 13C NMR , 16.11 (C-6″), 69.17 (C-5″), 70.18 (C-2″), 70.66 (C-3″), 72.34 (C-4″), 76.96 (C-3), 80.87 (C-2), 94.63 (C-8), 95,84 (C-6), 100.76 (C-1″), 101.18 (C-10), 113.87 (C-5′), 114.76 (C-2′), 118.04 (C-6′), 127.13 (C-1′), 144.91 (C-3′), 145.33 (C-4′), 163.06 (C-9), 164.62 (C-5), 167.36 (C-7), 192.47 (C-4). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [S,3R-taxifolin 3-O-α-l-rhamnopyranoside), the molecular formula was C21H22O11 , 0.84 , 2.45 , 3.05 , 3.19 , 3.46 , 4.21 , 4.47 , 4.77 , 4.79 , 5.55 , 5.92 , 5.95 , 6.72 , 6.84 , 8.89 , 9.00 , 10.95 , 11.76 ; 13C NMR , 16.37 (C-6″), 68.99 (C-5″), 70.60 (C-2″), 70.64 (C-3″), 71.94 (C-4″), 74.15 (C-3), 80.67 (C-2), 94.82 (C-8), 95.95 (C-6), 98.77 (C-1″), 100.39 (C-10), 113.83 (C-5′), 114.93 (C-2′), 117.97 (C-6′), 127.27 (C-1′), 144.96 (C-3′), 145.27 (C-4′), 163.09 (C-9), 164.75 (C-5), 167.36 (C-7), 192.94 (C-4). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [R,3S-taxifolin 3-O-α-l-rhamnopyranoside), the molecular formula was C21H22O11 , 1.04 , 3.11 , 3.26 , 3.38 , 3.90 , 3.96 , 4.49 , 4.51 , 4.71 , 4.75 , 5.29 ,5.88 , 5.91 , 6.78 , 7.32 , 9.63 10.95 , 11.82 ; 13C NMR , 16.45 (C-6″), 69.14 (C-5″), 70.38 (C-2″), 70.78 (C-3″), 72.40 (C-4″), 77.31 (C-3), 82.47 (C-2), 94.88 (C-8), 96.01 (C-6), 100.84 (C-10), 101.14 (C-1″), 115.05 (C-3′), 115.05 (C-5′), 127.22 (C-1′), 128.65 (C-2′), 128.65 (C-6′), 158.06 (C-4′), 162.74 (C-9), 164.12 (C-5), 167.16 (C-7), 194.67 (C-4). MS, 1H and 13C NMR spectral data of the isolated compound agree well with the data reported [21H22O10 -epicatechin, astilbin, neoastilbin, isoastilbin, neoisoastilbin, and engeletin in SGF was 1.77%, 18.10%, 11.04%, 5.03%, 4.09%, and 2.58%, respectively . Of thes+ radical scavenging capacity, as well as the total reducing power. As shown in 50 values 1.86 ± 0.22, 7.34 ± 0.22, 9.14 ± 0.23, 4.01 ± 0.18, 5.48 ± 0.22 μg/mL, respectively. However, engeletin had no obvious antioxidant activity in DPPH radical scavenging system. (−)-Epicatechin, astilbin, neoastilbin, isoastilbin, neoisoastilbin and engeletin showed strong ABTS+ radical scavenging activity with IC50 values 1.51 ± 0.13, 6.48 ± 1.13, 6.84 ± 0.55, 3.11 ± 0.90, 1.41 ± 0.55, 18.13 ± 1.72 μg/mL, respectively. Among them, (−)-epicatechin had the highest antioxidant activity  both in DPPH radical and in ABTS+ radical scavenging system; then followed by isoastilbin and neoisoastilbin. In order to evaluate the total reducing power of six flavonoids in SGF, we measured the ferric reducing antioxidant power (FRAP) using FeSO4 as the control. The results were consistent with the radicals scavenging experiments. As shown in 4 equivalent amount, respectively. However, engeletin had no obvious reducing power in this system. (−)-Epicatechin had the strongest reducing power , then followed by isoastilbin and neoisoastilbin.To evaluate the antioxidant activity of six flavonoids in SGF, we examined their DPPH radical and ABTSp < 0.01). To further clarify the anti-inflammatory mechanism of six flavonoids, we determined the protein expression of NF-κB p-p65 in RAW 264.7 cells. As shown in p < 0.01).The anti-inflammatory activity of six flavonoids in SGF on LPS-stimulated RAW264.7 cells was investigated by measuring IL-1β, IL-6 and NO concentrations in cell culture supernatant. As shown in S. glabra. However, the other stereoisomers of astilbin  are naturally low in content [S. glabra flavonoids extract (SGF), instead of S. glabra raw material. We isolated and prepared six flavonoids using PHPLC from SGF: astilbin, neoastilbin, isoastilbin, neoisoastilbin, engeletin and (−)-epicatechin. The content of these six flavonoids was 18.10%, 11.04%, 5.03%, 4.09%, 2.58% and 1.77%, respectively. All the purities of these flavonoids were more than 95% which determined by UPLC. The results showed that astilbin was still the main chemical component in SGF. However, the percentage of the other three stereoisomers of astilbin in SGF was higher than in the raw material.Astilbin is a flavonoids compound. Previous studies have shown that astilbin was the main compound in  content . Thus, tS. glabra. Excess radicals and oxidative stress caused a variety of diseases in the body. Previous investigations have indicated that reducing radicals could effectively prevent the occurrence of oxidative stress related diseases [R,3R), neoisoastilbin , isoastilbin  and neoastilbin . The trans-stereoisomers of astilbin  showed higher antioxidant activities than cis-stereoisomers . Moreover, structure-activity studies of flavonoids indicated that the substitution patterns of B-ring could affect the antioxidant capacity [+ radical scavenging and the FRAP evaluation, which indicated that 3′-OH on B-ring of flavonoids is more important to the antioxidant capacity.Some studies have shown that astilbin had good effects in the treatment of various diseases ,10,27. Hdiseases ,29,30. Tdiseases . The foucapacity . The cheS. glabra was often used in the treatment of inflammatory diseases. Therefore, we also assessed the anti-inflammatory activity of six flavonoids in SGF by determining the cytokines  and NO, and the expression of NF-κB p-p65/p65 in LPS-stimulated RAW 264.7 cells. The results showed that RAW 264.7 cells could release a variety of pro-inflammatory cytokines when stimulating by LPS, including IL-1β, IL-6 and NO, which were consistent with several literature reports [In some diseases, oxidative stress and inflammation often occur simultaneously . IL-1β,  reports ,35. More reports ,36,37.S. glabra was obtained from a traditional Chinese medicine factory , lot number 130101. It was identified by associate Prof. Chen Kongrong of Zhejiang Chinese Medical University. A voucher specimen was deposited at the Herbarium of College of Pharmaceutical Sciences at the Zhejiang Chinese Medical University (ZCPS7001). S. glabra flavonoids extract (SGF) was prepared as we described previously [The rhizome of eviously . (−)-Epi18 column ; column temperature, 30 °C; and the mobile phase: CH3CN (A) and H2O with 0.3% formic acid (B). Gradient elution: 14.8% A, 0–6 min, 14.8–100% A, 6-6.01 min, 100% A, 6.01–10 min, 100–14.8% A, 10–15 min. The injected sample volume flow was 1μL. The temperature of the auto-sampler was maintained at RT, and the volume flow rate was 0.35 mL/min. Detection wavelength was 290 nm.In order to optimize the isolation conditions of the six flavonoids in SGF, ultra-high-pressure liquid chromatography (UPLC) with ACQUITY UPLC H-Class system  was performed. The SGF was dissolved with methanol and centrifuged to obtain the supernatant. Chromatographic conditions: Ultra High-performance Liquid Chromatography System ; ACQUITY UPLC BEH C18 OBDTM 250 × 19 mm, 5 μm). The PHPLC condition for six flavonoids was as follows:Then, isolation and preparation of six flavonoids in SGF was executed on Prep 150 LC system  consisted of a Waters 2545Q preparative pump-equipped with a Waters 2489 UV/Visible Detector-, a Waters Fraction Collector III-, Waters 2707 Automatic Sampler and a preparative column (Waters Sunfire Prep C3CN (A) and H2O with 0.3% formic acid (B). Gradient elution: 10% A, 100 mL/min, 0–5 min; 10–18% A, 10-35 