Helicobacter pylori (HP) bacteria indicated to be responsible for various gastrointestinal diseases. Therefore, the spread of HP, currently regarded by the World Health Organization as a class 1 carcinogen, could be better controlled by targeting UE. It is in this line that we have synthesized three new derivatives (2–4) of the naturally occurring olean-12-en-3-one (1), which was previously isolated from the figs of Ficus vallis-choudae Delile (Moraceae). Among the synthesized compounds, 3 and 4 contain an indole moiety. Their structures were unambiguously assigned by spectroscopic and spectrometric techniques . The starting material and the synthesized compounds were screened for UE inhibition activity, and showed significant activities with IC50 values ranging from 14.5 to 24.6 μM, with compound (1) being the most potent as compared to the positive control thiourea (IC50 = 21.6 μM). Amongst the synthetic derivatives, compound 4 was the most potent (IC50 = 17.9 μM), while the others showed activities close to that of the control. In addition, molecular docking study of target compounds 2–4 was performed in an attempt to explore their binding mode for the design of more potent UE inhibitors.Urease enzyme (UE) has been reported to be a potent virulence factor for The online version of this article (10.1007/s13659-018-0193-7) contains supplementary material, which is available to authorized users. Helicobacter pylori (HP) infection is a worldwide problem and the first formally recognized bacterial carcinogen. It is one of the most successful human pathogens, as over half of the world’s population is colonized with this gram-negative bacterium  and carbons at 7′ [δC 128.5 (C-7′) for 3 and 4]. The formation of the hydrazone moiety was also confirmed by HMBC correlations observed between the broad singlet at position δH 11.14 (NH) and C-3 (δC 167.1)  and its synthetic derivatives (2-4) were screened for their UE inhibition activities. The IC50 values of the tested compounds are shown on Table 50 values ranging from 14.5 to 24.6 μM  and the moieties without the triterpene scaffold , together with thiourea were able to dock the GDP binding site, except for compound 2 and its moiety after removing the triterpene scaffold, 2′. The GlideScore (GS) standard precision (SP) docking scores and their various contributions; from rotatable bonds (GRB), lipophilicity (GSlip), H-bonding (GShb), van der Waals interactions (GSvdW), electrostatic interactions (GSele), along with the ligand efficiencies (LE) have been tabulated , Table 1) being tested to be the most active in the assays, the order of biological activities (1 > 4> 3 > 2) could be reproduced in the GS SP docking scores for the tested compounds (1 > 4> 3) and their moieties without the triterpene scaffold (1′ > 4′ > 3′), no docking poses having been observed for both 2 and 2′ , contrary to the native GDP to the s 1 > 4>  > 2 couls 1 > 4>  > 2 coulβ-amyrenone scaffold in order to obtain analogues with both improved activities and solubilities. Solubility predictions showed that the estimated solubilities of the tested compounds and the moieties without the triterpene scaffolds were about 2 log units less soluble than the native ligand (Supplementary Data). This implies the introduction of polar groups to the triterpene scaffold could both improve the solubility and binding by the improving the likelihood of formation of electrostatic interactions and H-bonds. Besides, docking of the moieties without the hydrophobic triterpene scaffold resulted in docking scores closer to those of thiourea and the native GDP. The former also showed less lipophilic and van der Waals contributions towards binding . Fractions were monitored by TLC using Merck pre-coated silica gel sheets (60 F254), and the detection of spots on the TLC plate was carried out using a vanillin spray (prepared by adding 15 g vanillin in 250 mL of ethanol followed by 2.5 mL of concentrated sulphuric acid) and heating the plate at about 80 °C. CDCl3 was used as solvent for 1H and 13C NMR experiments. Trifloroacetic acid, phenylhydrazine, 2,4-dinitrophenylhydrazine, and p-methoxyphenylhydrazine, all of analytical grade and purchased from Aldrich, USA, were used for synthesis. Olean-12-en-3-one commonly called β-amyrenone (1) was isolated from of the fruits of Ficus vallis-choudae Delile, collected from the North-West region of Cameroon , Rt= 20 min, 46.69 mg, 81.94%; mp 260 °C; c 0.0001, CHCl3); UV (CHCl3) λmax(logε) 364 (3.18) nm; IR (KBr) νmax 3450 (NH), 2922 (C–H); 1620 (C=C), 1592 (N–O), 1509 (C=N) cm−1; 1H-NMR: : δ = 2.50–2.39 , 2.64–2.57 , 5.19 , 0.83 , 1.17 , 1.04 , 1.12 , 1.02 , 1.3.0 , 0.85 , 9.10 , 8.27 , 7.94 , 11.14 ;13C-NMR: : δ = 38.1 (C-1), 20.7 (C-2), 167.1 (C-3), 42.2 (C-4), 55.4 (C-5), 19.7 (C-6), 32.3 (C-7), 39.8 (C-8), 46.7 (C-9), 36.7 (C-10),23.6 (C-11), 121.4 (C-12), 145.3 (C-13), 41.8 (C-14), 26.1 (C-15), 26.9 (C-16), 32.5 (C-17), 47.3 (C-18), 46.7 (C-19), 31.1 (C-20), 34.7 (C-21), 37.1 (C-22), 28.4 (C-23), 21.5 (C-24), 14.9 (C-25), 25.9 (C-26), 16.8 (C-27), 28.8 (C-28), 33.3 (C-29), 23.7 (C-30), 145.6 (C-1′), 128.9 (C-2′), 123.6 (C-3′), 137.5 (C-4′), 129.9 (C-5′), 116.5 (C-6′); EI-MS m/z (rel. int.%): 604 [M]+ (20), 589 (3), 386 (6), 305 (11), 218 (100), 203 (75), 109 (34) and 55 (47); HREIMS m/z 604.4022 ; Anal. Calcd. For C36H52N4O4: C, 71.49; H, 8.67; N, 9.26, O, 10.58. Found: C, 71.48; H, 8.67; N, 9.27, O, 10.58.3) was similar to that of (2) but with different Schiff base, temperature and reaction time. In this case, 2,4-dinitrophenylhydrazine was replaced by p-methoxyphenylhydrazine  and the reaction mixture was rather refluxed at 110 °C for 6 days. At completion of the reaction, the final mixture was treated as in the previous case and also successively chromatographed on silica gel column chromatography and HPLC eluting with the mixture of n-hexane-DCM (9.8:0.2) to yield 3 as an off white solid [n-hexane-DCM (8:2)], Rt= 50 min, 40 mg, 80.46%, mp 255; c 0.001, CHCl3); UV (CHCl3) λmax(logε) 283 (3.89) nm; IR(KBr) νmax: 3420 (N–H), 2922 (C-H), 1459 (C=C), 1211 (C–N) and 1086 (C–O) cm−1; 1H-NMR: : δ = 2.17 , 2.72 , 5.28 , 3.82 , 1.20  1.17 , 1.05 , 0.96 , 0.88 , 0.87 , 0.85 , 7.56 , 6.75 , 7.16 , 6.87 ; 13C-NMR: : δ = 36.9 (C-1), 106.9 (C-2), 141.9 (C-3), 32.5 (C-4), 53.1 (C-5), 19.3 (C-6), 32.1 (C-7), 39.9 (C-8), 47.3 (C-9), 38.0 (C-10), 23.6 (C-11), 121.9 (C-12), 145.0 (C-13), 41.9 (C-14), 26.1 (C-15), 27.0 (C-16), 34.0 (C-17), 46.3 (C-18), 46.8 (C-19), 31.1 (C-20), 34.7 (C-21), 37.1 (C-22), 31.0 (C-23), 23.7 (C-24), 15.8 (C-25), 25.8 (C-26), 16.6 (C-27), 28.5 (C-28), 33.4 (C-29), 23.4 (C-30), 131.1 (C-2′), 110.6 (C-3′), 110.9 (C-4′), 153.7 (C-5′), 100.3 (C-6′), 128.5 (C-7′), 56.0 (C-1″); EI-MS m/z (rel. int.%): 527 [M] + (100), 240 (9), 201 (51), 83 (40); HREIMS m/z 527.4099 ; Anal. Calcd. For C37H53NO: C, 84.19; H, 10.13; N, 2.65, O, 3.03. Found: C, 84.18; H, 10.13; N, 2.65, O, 3.03.The synthesis of compound (4) was also similar to that of compounds (2) and (3) with phenylhydrazine  as in the two previous reactions. Here, the reaction mixture was refluxed at 230 °C within 8 days. At completion of the reaction, the final mixture was treated as in the previous cases and equally successively chromatographed on silica gel column chromatography and HPLC eluting with the mixture of n-hexane-DCM (8:2) to yield 3 as an off white solid [n-hexane-DCM (8:2)], Rt= 20 min, 45.85 mg, 97.79%, mp 254.6; c 0.001, CHCl3); UV (CHCl3) λmax(logε) 283 (3.49) nm; IR (KBr) νmax: 3420 (N–H), 2916 (C-H), 1461 (C=C) and 1300 (C–N) cm−1; 1H-NMR: : δ = 2.19 , 2.77 , 5.28  1.30 , 1.21 , 0.96 , 1.05 , 1.17 , 0.85 , 0.87 , 0.88 , 7.70 , 7.28 , 7.09 , 7.04 , 7.40 ; 13C-NMR: : δ = 36.9 (C-1), 106.9 (C-2), 141.9 (C-3), 32.5 (C-4), 53.1 (C-5), 19.3 (C-6), 32.1 (C-7), 39.9 (C-8), 47.3 (C-9), 38.0 (C-10), 23.6 (C-11), 121.9 (C-12), 145.0 (C-13), 41.9 (C-14), 26.1 (C-15), 27.0 (C-16), 34.0 (C-17), 46.3 (C-18), 46.8 (C-19), 31.1 (C-20), 34.7 (C-21), 37.1 (C-22), 31.0 (C-23), 23.7 (C-24), 15.8 (C-25), 25.8 (C-26), 16.6 (C-27), 28.5 (C-28), 33.4 (C-29), 23.4 (C-30), 131.1 (C-2′), 110.6 (C-3′), 110.9 (C-4′), 153.7 (C-5′), 100.3 (C-6′), 128.5 (C-7′); EI-MS m/z (rel. int.%): 497 [M]+ (100), 279 (14), 210 (14), 171 (44); HREIMS: m/z 497.3993 ; Anal. Calcd. For C36H51N: C, 86.86; H, 10.33; N, 2.81. Found: C, 86.85; H, 10.33; N, 2.82.The synthesis of compound (1—4 (1 mM concentration) at 30 °C for 15 min in 96-well plates. In fact, 45 μL each of phenol reagent (1% w/v phenol and 0.005% w/v sodium nitroprusside) and 70 μL of alkali reagent (0.5% w/v NaOH and 0.1% active chloride NaOCl) were added to each well. The absorbance at 630 nm was recorded after 50 min, using a microplate reader . All reactions were performed in triplicate in a final volume of 200 μL. The results (change in absorbance per min) were processed by using Soft Max Pro software . All the assays were performed at pH 8.2 . Percentage inhibitions were calculated from the formula . Thiourea was used as the standard inhibitor of urease for this assay. Reaction mixtures with 25 μL of enzyme (Jack bean Urease bought from Sigma) solution and 55 μL of buffers containing 100 mM urea were incubated with 5 μL of compounds 7. Thiour50) using GraphPad Prism 5.0, USA.Urease inhibition percentages were used to determine the concentration that inhibit 50% of urease activity  was carried out for the determination of difference between groups . P > 0.05 was considered as significant.H. pylori urease have been deposited in the protein databank, e.g. PDB codes; 3O1Q , 3SF5 , 4HI0  and 4LPolution, ). Most otructure . This en1—4). Generally, the docked-pose of the native ligand in the active site of 4LPS showed similar interactions to the observed binding mode of the resolved H. pylori crystal structure (PDB: 4LPS) and the adenine (nitrogenous base) group was well accommodated into receptor active site. Key H-bond interaction between the adenine group and amino acid residue in the binding site could be reproduced, including those with Ser215, Thr61, Asp185, Gly56, Lys183, Arg89 and Glu135 via the Mg2+ cation  into the catalytic pocket of 4LPS. The binding pose of the co-crystallized ligands could be reproduced by the docking program with root mean square deviation (rmsd) values < 0.4 Å. The docking scores and various contributions towards binding have been shown on Table The ligands and protein–ligand complex used for the in silico studies were prepared as follows:H. pylori (PDB ID: 4LPS) was downloaded from the Protein Databank  [The high resolution crystal structure  of HypB csb.org) . All watcsb.org) . Proteincsb.org) , 17. Boncsb.org) .The default setting of the LigPrep tool implemented in Schrödinger’s software version 2017-1) was used to prepare the ligands for docking . All pos17-1 was Docking was done using the Glide program –26. The β-amyrenone has been carried out in order to derive three new compounds (two indoles and one Schiff base), having activities in the lower micromolar range in the H. pylori UE assay. The nitrogenous derivatives showed improved solubilities with respect to the parent compounds, but without a consequent improvement in biological activities. Docking the tested compounds and their less hydrophobic moieties towards the cofactor binding site of H. pylori UE showed that the binding of the tested compounds were more driven by hydrophobic and van der Waals interactions contrary to the native ligand. The exploitation of other regions of the parent compound scaffold would therefore be the most logical approach to improve activities. It is expected that the obtained results would serve as ground work for efforts towards the discovery of novel antimicrobials targeting HP infection from natural sources.The synthesis and testing of novel analogues of a naturally occurring compound Supplementary material 1 (PDF 6166 kb)Below is the link to the electronic supplementary material.