mL/min, 5-5.01 min; 18% A, 35 mL/min, 5.01–20 min; 18–95% A, 35 mL/min, 20–20.1 min; 95% A, 35 mL/min, 20.01–25 min; 95–10% A, 35-10 mL/min, 25–25.01 min; 10% A, 10 mL/min, 25–35 min. The detection wavelength and injection volume were 290 nm and 500 μL, respectively.CH1H-NMR and 13C-NMR, Bruker 600 MHz NMR spectrometer ) with those depicted in a previous study. The quantitative analysis and purity of six flavonoids in SGF were determined by ACQUITY UPLC H-Class system mentioned above.The isolated samples were lyophilized for further identification of structure. In this experiment, the six flavonoids were identified by confrontation of the spectral characteristics . MS conditions: electrospray ionization (ESI), negative ion mode, drying gas flow rate is 11 L/min, drying gas temperature is 300 °C, capillary voltage is 3500 V, the nebulizer pressure is 25 psig, the fragmentor voltage is 175 V, the skimmer voltage is 60 V. Six flavonoids isolated from SGF were dissolved in methanol to 1000 μg/mL and diluted to 100, 50, 25, 10, 5 and 1 μg/mL, respectively. Ascorbic acid was dissolved in deionized water to 1000 μg/mL and diluted to 100, 50, 25, 10, 5 and 1 μg/mL.i. Moreover, the absorbance of the mixture of different concentrations of sample solutions (2 mL) and absolute alcohol (2 mL) were determined as Aj. The absorbance of the mixture of DPPH radical solution (2 mL) and methanol (2 mL) were determined as Ac. Ascorbic acid standard was used for comparison.DPPH radical has the maximal absorbance at 517 nm, the absorption value could be decreased by the antioxidant compounds. The capacity of six flavonoids in SGF to remove DPPH radical was determined according to a previously described procedure with slight modifications . DPPH ra+ radical has the maximal absorbance at 734 nm, the absorption value could be decreased by the antioxidant compounds. The radical scavenging activity of six flavonoids in SGF for ABTS+ radical was determined by a previously described procedure [+ radicle was dissolved in deionized water to 7 mM, and the stock solution were generated by reacting ABTS+ radical stock solution with 2.45 mM potassium persulfate in the dark for 16 h at room temperature. The working solution was diluted in ethanol to an absorbance of 0.7 ± 0.02 at 734 nm. ABTS+ radical solution (3.6 mL) was mixed with different concentrations of sample solutions. Then, the mixture was incubated in the dark for 10 min at room temperature after vortex mixing, and the absorbance of the mixture was determined as Ai. Moreover, the absorbance of the mixture of ABTS+ radical solution (3.6 mL) and methanol (0.4 mL) was determined as Aj. Ascorbic acid standard was used for comparison.ABTSrocedure . ABTS+ r3·6H2O solution were mixed at 10:1:1.The FRAP value of six flavonoids in SGF was determined by a previously described procedure with minor modifications . FRAP wo4 standard curve: FeSO4 was dissolved in deionized water to 4000 μM and diluted to 1000, 800, 400, 200, 100 and 25 μM. Different concentrations of FeSO4 (0.1 mL) was mixed with FRAP working solution (3 mL) and deionized water (0.3 mL) at 37 °C water bath for 4 min, then determined the absorbance at 593 nm. The standard curve (R2 = 0.9993) was linear in the range 0–4000 µM.FeSO4 equivalent amount).FRAP values of samples: Take 0.1 mL 50 μg/mL samples, mixed with FRAP working solution (3 mL) and deionized water (0.3 mL) at 37 °C water bath for 4 min, then determined the absorbance at 593 nm. The total reducing power  was calculated according to the standard curve (µM FeSO5 cells/mL) was added to a 96-well plate (100 µL/well) at 37 °C, 5% CO2 for 24 h, then treated with 100 μM (−)-epicatechin, neoastilbin, astilbin, neoisoastilbin, isoastilbin or engeletin for 0.5 h, followed by stimulated with LPS (1 μg/mL) and incubated for another 6 h. Dexamethasone  as positive control [RAW 264.7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) contained with 10% fetal bovine serum (FBS), 100 µg/mL streptomycin and 100 U/mL penicillin. The cell suspension  were electrophoresed and separated in 10% polyacrylamide gel prior to being transferred onto PVDF membranes. After transferring the target proteins to PVDF membranes, the membranes were blocked with 5% BSA in TBST for 1 h, then incubated with anti-NF-κB p65 , anti-NF-κB p-p65  and anti-actin  at 4 °C for overnight . Finallyp < 0.05 was considered significant.The results were analyzed by one-way analysis of variance (ANOVA), followed by a post hoc test using the SPSS 20.0 software . The data are the mean ± SD of three independent experiments. + radicals scavenging capacitiesand in FRAP system. All the six flavonoids could inhibit the secretion of IL-1β, IL-6, NO and protein expression of NF-κB p-p65 in LPS-stimulated RAW264.7 cells. This study laid a foundation for future research on the potential pharmacological mechanisms and new product development of these six flavonoids and S. glabra.In summary, we identified six flavonoids from SGF: astilbin (18.10%), neoastilbin (11.04%), isoastilbin (5.03%), neoisoastilbin (4.09%), engeletin (2.58%) and (−)-epicatechin (1.77%). Moreover, we found that (−)-epicatechin, astilbin, neoastilbin, isoastilbin and neoisoastilbin had strong antioxidant activitiesboth in DPPH and ABTS


Jugulospora vestita was found to produce seven novel xanthone-anthraquinone heterodimers, xanthoquinodin A11 (1) and xanthoquinodins B10–15 (2–7), together with the already known compound xanthoquinodin B4 (8). The structures of the xanthoquinodins were determined by analysis of the nuclear magnetic resonance (NMR) spectroscopic and mass spectrometric data. Moreover, the absolute configurations of these metabolites were established by analysis of the 1H−1H coupling constants, nuclear Overhauser effect spectroscopy (NOESY) correlations, and Electronic Circular Dichroism (ECD) spectroscopic data. Antifungal and antibacterial activities as well as cytotoxicity of all compounds were tested. Xanthoquinodin B11 showed fungicidal activities against Mucor hiemalis [minimum inhibitory concentration (MIC) 2.1 µg/mL], Rhodotorula glutinis (MIC 2.1 µg/mL), and Pichia anomala (MIC 8.3 µg/mL). All the compounds 1–8 displayed anti-Gram-positive bacteria activity (MIC 0.2–8.3 µg/mL). In addition, all these eight compounds showed cytotoxicity against KB 3.1, L929, A549, SK-OV-3, PC-3, A431, and MCF-7 mammalian cell lines. The six novel compounds , together with xanthoquinodin B4, were also found in the screening of other strains belonging to Jugulospora rotula, revealing the potential chemotaxonomic significance of the compound class for the genus.During the course of a screening for novel biologically active secondary metabolites produced by the Sordariomycetes , the ex-type strain of  Hericium (Basidiomycota), which can be used to treat neurodegenerative diseases 20D + 491° ; UV (MeOH) λmax (log ɛ) 199 (4.6), 274 (3.8), 345 (4.4); CD  λmax (Δɛ) 228 (+14.90), 271 (+3.15), 326 (+28.27), 378 (+4.99); 1H-NMR and 13C-NMR see m/z 643.20 (M − H)− and 645.26 (M + H)+; high resolution electrospray ionisation mass spectrometry (HRESIMS) m/z 645.1965 (M + H)+ .Yellow, powder; 20D + 438° ; UV (MeOH) λmax (log ɛ) 198 (4.5), 274 (3.6), 354 (4.1); CD  λmax (Δɛ) 234 (–22.84), 263 (+2.78), 325 (+33.59), 353 (+19.33); 1H-NMR and 13C-NMR see m/z 643.19 (M − H)− and 645.24 (M + H)+; HRESIMS m/z 645.1966 (M + H)+ .Yellow, amorphous solid; [α]20D + 442° ; UV (MeOH) λmax (log ɛ) 199 (4.5), 275 (3.8), 355 (4.2); CD  λmax (Δɛ) 228 (+1.85), 264 (+2.74), 324 (+22.52), 359 (+12.43); 1H-NMR and 13C-NMR see m/z 659.20 (M − H)− and 661.24 (M + H)+; HRESIMS m/z 661.1915 (M + H)+ .Yellow, crystalline solid; [α]20D + 489° ; UV (MeOH) λmax (log ɛ) 199 (4.6), 274(3.9), 354 (4.3); CD  λmax (Δɛ) 228 (+12.74), 264 (+2.90), 323 (+15.04), 360 (+13.37); 1H-NMR and 13C-NMR see m/z 615.18 (M − H) − and 617.19 (M + H)+; HRESIMS m/z 617.1650 (M + H)+ .Yellow, amorphous solid; [α]20D + 508° ; UV (MeOH) λmax (log ɛ) 199 (4.5), 275 (4.0), 361 (4.1); CD  λmax (Δɛ) 233 (–12.96), 264 (+8.84), 323 (+26.05); 1H-NMR and 13C-NMR see m/z 643.20 (M − H)− and 645.24 (M + H)+; HRESIMS m/z 645.1967 (M + H)+ .Yellow, amorphous solid; [α]20D + 425° ; UV (MeOH) λmax (log ɛ) 199 (4.4), 207 (4.3), 276 (4.0), 361 (4.1); CD  λmax (Δɛ) 207 (–35.18), 231 (–17.20), 264 (+10.67), 323 (+37.35); 1H-NMR and 13C-NMR see m/z 661.19 (M − H)− and 663.27 (M + H)+; HRESIMS m/z 663.2072 (M + H)+ .Yellow, amorphous solid; [α]20D + 540.2° ; UV (MeOH) λmax (log ɛ) 202 (4.4), 274 (3.9), 352 (4.3); CD  λmax (Δɛ) 227 (+15.00), 263 (+1.74), 324 (+14.12), 357 (+14.60); 1H-NMR : δH 2.11 , 2.24 , 2.39 , 2.60 , 2.68 , 2.73 , 3.73 , 4.4 , 4.55 , 4.83 , 6.10 , 6.47 , 6.51 , 6.78 , 6.80 , 11.05 , 11:28 , 13.95 ;13C NMR : δC 22.0 , 23.8 , 27.7 , 35.3 , 39.0 , 42.8 , 53.2 , 71.6 , 73.5 , 85.3 , 101.6 , 105.0 , 105.3 , 111.1 , 114.1 , 115.4 , 119.0 , 122.1 , 131.9 , 132.9 , 140.6 , 147.9 , 148.0 , 160.0 , 161.7 , 169.9 , 178.5 , 183.9 , 186.7 , 188.2 ; ESI-MS: m/z 573.15 (M − H) − and 575.21 (M + H)+; HRESIMS m/z 575.1547 (M + H)+ .Yellow, amorphous solid; [α]Candida albicans, Mucor hiemails, Pichia anomala, Rhodotorula glutinis and Schizosaccharomyces pombe), four different Gram-positive bacteria , and three Gram-negative bacteria , using oxytetracycline as a positive control against Gram-positive and Gram-negative bacteria, while nystatin was used as an antifungal positive control. Besides, cytotoxicities of the compounds against seven mammalian cell lines  were determined by the microculture tetrazolium test (MTT) method, using epothilon B as the positive control. Both bioactivity assays were performed following our standard protocols + in the HRESIMS spectrum. The 1H- and 13C-NMR spectra , one methoxy (δC 53.3), five sp3 methylenes , three sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.5 and 84.3 (oxygenated)). In the 1H–1H correlation spectroscopy (1H–1H COSY) spectrum, there were three isolated spin systems . The heteronuclear multiple bond correlation (HMBC) spectrum showed correlations from H-21′ (δH 0.86) to C-20 and C-19′, from H-16′ (δH 2.38) to C-3′, C-4′, and C-5′, from H-3′ (δH 6.89) to C-1′, C-5′, C-7′, and C-16′, from H-5′ (δH 6.81) to C-3′, C-6′, C-7′, and C-16′, from H-1′ (δH 5.96) to C-2′, C-3′, C-7′, C-9′, C-13′, C-14′, C-15′, C-18′, and C-8′ (weak correlation), and from H-13′ (δH 6.05) to C-1′, C-9′, C-11′, C-14′, and C-10′ (weak correlation), above analysis, indicating an anthraquinone moiety (ABC-ring) with 1′-butyrate group. Moreover, the HMBC correlations from H-3 (δH 4.25) to C-2, C-4, C-5, and C-15, from H-5 (δH 2.65) to C-3, C-4, C-6, and C-7, from H-13 (δH 6.07) to C-9, C-11, and C-15′, and from 15-OCH3 (δH 3.67) to C-15 revealed the rest part as the xanthone moiety (DEF-ring). In addition, the key HMBC correlations from H-15′ (δH 2.74 and 2.68) to C-1′, C-9′, C-13′, C-14′, C-11, C-12, and C-13, and from OH-10 (δH 11.76) to C-9, C-14, C-11, and C-11′ (weak correlation) indicated that a methylene (C-15′) linked these two moieties at C-12 and C-14′, as well as C-11 connected to C-11′.Compound  spectra , accompa1 was elucidated as xanthone–anthraquinone heterodimer similar to xanthoquinodin A6 + in its HRESIMS spectrum. The 1H- and 13C-NMR spectra, accompanied with heteronuclear single quantum coherence (HSQC) correlations, revealed signals of two methyl , one methoxy (δC 53.6), five sp3 methylenes , three sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.4 and 84.5 (oxygenated)).Compound 3  located at the positions 2–7 in the F-ring, which indicated epimerization at C-2 or C-3. The small coupling constant (4.0 and 2.0 Hz) of the C-3 methine proton demonstrated an equatorial bond of H-3. Therefore, configurations of chiral carbons C-2 and C-3 were opposite, indicating that compound 2 and 3 were epimers at C-2. In addition, the experimental ECD spectra of compound 2 . The 1H- and 13C-NMR spectra, accompanied with HSQC correlations, revealed signals of two methyl , onrfde methoxy (δC 53.4), four sp3 methylenes , four sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.6 and 85.5 (oxygenated)). The NMR spectroscopic data of 4  located at the positions 3–6 in the F-ring, and C-5 (δC 65.9), which was oxygenated methine. Compared to 2, there were one more oxygen and hydrogen atom in 4 according to the molecular formulae. On the basis of the above analysis, 4 was proposed as a new xanthoquinodin compound that possesses one additional hydroxyl substituent at the C-5. The configurations at C-2, C-3, C-1′, C-11′, and C-14′ were assigned as the same as those of 1 and 2 because of the similar corresponding NMR data ) with Ha-4 . However, it barely showed correlations with Hb-4 , indicating the dihedral right-angle of H-5–C-5–C-4–Hb-4 and the bond of H-5 at the axial orientation consistent with the axial bond of Ha-4 , which showed large coupling constant (12.3 Hz) with H-3. Furthermore, compound 4 shared the similar experimental ECD curve with compound 2  according to the mass ion peak at m/z 617.1650 [M + H]+ in its HRESIMS spectrum. The 1H- and 13C-NMR spectra, accompanied with HSQC correlations, revealed signals of two methyl , one methoxy (δC 53.3), three sp3 methylenes , three sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.5 and 85.3 (oxygenated)). The NMR spectroscopic data of 5 . The 1H- and 13C-NMR spectra, accompanied with HSQC correlations, revealed signals of two methyl , one methoxy (δC 53.8), six sp3 methylenes , three sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.5 and 84.9 (oxygenated)). The NMR spectroscopic data of 6  based on the mass ion peak at m/z 663.2072 [M + H]+ in its HRESIMS spectrum. The 1H- and 13C-NMR spectra, accompanied with HSQC correlations, revealed signals of two methyl , one methoxy (δC 53.4), six sp3 methylenes , three sp3 methines , five aromatic methines , seventeen sp2 quaternary carbons, and two sp3 quaternary carbons (δC 41.4 and 87.3 (oxygenated)). The degrees of unsaturation and the observably different chemical shifts of C-2 to C-6 compared to compound 6 suggested the opening of the γ-lactone ring. The analysis of their NMR spectroscopic data  against the four tested fungi. A previous study has reported significant antifungal effects of xanthoquinodins A6 and ketoxanthoquinodin A6 against Co. truncatum and Cu. lunata, and of xathoquinodins B4 and B5 against Al. brassicicola, Co. gloeosporioides, Co. truncatum, Cu. lunata and Py. grisea [Only compound . grisea .1–8 were active against the three tested Gram-positive bacteria, i.e., Mi. luteus, B. subtilis, and S. aureus, with MIC values in a range of 0.20–8.30 µg/mL, but inactive against M. smegmatis. As for the tested Gram-negative bacteria, only compound 8 showed weak activity (MIC 66.70 µg/mL) against Ch. violaceum. This agrees with previous studies that demonstrated antibacterial activity of different xanthoquinodins against different Gram-positive bacteria, while activity against any Gram-negative bacteria tested was not observed [All the compounds observed ,24.1–8 were active against all seven mammalian cell lines (50) values in a range of 0.03–3.11 µM. Similarly, Anaya-Eugenio et al. reported that xanthoquinodin JBIR-99 exhibited high selective antiproliferative activity against PC-3 prostate cancer cells [1–3, and 5, which possess a F-ring (ρ-hydroxy hexatomic ring) and an ester group at C-1′, showed stronger cytotoxic activities against all tested cell lines, with IC50 values in a range of 0.03–1.46 µM, while compound 8 (possessing an OH group at C-1′ instead) displayed cytotoxic activities with IC50 values in a range of 0.10–4.70 µM. On the other hand, compounds 4, 6, and 7, which possess a hydroxylated F-ring, γ-lactone ring, and a ring-opening respectively, showed cytotoxic activities against the tested cell lines, with IC50 values in a range of 1.03–18.6 µM. Cytotoxicity activities have been observed in different xanthoquinodins [50 values in the range of 2.04–6.44 µM [p-hydroxy hexatomic F-ring plays a key role in the structure–activity relationship of xanthoquinodins.The cytotoxicity results demonstrated that compounds ll lines . Intereser cells . Compounuinodins ,23,24,26–6.44 µM . TherefoMoreover, some xanthoquinodins were reported to possess anticoccidial activity ,28,29, aApiosordaria globosa (CBS 110113) and A. hispanica (CBS 110112), now synonymyzed with J. rotula [J. vestita, with the six novel xanthoquinodins  and xanthoquinodin B4 also being present in both taxa. Moreover, we found the same novel compounds (except 4 and 7) in two strains of J. rotula (FMR 12690 and FMR 12781), both isolated from soil samples, which is also the same substrate from which the ex-type strain of J. vestita and those of A. globosa and A. hispanica have been isolated. Even though all taxa belonging to the genus Jugulospora included in our study produced similar chromatograms, the production of the compounds was variable  [Apiosordaria was synonymized with Triangularia and placed in the family Podosporaceae based on phylogenetic data [A. microcarpa, remain improperly taxonomically placed  . The maiariales) ,32. Eventic data . Howeveraced see . In thatequences .A. globosa and A. hispanica were synonymized with J. rotula. As we mentioned before, all these taxa now included in the genus Jugulospora produced similar chromatograms and compounds. However, the other species of Apiosordaria included in the screening study, i.e., A. backusii , producngularia ,33,34, hJugulospora), i.e., xanthoquinodins Al, A2, A3, Bl, B2, and B3 in Humicola sp. [Chaetomium elatum [Finally, it is important to mention that some of the xanthoquinodins reported to date were found in members also of the Sordariales (which includes the genus cola sp. ,29 and xm elatum .Jugulospora vestita and other strains belonging to the same genus, but not by other related genera of Sordariales, suggests that the production of secondary metabolites could be suitable as chemotaxonomic markers.The present study led to the isolation of eight cytotoxic and antimicrobial compounds, seven of which turned out to be new to science. Therefore, the potential of Sordariomycetes as prolific producers of bioactive secondary metabolites was again demonstrated here. Moreover, the production of these compounds by


In the first stage, the product of S-methylation-methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate was obtained. The subsequent alkylation with CH3I led to the formation of both O- and N-methylation products mixture-methyl 4-methoxy-2-(methylthio)quinoline-3-carboxylate and methyl 1-methyl-2-(methylthio)-4-oxo-1,4-dihydroquinoline-3-carboxylate with a predominance of O-methylated product. The structure of synthesized compounds was confirmed by means of elemental analysis, 1H-NMR, 13C-NMR, LC/MS, and single-crystal X-ray diffraction. The quantum chemical calculations of geometry and electron structure of methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate’s anion were carried out. According to molecular docking simulations, the studied compounds can be considered as potent inhibitors of Hepatitis B Virus replication. Experimental in vitro biological studies confirmed that studied compounds demonstrated high inhibition of HBV replication in 10 µM concentration.Consecutive alkylation of 4-hydroxy-2-thioxo-1,2-dihydroquinoline-3-carboxylate by CH Derivatives of 4-hydroxyquinoline-3-carboxylic acid are known as highly effective quinolone antibiotics . Their aNowadays, the main approach to creating anti-hepatitis B virus (HBV) agents is by developing a highly effective HBV replication depressor . HeteroaThose have demonstrated effective interactions with the corresponding HBV capsid and newly synthesized protein-nucleus and after the interaction of nucleus proteins, the protein could not be assembled properly.Formerly we elaborated and described a new HBV infection model ,18. Deri2 via condensation of methyl 2-isothiocyanatobenzoate 1 and methyl malonate + ion. Just S-alkylation was proved by the position of the singlet of SCH3 protons at 2.65 ppm in 1H-NMR spectrum of methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate 3 and SCH3 carbon at 15.6 ppm in 13C-NMR spectrum. The broad peak of the OH proton was present at 11.60 ppm, but the signal of the thioamide proton near 13 ppm, that is characteristic for cyclic thioamides + (90.0), 218.2 (100). Anal. calcd. for C12H11NO3S, %: C 57.82; H 4.45; N 5.62; S 12.86. Found, %: C 57.72; H 4.43; N 5.63; S 12.94.To the stirred solution of methyl 4-hydroxy-2-thioxo-1,2-dihydroquinoline-3-carboxylate 3  in anhydrous DMF (10 mL) 60% dispersion of NaH in mineral oil  was added. Then CH3I  was added, and the reaction mixture was heated at 60–80 °C for 1–8 h (3 (10 mL) twice. The extracts were washed with water (10 mL) thrice and dried over anhydrous Na2SO4. Then the mixture was purified by column chromatography (CHCl3) to give the products 4 and 5. The analytical data for representative compounds are shown below. 1H-NMR and 13C-NMR spectra of compounds 4 and 5 are presented in To the stirred solution of methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate or 1–8 h . The reaMethyl 4-methoxy-2-(methylthio)quinoline-3-carboxylate (4), yield of 0.42 g (80%), white solids, m.p. 100–101 °C. 1H-NMR spectrum δ, ppm : 8.09 , 7.89 , 7.80 , 7.56 , 4.03 , 3.94 , 2.62 . 