Their structures were elucidated by means of various spectroscopic analyses. Five of the identified compounds showed moderate cytotoxicities against the cell proliferation of SGC-7901, MCF-7, and BEL-7404.Five new taraxerene-type triterpenes, 2-nor-D-friedoolean-14-en-28-ol ( Davidia involucrata Baill., an ornamental tree known as the Chinese dove tree or handkerchief tree, is the only species in genus Davidia. D. involucrata is a relic deciduous tree species of the Tertiary period with important ecological, scientific and horticultural values + . The IR spectrum exhibited absorption bands due to hydroxyl (3560 cm−1) and olefinic (1640 cm−1) groups. Observed in the 1H-NMR  spectrum were signals for seven tertiary methyl groups at δH 0.87, 0.94, 0.94, 1.05, 1.07, 1.08 and 1.22 , while resonances at δH 3.10 and 3.23  were attributed to proton signals attached to an oxygenated methylene carbon. In addition, one olefinic proton at δH 5.51  of a trisubstituted double bond, coupled with the protons of a methylene at δH 1.68  and 2.13  as deduced from their coupling constants, was recognized in the 1H-NMR spectrum. The 13C-NMR  spectrum of 1 showed 29 carbon signals, which were classified from DEPT and HSQC data as seven methyls, ten methylenes, three methines, six quaternary carbons, one oxygen-bearing secondary carbon and a trisubstituted double bond. One of the six degrees of unsaturation came from a trisubstituted double bond at δC 116.0 and 158.5, and the remaining five degrees of unsaturation were therefore indicative of a pentacyclic skeleton. The characteristics of NMR data of compound 1 were comparable to those of myricadiol (6) +  with eight double-bond equivalents. The IR spectrum revealed the presence of hydroxyl (3437 cm−1), carbonyl (1726 cm−1) and olefinic (1635 cm−1) groups. The 1H-NMR  spectrum showed eight tertiary methyls at δH 0.78, 0.82, 1.02, 1.05, 1.06, 1.11, 1.29 and 1.32 , and a characteristic olefinic proton signal at δH 5.60  coupled with the protons of a methylene at δH 1.79  and 2.05 , as deduced from their coupling constants, indicative of the same D-friedoolean-14-en skeleton of 3 as compounds 1 and 2. In addition, an oxygenated methylene at δH 3.91  coupled with the protons of a methylene at δH 1.38  and 2.28  was observed in the 1H-NMR spectrum of 3. The 13C-NMR  spectrum, along with HSQC and DEPT data, resolved 29 carbons that came from eight methyls, seven methylenes, three methines, six quaternary carbons, an oxygenated secondary carbon, two keto carbonyls and a trisubstituted double bond. The HMBC correlations  and H3-24  to C-3 (δC 219.7) rationalized the existence of a keto group at C-3, while another keto group was assigned to C-21 as deduced from the HMBC correlations from H3-29  and H3-30  to C-21 (δC 219.7). An OH group was attached at C-6 as deduced from the key HMBC correlations of H-6  to C-5 (δC 58.8) and C-7 (δC 50.4), as well as the observation of an AMX spin system of H-5 , H-6 , H-7a  and H-7b  in the 1H-NMR spectrum. A Δ14 double bond was determined by the HMBC correlations from H3-26  to C-14 (δC 157.3), and from H-16a  and H-16b  to C-14 (δC 157.3) and C-15 (δC 116.5). The relative stereochemistry of 3 was established by the ROESY spectrum. The ROESY correlations . The IR spectrum exhibited the presence of hydroxyl (3500 cm−1), carbonyl (1700 cm−1) and conjugated carbonyl (1610 cm−1) functional groups. The 1H-NMR spectrum of compound 4 showed the presence of proton signals for seven tertiary methyls at δH 0.98, 1.06, 1.07, 1.22, 1.26, 1.35 and 1.36 , and resonances at δH 3.98  and δH 4.28  attributed to proton attached to two oxygenated methine carbons, as well as signals of two protons at δH 3.26 and 3.71  attached to an oxygenated methylene carbon. In addition, an olefinic proton signal at δH 5.96  was observed in the 1H-NMR spectrum. The 13C-NMR  including DEPT experiments showed that four of the nine degrees of unsaturation came from one trisubstituted double bond at δC 118.8 and 177.9, and three keto carbonyls at δC 206.3, 219.9 and 222.5. The remaining five degrees of unsaturation were therefore indicative of a pentacyclic skeleton. A Δ14 double bond was determined by the key HMBC correlations  and H3-27  to C-14 (δC 177.9). Key HMBC correlations from the olefinic proton at δH 5.96  to a carbonyl at δC 206.3 indicated that one keto group was placed at C-16. Another two keto groups were assigned to C-3 and C-21, which was confirmed by the observed key HMBC correlations from H3-23  and H3-24  to C-3 (δC 222.5), and correlations from H3-30  to C-21 (δC 219.9). Moreover, three hydroxyls were attached to C-6, C-11 and C-29 respectively, which was supported by the key HMBC correlations observed from H-5 , H-7a  and H-7b  to C-6 (δC 67.5), from H-9  and H-12a  to C-11 (δC 65.9), and from H3-30  to C-29 (δC 71.2). ROESY experiment was undertaken to establish the relative configuration of 4. The significant ROESY correlations  was determined as 6α,11α,29-trihydroxy-d-friedoolean-14-en-3,16,21-trione.Compound elations  from H3-elations  of H-11/5, obtained as a white amorphous powder, was assigned to have a molecular formula of C30H44O6 by HR-TOF-MS at m/z = 523.3030 [M + Na]+ , which was an isomer of 4. The NMR data of 5 were comparable to those of 4. Observed in the 1H-NMR spectra was the absence of proton signal of an oxymethine at H-11 , with the appearance of an additional oxygenated methylene proton resonances at δH 3.58  and 3.80  in the 1H-NMR spectra of 5. Correspondingly, an oxymethine carbon at δC 65.9 (C-11) was absent with the presence of an oxygenated secondary carbon at δC 72.5 in the 13C-NMR spectra of 5. The HMBC correlations from H-23a  and H-23b  to C-3 (δC 220.9), C-4 (δC 54.3) and C-5 (δC 54.2) indicated that a hydroxyl group was attached to C-23, which was also confirmed by the key ROESY correlations  was therefore determined as 6α,23,29-trihydroxy-d-friedoolean-14-en-3,16,21-trione.Compound elations  of H-23/1–2) and davinvolunones A–C (3–5), are all taraxerene-type triterpenoids with a d-friedoolean-14-en skeleton. To the best of our knowledge, compounds 1–3, as well as previously reported two ursane triterpenes, davinvolunic acid A and B from the same plant + ; 1H-NMR  and 13C-NMR  data (OD) data .Davinvolunol B (2): white amorphous powder;  = +104.5 ; IR (KBr) νmax: 3500, 2920, 1640, 1470 cm−1; HR-TOF-MS m/z = 451.3546 [M + Na]+ ; 1H-NMR  and 13C-NMR  data (OD) data .Davinvolunone A (3): white amorphous powder;  = +116.8 ; IR (KBr) νmax: 3437, 2930, 1726, 1635, 1456 cm−1; HR-TOF-MS m/z 463.3180 [M + Na]+ ; 1H-NMR  and 13C-NMR  data (l3) data .Davinvolunone B (4): white amorphous powder;  = +126.0 ; IR (KBr) νmax: 3500, 2930, 1700, 1610, 1456 cm−1; HR-TOF-MS m/z 523.3030 [M + Na]+ ; 1H-NMR  and 13C-NMR  data (OD) data .Davinvolunone C (5): white amorphous powder;  = +118.2 ; IR (KBr) νmax: 3489, 2920, 1700, 1610, 1453 cm−1; HR-TOF-MS m/z 523.3030 [M + Na]+ ; 1H-NMR  and 13C-NMR  data (OD) data .in vitro cytotoxic activity was determined by the MTT colorimetric method as described previously in our paper [−1 of penicillin-streptomycin at 37 °C in humidified atmosphere with 5% CO2. For the cytotoxicity tests, cells in exponential growth stage were harvested from culture by trypsin digestion and centrifuging at 180×g for 3 min, and then resuspended in fresh medium at a cell density of 5 × 104 per mL. The cell suspension was dispensed into a 96-well microplate at 100 μL and incubated for 24 h, and then treated with compounds at various concentrations. After 48 h of treatment, 50 μL solution of 1 mg·mL−1 MTT was added to each well, and the cells were further incubated for 4 h. Finally, supernatants were removed and the formazan crystals were dissolved by adding 100 μL DMSO and the optical density was recorded at 570 nm. All drug doses were tested with doxorubicin as positive control in triplicate.The ur paper . Three tDavidia involucrata has led to the isolation of five new taraxerene-type triterpenes, namely 2-nor-d-friedoolean-14-en-28-ol (1), 2-nor-d-friedoolean-14-en-3α,28-diol (2), 6α-hydroxy-2-nor-d-friedoolean-14-en-3,21-dione (3), 6α,11α,29-trihydroxy-d-friedoolean-14-en-3,16,21-trione (4), and 6α,23,29-trihydroxy-d-friedoolean-14-en-3,16,21-trione (5), in addition to five known compounds. This is the first report of the isolation of taraxerene-type triterpenes from the plant, and compounds 1–3 are rarely reported 2-nor pentacyclic triterpenoids with 29 carbons and a five-membered A-ring. Our studies suggested that pure triterpenoids isolated from the relic deciduous plant demonstrated moderate cytotoxic activities against the proliferation of SGC-7901, MCF-7, and BEL-7404 cell lines.The phytochemical study of the methanol extract from the branch barks of


R- and S-isomers of 5-bromo-2-chloro-N-(1-phenylethyl)pyridine-3-sulfonamide have been synthesized, and the stereostructures have been researched. Single crystals of both compounds were obtained for X-ray analysis, and the absolute configurations (ACs) have been further confirmed by electronic circular dichroism (ECD), optical rotation (OR) and quantum chemical calculations. The crystal structures and calculated geometries were extremely similar, which permitted a comparison of the relative reliabilities of ACs obtained by ECD analyses and theoretical simulation. In addition, the effect of stereochemistry on the PI3Kα kinase and anticancer activity were investigated. Compounds 10a and 10b inhibit the activity of PI3Kα kinase with IC50 values of 1.08 and 2.69 μM, respectively. Furthermore, molecular docking was performed to analyze the binding modes of R- and S-isomers.Pyridinesulfonamide is an important fragment which has a wide range of applications in novel drugs.  Sulfonamides were the first clinically available antibacterial agents, and they have been widely used in the design of drug candidates. Sulfonamides have been found to possess a large number of different biological activities, including antibacterial, antiviral, antidiabetic, diuretic, antitumor, and antithyroid activities ,2. Recen1, 2, 3 and 4   D = 58.2 . IR (KBr pellets): ν 3284.3 (NH), 3110.3, 3063.7, 3043.4, 3024.4 (ArH), 2980.5, 2933.4 (CH3), 1632.3 (C=N), 1601.0 (NH), 1585.3, 1540.2, 1493.5 (C=C), 1424.7 (CH3 asym), 1356.3 (CH3 sym), 1340.2 (SO2 asym), 1166.6 (SO2 sym), 1120.6 (C-N), 1109.6 (C-Br), 869.3, 851.7, 816.8, 728.3 (ArH), 768.2 (C-Cl), 703.8 (C-Br) cm−1. 1H-NMR  δ 8.36 , 7.96 , 7.10 , 7.08–7.04 , 5.55 , 4.58 , 1.55 . 13C NMR  δ 152.54 (C-9), 145.68 (C-4), 141.51 (C-12), 139.66 (C-1), 136.35 (C-3), 128.50 , 128.26 , 126.26 (C-11), 118.99 (C-2), 54.85 (C-5), 22.7 (C-13). MS (ESI+): m/z = 376.9 [M + H]+.A mixture of 5-bromo-2-chloropyridine-3-sulfonyl chloride  [10b by (S)-1-phenylethan-1-amine. m.p.: 174.5–175.5 °C. [α]D = −65.9 . IR (KBr pellets): ν 3284.3 (NH), 3110.2, 3063.6, 3043.4, 3024.4 (ArH), 2980.5, 2933.6 (-CH3), 1631.9 (C=N), 1600.9 (NH), 1585.2, 1540.1, 1493.5 (C=C), 1424.7 (CH3 asym), 1356.2 (CH3 sym), 1340.1 (SO2 asym), 1167.3 (SO2 sym), 1120.6 (C-N), 1109.5 (C-Br), 869.3, 851.7, 816.8, 728.2 (ArH), 768.1 (C-Cl), 703.7 (C-Br) cm−1. 1H-NMR  δ 8.36 , 7.96 , 7.10 , 7.08–7.03 , 5.51 , 4.58 , 1.55 . 13C-NMR  δ 152.54 (C-7), 145.67 (C-6), 141.51 (C-11), 139.65 (C-1), 136.35 (C-4), 128.50 , 128.27 , 126.26 (C-9), 119.00 (C-3), 54.86 (C-10), 22.7 (C-13). MS (ESI+): m/z = 376.9 [M + H]+.The similar procedure was conducted to afford The conformational analysis was firstly performed by arbitrarily fixing the absolute configuration of the target compound, using the Spartan 08 program  with MMF2, 1 mM MnCl2, and 0.1% NaN3) was added to each well. Various concentrations of compounds were diluted in 10 μL of 1% DMSO (v/v), with blank DMSO solution as the negative control. The kinase reaction was initiated by the addition of purified tyrosine kinase proteins diluted in 39 μL of kinase reaction buffer solution. The incubation time for the reactions was 30 min at 25 °C and the reactions were stopped by the addition of 5 μL of Streptavidin-XL665 and 5 μL Tk Antibody Cryptate working solution to all of wells. The plate was read using Envision  at 320 nm and 615 nm. IC50 values were calculated from the inhibition curves.The PI3Kα kinase activities were evaluated using homogeneous time-resolved fluorescence (HTRF) assays as previously reported protocol ,31. Brie10a and 10b were evaluated against human HepG2 cell line using the standard MTT assay in vitro. The cancer cell line was cultured in minimum essential medium (MEM) by supplement with 10 % fetal bovine serum (FBS). Approximate 4 × 103 cells, suspended in MEM medium, were plated onto each well of a 96-well plate and incubated in 5% CO2 at 37 °C for 24 h. The tested compounds at the indicated final concentrations were added to the culture medium and the cell cultures were continued for 72 h. Fresh MTT was added to each well at a terminal concentration of 5 mg/mL, and incubated with cells at 37 °C for 4 h. The formazan crystals were dissolved in 100 mL DMSO for each well, and the absorbancy at 492 nm (for absorbance of MTT formazan) and 630 nm (for the reference wavelength) were measured with an ELISA reader. All compounds were tested three times in each of the cell lines. The results expressed as IC50 (inhibitory concentration 50%) were the averages of three determinations and calculated by using the Bacus Laboratories Incorporated Slide Scanner  software.The anti-proliferative activities of compounds R- and S-isomers of 5-bromo-2-chloro-N-(1-phenylethyl)pyridine-3-sulfonamide have been synthesized. The ACs of the isomers were determined by X-ray diffraction, and the assignments of the ACs were further validated by comparing experimental and theoretical ECD spectra and OR. In addition, tests of the PI3Kα kinase and in vitro anticancer activities of the compounds have verified through docking analysis that the activity of the R isomer is better than that of S stereoisomer.


FL30r by using the OSMAC (one strain-many compounds) method. The structures of 1–3, including absolute configurations, were deduced based on MS, NMR, and time-dependent density functional theory (TD-DFT) calculations of specific ECD (electronic circular dichroism) spectra. Compounds 1 and 2 possessed a novel hexahydropyrimido isoindole moiety, and compound 3 exhibited weak radical scavenging activity against DPPH  with an IC50 value of 27.9 μM.Two new nitrogen-containing sorbicillinoids named sorbicillasins A and B ( Filamentous fungi are known as prolific microbial factories for the production of a wide range of metabolites having extensive biological activity . HoweverPhialocephala sp. FL30r was found to be an extensive producer of diverse polyketides, including monomeric sorbicillinol derivatives, bisorbicillinoids, and trisorbicillinoids  isoindole tricyclic skeleton. The radical scavenging activities of the new compounds against DPPH  were tested, and 3 showed weak activity with an IC50 value of 27.9 µM. Herein, we report the details of the isolation, structure elucidation, and biological activities of compounds 1–5.During our ongoing search for bioactive secondary metabolites from deep-sea-derived fungi , the funllinoids ,8,9,10. llinoids ,8,9,10. es (4–5) ,12. AmonPhialocephala sp. FL30r was cultured in mannitol-based medium (45.0 L) with agitation. The EtOAc extract (15.0 g) of fermentation was fractionated by silica gel column chromatography, Sephadex LH-20 column chromatography, medium-pressure preparation liquid chromatography , and semi-preparative HPLC to afford compounds 1 (4.5 mg), 2 (2.7 mg), 3 (7.0 mg), 4 (4.4 mg), and 5 (2.0 mg) (The fungus (2.0 mg) .1 and 2) were both obtained as yellow oils with the molecular formula C19H22N2O6 according to the protonated peak at m/z 375.1550  and the sodinated peak at m/z 397.1376  as analyzed by HRESIMS, respectively. The 1D NMR data  in the 13C-NMR spectrum. Consistent with this analysis, diagnostic HMBC correlations were observed from H3-14 to C-1, C-2, and C-3; from H3-15 to C-3, C-4, and C-5; from 3-OH to C-2, C-3, and C-4; and from 5-OH to C-6. The presence of a tetrahydro-pyrimidinone ring was postulated based on the COSY correlation (H-10/H-11) and the HMBC correlations from 8-NH to C-7, C-9, and C-10; from H2-10 to C-9 and C-11; and from H-11 to C-7, C-9, and C-10. The HMBC correlations from 8-NH and H2-16 to C-6 confirmed the connection of the tetrahydro-pyrimidinone structure and the benzene ring. The HMBC correlation from H2-16 to C-7 positioned the olefinic chain at C-7. Based on the key HMBC correlations from H-11 to C-13 and C-21, together with the chemical shift of C-11 (δC 48.2) and C-13 (δC 169.0), two carbonyls (C-13 and C-21) were connected to N-12 and C-11, respectively. Finally, when accounting for the molecular formula and the degree of unsaturation, C-1 was linked to C-13, and one hydroxyl group was attached to C-21, thus completing the planar structure of 1.Sorbicillasins A and B -1a using time-dependent density functional theory (TD-DFT). The DFT re-optimization of the initial MMFF (Merck molecular force field) minima was performed at the B3LYP/6-31+g(d) level with a polarizable continuum model (PCM) solvent for MeOH. The strong agreement between the calculated ECD spectra of -1a with experimental results suggested the absolute configuration of 1 as 7R, 11S -1b -1b .3 was obtained as a white amorphous powder, and the molecular formula was determined to be C10H12O5 by HRESIMS peaks at m/z 213.0764 . The 1D NMR data , two methylenes, one methine, and five non-protonated carbons. Comparison of the 1H and 13C NMR spectra  and the replacement of the 4-methoxyl group by a 4-hydroxyl group in 3  isoindole ring system in compounds 1 and 2, which compose a 6/5/6 tricyclic ring system, have not been found in nature, with only related synthetic structures reported + .Sorbicillasin A (2): pale yellow oil, λmax (log ε): 220 (2.65), 261 (2.01), 307 (1.80) nm; IR (KBr) νmax 3329, 2926, 1683, 1558, 1384, 1208, 1143, 838 cm−1, see λmax (Δε) 225 (+1.88), 250 (+4.32), 275 (−1.90) nm; 1D NMR data, see m/z 397.1376 [M + Na]+ .Sorbicillasin B (3): white amorphous powder; UV (MeOH) λmax (log ε): 231 (1.36), 285 (2.35) nm; IR (KBr) νmax 3104, 2954, 2705, 1743, 1407, 1170, 1001, 836 cm−1, see m/z 213.0764 [M + H]+ .Scirpyrone K  method  isoindole moiety. Pyrone 3 exhibited radical scavenging activity against DPPH.In summary, two new nitrogen-containing sorbicillinoids, one new 3,4,6-trisubstituted α-pyrone derivative, and two known biosynthetically related polyketides were isolated from the deep-sea-derived fungus


Their structures were elucidated by 1D and 2D NMR spectra and HRESIMS data. All compounds were evaluated for their antimicrobial and their antiplasmodial activities. Regarding antimicrobial activities, the best compounds are (2) and (3), with minimum inhibitory concentration (MIC) of 0.01 and 1 µg/mL, respectively, towards Vibrio natrigens, and (5), with MIC values of 1 µg/mL towards Vibrio carchariae. In addition the known 8-oxo-tryptamine (4) and the mixture of the (E)-6-bromo-2′-demethyl-3′-N-methylaplysinopsin (6) and (Z)-6-bromo-2′-demethyl-3′-N-methylaplysinopsin (7) showed moderate antiplasmodial activity against Plasmodium falciparum with IC50 values of 8.8 and 8.0 µg/mL, respectively.Chemical study of the CH Fascaplysinopsis have yielded several bioactive tryptophan alkaloids reported to exhibit cytotoxic activity against several cancer cell lines +. Analysis of the 1D and 2D 1H, and 13C NMR data for 1 -gelliusine F according to gelliusine F, reported in 1995 + .6-bromo-8,1′-dihydro-isoplysin A (2): yellow oil, c 0.5, MeOH); 1H and 13C NMR, see m/z 337.0483 [M + H]+ .5,6-dibromo-8,1′-dihydro-isoplysin A (3): yellow oil, c 0.5, MeOH); 1H and 13C NMR, see m/z 414.9630 [M + H]+ .Plasmodium falciparum chloroquine-sensitive 3D7 strains was assessed following the procedure already described in Frédérich et al. [50 of 0.006 ± 0.002 µg/mL.Activity against h et al. . The parh et al. . ArtemisRoseobacter litoralis (ATCC 495666), Shewanella putrefaciens (ATCC 8071), Vibrio carchariae (ATCC 35084), Vibrio natrigens (ATCC 14048) and Vibrio proteolyticus (ATCC 15338). Bacterial adhesion and growth rates were determined according to the methods of Thabard et al. [8 colony forming units/mL) were aseptically added to the microplate wells containing compound (0.01–10 μg/mL), and the plates were incubated for 48 h at 26 °C prior to assessment of bioactivity. Media only  was used as a blank. Bacterial growth was monitored spectroscopically at 630 nm. The minimal inhibitory concentration (MIC) for bacterial growth was defined as the lowest concentration which results in a decrease in OD, compared to the blank. The microplates were then emptied, and the bacterial adhesion assay was performed using aqueous crystal staining method [All compounds were tested against five marine bacterial strains commonly found on biofilms, d et al. , Messinad et al.  and Trepd et al. . Bacteri1), 6-bromo-8,1′-dihydro-isoplysin A (2) and 5,6-dibromo-8,1′-dihydro-isoplysin A (3), were isolated from Fascaplysinopsis reticulata together with four known alkaloids from the same family, 8-oxo-tryptamine (4), tryptamine (5), (E)-6-bromo-2′-demethyl-3′-N-methylaplysinopsin (6) and (Z)-6-bromo-2′-demethyl-3′-N-methylaplysinopsin (7). 6,6′-bis-(debromo)-gelliusine F (1) was a new alkaloid with a bis-tryptamine structure and 6-bromo-8,1′-dihydro-isoplysin A (2) and 5,6-dibromo-8,1′-dihydro-isoplysin A (3) were two new alkaloids with 1′,8-dihydroaplysinopsin structure. The 8-oxo-tryptamine (4) and the mixture of the known (E) and (Z)-6-bromo-2′-demethyl-3′-N-methylaplysinopsin  exhibited antiplasmodial activity against Plasmodium falciparum with IC50 values of 8.8 and 8.0 µg/mL respectively while 6,6′-bis-(debromo)-gelliusine F (1), 6-bromo-8,1′-dihydro-isoplysin A (2), 5,6-dibromo-8,1′-dihydro-isoplysin A (3) and tryptamine (5) did not show significant antimalarial activity. The two new 6-bromo-8,1′-dihydro-isoplysin A (2) and 5,6-dibromo-8,1′-dihydro-isoplysin A (3) showed promising antifouling activity against V. natrigens and trpyptamine (5) showed promising antifouling activity against V. carchariae. Further isolation, structure elucidation, and structure-activity relationship studies of this type of alkaloids are required for the development of new drugs.In conclusion, three new tryptophan derived alkaloids, 6,6′-bis-(debromo)-gelliusine F (


N-(methoxycarbonylthienylmethyl)thioureas were synthesised by a sequence of radical bromination of methylthiophenecarboxylic esters, substitution with trifluoroacetamide anion, deprotection, formation of the corresponding isothiocyanates and addition of ammonia. The interaction of these new thiophene-based thioureas with inducible and neuronal nitric oxide synthase was evaluauted. These novel thienylmethylthioureas stimulated the activity of inducible Nitric Oxide Synthase (iNOS).Two isomeric  There are three isoforms, endothelial NOS (eNOS) and neuronal NOS (nNOS) (constitutive Ca2+-dependent forms) and an inducible form (iNOS). Underactivity and overactivity of each of these isoforms can be associated with disease states. Excessive NO production by eNOS within blood vessel walls is thought to be the basis for conditions such as septic- and cytokine-induced circulatory shock -arginine to l-[U-14C]-citrulline conversion assay. 1400W 1 -citrulline from l-[U-14C]-arginine when the agent was added simultaneously with the substrate. The N-(thienylmethyl)thioureas 13 and 25 showed similar activity to that of 3, stimulating human iNOS activity but not modulating the activity of this isoform at all when the compounds were pre-incubated with the enzyme before addition of the substrate. Neither 13 nor 25 affected the activity of rat nNOS, with or without pre-incubation. As expected, 1400W eviously , stimula3), unless otherwise stated. IR spectra of samples were recorded as potassium bromide (KBr) discs, unless otherwise stated. Mass Spectra were recorded using a VG Analytical Mass Spectrometer in the FAB positive ion mode, unless otherwise stated. Elemental analyses was determined using Exeter Analytical Inc. CE-440 Elemental Analyzer. Solutions in organic solvents were dried with magnesium sulfate (MgSO4) and solvents were evaporated under reduced pressure. Experiments were conducted at ambient temperature, unless otherwise stated. Melting points were determined using a Reichert-Jung Thermo Galen Kofler block. NMR data were recorded on either JEOL/Varian GX 270 or EX 400 spectrometers, using solutions in deuteriochlorform (CDClEthyl 5-methylthiophene-2-carboxylate (6). 