13C-NMR spectrum, δ, ppm: 166.0 (2-CS), 159.8, 157.2, 148.3, 131.5, 127.5, 125.8, 122.7, 119.8, 114.6, 61.5 (4-OCH3), 52.9 (COOCH3), 13.0 (SCH3). LC/MS m/z (%): 264.2 [M + H]+ (100.0), 232.2 (50). Anal. calcd. for C13H13NO3S %: C 59.30; H 4.98; N 5.32; S 12.18. Found, %: C 59.22; H 5.01; N 5.26; S 12.11.Methyl 1-methyl-2-(methylthio)-4-oxo-1,4-dihydroquinoline-3-carboxylate (5), yield of 0.104 g (20%), white solids, m.p. 169–171 °C. 1H-NMR spectrum δ, ppm : 8.16 , 7.88–7.81 , 7.48 , 4.11 , 3.79 , 2.54 . 13C-NMR spectrum, δ, ppm: 172.4 (2-CS), 166.1, 148.1, 141.9, 133.2, 125.9, 125.3, 124.3, 124.2, 118.2, 52.1 (COOCH3), 36.9 (1-NCH3), 19.0 (SCH3). LC/MS m/z (%): 264.0 [M + H]+ (90.0), 232.0 (100.0). Anal. calcd. for C13H13NO3S %: C 59.30; H 4.98; N 5.32; S 12.18. Found, %: C 59.46; H 5.00; N 5.29; S 12.21.3  in corresponding solvent   was added. Then CH3I  was added, and the reaction mixture was heated at 60–80 °C for 1–8 h (3 (10 mL) twice. The extracts were washed with water (10 mL) thrice and dried over anhydrous Na2SO4. Then the mixture was purified by column chromatography (CHCl3) to yield the products 4 and 5.To the stirred solution of methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate F, DMSO)  (10 mL) or 1–8 h . The rea3  in a mixture of i-PrOH (5 mL) and 3 mL of 1 M water solution of NaOH  was refluxed for 2 h. After cooling, the reaction mixture was acidified to pH 6 by a solution of AcOH  in 10 mL of water. The precipitate that formed was filtered, washed twice with water (5 mL) and recrystallized from a mixture of DMF (5 mL) and water (10 mL). Yield 0.16 g (68%), white solids, m.p. 190 °C (decomposition). 1H-NMR spectrum δ, ppm : 16.44 , 11.10 , 8.22 , 8.07 , 7.85 , 7.53 , 2.52 . 13C-NMR spectrum, δ, ppm: 176.7 (2-CS), 166.8, 163.0, 139.2, 133.6, 125.7, 124.9, 121.8, 119.0, 105.2, 14.4 (SCH3). LC/MS m/z (%): 236.0 [M + H]+ (50.0), 218.0 (100). Anal. calcd. for C11H9NO3S, %: C 56.16; H 3.86; N 5.95; S 13.63. Found, %: C 55.97; H 3.88; N 5.93; S 13.59.The stirred solution of methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate 3 (C12H11NO3S) are monoclinic. At 293 °K a = 4.0161(9) Å, b = 17.850(4) Å, c = 15.891(6) Å, β = 96.13(3)°, V = 1132.6(5(2) Å3, Mr = 249.28, Z = 4, space group P21/c, dcalc = 1.462 g/cm3, μ(MoKα) = 0.280 mm−1, F(000) = 520. Intensities of 12582 reflections  were measured on the ”Xcalibur-3” diffractometer . The structure was solved by direct method using SHELXTL package [iso = nUeq (n = 1.5 for methyl group and n = 1.2 for other hydrogen atoms) of the carrier atom. Full-matrix least-squares refinement against F2 in anisotropic approximation for non-hydrogen atoms using 1960 reflections was converged to wR2 = 0.177 (R1 = 0.084 for 819 reflections with F > 4σ(F), S = 0.874). The final atomic coordinates, and crystallographic data for molecule 3 have been deposited to with the Cambridge Crystallographic Data Centre, 12 Union Road, CB2 1EZ, UK  and are available on request quoting the deposition numbers CCDC 1982129).Single crystals for X-ray diffraction study were grown from MeOH. The colorless crystals of 4 (C13H13NO3S) are monoclinic. At 293 °K a = 9.5401(7) Å, b = 11.8332(8) Å, c = 11.858(1) Å, β = 109.436(8)°, V = 1262.3(2) Å3, Mr = 263.3, Z = 4, space group P21/c, dcalc= 1.385 g/cm3, μ(MoKα) = 0.256 mm−1, F(000) = 552. Intensities of 13980 reflections  were measured on the ”Xcalibur-3” diffractometer . The structure was solved by direct method using SHELXTL package [iso = nUeq (n = 1.5 for methyl group and n = 1.2 for other hydrogen atoms) of the carrier atom. Full-matrix least-squares refinement against F2 in anisotropic approximation for non-hydrogen atoms using 2160 reflections was converged to wR2 = 0.123 (R1 = 0.049 for 1541 reflections with F > 4σ(F), S = 0.998). The final atomic coordinates, and crystallographic data for molecule 4 have been deposited to with the Cambridge Crystallographic Data Centre, 12 Union Road, CB2 1EZ, UK  and are available on request quoting the deposition numbers CCDC 1982132).Single crystals for X-ray diffraction study were grown from MeOH. The colorless crystals of 5 (C13H13NO3S) are triclinic. At 293 °K a = 7.8907(9) Å, b = 8.2795(9) Å, c = 9.8449(12) Å, α = 96.170(9)°, β = 102.146(10)°, γ = 98.598(9)°, V = 615.26(13) Å3, Mr = 263.3, Z = 2, space group Pcalc= 1.421 g/cm3, μ(MoKα) = 0.262 mm−1, F(000) = 276. Intensities of 6402 reflections  were measured on the “Xcalibur-3” diffractometer . The structure was solved by direct method using SHELXTL package [iso = nUeq (n = 1.5 for methyl group and n = 1.2 for other hydrogen atoms) of the carrier atom. Full-matrix least-squares refinement against F2 in anisotropic approximation for non-hydrogen atoms using 2164 reflections was converged to wR2 = 0.127 (R1 = 0.045 for 1820 reflections with F > 4σ(F), S = 1.037). The final atomic coordinates, and crystallographic data for molecule 5 have been deposited to with the Cambridge Crystallographic Data Centre, 12 Union Road, CB2 1EZ, UK  and are available on request quoting the deposition numbers CCDC 1982133).Single crystals for X-ray diffraction study were grown from MeOH. The colorless crystals of 3, 4, and 6). The human hepatoma cell line HepAD38 was chosen as the source of infectious viral particles to infect the HepG2 / NTCP cell line, carrying the stable integrated HBV virus gene under the control of a tetracycline-regulated promoter, and secreting viral particles into the culture medium in the absence of tetracycline. HBV preparation was obtained using the HepAD38 line according to the following protocol: HepAD38 cells were passaged in a DMEM medium containing 10% fetal calf serum, penicillin/streptomycin, and essential amino acids. The culture medium was taken once every 2 days, clarified by centrifugation  and stored at 4 °C for no longer than 7 days. Next, dry PEG 8000 was added to the culture media to a final concentration of 7.5% and incubated at 4 °C on a rotary platform overnight. The viral precipitate was separated by centrifugation , and the precipitate was suspended in 1/100 of the initial volume in OPTI-MEM medium. Thus obtained viral preparation was aliquoted and stored at −80 °C.The final stage of our investigation was the experimental study of the biological activity of synthesized molecules  were analyzed for viral antigen content using a commercial HBeAg ELISA 4.0 kit  according to the kit manufacturer’s protocol and the optical density of each analyzed well was measured at a wavelength of 450 nm using a plate densitometer.Infection was carried out as follows: The HepG2-NTCP cell suspension was distributed to 96-well plates at 2000 cells per well. After the cells were attached (on the same or the next day), the initial solution was removed by aspiration, and 50 μL of a solution of test compounds dissolved in OPTI-MEM medium  was added to each well or OPTI-MEM with 2% DMSO (in the wells of the positive and negative controls of the infection) and 50 µL of the HBV preparation diluted in OPTI-MEM with 2% DMSO (except negative infection control). After incubation for 24 h in a humidified atmosphere containing 5% CO3I primarily leads to the product of S-methylation methyl 4-hydroxy-2-(methylthio)quinoline-3-carboxylate. The subsequent alkylation with CH3I gives the mixture of products both O- and N-methylation – methyl 4-methoxy-2-(methylthio)quinoline-3-carboxylate and methyl 1-methyl-2-(methylthio)-4-oxo-1,4-dihydroquinoline-3-carboxylate with predominance of O-methylated product. The established regioselectivity of the reaction can be a good basis for further design and targeted synthesis of focused libraries of alkylated quinolines. According to Molecular Docking Simulations and experimental in vitro biological studies, the discussed compounds can be considered as potent inhibitors of HBV replication.Alkylation of 4-hydroxy-2-thioxo-1,2-dihydroquinoline-3-carboxylate by CH


The structures of intermediary and final substances (compounds 7a–j) were confirmed by 1H-NMR, 13C-NMR and IR spectra, as well as by elemental analysis. The in silico bioinformatic and cheminformatic studies evidenced an optimal pharmacokinetic profile for the synthesized compounds 7a–j, characterized by an average lipophilic character predicting good cell membrane permeability and intestinal absorption; low maximum tolerated dose for humans; potassium channels encoded by the hERG I and II genes as potential targets and no carcinogenic effects. The obtained compounds exhibited a higher antimicrobial activity against the planktonic Gram-positive Staphylococcus aureus and Bacillus subtilis strains and the Candida albicans fungal strain. The obtained compounds also inhibited the ability of S. aureus, B. subtilis, Escherichia coli and C. albicans strains to colonize the inert substratum, accounting for their possible use as antibiofilm agents. All the active compounds exhibited low or acceptable cytotoxicity levels on the HCT8 cells, ensuring the potential use of these compounds for the development of new antimicrobial drugs with minimal side effects on the human cells and tissues.In a drug-repurposing-driven approach for speeding up the development of novel antimicrobial agents, this paper presents for the first time in the scientific literature the synthesis, physico-chemical characterization, in silico analysis, antimicrobial activity against bacterial and fungal strains in planktonic and biofilm growth state, as well as the in vitro cytotoxicity of some new 6,11-dihydrodibenz[ Antibiotics represent one of the most important therapeutic discoveries in medical history, having contributed to reduce the mortality and morbidity caused by infectious diseases. They are also an essential tool for modern medicine, making possible common life-saving procedures such as transplantation, chemotherapy of cancer and surgery. Unfortunately, antibiotics have been liable to misuse, leading to the emergence and selection of resistant microorganisms. Presently, antibiotic resistance is one of the biggest public health challenges of our time, being responsible each year of at least 2 million cases of infection and at least 23,000 deaths in USA. The CDC AR Threats Report ranks the most dangerous 18 threats  into three categories based on the level of concern  about their threat to human health oxepins could represent promising candidates for developing new and efficient anti-pathogenic agents oxepin compounds, and the literature data showing that the carbamoyl and oximino pharmacophore groups are exhibiting anti-infective activity, have led us to combine in a single original molecule such biologically active fragments.Encouraging results of these investigations carried out on dibenzoxepinic nucleus by introducing arylcarbamoyloximino side-chains. In order to obtain biopharmacophore molecules unquoted in the literature we have conducted the chemical modeling of the position 11 of dibenzoxepin nucleus represents the basic structure for many drugs, mainly used in psychotropic therapy as antidepressants, anxiolytics, or antipsychotics , as analgesics, anti-inflammatory, antipyretics and antiphlogistics , as β-sympathomimetics (doxaminol) or as inhibitors of phospholipase A2 (olopatadine).The dibenzoxepin nucleus has also been used for the design of anti-cancer drugs. The 8--octanoic acid hydroxyamide has been shown to have histone deacetylase (HDAC) inhibitory activity and to induce cancer cells apoptosis oxepins were tested for their antibacterial and antifungal activity and proved to be active oxepin structure, isolated from solid cultures of Leptosphaeria sp., an ascomycete fungus, which showed antimicrobial and antifungal effects against the plant pathogens Fusarium nivale and Piricularia oryzae quinolin-7-one N,N-dimethylcarbamoyl oxime showed tuberculostatic activity, being active against Mycobacterium tuberculosis H37Rv oxepin-11(6H)one O-(arylcarbamoyl)oximes.This paper presents the synthesis, physico-chemical characterization, in silico analysis, antimicrobial activity against bacterial and fungal strains in planktonic and biofilm growth state, as well as the in vitro cytotoxicity of new 6,11-dihydro-dibenzoxepin-11(6H)-one (1a), and its 2-methyl- (1b) and 2-ethyl- (1c) derivatives, as intermediate substances. The mentioned ketones were obtained after cyclization of 2-(phenoxymethyl)benzoic acid (2a), 2-(4-methylphenoxymethyl)benzoic acid (2b), and 2-(4-ethylphenoxymethyl)benzoic acid (2c).In our synthesis we used 6,11-dihydro-dibenzoxepins were established through IR and NMR spectroscopy. The obtained compounds are white crystals, soluble in cold acetone and chloroform, soluble in hot inferior alcohols, ethyl acetate, benzene, toluene, DMSO. The structures of the new O-(arylcarbamoyl)-11-oximino-6,11-dihydro-dibenzoxepins are divided into two spectral regions corresponding to the oxepinic system and to the phenylcarbamoyl group attached to the oximino group. The presence of an oxygen in the 5th position makes possible the existence of E/Z isomers, evident in our spectra through the splitting of the proton and the carbon signals. The difference between chemical shifts of methylene group is insignificant, but visible in the spectra as two singlets.The b,e]oxepin scaffold in the 1H-NMR spectra, CH2O protons appeared as a singlet at 5.12–5.23 ppm as a minor signal and 5.17–5.22 ppm as a major signal in case a mixture of isomers was obtained or as a signal at 5.13–5.17 ppm in the case a single isomer was obtained. The hydrogen of the NH moiety appears in the 8.22–9.27 ppm range. The 13C-NMR spectra also displayed the characteristic signals of the suggested structures. The peak at about 155.64–158.36 ppm is attributed to C-11 and the signal of C-4a can be found in the 151.35–155.62 ppm range. Analyzing the chemical shifts of the dibenzoxepins with aryl isocyanates. We aimed to integrate in the same molecule the dibenzoxepinic nucleus and the biologically active oximino and phenylcarbamoyl groups. The structures of intermediary and final substances were confirmed through 1H-NMR and 13C-NMR spectra, IR spectra and by elemental analysis.In a drug-repurposing-driven approach for speeding up the development of novel antimicrobial agents, this paper presents for the first time in the scientific literature the synthesis, physico-chemical characterization, in silico analysis, antimicrobial activity against bacterial and fungal strains in planktonic and biofilm growth state, as well as the in vitro cytotoxicity of new 6,11-dihydro-dibenzoxepins are presented in The structure, molecular formula, molecular weight, melting point, yield and elemental analyses of the new O-(arylcarbamoyl)-11-oximino-6,11-dihydro-dibenzoxepins were synthesized according to the previously described procedure oxepin and its 2-methyl- and 2-ethyl-derivatives dissolved in 4 mL anhydrous THF with a solution of 0.01 mol of the corresponding aryl isocyanate in 1 mL anhydrous THF and refluxing in a round-bottom flask equipped with a condenser and drying tube. To obtain 7a–f the reaction mixture was refluxed 55 h and for 7g–j 80 h. Then the solvent was evaporated under reduced pressure. The resulting residue was purified from boiling ethyl acetate.The final compounds were obtained after treating 0.01 mol 11-hydroximino-6,11-dihydro-dibenzoxepin-11(6H)one O-(3-tolyl-carbamoyl)oxime (7a). 