2-Methylthiophene  was added to butyl lithium (220 mmol) in dry tetrahydrofuran (400 mL) at 0 °C. The mixture was stirred at 0 °C for 3 h before being added to ethyl chloroformate  in tetrahydrofuran (200 mL) at 0 °C. The mixture was stirred at 20 °C for 16 h, and then poured onto ice. The organic layer was washed with water, hydrochloric acid (1 M), aq. sodium hydrogen carbonate and water. Drying, evaporation and distillation gave 6  bp~1 120°C (lit.  δ 4.36 , 5.04 , 6.98 , 7.3 , 7.55 , 8.02 ; MS m/z 292.0644 (M + H) (C14H14NO4S requires 292.0640); Found: C, 57.50; H, 4.41; N, 4.63; C14H13NO4S requires C, 57.70; H, 4.50; N, 4.81%.Methyl 5-(benzyloxycarbonylaminomethyl)thiophene-2-carboxylate (10). Compound 9  was boiled under reflux with sulfuric acid (1.0 mL) in methanol (50 mL) for 2 d. Aq. sodium hydrogen carbonate was added until no further bubbling occurred. The suspension was filtered. The evaporation residue was washed with water and dried to afford 10  as a white solid: mp 80-83 °C; 1H-NMR δ 3.87 , 4.55 , 5.14 , 5.23 , 6.95 , 7.31-7.36 , 7.64 ; MS m/z 398 (M + glycerol + H), 306.0813 (M + H) (C15H16NO4S requires 306.0800), 259 (M - CO2Me); Found C, 58.99; H, 5.00; N, 4.52; C15H15NO4S requires C, 58.98; H, 4.95; N, 4.59%.Methyl 5-(aminomethyl)thiophene-2-carboxylate hydrobromide (11). Compound 10  was stirred with hydrogen bromide in AcOH  for 20 min. The evaporation residue was washed (10 ×) with diethyl ether and dried to afford 11  as a white solid: mp 212-215 °C; 1H-NMR [(CD3)2SO] δ 3.83 , 4.31 , 7.30 , 7.76 , 8.26 ; MS m/z 325 (M + mNBA + H), 172.0437 (M + H) (C7H10NO2S requires 172.0432); Found H, 4.01; N, 5.56; C7H9NO2S requires H, 4.00; N, 5.56%.Methyl 5-(isothiocyanatomethyl)thiophene-2-carboxylate (12). Compound 11  was stirred with calcium carbonate  and thiophosgene  in water (1.5 mL) and chloroform (10 mL) for 20 h. The solvent was evaporated from the organic layer. Chromatography (hexane / ethyl acetate 7:3) gave 12  as a colourless oil: IR (film) νmax 2074, 1711 cm-1; 1H-NMR δ 3.87 , 4.85 , 7.03 , 7.76 ; MS m/z 367 (M + mNBA + H), 214.0003 (M + H) (C8H8NO2S2 requires 213.9997), 182 (M - S); Found C, 44.90; H, 3.32; N, 6.55. C8H7NO2S2 requires C, 45.05; H, 3.31; N, 6.57%.Methyl 5-(thioureidomethyl)thiophene-2-carboxylate (13). Ammonia was passed through 12  in dichloromethane at 0°C for 20 min. The mixture was stirred for 3 h at 0 °C. Evaporation gave 13  as a white solid: mp 131-132 °C: IR νmax 3398, 1703, 1277 cm-1; 1H-NMR (CD3OD) δ 3.84 , 4.93 , 7.06 , 7.65 ; MS m/z 231.0255 (M + H) (C8H11N2OS2 requires 231.0262); Found C, 41.50; H, 4.32; N, 12.06; C15H15NO4S requires C, 41.72; H, 4.38; N, 12.16%.3-Methylthiophene-2-carboxaldehyde (15a) and 4-methylthiophene-2-carboxaldehyde (15b). 3-Methylthiophene 18  was added slowly to butyl lithium  in dry diethyl ether (175 mL) at 0 °C and the mixture was stirred for 3 h at 0 °C, before being added slowly to dimethylformamide  in dry diethyl ether (35 mL) at 0 °C. The mixture was stirred for 16 h and poured onto ice. The organic layer was washed with water, hydrochloric acid (1 M), aq. sodium hydrogen carbonate and water. Drying, evaporation and distillation (ca. 1 torr) gave a mixture of 15a and 15b  as a colourless liquid: 1H-NMR δ 2.32 , 2.58 , 6.97 , 7.36 , 7.58 , 7.63 , 9.86 , 10.04 .3-Methylthiophene-2-carboxylic acid (16a) and 4-methylthiophene-2-carboxylic acid (16b). Silver(I) oxide  was stirred with mixture 19a,b  and sodium hydroxide  in water (50 mL) for 16 h. The mixture was filtered, acidified (hydrochloric acid) and extracted with dichloromethane. Drying, evaporation and recrystallisation (hexane) gave a mixture of 16a and 16b  as white crystals: mp 118-120 °C. 1H-NMR δ 2.30 , 2.57 , 6.95 , 7.13 , 7.48 , 7.69 ; MS m/z 143.0169 (M + H) (C6H7O2S requires 143.0167).Methyl 3-methylthiophene-2-carboxylate (17a) and methyl 4-methylthiophene-2-carboxylate (17b). 3-Methylthiophene 14  was added slowly to butyl lithium  in dry tetrahydrofuran (100 mL) at 0 °C. The mixture was stirred for 3 h at 0 °C, before being added to methyl chloroformate  in dry tetrahydrofuran (20 mL) at 0 °C. The mixture was stirred for 16 h and poured onto ice. The organic layer was washed with water, hydrochloric acid (1 M), aq. sodium hydrogen carbonate and water. Drying, evaporation and distillation (ca. 1 torr) gave a mixture of 17a and 17b  as a colourless oil: 1H-NMR δ 2.28 , 2.56 , 3.86 , 6.91 , 7.13 , 7.38 , 7.60 .Methyl 4-bromomethylthiophene-2-carboxylate (18). Mixture 17a,b  was boiled under reflux with dibenzoyl peroxide  and N-bromosuccinimide  in tetrachloromethane (40 mL) for 17 h. Additional dibenzoyl peroxide  was added every 10 min for 1 h and then every 30 min (twice). Evaporation and chromatography (hexane/ethyl acetate 9:1) gave 18  as a pale buff oil; 1H-NMR δ 3.89 , 4.46 , 7.49 , 7.80 .Methyl 4-(trifluoroacetamidomethyl)thiophene-2-carboxylate (19) and N,N-bis((2-methoxycarbonyl-thien-4-yl)methyl)trifluoroacetamide (20). Trifluoroacetamide  in dry THF (2 mL) was stirred with KOBut  in dry THF (2 mL) for 1 h. Compound 18  in dry THF (2 mL) was added slowly and the mixture was stirred for 15 h. The evaporation residue, in CH2Cl2, was washed with H2O, aq. HCl (1 M) and H2O. Drying, evaporation and chromatography (CHCl3 / EtOAc 19:1) afforded 20  as a yellow solid: IR (film) νmax (NH), 1711 (CF3C=O), 1603 (amide I), 1541 (amide II) cm-1; 1H-NMR δ 3.89 , 3.90 , 4.50 , 4.54 , 7.34 , 7.36 , 7.57 , 7.60 ; 19F-NMR δ -68.67 ; MS m/z 422.0340 (M + H) (C16 H15F3NO5S2 requires 422.0344). Further elution gave 19  as a yellow solid: mp 80-83°C; IR νmax 3305 (NH), 1703 (CF3C=O), 1562 (amide) cm-1; 1H-NMR δ 3.89 , 4.53 , 6.61 , 7.45 , 7.71 ; 19F-NMR δ -76.2 ; MS m/z 268.0255 (M + H) (C9H9O3F3NS requires 268.0246).4-(Benzyloxycarbonylaminomethyl)thiophene-2-carboxylic acid (21). Compound 19  was boiled under reflux with NaOH  in H2O (3 mL) and MeOH (3 mL) for 16 h. The MeOH was evaporated. Benzyl chloroformate  was added at 0 °C and the mixture was stirred vigorously for 4 h, before being washed with EtOAc. The aqueous layer was acidified (aq. HCl) and extracted with EtOAc. Drying, evaporation and chromatography (EtOAc) afforded 21  as a white solid: mp 149-151 °C; 1H-NMR [(CD3)2SO] δ 4.08 , 5.02 , 7.33-7.36 , 7.57 , 7.60 , 7.82 ; MS m/z 315.0503 (M + Na) (13C112C13H13NNaO4S requires 315.0496), 314.0462 (M + Na) (12C14H13NNaO4S requires 3140463).Methyl 4-(benzyloxycarbonylaminomethyl)thiophene-2-carboxylate (22). Compound 21  was boiled under reflux with H2SO4 (1 mL) and MeOH (50 mL) for 2 d. Aq. NaHCO3 was added until no further bubbling occurred. The evaporation residue, in CHCl3 was washed with H2O. Drying and evaporation afforded 22  as a solid: 64-65ºC; 1H-NMR δ 3.88 , 4.35 , 5.13 , 7.34-7.38 , 7.71 ; MS m/z 306.0809 (M + H) (C15H16NO4S requires 306.0800).Methyl 4-(aminomethyl)thiophene-2-carboxylate hydrobromide (23). Compound 22  in AcOH (2.8 mL) was stirred with 30% HBr in AcOH (2.8 mL) for 20 min. The evaporation residue was washed with Et2O (10 ×) and dried to afford 23  as a white solid: mp 203-205 °C; NMR ((CD3)2SO) δΗ 3.85 , 4.08 , 7.95 , 7.97 , 8.13 ; MS m/z 172.0439 (M + H) C7H10NO2S requires 172.0432); Found C, 33.20; H, 3.88; N, 5.44; C7H9NO2S·HBr requires C, 33.33; H, 4.00; N, 5.56%.Methyl 4-(isothiocyanatomethyl)thiophene-2-carboxylate (24). Compound 23  was stirred with CaCO3  and thiophosgene  in H2O (2.0 mL) and CHCl3 (10 mL) for 20 h. The solvent was evaporated from the organic layer. Chromatography (CH2Cl2) afford 24  as a white solid: mp 69-70 °C: IR νmax 2115 (NCS), 1711 (C=O) cm-1; 1H-NMR δ 3.90 , 4.71 , 7.48 , 7.75 ; MS m/z 214 (M + H); Found C, 45.30; H, 3.37; N, 6.66. C8H7NO2S2 requires C, 45.03; H, 3.31; N, 6.57%.Methyl 4-(thioureidomethyl)thiophene-2-carboxylate (25). NH3 was bubbled through 24  in CH2Cl2 (3 mL) at 0°C for 20 min. The mixture was stirred for 3 hr at 0 °C. Evaporation afforded 25  as a white solid: mp 131-132 °C: 1H-NMR δ 3.87 , 4.64 , 5.98 , 6.77 , 7.46 , 7.72 ; MS m/z 231.0272 (M + H) C8H11N2O2S2 requires 231.0262); Found C, 41.39; H, 4.29; N, 11.96; C15H15NO4S requires C, 41.72; H, 4.38; N, 12.16%.Measurements of the inhibitory activity of the test compounds against nNOS was perfomed using an enzyme preparation from rat brain (in which the large majority of the NOS activity is nNOS), whereas the assay of activity against iNOS was performed using preparation of recombinant human iNOS (hiNOS) overexpressed in an HT1080 cell line as described previously by us . l-[U-14C]-arginine to l-[U-14C]-citrulline as described previously by us [l-[U-14C]-arginine (to start the enzymic reaction) and pre-incubation of the test compound with the enzyme preparation for 10 min before initiation of the enzymic reaction by addition of l-[U-14C]-arginine. The assay used the conversion of ly by us . CompounNG-nitroarginine (100 mM). Routinely, clones were grown for 48 h in the absence of puromycin and NG-nitroarginine prior to extracting the iNOS enzyme. Cells were grown to near confluence and were harvested by trypsinisation. Cells were then washed twice in cold phosphate-buffered saline and homogenised in five volumes of ice-cold buffer containing HEPES , sucrose (320 mM), EDTA (100 mM), dithiothreitol (50 mM), leupeptin (10 mg mL-1), soybean trypsin inhibitor (10 mg mL-1) and aprotinin (2 mg mL-1). The preparations were then sonicated using an MSE Soniprep 150 for 35 s at a nominal frequency of 23KHz and oscillation amplitude between 5 and 10 mm. Samples were placed in ice between each sonication. These suspensions were allowed to stand in ice for a further 10 min, then centrifuged at 9,000g for 15 min at 4 °C. The post-mitochondrial supernatant was treated with Dowex-50W  to remove endogenous arginine. The supernatant was incubated with the resin for 5 min and centrifuged at 10,000 rpm for 5 min to pellet the resin. This process was repeated twice, after which the cytosol was treated as free of endogenous arginine and was used for assays of inhibition, using the usual protocol with and without pre-incubation of the test compounds with the preparation. The results are shown in Briefly, for the studies with human iNOS, the enzyme was prepared as follows: an optimised mammalian expression vector  was designed to express human iNOS cDNA . Expression was linked with the selectable marker gene (pac) at the level of mRNA and antibiotic selection (puromycin) directly enforced expression of the cDNA. This vector was been used to transfect the human fibrosarcoma cell line, HT1080 and a series of iNOSexpressing clones were produced. To avoid loss of viability through the cytotoxic consequences of excessive NO production, clones were grown in the presence of a non-toxic dose of In this work, we have extended the range of compounds that cause this apparent stimulation of iNOS activity from benzylthioureas  into thiet al. for S-2-amino-5-pentanoic acid, S-2-amino-5-pentanoic acid and S-2-amino-5-(2-cyanoimidazol-1-yl)pentanoate on iNOS and nNOS [Actually, the biochemical origin and mechanism of this stimulation is unclear, although a similar but weaker stimulation was reported by Ulhaq and nNOS . On the and nNOS . Howeverand nNOS .N-hydroxyguanidines can act as artificial substrates for nitric oxide synthases [l-Arg to radiolabelled l-Cit, it is clear that the increased activity of the iNOS is a genuine stimulation. Therefore, one might speculate that these compounds interact with the enzyme through more than one binding site under different conditions; binding to one site would have a stimulatory effect while binding to the other would have inhibitory effect. The binding to the allosteric stimulatory site may be fast, whereas inhibitory binding of some known inhibitors (e.g. 1400W) to the catalytic haem site is known to be slow [Several synthetic guanidines and ynthases –20. Howe be slow .Recently, it was demonstrated that 1400W is not a slow-binding inhibitor but a time-dependent inactivator of iNOS . TherefoAlthough most effort in this area has gone into inhibiting biosynthesis of •NO, increasing biosynthesis may have some therapeutic value. Nitric oxide blocks the expression of NADP oxidase, an enzyme important in the etiology of cardiovascular disease . Also, sAlso, the thienylmethylthioureas and benzylthioureas reported here and previously  will be


All compounds were tested for their effect on cell viability by an MTT assay and by a Brine Shrimp Test.This study reports the synthesis and antioxidant activity of some new acetamide derivatives. The compounds’ structures were elucidated by NMR analysis and their melting points were measured. The  The syntheses of the compounds 30000-4 and 6  and 4-dimethylaminopyridine  and the mixture was stirred for 20 minutes at rt. After an appropriate 3-phenyl-propylamine (2 eq) was added and the mixture further stirred for 5 h at rt.The antioxidant activity is related with compounds capable of protecting a biological system against the potential harmful effects of oxidative processes. In the last years antioxidant compounds have received increased attention from nutritionists and medical researchers for their potential activities in preventing cancer, cardiovascular disorders, as well as aging . Several30005 (N-(3-phenylpropyl)acetamide (1 eq) and sodium iodide (2.2 eq) dissolved in acetonitrile (15 mL). Chlorotrimethylsilane (2.2 eq) was added to the solution which was stirred for 16 h at reflux. The synthesis of compound 30005  was achi30007 (N-(3-phenylpropyl) acetamide (1 eq) and zinc powder (10 eq) dissolved in EtOH (10 mL). Conc. HCl (1 eq) was added to the solution and cooled to -10 °C. The synthesis of compound 30007  was acco40000-4 and 6 -2-(4-methoxyphenyl)acetamide (1 eq) and sodium iodide (3 eq) dissolved in acetonitrile (35 mL). To the solution heated to 23 °C was added chlorotrimethylsilane (2.2 eq) and the mixture was stirred for 16 h at reflux. The synthesis of compound 40005  was achi40007 -2-(4-nitrophenyl) acetamide (1 eq) and zinc powder (10 eq) dissolved in EtOH (40 mL). Conc. HCl (1 eq) was added to the solution and it was cooled to -10 °C. The synthesis of compound 40007  was acco50 (in ppm). All compounds showed good activity in this test, in particular compounds 40006 and 40007, which showed interesting LD50 values of 3.0443 and 10.6444 ppm, respectively. None of the newly synthesized compounds were cytotoxic on macrophage cell line J774.A1, as revealed by a MTT test  1 h before and simultaneously to LPS significantly (P < 0.001) reduced NO production  reduced ROS production in J774.A1 macrophages stimulated with terz-butyl hyproperoxide (tBOH), while compound 40007 have no effect on ROS inhibition.As reported in 1H-NMR spectra. Melting points were taken on a Gallenkamp melting point apparatus and are uncorrected. The organic extracts were dried over sodium sulphate dry (Merck). 1H-NMR spectra were recorded with a Bruker Advance 300 MHz spectrometer, using CDCl3 as solvent. Mass spectrometry analysis ESI-MS was cwrried out on a Finningan LCQ Deca ion trap instrument.All reagents were purchased from Sigma-Aldrich s.r.l. . The reactions were monitored by thin-layer chromatography (TLC), using Whatman K6F silica gel on aluminum and alumina (Merck) plates with fluorescence indicators and appropriate solvents. A >95% purity could be inferred from the 1N,N-carbonyldiimidazole  and 4-dimethylaminopyridine  and the resulting mixture was stirred for 20 minutes at rt. After 3-phenylpropylamine  was added and the mixture stirred for 5 h at rt  = 363.9; [M +H+] = 364.9; 1H-NMR δ 7.39-7.04 , 5.60-5.53 , 4.88 , 3.53-3.37 , 2.68-2.60 , 1.89-1.77 ; Melting point: 135 °C; MS: [M – H+] = 407.1; [M + H+] = 409.1; 1H-NMR δ 7.50-7.46 , 7.37-7.10 , 5.57 , 4.48 , 3.40-3.30 , 2.64-2.59 , 1.96-1.82 . Compound30003: Yield: 0.640 g (71%); Melting point: 110 °C; MS: (M) 342.2, (m + H+) 345.2; 1H-NMR δ 7.62-7.21 , 7.18-7.10 , 5.57 , 4.90 , 3.38-3.30 , 2.63-2.57 , 1.88-1.77 . Compound30004: Yield: 0.640 g (78%); Melting point: 110 °C; MS: [M] = 359.5; [M+H+] = 360.5; 1H-NMR δ 7.64-7.20 , 6.92 , 5.58 , 4.88 , 3.87 , 3.40-3.30 , 2.64-3.57 , 1.89-1.76 . Compound30006: Yield: 0.600 g (78%); Melting point: 117 °C; MS: [M] = 374.4; [M +H+] = 375.4; 1H-NMR δ 8.40 ; 7.70 ; 7.45-7.10 ; 5.5 ; 4.87 ; 3.92-3.85 ; 3.35-3.28 ; 2.34-2.22 .2S3O3 and Brine, dried with anhydrous sodium sulphate, filtered and evaporated. The residue products were purified by column chromatography on silica . After recrystallization with DCM/Pe , white crystals were obtained . Melting point: 85 °C. m/z 347.2 (M++ 2); 345.2 (M+). 1H NMR δ 8.80-7.18 ; 7.10-7.00 ; 6.90-6.77 ; 5.67 ; 4.82 ; 3.89-3.80 ; 3.33-3.24 ; 2.30-2.20  (30005) .2-(4-Methoxyphenyl)-2-phenyl-N-(3-phenylpropyl)acetamide  and sodium iodide  were dissolved in acetonitrile (15 mL). Chlorotrimethylsilane  was added to the solution and stirred for 16 h at reflux. The mixture was poured into water and extracted with ethyl acetate, the organic layer was washed with Na2O, dried with anhydrous sodium sulphate, filtered and removed by evaporation. The residue products were purified by column chromatography on silica (Pe/EtOAc 1:1). A yellow-orange oil was obtained . m/z 346.4 (M++ 2); 344.4 (M+). 1H NMR  δ 7.37-7.19 ; 7.15 ; 7.05 , 6.68 ; 5.59 ; 4.48 ; 3.68 ; 3.36 ; 2.63 ; 2.10  (30007).2-(4-nitrophenyl)-2-phenyl-N-(3-phenylpropyl) acetamide  and zinc powder  were dissolved in EtOH (10 mL). HCl conc.  was added to the solution and cooled to -10 °C. The mixture was stirred and heated at 80 °C for 3h. After was poured into water and NaOH to neutralize the excess of acid, the organic layer was extracted with Et1-carbonyldiimidazole CDI  and 4-dimethylaminopyridin DMAP  were added to the solution and stirred for 20 minutes at rt. Thereafter was added 3,3-diphenylpropan-1-amine  and stirred for 24h at rt. To the mixture was added H2SO4 (2N) and extracted with ethyl acetate. The organic layer was dried with anhydrous sodium sulphate, filtered and removed by evaporation. The residue products were purified by column chromatography on silica . After recrystallization with DCM/Pe, white crystals were obtained . [M] = 329; [M+H+] = 330. 1H NMR  δ 8.0 , 7.41-7.06 , 4.10-4.05 , 3.44 , 3.33-3.20 , 2.19-2.11  (40000).An appropriate acid was dissolved in DCM dry (10 mL) and N+] = 364. 1H NMR  δ 7.64-6.94 , 5.27 , 3.91 , 3.47 , 3.27-3.16 , 2.29-2.18  (40001).After recrystallization with DCM/Pe, white crystals were obtained . [M] = 363; [M+H+] = 408. 1H NMR  β 7.80 , 7.32-7.12 , 6.96 , 5.33 , 3.90 , 3.46 , 3.27-3.16 , 2.29-2.19  (40002).After recrystallization with DCM/Pe, white crystals were obtained . [M] = 407; [M + H+] = 344.5. 1H NMR  δ 7.30-7.16 , 5.30 , 3.87 , 3.50 , 3.21 , 2.25  (40003).After recrystallization with DCM/Pe, white crystals were obtained . Melting point: 110 °C. [M] = 343.5; [M+H+] = 344.5. 1H NMR  δ 7.30- 7.16 , 5.30 , 3.87 , 3.50 , 3.21 , 2.25  (40004).After recrystallization with DCM/Pe, white crystals were obtained (80%). [M] = 343.5; [M+H+] = 344.5. 1H NMR  δ 7.30-7.16 , 5.30 , 3.87 , 3.50 , 3.21 , 2.25  (40006).After recrystallization with DCM/Pe, white crystals were obtained (80%). [M] = 343.5;  (MTT) to formazan and cells viability was assessed accordingly to the method of Mosmann [J774.A1, WEHI-164 and HEK-293  were added to the culture medium 24 h before and always simultaneously to dichlorofluorescein-diacetate. 2',7'-Dichlorofluorescein-diacetate  was added directly to the growth medium at a final concentration of 5 µM and the cells incubated for 1 h at 37 °C. H2DCF-DA is a non-fluorescent permeant molecule which diffuses passively into cells; the acetates are then cleaved by intracellular esterases to form H2DCF which is thereby trapped within the cell. In the presence of intracellular ROS, H2DCF is rapidly oxidized to the highly fluorescent DCF. Therefore, cells were washed twice with phosphate-buffered saline (PBS), placed in fresh medium and treated with terz-butyl hyproperoxide (tBOH) 3 mM for 30 min and than were placed in a fluorescent microplate reader . Fluorescence was monitored using an excitation wavelength of 490 nm and an emission wavelength of 520 nm. Results were expressed as Relative Fluorescence Units (RFU).The accumulation of ROS was evaluated by means of the probe 2',7'-dichlorofluorescein (DCF) according to the method described by Le Bel et al. . Briefly40006 showed the more interesting activity because of its inhibition of nitrite and ROS. Taken together, our data let us hypothesized that the inhibitory effect of compound 40006 on t-BuOH-induced NO formation in J774.A1 macrophages could be due to its free radical scavenger properties. Moreover, to have an antioxidant activity, is possible to modify the aromatic function binding to the amidic group, instead is necessary to keep unvaried the aromatic function binding to the alkylic chain. In fact the compound 40006 is more active than the compound 30006. Further studies need to characterize its pharmacological activity.Results show that these acetamide derivatives, obtained by simple and rapid chemical synthesis, had interesting biological activities and that none of the compounds are toxic on J774.A1, the cell line chosen for the other tests. Among the tested compounds,


N,N’,N’’-triacetic acid (NOTA) chelators as new bifunctional chelators are described. The syntheses are characterized by their simplicity and short reaction times, as well as practical purification methods and acceptable to very good chemical yields. The usefulness of these two synthetic pathways is demonstrated by the preparation of a set of mono- and bis-maleimide functionalized NOTA derivatives. In conclusion, these two methods can easily be expanded to the syntheses of further tailored maleimide-NOTA chelators for diverse applications.Two basic and simple synthetic routes for mono- and bis-maleimide bearing 1,4,7-triazacyclononane- Cx et al.  describein vivo binding affinity due to the multivalency effect thiourea}benzyl)-4,7-bis(carboxy-methyl)-1,4,7-triazacylononane}acetic acid (3a). HPLC method A1: tR = 11.4 min. HPLC method B1: tR = 14.5 min. Yield: 3.3 mg . ESI-MS m/z 591 [M+H]+, 613 [M+Na]+. 1H-NMR: 2.92–4.03 , 7.00 , 7.32 , 7.51 .(S)-2-propanamido]-hexanamido}-hexyl)thiourea]benzyl}-4,7-bis(carboxymethyl)-1,4,7-triazacylononane)acetic acid (3b). HPLC method A1: tR = 17.3 min. HPLC method B1: tR = 19.9 min. Yield: 4.5 mg . ESI-MS m/z 510 [M+H+Na]2+, 521 [M+2Na]2+, 997 [M+H]+, 1019 [M+Na]+, 1035 [M+K]+. 1H-NMR: 1.09–1.60 , 1.91 , 2.28–2.41 , 2.60–3.73 , 3.93 , 6.68 , 7.06 , 7.18 .2  in phosphate buffer  the corresponding N-hydroxysuccinimide ester functionalized maleimide  was added. The slightly yellow dispersion was incubated at 25 °C for 1.5 h. The reaction mixture was centrifuged and the supernatant was taken for HPLC purification. HPLC-fractions containing the product were lyophilized.To a solution of (S)-2-(2-){4-[3--pyrrole-1-yl)propanamido]benzyl}-4,7-bis(carboxy-methyl)-1,4,7-triazacylononane)acetic acid (4a). HPLC method A2: tR = 12.7 min. HPLC method B2: tR = 15.3 min.Yield: 2.4 mg . ESI-MS m/z 560 [M+H]+, 582 [M+Na]+. 1H-NMR: 2.53 , 2.60–3.70 , 3.74 , 6.70 , 7.10–7.30 .(S)-2--pyrrole-1-yl)butanamido]benzyl}-4,7-bis-(carboxymethyl)-1,4,7-triazacylononane)acetic acid (4b). HPLC method A2: tR = 13.6 min. HPLC method B2: tR = 14.8 min. Yield: 2.4 mg . ESI-MS m/z 574 [M+H]+, 596 [M+Na]+, 612 [M+K]+. 1H-NMR: 1.86 , 2.29 , 2.40–3.80 , 6.62 , 7.10–7.26 .(S)-2--pyrrole-1-yl)hexanamido]benzyl}-4,7-bis-(carboxymethyl)-1,4,7-triazacylononane)acetic acid (4c). HPLC method A2: tR = 14.8 min. HPLC method B2: tR = 16.3 min. Yield: 2.6 mg . ESI-MS m/z 602 [M+H]+, 624 [M+Na]+ hexanoic acid N-hydroxysuccinimide ester due to insufficient separation capacity).(S)-2-[2--pyrrole-1-yl)ethanamido]-4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61,64,67,70,73,76,79,81,84-octacosaoxa-heptaoctacontan}benzyl)-4,7-bis-(carboxymethyl)-1,4,7-triazacylononane]acetic acid (4d): To a solution of 2  in 200 μL phosphate buffer (pH = 6.6) Mal-OEG28-NHS ester  was added. The slightly yellow solution was incubated at 26 °C for 2.0 h. Product 4d was isolated by HPLC separation. HPLC-fractions containing product 4d were lyophilized to obtain 4d as a yellow-white powder. HPLC method A4: tR = 20.4 min. HPLC method B4: tR = 32.4 min. Yield: 4.7 mg . ESI-MS m/z 944 [M+H+Na]2+, 955 [M+2Na]2+. MALDI-TOF m/z 933 [M+2H]2+, 1864 [M+H]+, 1886 [M+Na]+, 1902 [M+K]+.4--pyrrole-1-yl)propanamido]hexanamido}hexylamino)-4-oxobutanoic acid (5). To a solution of the bis-maleimide starting compound Bis-mal-Oc-NH2*TFA  in anhydrous DMF-TEA  was added. Succinic anhydride  was dissolved in anhydrous DMF (70 μL) and added dropwise. The reaction mixture was incubated at 26 °C for 30 min. After the addition of TFA  to the colorless solution, the reaction mixture was fractionized by HPLC. HPLC-fractions containing product 5 were lyophilized to obtain 5 as a white powder. Yield: 8.1 mg . ESI MS (m/z): 647 [M+H]+, 669 [M+Na]+, 685 [M+K]+, 645 [M−H]−. HPLC method A2: tR = 13.0 min. HPLC method B2: tR = 17.5 min. 1H-NMR: 1.32–1.50 , 1.52–1.66 , 1.72–1.85 , 2.59–2.78 , 3.20–3.35 , 3.88–3.95 , 4.21 , 6.99 , 8.13  8.40 .(S)-2--propanamido]-hexanamido}-hexylamino)-4-oxobutanamido]benzyl}-4,7-bis-(carboxymethyl)-1,4,7-triazacylononane)acetic acid (6). Compound 5 (6.7 μmol) was dissolved in anhydrous DMF and TEA  was added. An anhydrous DMF solution (35 μL) of COMU  was added dropwise. After reaction time for a maximum of 40 min at 26 °C, a solution of 2 , dissolved in DMF (55 μL) and TEA , was added. The reaction mixture was incubated at 26 °C over night. After addition of TFA  the reaction mixture was fractionized by HPLC. HPLC-fractions containing product 6 were lyophilized to obtain 6 as a white powder. Yield: 1.46 mg . ESI MS (m/z): 520 [M + 2H]2+, 531 [M + H + Na]2+, 1038 [M + H]+, 1060 [M + Na]+. HPLC method A3: tR = 31.8 min. HPLC method B3: tR = 51.8 min. 1H-NMR: 1.20–1.34 , 1.37–1.48 , 1.58–1.74 , 2.45–2.78 , 2.80–4.02 , 4.1 , 6.85 , 7.30 , 7.40 .p-SCN-Bn-NOTA 1 and maleimide starting materials with primary amino groups under formation of thiourea bonds. In the second pathway, maleimide derivatives with reactive carboxylic acid ester residues and p-NH2-Bn-NOTA 2 were coupled to form amide bound products. The practicability of these two methods is demonstrated by the preparation of a set of different derivatives 3a–3b, 4a–4d and 6. The isolation of thiourea bond-bearing products 3a and 3b was initially difficult due to side-reactions after HPLC separation. The side-reactions were suppressed by increasing the pH value of both mobile phases during HPLC purification.We have developed two basic and simple synthetic pathways for the preparation of a variety of new mono- and bis-maleimido-NOTA chelators. The first route is based on nucleophilic addition reactions of 4d, which is characterized by a very long spacer consisting of 28 ethylene glycol units. Such compounds can be used to enable site-specific radiolabeling and optimization in terms of blood circulation half life, enhanced targeting rates, conjugate polarity, lower immunogenicity, or minimization of unspecific tissue uptake. Several maleimide compounds with reactive NHS ester and different OEG- or PEG-spacer lengths are commercially available to allow the preparation of suitable derivatives according to individual requirements.We also synthesized a derivative The described synthetic methods can be transferred to any other amino group or active ester bearing maleimide starting material. This enables access to tailored bifunctional NOTA-maleimide chelators for modification and subsequent radiolabeling of potential targeting molecules.in vivo behavior. Similarly, other chelating systems will be modified by utilization of the developed synthetic methods.Prospective investigations include the conjugation of selected NOTA-maleimide products with target-specific compounds as well as the evaluation of their radiolabeling ability and


H-pyrrole-2-carboxylates interact with 3-(arylamino)-1H-inden-1-ones to give the corresponding 1,1'-diaryl-3'-cinnamoyl-4'-hydroxy-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-triones in good yields.Methyl 1-aryl-3-cinnamoyl-4,5-dioxo-4,5-dihydro-1 The last step of the transformation is accompanied by the elimination of a water molecule and is completed by the substituted spiroquinoline-10,3'-pyrrole] derivative formation pyrrole-3,2'-pyrrole]-2,4,5'(1'H)-triones 3 in good yields  with iron(III) chloride for the presence of enol hydroxyl groups. The IR spectra of 3 have absorption bands inherent to stretching vibrations of the enolic hydroxy group , two lactam carbonyl groups C5'=O (1763–1781 cm−1) and C2=O (1715–1732 cm−1), and two ketone carbonyl moieties C4=O (1667–1680 cm−1) and C3'-C=O (1640–1647 cm−1). 1H-NMR spectra of 3 display signals of protons in the aromatic rings and substituents attached thereto, two doublets from the protons of the ethylene fragment of the cinnamoyl substituent (δ 7.64–7.66 and 7.72–7.75 ppm) with coupling constant (3J) values of about 16 Hz, and a broadened singlet from the enolic hydroxy proton (δ 13.31–13.45 ppm). In the 13C-NMR spectra of 3d we have observed carbon atom signals of the aromatic and aliphatic fragments, the carbonyl carbon atom of the cinnamoyl moiety (δ 182.46 ppm), ketone carbonyl carbon atom C4 (δ 183.74 ppm), lactam carbonyl carbon atoms C2 (δ 171.89 ppm) and C5' (δ 165.38 ppm), and spiro carbon atom (δ 68.34 ppm). The structure of 3c was unambiguously confirmed by single-crystal X-ray crystallography , H1-O4A 1.71(2), O1-O4A 2.644(3) Å, angle O1H1O4A 144(1)°] and O1A-H1A-O4 .Bond lengths and bond angles in compound 3 occurs due to the addition of the β-CH group of the enamino fragment of indenones 2 to the atom C5 of pyrrolediones 1, followed by the closure of the pyrrole ring through the intramolecular attack of the amino group of indenones 2 on the ester carbonyl group in the position 5 of the pyrroledione with simultaneous elimination of methanol  with DMSO-6d as solvent and TMS as internal reference, chemical shifts are expressed as δ ppm. All reactions were followed by TLC . X-Ray analyses were performed using an “Xcalibur 3” diffractometer.Melting points were recorded on a Gallenkamp apparatus. IR spectra  were recorded on an FMS-1201 spectrophotometer. The 3c. These data can be obtained free of charge via www.ccdc.cam.ac.uk/conts/retrieving.html .CCDC 909 605 contains the supplementary crystallographic data for compound 1 and enamine 2 in dry toluene (20 mL) was heated under reflux for 5–6 h (progress of the reactions was monitored using TLC with 5:1 benzene-ethyl acetate as eluent). The mixture was then cooled, the resulted precipitate was filtered off and recrystallized from toluene.A solution of 1.0 mmol of pyrroledione 1-Bromo-3'-cinnamoyl-4'-hydroxy-1'-phenyl-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3a): Red solid, yield 80%, mp 272–274 °C. IR νmax: 3173 (OH), 1768 (C2=O), 1726 (C5'=O), 1671 (C4=O), 1642 (C3'-C=O) cm−1. 1H-NMR δ: 6.51 , 7.13–7.89 , 7.66 , 7.75 , 13.35 .3'-Cinnamoyl-4'-hydroxy-1'-(4-methylphenyl)-1-phenyl-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3b): Red solid, yield 82%, mp 274–275 °C. IR νmax: 3174 (OH), 1781 (C2=O), 1725 (C5'=O), 1674 (C4=O), 1642 (C3'-C=O) cm−1. 1H-NMR δ: 2.29 , 6.38 , 7.02–7.69 , 7.65 , 7.74 , 13.40 .3'-Cinnamoyl-4'-hydroxy-1,1'-di(4-methylphenyl)-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3c): Red solid, yield 79%, mp 282–283 °C. IR νmax: 3174 (OH), 1763 (C2=O), 1727 (C5'=O), 1678 (C4=O), 1647 (C3'-C=O) cm−1. 1H-NMR δ: 2.28 , 2.43 , 6.39 , 7.01–7.69 , 7.64 , 7.74 , 13.31 . The structure of compound 3c was determined on a single-crystal X-ray diffractometer “Xcalibur 3” equipped with a CCD detector using the standard procedure  К, ω-scanning, scanning step 1°). The red crystals had dimensions 0.25 × 0.20 × 0.15 mm, crystal system monoclinic, the space group P21/n, unit cell parameters: a = 15.2533(6), b = 16.6750(12), c = 26.0000(19) Å, β = 101.496(4)°, V = 6480.4(7) Å3, Z = 8, calcd = 1.186 g/cm3. 23,715 reflections were collected in the range of angles of 2.57° < θ < 26.40°, from which 13,082 independent reflections (Rint = 0.0361) and 5076 with I > 2σ(I). The completeness of the experiment at the corners of θ ≤ 26.0° is 98.8%. The structure was solved by direct methods and refined by full-matrix least-squares procedure on F2 with the SHELXTL-97 pyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3d): Red solid, yield 84%, mp 275–276 °C. IR νmax: 3180 (OH), 1777 (C2=O), 1732 (C5'=O), 1680 (C4=O), 1640 (C3'-C=O) cm−1; 1H-NMR δ: 2.29 , 3.87 , 6.40 , 7.02–7.69 , 7.65 , 7.74 , 13.35 . 13C-NMR δ ppm: 20.54 (Me), 55.47 (OMe), 68.34 (C-3), 108.08 (C-3a), 115.03 (CmAr2), 117.72 (C-3'), 120.52–138.40, 142.74 (CO-CH=CH-Ph), 155.63 (C-8b), 159.89 (CipAr2), 165.38 (C-5'), 171.89 (C-2), 177.22 (C-4'), 182.46 (C3'-C=O), 183.74 (C-4).3'-Cinnamoyl-4'-hydroxy-1'-(4-methoxyphenyl)-1-phenyl-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3e): Red solid, yield 81%, mp 269–270 °C. IR νmax: 3180 (OH), 1773 (C2=O), 1715 (C5'=O), 1669 (C4=O), 1640 (C3'-C=O) cm−1. 1H-NMR δ: 3.74 , 6.38 , 7.36–7.69 , 7.65 , 7.73 , 13.40 .3'-Cinnamoyl-4'-hydroxy-1'-(4-methoxyphenyl)-1-(4-methylphenyl)-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3f): Red solid, yield 80%, mp 252–253 °C. IR νmax: 3161 (OH), 1773 (C2=O), 1725 (C5'=O), 1671 (C4=O), 1644 (C3'-C=O) cm−1. 1H-NMR δ: 2.43 , 3.74 , 6.39 , 7.03–7.69 , 7.65 , 7.73 , 13.35 .3'-Cinnamoyl-4'-hydroxy-1,1'-di(4-methoxyphenyl)-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3g): Red solid, yield 79%, mp 259–260 °C. IR νmax 3161 (OH), 1773 (C2=O), 1717 (C5'=O), 1667 (C4=O), 1642 (C3'-C=O) cm−1. 1H-NMR δ: 3.74 , 3.86 , 6.40 , 7.03–7.73 , 7.65 , 7.73 , 13.30 .1-Bromo-3'-cinnamoyl-4'-hydroxy-1'-(4-methoxyphenyl)-1H-spiropyrrole-3,2'-pyrrole]-2,4,5'(1'H)-trione (3h): Red solid, yield 83%, mp 256–257 °C. IR νmax 3188 (OH), 1777 (C2=O), 1732 (C5'=O), 1673 (C4=O), 1644 (C3'-C=O) cm−1. 1H-NMR δ: 3.74 , 6.51 , 7.02–7.87 , 7.64 , 7.72 , 13.45 .H-pyrrole-2,3-diones with 3-(arylamino)-1H-inden-1-ones. Our work presents a very simple reaction performed under neutral conditions and in the absence of any catalyst. From a structural viewpoint, the products are polycarbonyl compounds suitable for further elaboration. High yields and simple reaction and purification procedures are the key advantages of this approach.The described reaction presents a rare example of a regioselective synthesis of a hardly accessible spiroheterocyclic system—spiropyrrole-3,2'-pyrrole] including various functional substituents in both heterocyclic fragments. Overall, we have succeeded in developing a method for synthesis of new functionalized spiropyrrole-3,2'-pyrroles derivatives of potential synthetic and pharmacological interest from the reaction of 1


H-1,2,3-triazol-1-yl)methyl)quinoline (1), 2-((4-(m-tolyl)-1H-1,2,3-triazol-1-yl)methyl)quinoline (2) and 2-((4-(p-tolyl)-1H-1,2,3-triazol-1-yl)methyl)quinoline (3) have been prepared with CuAAC click reactions and used as a model series to probe the relationship between lattice H-bonding interaction and crystal direction of growth. Crystals of 1–3 are 1D tape and prism shapes that correlate with their intermolecular and solvent 1D lattice H-bonding interactions. All compounds were thermally stable up to about 200 °C and blue-green emissive in solution. Quinoline-triazoles 2-((4-(diethoxymethyl)-1 Each reaction was stirred at 50 °C for 20 h on a MultiMax reactor. The alkynes, 3,3-diethoxy-1-propyne, 3-ethynyltoluene and 4-ethynyltoluene were used for the synthesis of 1–3, respectively. methyl)quinoline (1), C17H20N4O2, 312.37. Yield: 560 mg, 90%. ESI-MS : [1 + H]+ . Main IR bands (cm−1): 3111 (m), 3069 (m), 2976 (m), 2926 (m), 2904 (m), 2878 (m), 1618 (m), 1598 (m), 1567 (m), 1506 (m), 1455 (m), 1426 (m), 1385 (m), 1345 (m), 1320 (m), 1309 (m), 1271 (m), 1225 (m), 1178 (m), 1117 (m), 1097 (m), 1061 (s), 1050 (s), 1038 (m), 1021 (m), 1004 (m), 910 (m), 873 (m), 847 (m), 823 (m), 803 (m), 778 (m), 745 (m), 734 (m), 618 (m), 491 (m), 479 (m) and 432 (m). The bands among 1004–1117 cm−1 should be attributed to the C−O stretching vibrations. 1H-NMR  δ 8.15 and 8.13 , 8.07 and 8.05 , 7.82 and 7.80 , 7.76–7.73 , 7.58–7.55 , 7.28 and 7.27 , 5.83 , 5.71 , 3.73–3.56 , 1.22, 1.21 and 1.19 . 13C-NMR  δ 154.6, 147.85, 147.77, 137.8, 130.3, 129.3, 127.8, 127.7, 127.3, 122.8 and 119.7 (quinoline and triazole groups), 96.9, 61.8, 56.5 (quinoline-CH2-triazole) and 15.2.2-(Chloromethyl)quinoline hydrochloride , Nactively.  On complm-Tolyl)-1H-1,2,3-triazol-1-yl)methyl)quinoline (2), C19H18N4O (2·H2O), 318.37. Yield: 510 mg, 80%. ESI-MS : [2 + H]+ . Main IR bands (cm−1): 3134 (m), 2944 (m), 2918 (m), 1664 (m), 1615 (m), 1600 (m), 1568 (m), 1508 (m), 1488 (m), 1460 (m), 1432 (m), 1420 (m), 1360 (m), 1322 (m), 1230 (m), 1218 (m), 1179 (m), 1142 (m), 1121 (m), 1082 (m), 1045 (m), 980 (m), 961 (m), 894 (m), 847 (m), 821 (m), 797 (s), 784 (m), 760 (m), 749 (m), 727 (m), 697 (m), 618 (m), 477 (m), 442 (m) and 410 (m). 1H-NMR  δ 8.17–8.10 , 7.94–7.93 , 7.81–7.79 , 7.77–7.73 , 7.63 , 7.58, 7.56 and 7.55 , 7.34–7.30 , 7.25 and 7.23 , 7.10 and 7.08 , 5.90–5.88  and 2.34 . 13C-NMR  δ 154.5, 148.6, 147.1, 138.6, 138.5, 130.7, 130.4, 129.1, 128.82, 128.78, 127.9, 127.7, 127.5, 126.5, 123.0, 120.4 and 119.9 (quinoline and triazole groups), 56.0 (quinoline-CH2-triazole) and 21.5 (CH3).2-((4-(p-Tolyl)-1H-1,2,3-triazol-1-yl)methyl)quinoline (3), C19H16N4, 300.36. Yield: 490 mg, 82%. ESI-MS : [3 + H]+ . Main IR bands (cm−1): 3132 (m), 3111 (m), 2939 (m), 2920 (m), 1617 (m), 1601 (m), 1568 (m), 1499 (m), 1460 (m), 1425 (m), 1356 (m), 1323 (m), 1313 (m), 1225 (m), 1187 (m), 1141 (m), 1118 (m), 1076 (m), 1049 (m), 1022 (m), 976 (m), 890 (m), 821 (m), 812 (s), 778 (m), 763 (m), 746 (m), 730 (m), 616 (m), 517 (m), 477 (m) and 415 (m). 1H-NMR  δ 8.17 and 8.16 , 8.11 and 8.09 , 7.89 , 7.84 and 7.82 , 7.78–7.75 , 7.71–7.70 , 7.60–7.57 , 7.34 and 7.32 , 7.22 and 7.20 , 5.88  and 2.36 . 13C-NMR  δ 154.9, 148.7, 147.8, 138.2, 137.9, 130.4, 129.6, 129.4, 127.9, 127.8, 127.7, 127.3, 125.8, 120.0 and 119.8 (quinoline and triazole groups), 56.6 (quinoline-CH2-triazole) and 21.4 (CH3).2-((4-(1–3 were synthesized using 2-(chloromethyl)quinoline hydrochloride, sodium azide and an alkyne  in good to high yields (80–90%). The structures of 1–3 were characterized by 1H-NMR and 13C-NMR spectroscopy  align with the dominant crystal belt direction (b axis). Compound 3 bearing a pendant p-tolyl group is isostructural with 2 in generating a prism shaped crystal along the dominant b axis. The bulk crystal does not capture lattice solvent, but displays 1D H-bond interactions as in 3,  in 2 probably decreases the possibility of forming intermolecular H-bonding interaction.Quinoline-triazoles troscopy , electroas in 3,  which sib) in all three compounds 1–3. We are currently investigating if such correlation can be extended to related systems, and the potential of small molecule  entrapment to control crystal shape, size and function. This series of molecules show some correlation between compound lattice structures and the resulting shape of bulk single crystals. The lattice H-bonding directions are aligned with the direction of material growth, which also align with the shortest axis  before 115 °C in 2 was assigned to the loss of lattice water molecules (5.7%). All three compounds continue to degraded and reach residual weights at about 640 °C.The experimental powder X-ray diffraction patterns for compounds e purity a. Thermo1 in ethanol displayed two broad absorption bands centred at 205 and 230 nm  AXS GADDS X-ray diffractometer with Cu-Kα radiation (λ = 1.54056 Å). Single-crystal X-ray diffraction data were collected using a Bruker AXS SMART APEXII CCD diffractometer using Mo-Kα radiation (λ = 0.71073 Å) , 1453357(2 H2O) and 1453358(3) contain the supplementary crystallographic data for this paper. These data can be obtained free of charge via http://www.ccdc.cam.ac.uk/conts/retrieving.html .All starting chemicals were used as received. Electrospray ionization mass spectra (ESI-MS) were recorded in positive ion mode using a Shimadzu LCMS-IT-TOF mass spectrometer. NMR spectra were measured at room temperature using a JEOL 500 MHz NMR spectrometer. Thermogravimetric analysis (TGA) was carried out in an air stream using a 71073 Å) . Data inr SHELXL . Structu1–3, conveniently prepared by copper-catalyzed azide-alkyne cycloaddition reactions. Lattice H-bonding interactions observed from their single-crystal structures align with the shortest axis. They are also the primary driving force for the direction of bulk crystal growth. This trend was seen for all three compounds and we are investigating the predictive capabilities of this correlation between intermolecular interactions and the direction of crystal growth with a view to more rational crystal engineering.This work reports a series of new, luminescent quinoline-triazoles


Z)-neomanoalide-24-acetate, two diastereomers of 24-O-methylmanoalide, luffariolide B, manoalide, (6E)- and (6Z)-neomanoalide, seco-manoalide, scalarafuran, 12-acetylscalarolide, 12-epi-O-deacetyl-19-deoxyscalarin, 12-epi-scalarin, and 12-O-deacetyl-12-epi-scalarin, three indole alkaloids, 5-hydroxy-1H-indole-3-carbaldehyde, hyrtiosine A, and variabine B, and one norterpene, cavernosine were isolated from the marine sponge Hyrtios erectus. Their structures were determined by means of spectroscopic methods and the absolute configurations of the asymmetric centers were determined using the modified Mosher’s method. The cytotoxic activities for the isolated compounds have been reported.Four sesterterpenes, erectusolides B, C, D, and seco-manoalide-25-methyl ether, two 2-furanone derivatives, erectusfuranones A and B, together with thirteen known sesterterpenes, (6 Hyrtios (order Dictyoceratida family Thorectidae) have yielded scalarane  oct-2-ene (C4–C11 and C-24) in the molecule. The HMBC correlations of 1 (H 5.86 (H-2) correlated to methine carbon at δC 51.0 (C-4), H-4 at δH 2.74 correlated to quaternary carbon at δC 169.7 (C-3), methine carbon at δC 115.3 (C-2), and methylene carbon at δC 73.2 (C-25) allowed the connectivity of the butenolide with the bicyclic ring through the linkage of C-3 and C-4. The linkage of the polyalkylated-cyclohexene and CH3-23 at quaternary C-11 on the molecule were deduced from the HMBC correlations of H3-23 (δH 1.13) with C-4, C-10 (δ 37.0), C-11 (δ 43.0), and C-12 (δ 36.6) and the correlations of H2-12 (δH 1.28–1.36 and 1.58–1.66) with C-4, C-10, C-14 (δ 136.3), and C-23 (δ 21.5). The large trans-diaxial coupling constant between Hax-9 and Hax-10 (J = 12.2 Hz) on the cyclohexene ring suggested the β-configuration of Hax-10. The relative stereochemistry of 1 was established by analysis of the (nuclear overhauser effect spectroscopy (NOESY) spectrum  are on the same face while the methyl group (CH3-23) and H-4 on the opposite face of the molecule. Structure 1 with relative stereochemistry as shown was thus assigned to erectusolide B, which contained fused cyclohexene and cyclobutane rings. It could derive from luffariolide A  by HRAPCIMS. Infrared (IR) absorption bands of compound 2 suggested the presence of β-substituted α,β-unsaturated γ-lactone at 1779 and 1746 cm−1 and α,β-unsaturated carbonyl group at 1682 cm−1. The 1H and 13C NMR spectroscopic data of 2  for 2 in place of the hydroxylmethyl group  for 1. This was further confirmed by the HMBC correlations (H 6.74) and aldehydic carbon (δC 192.9) and between proton aldehyde (δH 9.45) and C-7 (δC 143.7) and C-8 (δC 19.4). The NOESY spectrum of 2 was similar to that observed of 1 indicating the same relative stereochemistry. Thus, compound 2 was suggested to be the formaldehyde analog of 1, and named erectusolide C. This compound has low optical rotation , infrared (IR), 1H, and 13C NMR spectra (E) seco-manoalide (14) [3 indicated a pseudo molecular ion peak at m/z 465.2414 [M + Cl]− , consistent with the molecular formula C26H38O5. This difference of 14 amu compared to the molecular formula of 14, and the appearance of a signal of methoxy group in 1H, and 13C NMR spectra of 3 at δH 3.65 and δC 57.9. It is suggested that the hydroxyl group at C-25 was replaced by a methoxyl group. This was further confirmed by the HMBC correlations (H 3.65) and hemiacetal carbon (δC 103.3) and between H-25 (δH 5.84) and methoxyl carbon (δC 57.9). The absolute stereochemistry at C-4 of 3 was determined by the modified Mosher’s method  to afford 21 (1.4 mg). F7 (508.1 mg) was subjected to column chromatography on Sephadex LH-20, using 100% MeOH to give a mixtures compound (12.1 mg) that was separated using on a HPLC column  to afford compound 22 . F8 (298.1 mg) was repeatedly fractionated by Sephadex LH-20, using CH2Cl2–MeOH (1:1) to give a mixture compound (127.6 mg) that was separated using a HPLC column  to afford compounds 4 , and 13 . F9 (249.2 mg) was subjected to repeated chromatography on Sephadex LH-20, using CH2Cl2–MeOH (1:1) to give a mixtures compound (20.3 mg) that was separated using a HPLC column , flowrate 12 mL/min, λ 234 nm) to afford compound 23 . The flesh sponge Hyrtios erectus collection CRI 588 (14.0 kg) was cut into small pieces and extracted repeatedly with MeOH (20 L) three times (3 × 20 L). After the evaporation of the solvent, the concentrated MeOH extracts was partitioned between EtOAc and water and the EtOAc fraction was chromatographed on a silica gel column with CH2Cl2–hexane (1:1) containing increasing proportions of acetone as eluent, to give 7 fractions (A–G). Fraction C (4.0 g) was fractionated by Sephadex LH-20 with MeOH–CH2Cl2 (1:1) as eluent, to give 2 fractions (C1–C2). C2 (230.3 mg) was repeatedly chromatography on Sephadex LH-20, using CH2Cl2–MeOH (4:1) to give a mixtures compound (200 mg) that was separated using a HPLC column  with MeOH–H2O (93:7), flowrate 2.5 mL/min, λ 224 nm, to afford compounds 5 (4.1 mg at 45.5 min), and 6 (7.2 mg at 48.2 min). Fraction D (9.0 g) was further fractionated on a Sephadex LH-20 column chromatography with MeOH–CH2Cl2 (1:1) to yield 4 fractions (D1–D4). D4 (400 mg) was further purified by HPLC column  with CH3CN–H2O (80:20), flowrate 8 mL/min, λ 224, 236 nm, to afford compounds 1 , 2 , and 3 . Fraction E (25.0 g) was further fractionated by vacuum liquid chromatography eluted with CH2Cl2–hexane (1:1) containing increasing proportions of acetone as eluent, to give 4 fractions (E1–E4). Fraction E1 (2 g) was further purified by a HPLC column  with CH3CN–H2O (85:15), flowrate 8 mL/min, λ 224, 236 nm, to afford compounds 7 , 8 , and 9 . Fraction E3 (3 g) was further purified by a HPLC column  with CH3CN–H2O (80:20), flowrate 8 mL/min, λ 224, 236 nm, to afford compounds 10 , and 11 . Fraction E4 (1.4 g) was further purified by HPLC column  with CH3CN–H2O (82:18), flowrate 8 mL/min, λ 224, 236 nm, to afford compounds 13  and 14 . Fraction G (4.0 g) was further filtrated on a Sephadex LH-20 column chromatography with MeOH to give a residue (655 mg), which was further purified by HPLC column  with CH3CN–H2O (60:40), flowrate 8 mL/min, λ 224, 236 nm, to afford compound 12 .A frozen sample (2.6 kg) of S)- and (R)-MTPA Esters of compound 3: To a solution of 3 (3.1 mg) in pyridine (0.9 mL) was added (R)-MTPA-chloride (17 µL). The mixture was stirred at room temperature for 3 h, checked with thin layer chromatography (TLC) to make sure that the reaction was complete, quenched by the addition of 2 mL of H2O, and the mixture was subsequently extracted with CH2Cl2 (2 mL) three times (3 × 2 mL). The CH2Cl2 soluble layers were combined, dried over anhydrous MgSO4, and evaporated. The residue was subjected to short silica gel column chromatography using hexane–EtOAc (4:1) to give the (S)-MTPA ester 3a (1.6 mg).Preparation of (1H NMR (CDCl3 600 MHz) of 3a: δH 9.32, s, H-24; 6.28, t (J = 7.1 Hz), H-6; 5.86, brs, H-2; 5.79, s, H-25; 5.74, dd , H-4; 5.09, t (J = 7.1 Hz), H-10; 3.03, ddd , Ha-5; 2.90, dt , Hb-5; 2.24, t (J = 7.6 Hz), H2-8; 2.02, m, H2-9. Selected R)-MTPA ester 3b (2.5 mg from 3.9 mg of 3) with (S)-MTPA chloride.The same procedure was used to prepare the (1H NMR (CDCl3 600 MHz) of 3b: δH 9.21, s, H-24; 6.13, t (J = 7.1 Hz), H-6; 6.00, brs, H-2; 5.81, s, H-25; 5.74, dd , H-4; 5.07, t (J = 6.9 Hz), H-10; 3.00, ddd , Ha-5; 2.87, dt , Hb-5; 2.20, t (J = 7.7 Hz), H2-8; 1.99, m, H2-9Selected S)-MTPA esters 5a (1.3 mg from 1.4 mg of 5) and 6a (1.8 mg from 1.4 mg of 6) and (R)-MTPA esters 5b (1.5 mg from 1.4 mg of 5), 6b (1.7 mg from 1.2 mg of 6) were prepared by the method mentioned above.  of 5a: δH 5.77, brs, H-3; 2.72, dd , Ha-1′; 2.62, dd , Hb-1′; 5.28, m, H-2′; 1.67, m, H2-3′; 4.56, dd , Ha-5; 4.25, brd (J = 17.6 Hz), Hb-5. Selected 1H NMR  of 5b: δH 5.86, s, H-3; 2.78, dd , Ha-1′; 2.69, dd , Hb-1′; 5.29, m, H-2′; 1.63, m, H2-3′; 4.69, brd (J = 17.4 Hz), Ha-5; 4.52, brd (J = 17.4 Hz), Hb-5.Selected 1H NMR  of 6a: δH 5.77, brs, H-3; 2.72, dd , Ha-1′; 2.63, dd , Hb-1′; 5.28, m, H-2′; 1.71, m, Ha-3′; 1.59, m, Hb-3′; 1.29, m, H2-4′; 4.57, dd , Ha-5; 4.26, brd (J = 16.7 Hz), Hb-5.Selected 1H NMR  of 6b: δH 5.86, brs, H-2; 2.78, dd , Ha-1′; 2.69, dd , Hb-1′; 5.29, m, H-2′; 1.67, m, Ha-3′; 1.55, m, Hb-3′; 1.23, m, H2-4′; 4.69, d (J = 17.5 Hz), Ha-5; 4.52, d (J = 17.5 Hz), Hb-5.Selected 1): Colorless powder; c 0.56, CHCl3); UV λmax (MeOH) nm (log ε) 202 (3.9), 224 (3.4), IR (ATR) νmax 3443 (br), 2927, 2861, 1779, 1746, 1629, 1456, 1379, 1266, 1143, 1041, 888, 851, 735, 702 cm−1; 1H and 13C NMR data see m/z 385.2727 [M + H]+ .Erectusolide B (2): Colorless powder; c 0.16, CHCl3); UV λmax (MeOH) nm (log ε) 234 (4.3); IR (ATR) νmax 2924, 2855, 1779, 1748, 1682, 1632, 1458, 1378, 1261, 1142, 1040, 887 cm−1; 1H and 13C NMR data see m/z 405.2390 [M + Na]+ .Erectusolide C (3): Pale yellow solid; c 1.86, CHCl3); UV λmax (MeOH) nm (log ε) 203.5 (4.3), 232 (sh); IR (ATR) νmax 3446 (br), 2930, 2863, 1762, 1687, 1457, 1371, 1203, 1120, 1070, 960, 898, 870, 735, 702 cm−1; 1H and 13C NMR data see m/z 465.2414 [M + Cl]− .seco-Manoalide-25-methyl ether (4): Pale yellow gum; c 1.64, CHCl3); UV λmax (MeOH) nm (log ε) 202 (4.3), IR (ATR) νmax 3424 (br), 2928, 2857, 1739, 1642, 1456, 1382, 1258, 1172, 1063, 898 cm−1; 1H and 13C NMR data see m/z 453.2418 [M + Cl]− .Erectusolide D (5): Colorless powder; c 0.32, CHCl3); UV λmax (MeOH) nm (log ε) 206 (3.9); IR (ATR) νmax 3442 (br), 2923, 2853, 1777, 1739, 1637, 1457, 1378, 1175, 1027, 888, 720 cm−1; 1H and 13C NMR data see m/z 367.3213 [M + H]+ .erectusfuranone A (6): Colorless powder; c 0.61, CHCl3); UV λmax (MeOH) nm (log ε) 206 (3.9); IR (ATR) νmax 3402 (br), 2916, 2851, 1779, 1756, 1736, 1634, 1469, 1025, 892, 720 cm−1; 1H and 13C NMR data see m/z 353.3053 [M + H]+ .erectusfuranone B -2,5-diphenyltetrazoliumbromide (MTT) assay ,26. For H. erectus were studied. Three new sesterterpenes, erectusolides B (1), C (2), and seco-manoalide-25-methyl ether (3), two new 2-furanone derivatives, erectusfuranones A (5) and B (6) together with eight known sesterterpenes, 7–14 were isolated from H. erectus collection CRI 588. One additional new sesterterpene, erectusolide D (4), and ten known compounds  were also obtained from the H. erectus collection CRI 572. Three sesterterpenes 8, 9, and 20 showed good cytotoxic activity against the MOLT-3 cell line with IC50 values of 1.30–7.64 μM and with SI (selectivity index) values 7–9.Two separate collections of