1H-NMR : 8.26 ; 8.20 ; 7.74 ; 6.91–7.60 ; 5.23 ; 5.17 ; 2.36  2.30 . 13C-NMR : 160.78 (C-12); 157.54 (C-11); 151.86 (C-4a); 138.10 (C-13); 136.78 (C-7a); 133.96 (C-15); 132.73 (C-3); 132.42 (C-10a); 130.59 (C-8); 130.36 (C-1); 129.98 (C-17); 128.95 (C-9); 128.44 (C-7); 128.35 (C-10); 121.36 (C-2); 120.35 (C-16); 119.82 (C-1a); 119.62(C-14); 117.45 (C-4); 116.85 (C-18); 70.64 (C-6m); 70.56 (C-6M); 21.48 (CH3M); 21.05 (CH3m). FT-IR : 3262m; 3149w; 3068w; 2982w; 2869w; 1730s; 1599s; 1558vs; 1529s; 1483m; 1442m; 1333w; 1301s; 1218vs; 1158m; 1108w; 1021s; 975vs; 942m; 903m; 801w; 774m; 751s; 691m; 635m; 583w; 514w; 442w.6,11-Dihydrodibenzoxepin-11(6H)one O-(4-fluorophenyl-carbamoyl)oxime (7b). 1H-NMR : 8.30 ; 7.73 ; 7.35–7.62 ; 7.06 ; 7.04 ; 6.94 ; 5.16 ; 5.22 (s H-6m). 13C-NMR : 159.02 ; 157.52 (C-12); 156.2 (C-11); 152.04 (C-4a); 133.85 (C-13); 132.75 (C-3); 132.37 (C-7a); 130.95 (C-10a); 130.61 (C-8); 130.28 (C-1); 129.22 (C-9); 128.92 (C-7); 128.35 (C-10); 121.5 ; 121.32 (C-2), 120.28 (C-4); 119.49 (C-1a); 115.78 ; 70.51 (C-6M); 70.29 (C-6m). FT-IR : 3268m; 3080w; 2930w; 2875w; 1740vs; 1601m; 1480w; 1464w; 1441m; 1423m; 1325s; 1305m; 1269m; 1250s; 1210s; 1161s; 1147s; 1133m; 1108w; 1077s; 1051m; 1006w; 981m; 846m; 752s; 731w; 707w; 675w; 631w; 548w.6,11-Dihydrodibenzoxepin-11(6H)one O-(3-chlorophenyl-carbamoyl)oxime (7c). 1H-NMR : 8.37 ; 7.72 ; 7.58 ; 7.30–7.48 ; 7.25 ; 6.94–7.15 ; 6.92 ; 5.12 ; 5.20 (s H-6m)13C-NMR : 161.25 (C-12); 157.52 (C-11); 151.57 (C-4a); 138.16 (C-13); 134.74 (C-7a); 134.40 (C-15); 132.82 (C-16); 132.40 (C-10a); 130.65 (C-3); 130.32 (C-1); 130.09 (C-8); 129.24 (C-14); 128.88 (C-9); 128.36 (C-7); 127.95 (C-10); 126.91 (C-17); 121.62 (C-2); 119.66 (C-18); 119.42 (C-1a); 117.63 (C-4); 70.49 (C-6M); 70.29 (C-6m)FT-IR : 3271m; 3081w; 3001w; 2932w; 2900w; 2885w; 1740vs; 1592m; 1481s; 1461m; 1375w; 1342w; 1310m; 1283m; 1246vs; 1231s; 1216s; 1173m; 1155m; 1089m; 1065s; 1020s; 997vs; 919m; 8852w; 859m; 821m; 763m; 746s; 694w; 671w; 656w; 552w.6,11-Dihydrodibenzoxepin-11(6H)one O-(4-chlorophenyl-carbamoyl)oxime (7d). 1H-NMR : 8.30 ; 7.71 ; 7.46 ; 7.31; 7.28–7.50 ; 7.01 ; 6.95 ; 5.17 .13C-NMR : 161.18 (C-12); 157.58 (C-11); 151.71 (C-4a); 135.50 (C-13); 133.86 (C-7a); 132.85 (C-3); 132.42 (C-10a); 130.96 (C-16); 130.69 (C-8); 130.31 (C-1); 129.19 ; 129.12 (C-9); 128.98 (C-7); 128.40 ; 126.93 (C-10); 121.39 (C-2); 120.93 (C-1a); 120.35 (C-4); 70.56 (C-6).FT-IR : 3259s; 3193w; 3124w; 3069w; 2963w; 2904w; 2862w; 1733s; 1631m; 1598s; 1542vs; 1488vs; 1441m; 1399m; 1333w; 1306m; 1257m; 1208vs; 1155w; 1086s; 1045m; 1016vs; 974vs; 922w; 853w; 797s; 774m; 752m; 634m; 504m; 450w.6,11-Dihydrodibenzoxepin-11(6H)one O-oxime (7e). 1H-NMR : 9.25 ; 8.24 ; 7.76 ; 7.59 ; 7.41 ; 7.30–7.52 ; 7.26 ; 7.03 ; 6.94 ; 5.17 13C-NMR : 161.34 (C-12); 157.90 (C-11); 151.35 (C-4a); 133.78 (C-13); 133.61 (C-7a); 132.79 (C-3); 130.72 (C-1); 130.65 (C-8); 130.12 (C-15); 129.21 (C-10a); 128.37 (C-9); 128.27 (C-7); 128.1 (C-10); 127.89 (C-17); 126.95 (C-16); 121.48 (C-14); 121.43 (C-2); 120.57 (C-18); 120.45 (C-4); 119.61 (C-1a); 70.70 (C-6)FT-IR : 3262m; 3072w; 2961m; 2930w; 2871w; 1746vs; 1604s; 1569m; 1477m; 1475w; 1439s; 1330m; 1305s; 1235vs; 1209s; 1181s; 1153m; 1112w; 1065vs; 1044vs; 1007s; 976vs; 889s; 851m; 797w; 744s; 695m; 630m; 547w.6,11-Dihydrodibenzoxepin-11(6H)one O-(3-trifluoromethylphenyl-carbamoyl)oxime (7f). 1H-NMR : 8.43 ; 8.39 ; 7.33–7.80 ; 7.06 ; 6.93 ; 5.22 ; 5.17. 13C-NMR : 161.54 (C-12); 157.60 (C-11); 151.70 (C-4a); 137.56 (C-13); 136.78 (C-7a); 133.80 (C-15); 132.93 (C-3); 132.43 (C-10a); 130.75 (C-8); 130.32 (C-1); 129.77 (C-17); 129.31 (C-9); 128.67 (C-7); 128.43 (C-10); 122.74 (CF3); 121.43 (C-2); 121.24 (C-18); 120.91 (C-16); 120.37 (C-4); 119.40 (C-1a); 116.38 (C-14); 70.54 (C-6).FT-IR : 3267m; 3162w; 3105; 2964w; 2869w; 1740s; 1618m; 1564vs; 1492m; 1442m; 1335vs; 1310s; 1246m; 1204vs; 1161s; 1068s; 1023s; 982vs; 924w; 884s; 796m; 775m; 752m; 693m; 516w; 439w.2-Methyl-6,11-dihydro-dibenzoxepin-11(6H)one O-oxime (7g). 1H-NMR : 9.22 ; 8.23 ; 7.58 ; 7.42–7.51 ; 7.42 ; 7.29 ; 7.19 ; 6.84 ; 5.14 ; 2.33 13C-NMR : 160.98 (C-12); 155.71 (C-11); 151.39 (C-4a); 133.73 (C-13); 133.41 (C-3); 132.87 (C-10a); 132.77 (C-2); 133.7 (C-7a); 130.76 (C-8); 130.69 (C-15); 130.04 (C-1); 128.98 (C-9); 128.83 (C-7); 128.4 (C-10); 128.35 (C-17); 128.08 (C-16); 123.52 (C-14); 121.11 (C-18); 120.18 (C-4); 118.86 (C-1a); 70.62 (C-6); 20.3 (CH3)FT-IR : 3271m; 3142w; 3089w; 3063w; 2964w; 2918w; 2869w; 1754vs; 1618w; 1565vs; 1491m; 1335s; 1310s; 1226s; 1161s; 1020s; 982vs; 775m; 752m; 742m; 516w.2-Ethyl-6,11-dihydro-dibenzoxepin-11(6H)one O-(phenyl-carbamoyl)oxime (7h). 1H-NMR : 8.22 ; 7.59 ; 7.53 ; 7.50 ; 7.49–7.39 ; 7.35 ; 7.21 ; 7.12 ; 6.85 ; 5.14 ; 2.65 ; 1.26 13C-NMR : 161.21 (C-12); 155.64 (C-11); 151.85 (C-4a); 137.20 (C-13); 137.04 (C-7a); 134.06 (C-2); 132.56 (C-10a); 132.48 (C-3); 130.51 (C-8); 129.17 ; 129.09 (C-1); 128.86 (C-10); 128.42 (C-9); 128.34 (C-7); 124.36 (C-16); 120.22 (C-4); 119.76 ; 119.26 (C-1a); 70.55 (C-6); 27.85 (2CH-CH3); 15.65 (CH2-3CH)FT-IR : 3269m; 3198w; 3137w; 3081w; 2967w; 2932w; 2878w; 1728vs; 1601s; 1574vs; 1545s; 1499s; 1488s; 1415m; 1376w; 1326m; 1315s; 1302m; 1249m; 1204vs; 1196vs; 1151m; 1015m; 991s; 968s; 929m; 829w; 807m; 755s; 694m; 655m; 507w.2-Ethyl-6,11-dihydro-dibenzoxepin-11(6H)one O-(4-fluorophenyl-carbamoyl)oxime (7i). 1H-NMR : 8.24 ; 7.58 ; 7.52 ;7.45–7.35 ; 7.22 ; 7.06 ; 7.04 ; 6.86 ; 5.13 ; 2.64 ; 1.25 13C-NMR : 161.4 (C-16); 160.79 (C-12); 158.36 (C-11); 155.62 (C-4a); 152.11 (C-13); 137.19 (C-7a); 133.99 (C-2); 132.54 (C-3); 132.51 (C-10a); 130.55 (C-1); 129.06 (C-8); 128.86 (C-9); 128.37 (C-7); 128.35 (C-10); 121.61 ; 120.25 (C-4); 119.16 (C-1a); 115.89 ; 70.52 (C-6); 27.84 (2CH-CH3); 15.67 (CH2-3CH)FT-IR : 3303m; 3077w; 2959w; 2931w; 2878w; 1731s; 1612m; 1521vs; 1456m; 1407m; 1376vs; 1303s; 1251m; 1191vs; 1156s; 1126s; 1102s; 999s; 957w; 920w; 874s; 836w; 812w; 769m; 698m; 641w; 619w; 551w; 519w.b,e]oxepin-11(6H)one O-oxime2-Ethyl-6,11-dihydro-dibenzoxepin derivatives and performed chemical modulations in order to integrate in the same molecule the dibenzoxepinic nucleus and the biologically active oximino and phenylcarbamoyl groups. The new compounds were characterized by their main physical constants and 1H-NMR, 13C-NMR, and IR spectral analysis and by elemental analysis, which confirmed the structure of the obtained compounds.In the purpose of obtaining compounds with antimicrobial activity we have synthesized new dibenz[7a–j suggested an optimal pharmacokinetic profile with an average lipophilic character that allows both cell membrane crossing and intestinal absorption.The predicted values for the synthesized compounds C. albicans, representing thus potential candidates for the design of new therapeutic agents for microbial infections, especially those produced by multiresistant strains. The tested substances also inhibited the ability of S. aureus, B. subtilis, E. coli and C. albicans strains to colonize the inert substratum, accounting for their possible use as antibiofilm agents. All the active compounds exhibited very low cytotoxicity levels on the HCT8 cells, aspect that could represent an advantage for the development of new antimicrobial drugs with minimal side effects on the human cells and tissues, excepting the compound with high number of F that seems to have antitumor potential. The in vitro assay of the antimicrobial activity of the tested compounds showed that they exhibited a higher antimicrobial activity against the Gram-positive bacterial strains and


Botrytis cinerea were prepared using biocatalytic methods. As a result of the biotransformation of racemic 6-chloroindanol (1) and 5-chloroindanol (2) by the fungus B. cinerea, the compounds anti-(+)-6-chloroindan-1,2-diol (anti-(+)-7), anti-(+)-5-chloroindan-1,3-diol (anti-(+)-8), syn-(+)-5-chloroindan-1,3-diol (syn-(+)-8), syn-(-)-5-chloroindan-1,3-diol (syn-(-)-8), and anti-(+)-5-chloroindan-1,2-diol (anti-(+)-9) were isolated for the first time. These products were characterized by spectroscopic techniques and their enantiomeric excesses studied by chromatographic techniques. The results obtained in the biotransformation seem to suggest that the fungus B. cinerea uses oxidation reactions as a detoxification mechanism.Indanols are a family of chemical compounds that have been widely studied due to their broad range of biological activity. They are also important intermediates used as synthetic precursors to other products with important applications in pharmacology. Enantiomerically pure chloroindanol derivatives exhibiting antifungal activity against the phytopathogenic fungus  S, 2R)-1-aminoindan-2-ol has attracted interest due to its usefulness in the synthesis of indinavir®, a potent protease inhibitor of the human immunodeficiency virus (HIV) c 0.2, CHCl3), 24% ee), in favor of configuration R. Based on the results of the antifungal assays, this result appears to indicate that the S-enantiomer is detoxified first because the S-enantiomer is more active against B. cinerea UCA992.In addition to the starting material, the biotransformation of 6-chloroindanol  and 4.38 ppm  corresponding to an anti-position. This hypothesis was confirmed by means of nOe’s experiments. The stereochemistry at C-1 is tentatively given as S configuration considering that the compound 1 recovered from the biotransformation was enriched in the R enantiomer. It is therefore likely that the enantiomer obtained from the biotransformation has an S configuration at C-1. The stereoselectivity of the reaction leading to diol 7 was very high, affording only one diastereoisomer with good enantioselectivity (73% ee). Compound anti-(+)-6-chloroindan-1,2-diol (anti-(+)-7) is reported here for the first time.Compound pic data  were coma  indicated that hydroxyl groups were at C-1 and C-3. The main difference between anti- or syn- is observed for H-2 , both H-2 for syn-diastereoisomer are shown as a single signal (δH 2.37 ppm), indicating the symmetry in the molecule. Their relative configuration was also determined by nOe’s experiments. Stereochemistry at C-3 was also tentatively given as S configuration as it was for the compound anti-(+)-7. In this case, owing to the position of the chlorine atom, C-3 would be the equivalent carbon to C-1 in compound 7. Compounds anti-(+)-5-chloroindan-1,3-diol (anti-(+)-8) and syn-(-)-5-chloroindan-1,3-diol (syn-(-)-8) are described here for the first time.Compound NMR data . The NMR for H-2 . Whereas2) by B. cinerea UCA992 afforded the known compounds 5-chloroindanone (4) and 5-chloro-3-hydroxyindanone (10) c 0.2, CHCl3), 55% ee), in favor of the configuration R.The biotransformation of 5-chloroindanol (one (10) , and thr9 was isolated as a white solid with a molecular formula of C9H9O2Cl deduced from its HRESIMS and corroborated by 13C NMR data -5-chloroindan-1,2-diol (anti-(+)-9) is reported here for the first time.Compound NMR data . Its speNMR data  were com8 was obtained as two diastereoisomers in favor of the syn-configuration (24% de). The spectroscopic data of both diastereoisomers corresponded to the products obtained from the biotransformation of 6-chloroindanol (1). Comparison of their specific rotation values showed that anti-8 was the same enantiomer for both biotransformations (anti-(+)-8), but the enantiomer isolated from the biotransformation of 5-chloroindanol (2) was the opposite (syn-(+)-8, [α]1 (syn-(-)-8, [α]S configuration at C-1.Diol 10 was established as 5-chloro-3-hydroxyindanone (10), a known compound, by comparing its spectroscopic data with data found in the literature c 1, CHCl3): >99% ee [R)-2: [α]c 1, CHCl3): >99% ee +; found: 183.0210 [M-1]+. HPLC : Rt 25.9 min (minor); 27.2 min (major).antianti-(+)-8).-(+)-5-chloroindan-1,3-diol . IR νmax (cm−1): 3300, 2930, 1471, 1286, 1073, 1051, 897, 827. 1H-NMR : δ 7.43 , 7.37 , 7.32 , 5.04 , 2.90 , 1.90 . 13C-NMR : δ 46.8 , 72.7 , 72.8 , 124.7 , 126.6 , 129.2 , 134.8 , 142.7 , 146.2 . HRESIMS m/z: calcd. for C9H9O2Cl: 207.0189 [M + Na]+; found: 207.0181 [M + Na]+. HPLC : Rt 26.7 min (minor); 30.0 min (major).synsyn-(-)-8).-(-)-5-chloroindan-1,3-diol . IR νmax (cm−1): 3300, 2930, 1471, 1286, 1073, 1051, 897, 827. 1H-NMR : δ 7.40 , 7.35 , 7.32 , 5.39 , 2.37 . 13C-NMR : δ 46.8 , 73.6 , 73.9 , 124.9 , 125.9 , 129.3 , 134.9 , 142.8 , 146.5 . HRESIMS m/z: calcd. for C9H9O2Cl: 207.0189 [M + Na]+; found: 207.0179 [M + Na]+. HPLC : Rt 26.3 min (major); 39.2 min (minor).2 as a substrate: 5-chloroindanone (4) , recovered (R)-5-chloroindanol ((R)-2)  , 55% ee), anti-(+)-5-chloroindan-1,2-diol (anti-(+)-9)  , 100% de, 37% ee), anti-(+)-5-chloroindan-1,3-diol (anti-(+)-8)  , syn-(+)-5-chloroindan-1,3-diol (syn-(+)-8)  , and 5-chloro-3-hydroxyindanone (10)  .In addition, the following compounds were isolated using antianti-(+)-9).-(+)-5-chloroindan-1,2-diol . IR νmax (cm−1): 3300, 2929, 1470, 1285, 1073, 1051, 897, 827. 1H-RMN : δ 7.35 , 7.25 , 7.23 , 4.99 , 4.54 , 3.12 , 2.95 . 13C-RMN : δ 38.6 , 73.5 , 75.4 , 125.6 , 126.2 , 127.5 , 134.6 , 140.5 , 142.0 . HRESIMS m/z: calcd. for C9H9O2Cl: 183.0213 [M-1]+; found: 183.0730 [M-1]+. HPLC : Rt 25.8 min (minor); 27.4 min (major).antianti-(+)-8).-(+)-5-chloroindan-1,3-diol . HPLC : Rt 26.7 min (minor); 30.0 min (major).synsyn-(+)-8).-(+)-5-chloroindan-1,3-diol . HPLC : Rt 26.3 min (minor); 39.2 min (major).5-chloro-3-hydroxyindanone (10) D26: