Elucidation of their structures was performed based on high-resolution electrospray ionisation mass spectrometry (HR-ESI-MS), infrared spectra (IR), and nuclear magnetic resonance (NMR) spectroscopic data. The anti-inflammatory activity of the isolates toward lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages was investigated. The results demonstrated that compounds 3, 5, and 6 inhibited the expression of inducible nitric oxide synthase (iNOS) protein in comparison with LPS stimulation in RAW264.7 cells.Seven new triterpenoid saponins ( Ilex pubescens, belonging to the plant family Aquifoliaceae, is widely distributed throughout the south of China. The roots and leaves of Ilex pubescens have been used in folk medicine for treating thromboangiitis obliterans -urs-19-oxo-18,19-secoursa-11,13(18)-dien-28-oic acid (Pubescenoside E (1)  was isolR-ESI-MS . The sug 13C NMR  showed 1oic acid , their smoieties . The rotntations . Along woic acid .F (2), a white amorphous powder, was determined to be C53H84O22 by the HRESIMS ion at m/z 1071.5388 [M − H]−  and NMR data. The IR spectrum revealed the existence of hydroxyl, olefinic, and carboxyl absorption bands. The sugar components of acid-hydrolyzed 2 included d-Xylose, d-glucoses, and l-rhamnose, as identified by TLC and HPLC analyses. The 13C NMR spectrum (δC 212.3 (C-19), one carboxyl at δC 175.1 (C-28), and four anomeric carbons. The 1H NMR spectrum , 1.32, and 2.13 and one methyl doublet at δH 1.04. The structure of 2 resembled that of 1, except for an additional glucose unit at C-28. The HMBC from Glc-H-1 to C-28 and inner-Xyl-H-1 to C-3 demonstrated glycosylation sites at the 28-O- and 3-O- positions. Eventually, compound 2 was elucidated as β-d-glucopyranosyl 3β-[α-l-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl-(1→2)-β-d-xylopyranosyl]-urs-19-oxo-18,19-secoursa-11,13(18)-dien-28-oate (The molecular formula of Pubescenoside spectrum  showed 5spectrum  of 2 dis-28-oate .G (3) displayed a molecular ion peak [M + COOH]− at m/z 1103.5642 , indicating that its molecular formula was C53H86O21. The IR data also manifested absorption bands for hydroxyl, alkyl, carbonyl, and double bond groups. The 1H NMR and 13C NMR data , one ester carbonyl carbon at C-28 (δC 176.8), four anomeric carbon signals, and seven methyl singlets at δH 0.82, 0.87, 0.91, 1.07 (6H), 1.24, and 1.33. The NMR data of 3 and 3-β-O-{[O-β-d-galactopyranosyl-(1→3)-β-d-glucopyranosyl-(1→2)]-β-d-glucooyranosyl}oleanolicacid 28-β-d-glucopyranosyl ester -olean-12-en-28-oate  indicated that the molecular formula of Pubescenoside H (4) was C53H86O22. The IR spectrum demonstrated the existence of hydroxyl, olefinic, and carboxyl absorption bands. The configurations of the sugar units were determined by hydrolysis to be d-Xylose, d-glucoses, and l-rhamnose. The 1H NMR and 13C NMR spectrum , 1.09, 1.25, and 1.32; one double bond ; one quaternary carbon at C-29 (δC 74.0); one ester carbonyl carbon at C-28 (δC 176.8); and anomeric protons of the four sugar units . The NMR data for compound 4 were almost same as for compound 3, except for the additional hydroxyl group at C-29. The linkages were confirmed by observation of HMBC from H-29 (δH 3.56) to C-19 (δC 41.2), C-20 (δC 36.7), and C-21 (δC 29.1). Based on these results, the structure of 4 was established as β-d-glucopyranosyl 3β-[α-l-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl-(1→2)-β-d-xylopyranosyl]-29α-hydroxyurs-olean-12-en-28-oate (The HR-ESI-MS data ([M − H]spectrum  gave six-28-oate .I (5) was obtained as a white amorphous powder with the molecular formula C53H84O21 (HR-ESI-MS: m/z 1101.5488 [M + COOH]−). In the IR spectrum, absorption bands for hydroxyl (3396 cm−1), alkyl (2930 cm−1), carbonyl (1729 cm−1), and double bond groups (1641 cm−1) were observed. The configuration of the sugar units was ascertained by hydrolysis to be d-Xylose, d-glucoses, and l-rhamnose. The 13C NMR data ; another tri-substituted double bond ; one ester carbonyl carbon at C-28 (δC 176.5); four anomeric signals ; five tertiary methyl at δH 0.81, 0.98, 0.99, 1.04, and 1.27 and two methyl doublets at δH 1.32  and δH 0.76 . The results described above indicated that Pubescenoside I (5) was highly similar to ilexsaponion L -urs-9(11),12-dien-28-oate  was obtained as white amorphous powder and had a molecular formula of C41H64O12, deduced from an ion peak in the HR-ESI-MS at m/z 793.4462 [M + COOH]− . The IR spectrum of 6 showed hydroxyl, alkyl, and carbonyl moieties at 3385 cm−1, 2941 cm−1, and 1729 cm−1, respectively. The sugar components of acid-hydrolyzed 6 included d-xylose and d-glucose, as identified through TLC and HPLC analyses. The 1H NMR and 13C NMR spectrum ; one carboxylic acid ; and two anomeric signals . The HMBC analysis was as follows: from H-12 to C-9, C-11 and C-14; from H-18 to C-12, C-13, C-14, C-16, C-17, C-20, C-28 and C-30; from H-25 to C-1, C-5, and C-9; and from H-30 to C-18, C-19 and C-20. A precise comparison of its 1H and 13C NMR data with those of ilexsaponin I − ions at m/z 845.4052 . The IR spectrum also showed absorption signals for hydroxyl, double bond, and ester groups. The 1H NMR data , and 1.73, and one methyl doublet at δH 1.11. Furthermore, signals for one tri-substituted double bond , one ester carbonyl carbon at C-28 (δC 177.3), one xylopyranose linked to C-3 of the aglycone, and one β-d-glucopyranose linked to C-28 of the aglycone were observed. The HMBC analysis results were as follows: from H-12 to C-9, C-14, and C-18; from H-18 to C-16, C-17, C-19, and C-20; from H-23 to C-3, C-4 and C-5; from H-25 to C-1, C-5, C-9 and C-10; from H-30 to C-19, C-20, and C-21; and from inner-Xyl-H-1 to C-3. The difference between compound 7 and ilexpublesnin E -19α-hydroxy-urs-12-en-28-oate (The molecular formula of Pubescenoside NMR data  of 7 sho-28-oate .8–10) (I (8) 3OH); IR (KBr) vmax: 3426, 2938, 2874, 1703, 1644, 1357, 1041 cm−1; NMR spectroscopic data , see m/z 955.4910 [M + COOH]− .White amorphous powder; [α]3OH); IR (KBr) vmax: 3396, 2930, 2877, 1700, 1635, 1360, 1074 cm−1; NMR spectroscopic data , see m/z 1071.5388 [M − H]− .White amorphous powder; [α]3OH); IR (KBr) vmax: 3408, 2930, 2870, 1738, 1646, 1384, 1077 cm−1; NMR spectroscopic data , see m/z 1103.5642 [M + COOH]− White amorphous powder; [α]3OH); IR (KBr) vmax: 3373, 2933, 2870, 1735, 1644, 1452, 1074 cm−1; NMR spectroscopic data , see m/z 1073.5614 [M − H]− .White amorphous powder; [α]3OH); IR (KBr) vmax: 3396, 2930, 2877, 1729, 1641, 1360, 1077 cm−1; NMR spectroscopic data , see m/z 1101.5488 [M + COOH]− . White amorphous powder; [α]3OH); IR (KBr) vmax: 3385, 2941, 2874, 1729, 1074 cm−1; NMR spectroscopic data , see m/z 793.4462 [M + COOH]− .White amorphous powder; [α]3OH); IR (KBr) vmax: 3458, 2938, 2880, 1735, 1646, 1230, 1065 cm−1; NMR spectroscopic data , see m/z 845.4052 [M − H]− .White amorphous powder; [2O (4:3:1) was chosen as the solvent system, and the spots were observed after spraying the plates with H2SO4 and heating at 105 °C for 1 min. The fR values of xylose, glucose, and rhamnose via TLC, which were hydrolyzed from Pubescenoside E–K, were 0.47, 0.60, and 0.71, respectively. The residue was dissolved in pyridine, and 2 mg of l-cysteine methyl ester hydrochloride was added. The mixture was stirred at 60 °C for 2 h, 5 μL O-tolyl isothiocyanate was added, and the mixture was stirred at 60 °C for another 2 h. After cooling, the reaction mixture was analyzed by reversed phase HPLC . The retention time of d-xylose , d-glucose , and l-rhamnose  was determined by comparison with standards.Each solution of the seven new compounds (2 mg) was stirred in 2N HCl (5 mL) at 80 °C in a stoppered reaction vial for 5 h. The solution was evaporated to dryness under vacuum, and then the residue was compared with standard sugars by silica gel TLC. N-butanol-acetone-Hl-glutamine (2 mM) in a humidified incubator containing 5% CO2 at 37 °C. Compounds 1–10 were completely dissolved in dimethyl sulfoxideto a final concentration of 30 mM, and the working concentration was 30 μΜ. RAW264.7 cells (8 × 104 cells/wells) were seeded in 24-well plates for 24 h, pretreated with compounds 1–10 or dexamethasone for 1 h, and then stimulated with LPS (100 ng/mL) for another 18 h. Dexamethasone at a concentration of 0.5 μΜ was selected as a positive control for inhibition of iNOS and COX-2 expression. LPS-stimulated cells without any intervention were used as the model control, and cells cultured in DMEM medium were used as the normal control. The Western blot methods were based on our previous research [RAW264.7 cells were purchased from American Type Culture Collection  and cultured in Dulbecco’s Modified Eagle’s Medium containing 10% heat-inactivatedfetal bovine serum, penicillin G (100 units/mL), streptomycin (100 mg/mL), and research .


All compounds were fully characterized by elemental analyses, 1H-NMR, 13C-NMR and IR spectroscopy techniques. All compounds studied in this work were screened for their in vitro antimicrobial activities against the standard strains: Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and the fungi Candida albicans and Candida tropicalis. The new imidazolidin-2-ylidene silver complexes have been found to display effective antimicrobial activity against a series of bacteria and fungi. A series of imidazolidinium ligand precursors are metallated with Ag With metallopharmaceuticals playing a significant role in therapeutic and diagnostic medicine, the discovery and development of new metallodrugs remain an ever-growing area of research in medicinal inorganic chemistry ,2,3. MetN-heterocyclic carbine (NHC) [N-Heterocyclic carbene complexes of Ag are commonplace in the organometallic literature. The interest in Ag-NHC complexes is largely due to their ease of synthesis and their ability to serve as useful to other NHC-metal complexes by NHC transfer reactions [The biomedical applications of metal complexes based on ne (NHC) ,16,17,18ne (NHC) ,20,21. Neactions .In additeactions ,26,27,28eactions , liquid eactions  and orgaeactions  also conAg-carbene complexes derived from imidazolium salts were synthesized and characterized for the first time by Arduengo in 1993 . These c1) isolated high yield, by complete elemental analyses, FT-IR, 1H and 13C-NMR spectroscopy. The finding silver based antimicrobials active against bacteria, we report the preparation and characterization of the imidazolidin-2-ylidene silver(I)  the reaction of a free NHC with silver salts , ii) theemployed . Soon afemployed , the Ag22O in CH2Cl2 afforded quantitavely after 24 hours the expected carbenes 1a-1f (1a‑f were obtained as white solids in 91–95% yields. The silver carbene complexes (1a–f) were soluble in halogenated solvents and insoluble in non-polar solvents.The ligand precursor 1,3-dialkylimidazolidinium chloride was obtained according to literature . Treatmees 1a-1f . Silver-1H and 13C-NMR spectra are consistent with the proposed formula. In the 1H-NMR and 13C-NMR spectra of this solid product in DMSO-d6 and CDCl3 loss of the imidazolidinium proton (NCHN) and imidazolidinium carbon (NCHN) signal suggests the formation of the silver-NHC complexes. The 13C-NMR spectra exhibit singlets at 203.1, 202.1 and 202.9 ppm for 1b, 1c and 1f respectively characteristic of the carbenic carbon resonance. The chemical shift is well consistent with those of the knows silver-NHC complexes in the range of 213-163 ppm [(NCN) band typically at 1,501, 1,500, 1,515, 1,502, 1,507 and 1,508 cm-1,respectively, for 1a-f.Their -163 ppm ,42. In t-163 ppm ,44,45. A1a-f as antimicrobial agents was evaluated. Minimal inhibitory concentrations for each compound were investigated against standard bacterial strains; Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and the fungal strains Candida albicans and Candida tropicalis. The test organism was laboratory strains used to test a range of concentration of the silver compounds for minimum inhibitory concentration determination. Antimicrobial activities of the Ag (I)-NHC complexes were determined by using agar dilution procedure and were tested with different concentrations of the compounds. The minimum inhibitory concentration (MIC) of synthesized silver complexes against Gram positive, Gram negative bacteria and fungus are summarized in The antimicrobial activity was reported in terms of the minimum inhibitory concentration (MIC) values, which are defined as the lowest concentration of an antimicrobial that visibly inhibits the growth of the bacteria after an overnight incubation . The use1a, 1c and 1f shoved high activity against the fungi C. albians and C. tropicalls with a range of MICs between 6.25–50 µg/mL. Incorporation of same groups at positions 2 and 3 on aromatic group (1a and 1b) exhibit difference in activity (1a is more active than 1b). In case of incorporation of an i-propyl group at position 4 on aromatic ring (1d) decreases the activity. The ethyl group on aromatic ring (1c) particularly enhanced the antifungal activity. From the data obtained in this work, it is suggested that the substituents on the N-atom may play a crucial role in the antimicrobial activity.As shown in the table, antimicrobial activity against bacteria and fungi was observed in the silver-NHC complexes tested at 200–6.25 μg/mL concentrations. The new complexes showed effective activities against Gram-positive, Gram-negative bacteria and fungi. The complexes were found effective in inhibiting the growth of Gram-positive and Gram-negative bacteria with MICs values between 100–200 µg/mL. The tested compounds showed antifungal activity with a range of the MICs between 6.25 and 100 μg/mL. However, among the silver complexes tested 2O , CH2Cl2 (P4O10), hexane, toluene (Na). Melting points were determined in glass capillaries under air with an Electrothermal-9200 melting point apparatus. FTIR spectra were recorded as KBr pellets in the range 400–4000 cm–1 with an ATI UNICAM 1000 spectrometer. 1H-NMR and 13C-NMR spectra were recorded with a Varian AS 400 Merkur spectrometer operating at 400 MHz (1H), 100 MHz (13C) in CDCl3 and DMSO-d6 with tetramethylsilane as an internal reference. Elemental analyses were performed by Turkish Research Council  Microlab. All reactions for the preparation of imidazolidinium salts and silver (NHC) complexes were carried out under argon in flame-dried glassware using standard Schlenk techniques. The solvents used were purified by distillation over the drying agents indicated and were transferred under Ar: Et2O (0.5 mmol) and activated 4Ǻ molecular sieves in dichloromethane (30 mL) was stirred room temperature for 24 hours. The reaction mixture was filtered through celite and the solvent removed under reduced pressure. The crude product was recrystallized from dichloromethane/hexane at room temperature.A solution of imidazolidinium salt (1.0 mmol), AgChloro-1,3-bis(2-methylbenzyl)imidazolidin-2-ylidenesilver (I) (1a). Yield: 0.20 g; 90%, m.p.: 155–156 °C; υ(CN) = 1,501 cm -1. 1H-NMR (CDCl3) δ: 2.37 -2), 3.44 , 4.78 -2), 7.11–7.29 -2). 13C{H}-NMR (CDCl3) δ: 19.2 (CH2C6H2(CH3)-2), 48.6 (NCH2CH2N), 53.5 (CH2C6H4(CH3)-2), 127.0, 128.4, 128.5, 131.0, 136.7 ve 138.1 (CH2C6H4(CH3)-2). Anal. Calcd for C19H23AgClN2: C, 53.98; H, 5.48; N, 6.63%; found: C, 53.90; H, 5.43; N, 6.59. Chloro-1,3-bis(4-methylbenzyl)imidazolidin-2-ylidenesilver (I) (1b). Yield: 0.21 g; 95%, m.p.: 230–231 °C; υ(CN) = 1,500 cm -1. 1H-NMR (DMSO-d6) δ: 2.30 -4), 3.49 , 4.66 -4), 7.09–7.24 -4). 13C{H}-NMR (DMSO-d6) δ: 21.2 (CH2C6H2(CH3)-4), 48.9 (NCH2CH2N), 54.2 (CH2C6H4(CH3)-4), 128.2, 129.8, 133.6, 137.6 (CH2C6H4(CH3)-4), 203.1 (carbC). Anal. Calcd for C19H23AgClN2: C, 53.98; H, 5.48; N, 6.63%; found: C, 53.96; H, 5.40; N, 6.62%. Chloro-1,3-bis(4-ethylbenzyl)imidazolidin-2-ylidenesilver (I) (1c). Yield: 0.20 g; 92%, m.p.: 97–98 °C; υ(CN) = 1,515 cm -1. 1H-NMR (DMSO-d6) δ: 1.12 -4), 2.54 -4), 3.46 , 4.52 -4), 7.07 ve 7.22 -4). 13C{H}-NMR (DMSO-d6) δ: 14.6 (CH2C6H2(CH2CH3)-4), 27.3 (CH2C6H2(CH2CH3)-4), 46.3 (NCH2CH2N), 50.5 (CH2C6H4(CH2CH3)-4), 127.3, 128.2, 128.7, 143.2 (CH2C6H4(CH2CH3)-4), 202.1 (carbC). Anal. Calcd for C21H27AgClN2: C, 55.95; H, 6.04; 7.86; N, 6.21%; found: C, 55.90; H, 5.98; N, 6.26%. Chloro-1,3-bis(4-i-propylbenzyl)imidazolidin-2-ylidenesilver (I)  (1d). Yield: 0.20 g; 94%, m.p.: 194–195 °C; υ(CN) = 1,503 cm -1. 1H-NMR (DMSO-d6) δ: 1.88 2)-4), 2.87 2)-4), 3.56 , 4.6 2)-4), 7.18 ve 7.29 2)-4). 13C{H}-NMR (DMSO-d6) δ: 22.6 (CH2C6H2(CH(CH3)2)-4), 33.4 (CH2C6H2(CH(CH3)2)-4), 46.6 (NCH2CH2N), 53.0 (CH2C6H4(CH(CH3)2)-4), 126.2, 127.9, 129.8, 148.9 (CH2C6H4(CH3)-4). Anal. Calcd for C23H31AgClN2: C, 57.69; H, 6.53; N, 5.85%; found: C, 57.63; H, 6.50; N, 5.83%.Chloro-1,3-bis(4-diethylaminobenzyl)imidazolidin-2-ylidenesilver (I)  (1e). Yield: 0.19g; 91%, m.p.: 163–165 °C; υ(CN) = 1,507 cm -1. 1H-NMR (CDCl3) δ: 1.15 2-4), 3.36 2-4), 3.69 , 4.57 2-4), 6.61 ve 6.64 2-4). 13C{H}-NMR (CDCl3) δ: 12.5 (CH2C6H2N(CH2CH3)2-4), 44.3 (CH2C6H2N(CH2CH3)2-4), 48.2 (NCH2CH2N), 52.0 (CH2C6H4N(CH2CH3)2-4), 111.7, 121.1, 130.3, 147.7 (CH2C6H4N(CH2CH3)2-4). Anal. Calcd for C25H37AgClN4: C, 55.93; H, 6.95; N, 10.44%; found: C, 55.90; H, 6.93; 6.63; N, 10.42%. Chloro-1,3-bisimidazolidin-2-ylidenesilver (I) (1f). Yield: 0.19 g; 93%, m.p.:134–135 °C; υ(CN) = 1,508 cm -1. 1H-NMR (DMSO-d6) δ: 3.36 , 3.53 2-3), 3.75 2-4), 4.63 2-3,4), 6.76-7.012-3,4). 13C{H}-NMR (DMSO-d6) δ: 49.0 (NCH2CH2N), 54.2 (CH2C6H3(OCH3)2-3), 55.9 (CH2C6H3(OCH3)2-4), 56.0 (CH2C6H3(OCH3)2-3,4), 112.1, 112.4, 120.4, 129.0, 148.9, 149.3 (CH2C6H3(OCH3)2-3,4), 202.9 (carbC). Anal. Calcd for C21H27AgClN2O4: C, 49.00; H, 5.29; N, 5.44; O, 12.43%; found: C, 49.02; H, 5.30; N, 5.40; O, 12.44%. N-Heterocyclic carbene ligand were determined using the agar dilution procedure recommended by the Clinical and Laboratory Standards Institute [Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 were obtained from American Type Culture Collection  and the fungal strains Candida albicans and Candida tropicalis obtained from the Department of Microbiology, Faculty of Medicine, Ege University (Turkey). Bacterial strains were subcultured on Muller Hinton Broth (HiMedia Laboratories Pvt. Ltd. Mumbai-India) and fungal strains were also on RPMI 1640 Broth . Their turbidities matched that of a McFarland no. 0.5 turbidity standard [6 CFUs/mL) was spread over the surface of agar plates. All the inoculated plates were incubated at 35 °C and results were evaluated after 16–20 h of incubation for bacteria and 48 h for yeasts. The lowest concentration of the compounds that prevented visible growth was considered to be the minimal inhibitory concentration (MIC).Antimicrobial activities of the Ag (I) complexes with nstitute ,48. Ministandard . The sto1H-NMR, 13 C- NMR, IR and elemental analysis. Also, antimicrobial activities of the new complexes are reported. The silver complexes 1a, 1c and 1f showed better antimicrobial activity against the fungi than other complexes, even at the much lower concentrations. Although the mechanism of antimicrobial activity is not known, it was found that substituents on the N-atom have an effect on antimicrobial activity in this work. Detailed investigations focusing on new silver-NHC complexes and other biomedical applications are ongoing. In summary, six Ag(I)-NHC complexes were synthesized starting from 1,3-dialkylimidazolidinium chloride, following a common procedure. The silver-NHC complexes characterized by


Their structures and relative configurations were determined by extensive 1D/2D NMR and mass spectrometric data, and the absolute configurations of 3–6 were assigned by analysis of the ECD spectra aided by quantum chemical computations. Compounds 1, 2, 5, and 6 showed growth inhibition of some marine phytoplankton species and pathogenic bacteria.One new bisabolane sesquiterpene, bisabolan-1,10,11-triol ( Trichoderma strains associated with marine algae have an excellent ability to produce halogenated organic molecules +), requiring one degree of unsaturation. The 1H NMR spectrum , implying three degrees of unsaturation. The 1H NMR spectrum , suggesting three degrees of unsaturation. The EI mass spectrum showed fragment ion peaks at m/z 166 and 138, indicating the presence of hydroxy and carboxyl groups. In combination with HSQC data, the 1H NMR spectrum . Its 1H and 13C NMR as well as EIMS data closely resembled those of 3, and COSY and HMBC correlations , one more oxygen atom than trichodenone C + (7), 223 (100), 205 (30), 163 (41), 109 (49), 95 (60), 81 (73), 69 (61), 59 (58), 55 (71); HREIMS m/z 258.2191 [M]+ .Isabolan-1,10,11-triol (2): Colorless oil; c 0.08, MeOH); IR (KBr) vmax 3402, 2962, 2931, 1712, 1647, 1381, 1261, 1103, 1030 cm−1; 1H and 13C NMR data, m/z (%) 300 [M]+ (<1), 153 (20), 128 (16), 113 (58), 110 (100), 109 (21), 95 (32), 86 (27), 71 (17); HREIMS m/z 300.1957 [M]+ .12-nor-11-Acetoxybisabolen-3,6,7-triol (S)-1-Hydroxy-3-p-menthen-9-oic acid (3): Colorless oil; c 0.016, MeOH); 1H and 13C NMR data, m/z (%) 184 [M]+ (13); 166 (80), 138 (60), 123 (62), 95 (59), 81 (60), 58 (70), 44 (100); HREIMS m/z 184.1100 [M]+ .(7R)-1-Hydroxy-3-p-menthen-9-oic acid (4): Colorless oil; c 0.040, MeOH); 1H and 13C NMR data, m/z (%) 184 [M]+ (13); 167 (42), 166 (100), 138 (71), 123 (73), 121 (65), 95 (71), 81 (74), 58 (78), 44 (100); HREIMS m/z 184.1097 [M]+ .(75): Colorless oil; IR (KBr) vmax 3410, 2978, 2927, 1697, 1612, 1435, 1265, 1153, 1080, 864 cm−1; 1H and 13C NMR data, Dechlorotrichodenone C (6): Colorless oil; c 0.040, MeOH); IR (KBr) vmax 3398, 2924, 2854, 1720, 1628, 1388, 1072 cm−1; 1H and 13C NMR data, m/z (%) 176 [M]+ (4), 177 (58), 175 (55), 132 (31), 121 (100), 57 (40); HREIMS m/z 176.0236 [M]+ .3-Hydroxytrichodenone C (7): White powder; c 0.050, MeOH); IR (KBr) vmax 3278, 2947, 2873, 1655, 1412, 1095 cm−1; 1H and 13C NMR data, m/z (%) 240 [M]+ (6), 209 (40), 184 (55), 152 (100), 100 (41), 86 (65), 68 (45); HREIMS m/z 240.1488 [M]+ .Methylcordysinin A (8): Colorless oil; 1H and 13C NMR data, m/z (%) 141 [M]+ (28), 123 (20), 97 (28), 96 (100), 95 (80), 54 (38), 42 (43), 40 (65); HREIMS m/z 141.0428 [M]+ .4-Oxazolepropanoic acid  and four new naturally occurring ones (3–5 and 8), involving two sesquiterpenes (1 and 2), two monoterpenes (3 and 4), two trichodenones (5 and 6), and two nitrogen-bearing metabolites (7 and 8). It is worth mentioning that 3-hydroxytrichodenone C (6) is a new chlorine-containing molecule, and it along with 1, 2, and 5 can inhibit the four marine phytoplankton species and four marine-derived Vibrio bacteria tested.A chemical survey on a marine-alga-endophytic isolate (cf44-2) of the fungus


Phosphonate reagents were developed for the two-carbon homologation of aldehydes or ketones to unbranched- or methyl-branched α,β-unsaturated aldehydes. The phosphonate reagents, diethyl methylformyl-2-phosphonate dimethylhydrazone and diethyl ethylformyl-2-phosphonate dimethylhydrazone, contained a protected aldehyde group instead of the usual ester group. A homologation cycle entailed condensation of the reagent with the starting aldehyde, followed by removal of the dimethylhydrazone protective group with a biphasic mixture of 1 M HCl and petroleum ether. This robust two-step process worked with a variety of aldehydes and ketones. Overall isolated yields of unsaturated aldehyde products ranged from 71% to 86% after the condensation and deprotection steps. E) double bond, an α-alkyl branch (size depending on the choice of phosphonate reagent), and a terminal ester group. The ester is then reduced to an α,β-unsaturated alcohol, and the alcohol is partially oxidized to the desired α,β-unsaturated aldehyde. One way to shorten this scheme would be to use a homologating reagent that keeps the oxidation state of an aldehyde throughout.A recurring theme in the synthesis of various polyketide natural products and other compounds is two-carbon homologation of aldehydes or ketones to unbranched or branched  α,β-unsaturated aldehydes. A typical approach based on the Horner-Wadsworth-Emmons condensation  uses a pE)-2-methyl-2-butenal -phosphonic acid diethyl ester.and diethyl ethylformyl-2-phosphonate dimethylhydrazone can also be named [2-(dimethyl-hydrazino)-1-methyl-ethyl]-phosphonic acid diethyl ester. Alternative names for the aldehyde phosphonates and aldehyde phosphonate dimethylhydrazones 1 and 2 were readily converted to their corresponding anions with the base LDA and reacted smoothly with aldehydes or ketones, added in excess, to give unbranched- or methyl-branched, unsaturated aldehyde dimethylhydrazones with chains having two additional carbons  ratio of 2E to 2Z products from aldehyde starting materials and a ratio (2.0:1 to 2.2:1) of 2E to 2Z products from ketone starting materials. Homologation of cycloheptanone with phosphonate 2 resulted in an improved synthesis of cycloheptylidene-acetaldehyde 26. The overall yield for the preparation of 26 increased from 42% [Using these reagents led to a two-step aldehyde homologation cycle (condensation and deprotection) instead of the previous three-step cycle. Overall yields, of unsaturated aldehyde products, for the two-step sequence ranged from 71% to 86%. Homologation of unbranched aldehyde starting materials with phosphonate from 42%  to 74%.4, 6, 9, 11, 14 and 16  NMR methods. Irradiation at either H-1 or H-3 caused a nuclear Overhauser enhancement of 2.1% to 4.3% at H-1 or H-3 for compounds 5, 7, 10, 12, 15 and 17, but only a nOe of 0% to 0.2% was observed for the methyl group protons for any of the compounds. Irradiation of the methyl group protons caused a nOe of just 0% to 0.5% for either H-1 or H-3. The data support the assignment of 2E configuration for compounds 5, 7, 10, 12, 15 and 17, because the H-1 and H-3 protons are closer to each other than to the protons of the methyl group, attached to C-2.The major isomer configuration, at the α,β-double bond of disubstituted alkene compounds 4 and 16  was readZ-isomer product was identified by its shorter GC retention time and nearly identical mass spectra, compared to the 2E-isomer [19, and 20. The major isomer configuration of the trisubstituted double bonds, of compounds 19, and 20 was determined to be E because of the later GC retention time, previously observed for (2E)-N,N-Dimethyl-N’-(dec-2-enylidene)-hydrazine and (2E)-2-Decenal [E-isomer of 20 reported previously [The proportion of 2E-isomer , for comeviously . We coul22 and 23, derived from methyl ketone starting materials, 6, and 7 was determined by nuclear Overhauser enhancement (nOe) NMR methods. A moderate nOe enhancement was observed (2D NOESY experiments) between the methyl group protons and H-2, in the case of the minor isomer for both compounds. Only weak nOe enhancement was observed between the methyl group protons and H-2, in the case of the major isomer of compound 6 and no nOe enhancement was observed between the methyl group protons and H-2, in the case of the major isomer of compound 23. The data support the assignment of 2Z configuration for the minor isomer of compounds 22 and 23 because the H-2 protons are closer to the protons of the methyl group, attached to C-3. Therefore, the major isomer has the E-configuration. The GC retention time for the major isomer was also later than that observed for the minor isomer, consistent with the assignment of E-configuration for the major isomer. Compound 23 has been prepared previously [E-isomer and δ 26.4 for the minor Z-isomer, consistent with reported 13C-NMR data [The major isomer configuration of the trisubstituted double bonds, of compounds eviously ,25 and tDiethyl-2,2-(diethoxy)ethylphosphonate, diethyl methylphosphonate and diethylethylphosphonate were purchased from Alfa Aesar . Diethyl ethylphosphonate can also be prepared in 98% yield by refluxing a mixture of triethyl phosphite and ethyl iodide for 3 h . THF wasE and 2Z isomers were considered to be the product for all product compounds; however, the 2E:2Z isomer ratios are reported.Progress of synthetic reactions was monitored by gas chromatography (GC). All reaction products were identified by GC/MS and the structures were additionally verified by NMR. Yields were corrected for purity. Both 29 and 10 because considerable degradation of unsaturated aldehyde dimethylhydrazones was observed during GC runs with higher inlet temperatures.The Hewlett-Packard (HP) 5890 Series II gas chromatograph was equipped with flame ionization detector and split/splitless inlet and was interfaced to an HP ChemStation data system. The column was a DB-5 capillary . Carrier gas was He. The oven temperature was programmed from 50 to 280 °C at 10 °C/min, and the detector temperature was 280 °C. The inlet temperature was 220 °C, and 1.0 µL sample injections were made in splitless mode. The inlet temperature was 120 °C, and 1.0 µL sample injections were made in splitless mode for analysis for compounds 9 and 10.Electron impact mass spectra (70 eV) were obtained with an HP 5973 MSD instrument, interfaced to an HP 6890 GC, equipped with a splitless inlet. Several columns were used, but gave comparable results to that used for GC. The oven temperature was programmed from 50 to 250 °C at 10 °C/min; the inlet temperature was 220 °C and the transfer line temperature was 250 °C. The inlet temperature was 120 °C for compounds 1H-NMR, 13C-NMR and 2D NMR spectra were collected on a Bruker  Avance 500 spectrometer using a 5 mm broadband probe. Samples were dissolved in either CDCl3 or C6D6 (where indicated) and all spectra  were acquired at 300° K. Trisubstituted double-bond “E” or “Z” assignments were based on 1D PFG-NOE experiments and 2D PFG-NOESY experiments. Chemical shifts are reported as parts per million from tetramethylsilane based on the lock solvent. Coupling constants (J) are in Hertz. Assignments were made with the help of NMR prediction software from Advanced Chemistry Development, Inc. (ACD/Labs) [6, 7, 20, 22, 23 and 26 are known, spectral data are given because MS and NMR data are very difficult to find in the older chemical literature. Methyl branches are reported by the point of attachment to the carbon chain .CD/Labs)  and by aDiethyl methylformylphosphonate dimethylhydrazone (1). Diethyl-2,2-(diethoxy)ethylphosphonate  was added to acetone (200 mL), followed by water (3 mL) and p-toluenesulfonic acid . The mixture was stirred at r.t., under an argon atmosphere for 4 days. A solution of NaHCO3  was added with stirring to quench the reaction and neutralize acid present. The pH was checked and found to be 7. After removal of solvent, aqueous oil remained. The oil was saturated by the addition of NaCl (25 g). The aqueous oil was repeatedly extracted with CH2Cl2 (100 mL then 2 × 50 mL), and the combined CH2Cl2 extracts were dried over anhydrous Na2SO4 and filtered. Solvent was removed to afford 5.59 g of crude diethyl methylformyl-2-phosphonate. Without further purification, the crude product was converted to the dimethylhydrazone (DMH) derivative by adding it to a mixture of CH2Cl2 (75 mL) containing anhydrous MgSO4  then N,N-dimethylhydrazine  was added in one portion. The mixture was stirred (48 h) until analysis by GC showed complete protection of the aldehyde as the DMH derivative. Filtration, removal of solvent, and Kugelrohr distillation  afforded 4.41 g of oil containing 1 . The oil also contained 8% starting material, diethyl-2,2-(diethoxy)ethylphosphonate. MS of product: (EI) m/z (%) 222 , 180 (9), 152 (91), 125 (100), 122 (74), 108 (27), 97 (37), 85 (85), 71 (15), 58 (8), 44 (91). 1H-NMR δ 1.30 , 2.77 , 2.81 , 4.09, , 6.47 . 13C-NMR δ 16.4 (CH3–CH2–O), 31.3 (C-2), 42.9 (N–CH3), 61.9 (CH3–CH2–O), 126.1 (C-1). The MS and NMR spectral data were consistent with the reference spectral data from previous work [ous work .Alternative preparation of diethyl methylformylphosphonate dimethylhydrazone (1). A solution of n-butyllithium  was added to dry THF (80 mL) and the mixture was cooled in an argon atmosphere to dry ice-ethanol bath temperature (approx −78 °C). Diethyl methylphosphonate  was added dropwise and the mixture was stirred for an additional hr before dry dimethylformamide  was added. After warming to 20 °C, ice cold 3 M aqueous HCl (300 mL) was added and the mixture was stirred 5 min. A fine white precipitate formed initially, and then the solution cleared and separated into two phases . The phases were separated, and the organic phase was washed with 10 mL of alkaline brine solution (0.5 % NaHCO3 in saturated aqueous NaCl), dried over anhydrous MgSO4, and filtered. The acidic aqueous phase was repeatedly extracted with CH2Cl2 (5 × 50 mL), and the combined CH2Cl2 extracts were washed with alkaline brine solution (2 × 10 mL). The alkaline brine washes were combined and extracted again with CH2Cl2 (4 × 15 mL). All CH2Cl2 extracts were combined, dried over anhydrous MgSO4, and filtered. Solvent was removed from the dried organic phase and all dried CH2Cl2 extracts to afford 9.8 g of crude diethyl methylformyl-2-phosphonate, which still contained some DMF. Without further purification, the crude product was converted to the dimethylhydrazone (DMH) derivative by adding it to a mixture of CH2Cl2 (200 mL) containing anhydrous MgSO4  then N,N-dimethylhydrazine  was added in one portion. The mixture was stirred (48 h) until analysis by GC showed complete protection of the aldehyde as the DMH derivative. Filtration, removal of solvent, and Kugelrohr distillation  afforded 7.1 g of 1 . MS (EI) m/z (%) 222 , 180 (9), 152 (91), 125 (100), 122 (74), 108 (27), 97 (37), 85 (85), 71 (15), 58 (8), 44 (91). EI-HRMS C8H19N2O3P calcd. for 222.1133 (obs. 222.1133). 1H-NMR δ 1.30 , 2.77 , 2.81 , 4.09, , 6.47 . 13C-NMR δ 16.4 (CH3–CH2–O), 31.3 (C-2), 42.9 (N–CH3), 61.9 (CH3–CH2–O), 126.1 (C-1).Diethyl ethylformyl-2-phosphonate dimethylhydrazone (2). A solution of n-butyllithium  was added to dry THF (250 mL) and the mixture was cooled in an argon atmosphere to dry ice-ethanol bath temperature (approx −78 °C). Diethyl ethylphosphonate  was added dropwise and the mixture was stirred for an additional hr before dry DMF  was added. After warming to 20 °C, ice cold 3 M aqueous HCl (300 mL) was added and the mixture was stirred 5 min. A fine white precipitate formed initially, and then the solution cleared and separated into two phases . The phases were separated, and the organic phase was washed with 30 mL of alkaline brine solution (0.5% NaHCO3 in saturated aqueous NaCl), dried over anhydrous MgSO4, and filtered. The acidic aqueous phase was repeatedly extracted with CH2Cl2 (5 × 150 mL), and the combined CH2Cl2 extracts were washed with 30 mL of alkaline brine solution. The alkaline brine washes were combined and extracted again with CH2Cl2 (4 × 15 mL). All CH2Cl2 extracts were combined, dried over anhydrous MgSO4, and filtered. Solvent was removed from the dried organic phase and all dried CH2Cl2 extracts to afford 44 g of crude diethyl ethylformyl-2-phosphonate, which still contained some DMF. Without further purification, the crude product was converted to the dimethylhydrazone (DMH) derivative by adding it to a mixture of CH2Cl2 (600 mL) containing anhydrous MgSO4  then N,N-dimethylhydrazine  was added in one portion. The mixture was stirred (48 h) until analysis by GC showed complete protection of the aldehyde as the DMH derivative. Filtration, removal of solvent, and Kugelrohr distillation  afforded 47.6 g of 3 . Isolated yields >95% were routinely obtained. MS (EI) m/z (%) 236 , 194 (8), 166 (82), 136 (42), 122 (10), 111 (12), 99 (100), 53 (12), 81 (12), 72 (11), 56 (12), 44 (36). EI-HRMS C9H21N2O3P calcd. for 236.1290 (obs. 236.1293) 1H-NMR δ 1.19 and 1.20 , 1.26 , 2.76 , 2.66 , 3.99 and 4.00, , 6.38 . 13C-NMR δ 12.7 (C-3), 16.4 (CH3–CH2–O), 36.0 and 37.1 , 43.1 (N–CH3), 62.0 (CH3–CH2–O), 132.6 (C-1).(2E)-N,N-Dimethyl-N’-(dec-2-enylidene)-hydrazine (4). A commercial 2.0 M solution of LDA  was added dropwise (reaction exothermic) to a 250 mL round-bottomed flask containing dry THF (25 mL), 1  and a few mg of ethyl(triphenyl)phosphonium bromide, as an indicator, and the mixture was stirred in an argon atmosphere at r.t. for 1.5 h to ensure complete formation of the phosphonate ylide. A persistent red color developed in the solution when sufficient base was added to convert the phosphonate to its anion. Then, n-octanal  was added dropwise; final color was yellow-orange. The mixture was stirred at r.t. for 5 h. Consumption of the phosphonate 1 was monitored by GC. Water (6 mL) was added to quench the reaction (excess water should be avoided because it leads to problematic emulsions during subsequent extraction steps). The solvent was removed by rotary evaporation to give an oily residue, which was extracted with ethyl acetate (5 × 25 mL). The combined extracts were washed with saturated aqueous NaCl solution (15 mL), dried over anhydrous Na2SO4 and filtered. Removal of solvent left a yellow oil. Kugelrohr distillation  afforded 1.80 g of 4 . By GC and GC-MS, the 2E/2Z isomer ratio was 8.5 to 1. MS (EI) m/z (%) 196 , 181 (8), 167 (2), 152 (19), 140 (7), 125 (14), 111 (100), 82 (40), 59 (21), 44 (32), 42 (19). EI-HRMS C12H24N2 calcd. for 196.1939 (obs. 196.1940). 1H-NMR δ 0.89 , 1.27-1.30 , 1.42 , 2.15 , 2.84 , 5.87 , 6.20 , 7.11 . 13C-NMR δ 13.3 (C-10), 22.6 (C-9), 29.0 , 31.7 (C-8), 32.6 (C-4), 43.0 (N–CH3), 128.6 (C-2), 136.4 (C-3), 137.6 (C-1).(2E)-2-Decenal (6). Deprotection was initiated by stirring 4  with 1 M HCl (50 mL) at room temperature for 5 min , and then petroleum ether  was added and stirring continued. After 4 h, the phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (30 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. The combined petroleum ether phases were washed with alkaline brine solution (2 × 10 mL), and the brine phase was back-extracted once with petroleum ether (20 mL). The combined organic phases were dried, filtered, and removal of solvent afforded 0.75 g of the free aldehyde 6. Purity by GC was 92%, and yield corrected for purity was 90%. The 2E:2Z isomer ratio was 25 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [(2E)-N,N-Dimethyl-N’-(2-methyl-dec-2-enylidene)-hydrazine (5). Compound 5 was prepared in a way similar to compound 4. The phosphonate anion was prepared from 2  in THF (25 mL) by treatment with LDA , and n-octanal  was subsequently added and the mixture was stirred at r.t. for 20 h. Water (3 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 2.04 g of 5. . By GC and GC-MS, the 2E/2Z isomer ratio was 8.2 to 1. MS (EI) m/z (%) 210 , 195 (24), 180 (3), 166 (82), 125 (100), 111 (13), 96 (29), 82 (37), 59 (18), 44 (31), 42 (15). EI-HRMS C13H26N2 calcd. for 210.2096 (obs. 210.2093). 1H-NMR δ 0.90 , 1.28-1.31 , 1.41 , 1.84 , 2.20 , 2.83 , 5.61 , 7.15 . 13C-NMR δ 11.6 (CH3 attached to C-2), 14.1 (C-10), 22.6 (C-9), 28.1 (C-4), 29.3 (C-6 and C-7), 29.9 (C-5), 31.9 (C-8), 43.0 (N–CH3), 132.4 (C-2), 134.2 (C-3), 141.2 (C-1).(2E)-2-Methyl-2-Decenal (7). Deprotection procedure was similar to that used for 6. A solution containing 5  and 1 M HCl (60 mL), to which petroleum ether  was added after 5 min, was stirred at room temperature for 6 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (40 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. Workup of the reaction as for 6 resulted in 0.85 g of the free aldehyde 7 . The 2E:2Z isomer ratio was 24 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [ous work .-N’--N,N-dimethyl-hydrazine (9). Compound 9 was prepared in a way similar to compound 4. The phosphonate anion was prepared from 1  in THF (25 mL) by treatment with LDA , and (2E)-2-nonenal  was subsequently added and the mixture was stirred at r.t. for 3 h. Water (6 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 1.62 g of 9. . By GC and GC-MS, the 2E/2Z isomer ratio was 16 to 1. MS (EI) m/z (%) 208 , 137 (3), 123 (100), 108 (2), 94 (8), 80 (11), 67 (5), 44 (7), 42 (3). EI-HRMS C13H24N2 calcd. for 208.1939 (obs. 208.1944). 1H-NMR δ 0.90 , 1.29–1.31 , 1.41 , 2.12 , 2.87 , 5.75, , 6.13 , 6.28 , 6.28 , 7.05 . 1H-NMR in C6D6 δ 0.98 , 1.29–1.31 , 1.41 , 2.10 , 2.63 , 5.77, , 6.22 , 6.41 , 6.72 , 7.14 . 13C-NMR δ 14.0 (C-11), 22.6 (C-10), 28.9 (C-8), 29.2 (C-7), 31.6 (C-9), 32.8 (C-6), 42.9 (N–CH3), 128.6 (C-2), 130.3 (C-4), 133.2 (C-3), 135.8 (C-5), 136.2 (C-1).-2,4-undecadienal (11). Deprotection procedure was similar to that used for 6. A solution containing 9  and 1 M HCl (50 mL), to which petroleum ether  was added after 5 min, was stirred at room temperature for 24 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (40 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. Workup of the reaction as for 6 resulted in 0.81 g of the free aldehyde 11 . The 2E:2Z isomer ratio was 9.5 to 1. MS (EI) m/z (%) 166 , 137 92), 123 (4), 109 (5), 95 (12), 81 (100), 67 (15), 55 (11), 43 (10), 41 (18). 1H-NMR δ 0.91 , 1.30-1.32 , 1.47 , 2.24 , 6.09 , 5.79 , 6.31, , 6.31 , 7.10 , 6.93 , 9.55 , 10.19 . The sample also contained about 6% of the 2E,4Z isomer: δ 6.16 , 6.30 , 7.45 1H, dd, J3-4 = 9.8 and J2-3 = 15.1, =CH, H-3), 9.63 . 1H-NMR in C6D6 δ 0.99 , 1.27–1.32 , 1.91 , 5.79, , 5.90 , 6.03 , 6.55 , 9.54 . 13C-NMR δ 14.0 (C-11), 22.5 (C-9), 28.5 (C-7), 28.8 (C-8), 31.6 (C-10), 33.2(C-6), 128.6 (C-4), 130.0 (C-2), 147.4 (C-5), 152.8 (C-3), 193.9 (C-1).-N’--N,N-dimethyl-hydrazine (10). Compound 10 was prepared in a way similar to compound 4. The phosphonate anion was prepared from 2  in THF (25 mL) by treatment with LDA , and (2E)-2-nonenal  was subsequently added and the mixture was stirred at r.t. for 20 h. Water (3 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 2.09 g of 10. . By GC and GC-MS, the 2E/2Z isomer ratio was 39 to 1. MS (EI) m/z (%) 222 , 137 (100), 108 (7), 94 (13), 44 (4), 43 (5). EI-HRMS C14H26N2 calcd. for 222.2096 (obs. 222.2102). 1H-NMR δ 0.91 , 1.29-1.31 , 1.43 , 1.96 (CH3 attached to C-2), 2.16 , 2.87 , 5.77, , 6.12 , 6.44 , 7.08 . 13C-NMR δ 11.8 (CH3 attached to C-2), 14.1 (C-11), 22.6 (C-10), 28.9 (C-8), 29.4 (C-7), 31.7 (C-9), 33.2 (C-6), 43.2 (N–CH3), 126.7 (C-4), 131.2 (C-2), 133.2 (C-3), 135.8 (C-5), 140.0 (C-1).-2-Methyl-2,4-undecadienal (12). Deprotection procedure was similar to that used for 6. A solution containing 10  and 1 M HCl (50 mL), to which petroleum ether  was added after 5 min, was stirred at room temperature for 48 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (40 mL) was added and the mixture was stirred at rt for an additional 1 h before the phases were separated. Workup of the reaction as for 6 resulted in 0.91 g of the free aldehyde 12 . The 2E:2Z isomer ratio was 30 to 1. MS (EI) m/z (%) 180 , 109 (6), 95 (100), 81 (10), 79 (9), 67 (12), 55 (5), 53 (5), 43 (5), 41 (10). 1H-NMR δ 0.91 , 1.32–1.34 , 1.48 , 1.85 (CH3 attached to C-2), 2.26 , 6.26, , 6.54 , 6.84 , 9.44 , 10.28 , The sample also contained a trace amount of the 2E,4Z isomer: δ 6.13 , 6.85 , 9.51 . 13C-NMR δ 14.0 (C-11), 22.5 (C-9), 28.5 (C-7), 28.8 (C-8), 31.6 (C-10), 33.2(C-6), 128.6 (C-4), 130.0 (C-2), 147.4 (C-4), 152.8 (C-3), 193.9 (C-1).(2E)-N,N-Dimethyl-N’--hydrazine (14). Compound 14 was prepared in a way similar to compound 4. The phosphonate anion was prepared from 1  in THF (25 mL) by treatment with LDA , and benzaldehyde  was subsequently added and the mixture was stirred at r.t. for 5 h. Water (3 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 1.62 g of 14. . By GC and GC-MS, the 2E/2Z isomer ratio was 50 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [ous work .(2E)-Cinnamaldehyde (16). Deprotection procedure was similar to that used for 6. A solution containing 14  and 1 M HCl (50 mL), to which petroleum ether  was added after 5 min, was stirred at room temperature for 18 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (50 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. Workup of the reaction as for 6 resulted in 0.61 g of the free aldehyde 16 . The 2E:2Z isomer ratio was 170 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [(2E)-N,N-Dimethyl-N’--hydrazine (15). Compound 15 was prepared in a way similar to compound 4. The phosphonate anion was prepared from 2  in THF (25 mL) by treatment with LDA , and benzaldehyde  was subsequently added and the mixture was stirred at r.t. for 20 h. Water (3 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 1.82 g of 15. . By GC and GC-MS, the 2E/2Z isomer ratio was 25 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [ous work .(2E)-Methyl cinnamaldehyde (17). Deprotection procedure was similar to that used for 6. A solution containing 15  and 1 M HCl (50 mL), to which petroleum ether  was added after 5 min, was stirred at r.t. for 48 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Petroleum ether (40 mL) was added and the mixture was stirred at r.t. for an additional 1 hr before the phases were separated. Workup of the reaction as for 6 resulted in 0.78 g of the free aldehyde 17 . The 2E:2Z isomer ratio was 56 to 1. The MS and NMR spectral data were consistent with the reference spectral data from previous work [N,N-Dimethyl-N'-(3-methyl-non-2-enylidene)-hydrazine (19). A commercial 2.0 M solution of LDA,  was added dropwise (reaction exothermic) to a 250 mL round-bottomed flask containing dry THF (24 mL), 1  and a few mg of ethyl(triphenyl)phosphonium bromide, as an indicator, and the mixture was stirred in an argon atmosphere at r.t. for 1.5 h to ensure complete formation of the phosphonate ylide. A persistent red color developed in the solution when sufficient base was added to convert the phosphonate to its anion. Then, 2-octanone  was added dropwise; final color was yellow-orange. The mixture was stirred at r.t. for 22 h. Consumption of the phosphonate 1 was monitored by GC. Water (6 mL) was added to quench the reaction (excess water should be avoided because it leads to problematic emulsions during subsequent extraction steps). The solvent was removed by rotary evaporation to give an oily residue, which was extracted with ethyl acetate (5 × 25 mL). The combined extracts were washed with saturated aqueous NaCl solution (15 mL), dried over anhydrous Na2SO4 and filtered. Yellow oil remained after removal of solvent. Kugelrohr distillation  afforded 1.95 g of 19 . By GC and GC-MS, the 2E/2Z isomer ratio was 2.2 to 1. MS (EI) m/z (%) 197, , 196 , 181 (27), 166 (7), 154 (17), 139 (13), 125 (58), 110 (21), 96, (75), 94 ((48), 82 (36), 67 (18), 59 (38), 44 (41). 1H-NMR, major 2E-isomer, δ 0.89 , 1.29  1.45 , 1.83 , 2.09 , 2.85 , 5.99 , 7.26 . 13C-NMR δ 14.1 (C-9), 17.0 (C-10), 22.6 (C-3), 27.6 (C-5), 28.9 (C-6), 31.7 (C-7), 40.0 (C-4), 43.3 (C-11), 122.5 (C-2), 128.2 (C-1), 141.3 (C-3). 1H-NMR, minor 2Z-isomer, δ 0.89 , 1.29  1.45 , 1.81 , 2.22 , 2.84 , 5.99 , 7.16 . 13C-NMR δ 14.1 (C-9), 22.6 (C-8), 24.0 (C-10), 28.0 (C-5), 29.1 (C-6), 31.7 (C-7), 32.5 (C-4), 43.3 (C-11), 123.5 (C-2), 128.4 (C-1), 135.2 (C-3).3-Methyl-non-2-enal (20). Deprotection was initiated by stirring 19  with 1 M HCl (100 mL) at room temperature for 5 min , and then hexane (100 mL) was added and stirring continued. After 3 h, the phases were separated and the aqueous phase was returned to the reaction vessel. Hexane (100 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. The combined hexane phases were washed with alkaline brine solution (2 × 10 mL), and the brine phase was back-extracted once with hexane (20 mL). The combined organic phases were dried, filtered, and removal of solvent afforded 1.04 g of the free aldehyde 20. Purity by GC was 95%, and yield corrected for purity was 86%. The 2E:2Z isomer ratio was 2.0 to 1. MS (EI) m/z (%) 154 , 139 (17), 125 (11), 111 (5), 97, (100), 84 (61), 69 (22), 55 (23), 41 (36). 1H-NMR, major 2E-isomer, δ 0.89 , 1.29 , 1.30 , 1.50 , 2.16 , 2.21 , 5.88 , 9.99 . 13C-NMR δ 14.1 (C-9), 17.5 (C-10), 22.5 (C-8), 27.1 (C-5), 28.8 (C-6), 31.5 (C-7), 40.6 (C-4), 127.3 (C-2), 164.4 (C-3), 191.1 (C-1). 1H-NMR, minor 2Z-isomer, δ 0.89 , 1.29 , 1.30 , 1.32 , 1.54 , 1.97 , 2.57 , 5.87 , 9.96 . 13C-NMR δ 14.0 (C-9), 22.5 (C-8), 25.0 (C-10), 28.8 (C-5), 29.0 (C-6), 31.5 (C-7), 32.6 (C-4), 128.4 (C-2), 164.9 (C-3), 190.7 (C-1). The MS and NMR spectral data were consistent with the reference spectral data from previous work [(61), 69 , 55 (23)N,N-Dimethyl-N'-(3-phenyl-but-2-enylidene)-hydrazine (22). Compound 22 was prepared in a way similar to compound 19. The phosphonate anion was prepared from 1  in THF (25 mL) by treatment with LDA , and acetophenone  was subsequently added and the mixture was stirred at r.t. for 22 h. Water (6 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 1.64 g of 22. . By GC and GC-MS, the 2E/2Z isomer ratio was 2.1 to 1. MS (EI) m/z (%) 189 , 188 , 173 (15), 158 (15), 144 (84), 128 (22), 115 (61), 103 (9), 91 (18), 77 (15), 58 (15), 42 (13). 1H-NMR, major 2E-isomer, δ 2.25 , 2.85 , 6.68 , 7.12 , 7.26 , 7.35 , 7.51 . 13C-NMR δ 16.0 (C-10), 43.0 (C-11), 125.0 (C-2), 125.4 (C-5), 127.1 (C-7), 128.3 (C-6), 135.6 (C-1), 139.4 (C-4), 142.6 (C-3). 1H-NMR, minor 2Z-isomer, δ 2.18 , 2.78 , 6.30 , 7.26 , 7.35 , 7.41 , 7.51 . 13C-NMR δ 25.6 (C-10), 43.0 (C-11), 125.4 (C-2 and C-5), 127.2 (C-7), 128.3 (C-6), 133.3 (C-1), 139.4 (C-4), 141.4 (C-3). This compound has been prepared previously [eviously .3-Phenyl-but-2-enal (23). The deprotection procedure was similar to that used for 19. A solution containing 22  and 1 M HCl (100 mL), to which hexane (100 mL) was added after 5 min, was stirred at room temperature for 3 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Hexane (100 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. Workup of the reaction as for 19 resulted in 0.95 g of the free aldehyde 23 . The 2E:2Z isomer ratio was 2.1 to 1. MS (EI) m/z (%) 146 , 145 (100), 131 (24), 115 (42), 103 (15), 91 (17), 77 (11), 63 (6), 51 (10). 1H-NMR, major 2E-isomer, δ 2.59 , 6.42 , 7.42 , 7.43 , 7.43 , 10.20 . 13C-NMR δ 16.4 (CH3 attached to C-3), 126.3 (C-5) 127.3 (C-2), 128.8 (C-6), 130.1 (C-7), 140.6 (C-4), 157.6 (C-3), 191.1 (C-1), 1H-NMR, minor 2Z-isomer, δ 2.33 , 6.15 , 7.31 , 7.42 , 9.49 . 13C-NMR δ 26.4 (CH3 attached to C-3), 128.4 (C-5 and C-6), 129.2 (C-2 and C-7), 138.4 (C-4), 162.1 (C-3), 193.4 (C-1). The MS and NMR spectral data were consistent with the reference spectral data from previous work [00), 131 , 115 (42ous work .N'-(2-Cycloheptylidene-ethylidene)-N,N-dimethyl-hydrazine (25). Compound 25 was prepared in a way similar to compound 19. The phosphonate anion was prepared from 1  in THF (25 mL) by treatment with LDA , cycloheptanone  was subsequently added and the mixture was stirred at r.t. for 22 h. Water (6 mL) was added to quench the reaction and the reaction was worked up as before. Kugelrohr distillation  afforded 1.40 g of 25.  MS (EI) m/z (%) 181 , 180 , 165 (64), 138 (38), 136 (37), 123 (22), 112 (41), 94 (41), 80 (50), 67 (26), 59 (26), 44 (30). 1H-NMR δ 1.52 , 1.54 , 1.62 , 1.64 , 2.32 , 2.46 , 2.85 , 5.98 , 7.30 . 13C-NMR δ 27.1 (C-8), 28.8 (C-5), 29.0 (C-6), 29.7 (C-7), 30.6 (C-9), 43.2 (C-10), 123.1 (C-2), 128.2 (C-1), 146.9 (C-3).Cycloheptylidene-acetaldehyde (26). The deprotection procedure was similar to that used for 19. A solution containing 25  and 1 M HCl (100 mL), to which hexane (100 mL) was added after 5 min, was stirred at room temperature for 3 h. The phases were separated and the aqueous phase was returned to the reaction vessel. Hexane (100 mL) was added and the mixture was stirred at r.t. for an additional 1 h before the phases were separated. Workup of the reaction as for 19 resulted in 0.76 g of the free aldehyde 26 . MS (EI) m/z (%) 138 , 137 (12), 123 (18), 110 (48), 109 (51), 95, (100), 81 (66), 67 (66), 55 (35), 41 (46).. 1H-NMR δ 1.55 , 1.57 , 1.69 , 1.73 , 2.45 , 2.84 , 5.85 , 9.9 . 13C-NMR δ 27.4 , 28.6 (C-6), 29.3 (C-7), 30.5 (C-9), 38.9 (C-4), 127.6 (C-2), 170.2 (C-3), 191.1 (C-1), The MS and NMR spectral data were consistent with the reference spectral data from previous work [ous work .1 and 2 were prepared without difficulty from sufficiently inexpensive starting materials and were stable enough to be distilled. The starting materials are thus inexpensive and the homologation reactions are sufficiently robust, so use of this two-step process can be commercially viable. Subsequent removal of the dimethylhydrazone protective group could be easily accomplished, even for delicate product aldehydes. Furthermore, the new method eliminates the use and disposal of some metal-containing reagents . This more concise homologation chemistry could be applied to the practical synthesis of a variety of natural products, pharmaceuticals, and other compounds. This new protocol should replace the usual three-step Horner-Wadsworth-Emmons [The new phosphonate reagents


The formation of  The relative amounts of these two building blocks determine the negative charge of the framework. This charge is compensated by the presence of counterions such as sodium, calcium or proton. Protonic dealuminated zeolites have found important applications as petroleum cracking catalysts. They contain Brønsted-type acid sites associated to framework Al  cycloaddition and electron transfer. One dimer (6e in our work) was reported as the only one formed from trans-anethole; whereas a different dimer (6c in our work) was formed from trans-anethole only in the presence of electronic acceptors . We present results on the study of UV-Vis-induced isomerization and dimerization of trans-anethole in toluene, which indicate that dimer formation could take place without the addition of electronic acceptors. d Kojima ,10 studitrans-anethole to HY zeolites at 30, 60 and 90 ºC, for 5 h. The distinction between geometrical isomers was based on the comparison of their molecular ion abundances in their mass spectra, and some characteristic ion-fragments. (E)-geometry was associated with the more stable molecular ion of the two isomers. Since there were no standard substances available, dimer structure assignment relied strongly on the comparison of typical fragment losses found in their mass spectra and on the structures which appear in trans-anethole and its transformation products. The structural formulae of dimers 2a–5b appear in 5b (20–63%) was the most abundant compound obtained under the conditions presented in cis-Anethole , the dimers  2a (0.4–2.7%), 2b (0.7–4%), dimers 3a and 3b (1–23%), 4a (0.4–5%), 4b (0.71–11%) and 5a (<0.1%) were also found as transformation products.A total of nine compounds (> 0.1%) were tentatively identified by means of GC-MS of the mixture obtained after exposure of cis-anethole was formed in higher amount at 60 °C than at either 30 or 90 °C. This suggests that competition between isomerization and dimerization exists and it is consistent with the existence of a common intermediate, i.e. protonated anethole, for both processes. The increase of the dimers as the temperature was incremented could be explained by a higher reaction rate for dimerization than for isomerization. The relative amount of dimers increased with temperature , whereas3a–5b appeared in studies on biological activity and synthetic estrogens carried out in the first half of the last century  cycloaddition leads to dimers 6c (anti head-to-head) and 6e (syn head-to-head). The structure of dimer 6e, the most abundant product (peak 7) in the chromatograms  product reported by Lewis and Kojima  reactions with singlet 9-cyanophenanthrene. The dimer structures proposed here maintain the cis-anethole stereochemistry. Dimerizations result from 1t-A*-1t-A, and to a lesser extent, from 1t‑A*-1c-A interactions. The latter happens with lower frequency due to the low cis-anethole concentration. trans-anethole toluene solution irradiation for 2 h at -5, 5 and 15 °C. cis-Anethole and dimer 6e were the most abundant products. While trans-anethole conversion increased with temperature, the selectivity towards cis-anethole decreased, whereas those of 6c and 6e dimers increased. This shows the competition that exists between isomerization and dimerization.Caldwell et al.  observedcis-anethole  and 6c  and 6e  dimers at ‑5, 5 and 15 °C, were calculated as the ratio of reaction rate over flux of absorbed photons  with trans-anethole than that observed for its reaction with cis-anethole. Since cis-anethole is being formed, encounters of 1t-A* with cis-anethole are much less likely than those with trans-anethole. Additionally, cis-anethole has been reported as less reactive than trans-anethole in the photocycloaddition with 9-cyanophenanthrene and 6-methylphenantridine + 267(7), [M-C2H5-C7H8O]+ 159(28), [M-C12H15O]+ 121(100). (E)-1,4-bis(4-methoxyphenyl)-2,3-dimethylbut-1-ene (b2). WM+· (5.8%). m/z (I%): M+· 296(13), [M-C2H5]+  267(5), [M-C2H5-C7H8O]+ 159(24), [M-C10H12O]+ 148(1), [M-C12H15O]+ 121(100).(Z)-3,5-bis(4-methoxyphenyl)-4-methylpent-2-ene (a3). WM+· (1.7%). m/z (I%): M+·296(4), [M-C2H5-C7H8O]+ 159(2), [M-C10H12O]+ 148(100), [M-C12H15O]+ 121(38).(E)-3,5-bis(4-methoxyphenyl)-4-methylpent-2-ene (b3). WM+· (1.3%). m/z (I%): M+· 296(3), [M-C2H5-C7H8O]+ 159(3), [M-C10H12O]+ 148(100), [M-C12H15O]+ 121(34).(E)-1,3-bis(4-methoxyphenyl)-2-methylpent-1-ene (a4). WM+· (7.6%). m/z (I%): M+· 296(28), [M-C2H5]+ 267(100), [M-C2H5-C7H8O]+ 159(28), [M-C10H12O]+ 148(2), [M-C12H15O]+ 121(17).(Z)-1,3-bis(4-methoxyphenyl)-2-methylpent-1-ene (b4). WM+· (5.0%). m/z (I%): M+· 296(24), [M-C2H5]+ 267(100), [M-C2H5-C7H8O]+ 159(31), [M-C10H12O]+ 148(1), [M-C12H15O]+ 121(19).(Z)-3,4-bis(4-methoxyphenyl)hex-2-ene (a5). WM+· (6.5%). m/z (I%): M+· 296(34), [M-C2H5]+ 267(100), [M-C2H5-C7H8O]+ 159(82), [M-C12H15O]+ 121(42).(E)-3,4-bis(4-methoxyphenyl)hex-2-ene (b5). WM+· (5.2%). m/z (I%): M+· 296(28), [M-C2H5]+ 267(100), [M-C2H5-C7H8O]+ 159(98), [M-C10H12O]+ 148(3), [M-C12H15O]+ 121(30).-1,2-bis(4-methoxyphenyl)-3,4-dimethylcyclobutane (a6). WM+· (6.6%). m/z (I%): M+· 296(4), [M-CH3]+ 267(1), [M-C4H8-H]+ 249(1), [M-CH3-C7H8O]+ 175(4), [M-C10H12O]+· 148(100), [M-C12H15O]+ 121(12). -1,2-bis(4-methoxyphenyl)-3,4-dimethylcyclobutane (b6). WM+· (4.7%). m/z (I%): M+· 296(81), [M-CH3]+ 267(9), [M-C4H8-H]+ 249(15), [M-CH3-C7H8O]+ 175(36), [M-C10H12O]+· 148(67), [M-C12H15O]+ 121(100).-1,2-bis(4-methoxyphenyl)-3,4-dimethylcyclobutane (c6). WM+· (0.6%). m/z (I%): M+· 296(<1), [M-C10H12O]+· 148(100), [M-C12H15O]+ 121(4).-1,2-bis(4-methoxyphenyl)-3,4-dimethylcyclobutane (d6). WM+· (1.2%). m/z (I%): M+· 296(88), [M-CH3]+ 267(8), [M-C4H8-H]+ 249(9), [M-CH3-C7H8O]+ 175(38), [M-C10H12O]+· 148(34), [M-C12H15O]+ 121(100).-1,2-bis(4-methoxyphenyl)-3,4-dimethylcyclobutane (e6). WM+· (0.5%). m/z (I%): M+· 296(<1), [M-C10H12O]+· 148(100), [M-C12H15O]+ 121(4).NaY zeolite (10 g) was suspended at 60 ºC for 2 h in an aqueous ammonium nitrate solution  under continuous stirring . It was dried afterwards  and placed in a quartz tube (1 cm I.D.) in a cylindrical oven and heated to 550 °C under a steam flow at 1.38 bar. Heating periods of 2, 4 and 6 h led to the formation of zeolite samples HY1, HY2 and HY3, respectively. They were subsequently treated with HCl  to remove extra-framework aluminum. Dealuminated HY zeolites were characterized by means of elemental analysis , infrared spectroscopy  and X-ray diffraction . trans-Anethole toluene solutions  were placed into three screw-capped test tubes (10 mL), each one containing a different zeolite . Zeolites had been previously activated . Reactions were performed by heating the tubes in a water bath . After chromatographic analysis of the resultant mixtures, conversion and selectivities were calculated as follows:M+, were calculated from the dimers’ mass spectra and used to distinguish configuration isomers: according to equation 4, the molecular ion abundance is divided by the sum of abundances of all fragment ions > m/z 50:Molecular ion stabilities, W2O  and heated to 60 °C for 6 h. It was then vacuum filtered and dried . An infrared spectrum of the resulting material was acquired (Bruker Tensor FT-IR spectrometer). This zeolite sample was then suspended in a test tube  with a trans-anethole toluene solution  and heated to 60 °C for 5 h under continuous stirring. The tube was filled under dry nitrogen flow. The mixture was decanted and analyzed by GC-MS in selected ion monitoring mode (SIM). The selected ions corresponded to the molecular ion and its main fragments, together with their deuterated analogs . Zeolite F1 (10 mg) was suspended in DCl/Dtrans-anethole toluene solution  was irradiated in an Ace Glass  7880 photoreactor assembly with a 5.5 Watt Pen-Ray gaseous Hg discharge UV-Vis lamp, at 5 °C for 120 min under constant stirring (100 rpm). trans-Anethole toluene solution aliquots (10 µL) were withdrawn every 20 min and analyzed by GC-FID, using the external standard (trans-anethole) method for quantification. A cis-anethole, dimer 2c, or dimer 2e:The Pen Ray UV lamp emission spectra were recorded with a high resolution HRG000c6-UV-NIR spectrometer  at the Atomic and Molecular Spectroscopy Laboratory of the Industrial University of Santander (Colombia). Discrete emission lines were found at 255, 313, 366, 405, 437, 548, and 579 nm. Quantum yields were determined as the quotient between the time derivative of the concentration and the flux, according to the following equation, in which X stands for trans-anethole per unit concentration, per unit time, according to the IUPAC recommended procedure [Potassium ferrioxalate was used for the actinometric determination of Ia, the number of quanta absorbed by rocedure . trans-anethole transformation observed during its exposure to acid sites in a zeolite or UV-Vis irradiation. Proton transfer from the zeolite to trans-anethole generates cationic intermediaries which lead the formation of eight different anethole dimers (methoxyphenyl-disubstituted hexenes). Their relative amounts agreed with those expected from the relative stability of the intermediary cationic species. The use of deuterated zeolites showed that the proton eliminated during the isomerization process is not necessarily the same proton transferred to the trans-anethole molecule when the cationic intermediary is formed. cis-Anethole and five isomeric methoxyphenyl-disubstituted cyclobutanes were observed as products of the transformation induced by the UV-Vis irradiation of trans-anethole. Isomerization and dimerization are concurrent processes in the


H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) assay. The antiproliferative activity of the tested acetylenic quinolinesulfonamides is comparable to that of cisplatin. The bioassay results demonstrate that most of the tested compounds show potent antitumor activities, and that some compounds exhibit better effects than the positive control cisplatin against various cancer cell lines. Among these compounds, 4-(3-propynylthio)-7-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline shows significant antiprolierative activity against T47D cells with IC50 values of 0.07 µM. In addition, 2-(3-Propynylthio)-6-[N-methyl-N-(3-propynyl)sulfa-moyl]quinoline and 2-(3-propynylseleno)-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline display highly effective atitumor activity against MDA-MB-231 cells, with IC50 values of 0.09 and 0.50 µM, respectively. Furthermore, most of the tested compounds show a weak cytotoxic effect against the normal HFF-1 cell line. Additionally, in order to suggest a mechanism of action for their activity, all compounds are docked into the binding site of two human cytochrome P450 (CYP) isoenzymes. These data indicate that some of the title compounds display significant cytotoxic activity, possibly targeting the CYPs pathways.In this study, a series of regioisomeric acetylenic sulfamoylquinolines are designed, synthesized, and tested in vitro for their antiproliferative activity against three human breast cacer cell lines  and a human normal fibroblast (HFF-1) by 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2 Interest in cancer has grown during the past century as infectious diseases have increasingly been controlled as a result of improved sanitation, vaccination, and antibiotics. Colon, stomach, liver, lung, and breast cancers are the most common causes of cancer death every year. Currently, breast cancer comprises 12% of all cancers in women worldwide. Moreover, the incidence of cancer continues to rise, and in many cases the applied treatment is ineffective pyrene, and physiologically relevant compounds such as estradiol . AromatiThe expression of the CYP1 family in malignant tumors has important consequences for the treatment of cancer. The expression of CYP1, especially extrahepatic CYPs 1A1 and 1B1 in tumor cells provides a molecular target for the development of new anticancer drugs, especially in the case of hormone-dependent cancer diseases. The generally accepted mechanism of inhibition by the acetylenes involves two possible pathways. The first pathway involves the oxidation of the internal carbon of the triple bond, which leads to heme destruction and enzyme deactivation through the formation of an Fe–O–CR=C(•/+)–Heme complex where a radical or positive charge is localized on the terminal carbon. The second pathway involves the oxidation of the terminal carbon of the triple bond, which results in the formation of a reactive ketene intermediate. This ketene intermediate can covalently bind to a nucleophilic amino acid residue in the enzyme’s active site, leading to irrversible inhibition without destruction of the heme .Against this background and in the continuation of efforts to synthesize potentially useful quinoline-based molecules ,19,20,211a–f, were obtained from their respective dihalogenoquinolines, as described previously by [N-methyl-N-(propynyl)sulfamoyl]quinolines 2a–f was produced by amination of chloroquinolinesulfonyl chlorides 1a–f with N-methylpropargylamine in chloroform with an excess of trethylamine  alkylation of sodium thiolates (or selenoates) with propargyl bromide resulted in the synthesis of compounds 5a–f and 6a–f   . This pr excess) , which weviously .7 with sodium phenylmethanethiolate (generated in situ from S-phenylmethylisothiouronium sulfate and sodium hydroxide) proceeded mainly by sustitution of the 3-bromine substituent to give the respective 6-chloro-3-phenylmethylsulfanylquinoline 10. Phenylmethylsulfanylquinoline 10 was subjected to oxidative chlorination with a system of 6% sodium hypochlorite/10% hydrochloric gaseous chlorine, which proceeded by splitting of the benzyl group and oxidative chlorination of the 3-thioquinoline fragment to provide the expected 3-quinolinesulfonyl chloride 13. Compounds 9 and 13 were then converted to the respective quinolinesulfonamides 12 and 14 by amination with N-methylpropargylamine with the presence of trimethylamine in chloroform with good yields (The reaction of 3-bromo-6-chloroquinoline d yields .1H-NMR), carbon nuclear magnetic resonance (13C-NMR), infrared spectroscopy (IR), and high-resolution mass spectrometry (HRMS). The 1H-NMR spectra of propargylsulfamoylquinolines 2, 5, 6, 11, 12, and 14 show the typical proton signals for the propargyl group at 1.97–2.35 ppm (trplet) and 3.61–4.40 ppm (doublet) with a coupling constant of 4J = 2.7 Hz. In the 13C-NMR spectra of compounds 2, 5, 6, 11, 12, and 14, the signals at 10.7–40.6 ppm , 70.8–75.0 ppm (-C≡), and 75.7–80.8 ppm (≡CH) are also attributed to the propargyl group. Moreover, in the 1H-NMR spectra of disubstituted quinolines 2, 5, 6, 11, 12, and 14, protons of the pyridine ring of the quinoline moiety formed an AX spin system, and protons of the benzene ring formed an AMX spin system. The assignments of both the 1H- and 13C-NMR spectra of the quinoline unit were based on the analysis of coupling patterns and the following two-dimensional nuclear magnetic resonance spectroscopy (2D NMR) experiments: correlation spectroscopy (COSY), heteronuclear multple-quantum correlation spectroscopy (HMQC), and heteronuclear multiple bond correlation spetroscopy (HMBC).All of the final compounds were pure and stable, as characterized by proton nuclear magnetic resonance . Cisplatin was used as a positive control to induce cell death.Encouraged by promising results in the field of antiproliferative derivatives of acetylenic sulfanylquinoline derivatives ,19,20,212a–f). Next, we introduced in positions 2- or 4- of the quinoline moiety the thipropargyl or the selenopropargyl group. A total of 21 compounds were synthesized. The results of the cytotoxicity studies of these compounds are summarized in 5e > 6f > 2e > 2c > 12 > 11 > 6b. As shown in N-methyl-N-propargylsulfamoyl]quinoline 5e had the highest antiproliferative activity (0.27 µM), which could be due to the presence of the thiopropargyl group in the 4- and sufamoyl group in the 7-quinoline positions. A structure–activity relationship was observed for the title compounds, and indicated that the replacement of a sulfur atom by a selenium atom retains activity against the MCF-7 cell line. In compound 6f, which contains a selenopropargyl group at the 4-quinoline position, the IC50 values were comparable to those obtained for compound 5e. Moreover, the MCF-7 cell line was found to be sensitive to 4-chloro-7-sulfamoyl- and 4-chloro-3-sulfamoyl-substituted quinolines 2e and 2c . It is iportant to note that the MCF-7 cell line was found to be sensitive to 4-substituted quinolines. For the tested compounds, the ranked order of the anticancer activity against the T47D cell lines is as folows: 5e > 5f ≈ 2b > 2c > 12 > 6b. As shown in 5e and 4-propargylthio-8-sulfamoylquinoline 5f had higher antiproliferative activity which could be due to the presence of the sulfamoyl group in the 7- or 8-quinoline position of the quinoline units. It is also noteworthy that compounds 5e and 5f showed higher cytotoxic activity than cisplatin in the tested cancer cell line, while their toxicity in normal human fibroblasts was low. However, the MCF-7 cell line was also found to be sensitive to 2,6-disubstituted quinoline 2b.The SAR analysis started by varying chlorine substituent of the 2- or 4- position of quinoline moiety with the 3-, 6-, 7-, and 8-position being substituted by sulfamoyl group. We first synthesized eight compounds .The data indicated that the cytotoxic activity of the compounds against the MDA-MB-231 cell line was ranked in the following order: 2b, 2c, 5e, 5f, 5b, 6b, 11, and 12 exhibited promising activity against the different cell lines used.It is noteworthy that compounds Molecular docking techniques were applied to investigate the binding mechanism for the series of acetylenic quinolinesulfonamides with human CYPs. Both CYP1A1 and CYP1B1 macromolecules are composed of 12 canonical α-helices (labeled from A to L), several short α-helices , and four canonical β-sheets (labeled from 1 to 4) with several β-strands ,36. Base5b > 11 > 6c > 6d > 6b > 6f > 12 (for CYP1A1) and 6d > 6b > 6f > 12 > 5f > 11 > 5b (for CYP1B1). Steric interactions make the biggest contribution in the formation of all complexes. Hydrogen bonds occur very rarely and are only possible when compounds 6e, 11 and 5e bind to CYP1A1. In summary, the binding affinity of acetylenic quinolinesulfonamides to CYPs is positively related to their molecular weight and steric hindrance. Thiopropargyl or selenopropargyl compounds substituted in the positions: 3,2-, 3,4- and 4,8- of the quinoline moiety or its chloro derivtives should not form stable complexes with CYPs in vivo.As a result of docking the computational models of quinolinesulfonamide derivatives to the active sites of CYP1A1 and CYP1B1, 42 possible ligand–protein complexes were obtained. All docked compounds were located in similar spaces inside the active sites, close to the heme-iron center. The superposition of studied compounds inside CYP1A1 and CYP1B1 (poses) were scored according to their intramolecular conformations (unfavorable ligand contributions) and binding interactions with amino acids (favorable ligand-protein contributions). Docking solutions with the lowest total score correspond to the highest binding affinities and present the most probable systems in vivo. The interaction energies between quinolinesulfonamides and CYPs are summarized in 5b, 5e, and 5f) or selenopropargyl (6a) groups. The orientation of 3,6-di(3-propynylsulfanyl)quinoline (11) differs depending on the type of enzyme, in the active site of CYP1A1 ferrous ion is very close to the quinoline moiety of compound 11, but in the active site of CYP1B1 its thiopropargyl group is oriented towards the heme.In order to find the best in silico orientation of docked compounds inside the CYP1A1 and CYP1B1 active sites to allow the inhibition of enzymes by ligands, the distances between the ligands’ functional groups and ferrous ion in heme molecules have been calculated see . Most of5b, 6b, and 11  shows that bounded acetylenic quinolinesulfonamides have deeply penetrated the strong hydrophobic matrix of these proteins. 2a and 2c, which form a halogen non-bond interaction in the active site of CYP1B1 with negatively charged Asp333 residue. Detailed information about the non-bond interactions of acetylenic quinolinesulfonamides with the residues of CYP1A1 and CYP1B1 are summarized in The quinoline moiety of all docked acetylenic quinolinesulfonamides is stably anchored to the CYPs’ active sites via single or double π–π stacked and π–π T-shaped type interactions, with the benzyl side chain of phenylalanines in positions Phe123, Phe224, and Phe258 (CYP1A1) or Phe134, Phe231, and Phe268 (CYP1B1). The above-described highly conjugated system is additionally strengthened by single or double amide–π stacked interactions between the quinoline part and amide group, between residues of Gly316 and Ala317 (CYP1A1) or Gly329 and Ala330 (CYP1B1). Additionally, the quinoline moiety may form hydrophobic π–δ interactions with Phe123, Gly316, and Ala317 (CYP1A1) or Ala330 (CYP1B1). A number of π–alkyl interactions between quinoline rings and aliphatic side chains of Ile115, Leu312, Ala317 (CYP1A1), or Val126, Ala133, and Ala330 (CYP1B1) have also been observed. The differences in the mechanism of binding acetylenic quinlinesulfonamides to active sites of human CYPs are related to the positions of their sulfamoyl and/or thiopropargyl or selenopropargyl groups. Chlorine atoms in compounds from series 2 do not paticipate in the formation of complexes with CYPs, except for compounds 5b, 6b, and 11 are visualized in their optimized orientations inside CYP1A1 and CYP1B1, between the interacting amino acids and heme. The thiopropargyl group of molecule 5b is oriented towards the ferrous ion of heme, inside the cavity of CYP1A1 or CYP1B1, and its superposition is stabilized by hydrophobic interactions with Val382 or Val395 and Leu496 or Leu509 aliphatic side chains  of CYP1A1, and Phe134 (length 5.82 Å) of CYP1B1 (11) docked to CYPs is oriented with the first thipropargyl group towards heme, and the second towards a highly conjugated system combined of three phenylalanines: Phe224, Phe258, and Phe319 in CYP1A1, or Phe134, Phe231, and Phe268 in CYP1B1  in the formation of a complex between compound 11 (nitrogen atom acts as acceptor) and CYP1A1 (hydroxyl group of Ser 122 residue acts as a donor). 11 and Ser 122 in the active site’s cavity of CYP1A1.Only one conventional hydrogen bond (length 2.56 Å) has a small contribution  in deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6); chemical shifts (δ) are reported in ppm and J values in Hz. The peak multiplicity is designated by a singlet (s), doublet (d), triplet (t), doublet of doublets (dd), doublet of triplets (dt), and multiplet (m). The HR MS analysis was performed on a Bruker Impact II instrument (Bruker). The IR spectra were recorded on an IRAffinity-1 Shimadzu spectrophotometer . Thin layer chrmatography (TLC) was carried out on silica gel 60 254F plates  using ethyl acetate as an eluent, and the spots were visualized by UV light (254 nm). All new compounds were purified by column chromatography. Silica gel 60 was used as a solid phase, and ethyl acetate was used as the eluent. The starting compounds, chloro-quinolinesulfochlorides 1a–f and 3,6-dichlorosulfonylquinoline 9, were obtained according to previously described methods quinoline (2a). Yield: 89%, m.p. 110–111 °C. 1H-NMR  δ: 2.20 , 3.08 , 4.22 , 7.69 , 7.89–7.99 , 8.10 , 8.98 . 13C-NMR  δ: 34.7 (CH3), 39.7 (CH2CCH), 73.9 (CH2CCH), 76.8 (CH2CCH), 125.7 (C-4a), 128.4 (C-8), 128.5 (C-6) 128.9 (C-5), 130.8 (C-3), 133.5 (C-7), 142.9 (C-4), 145.3 (C-2), 148.4 (C-8a). IR  νmax: 3292 (≡C-H), 3012 and 2966 , 2121 (C≡C), 1334 (S=O), 1153 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0119.2-Chloro-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (2b). Yield: 95%, m.p. 140–141 °C. 1H-NMR  δ: 1.97 , 2.92 , 4.14 , 7.54 , 8.08–8.17 , 8.26 , 8.40, . 13C-NMR  δ: 34.5 (CH3), 39.8 (CH2CCH), 74.4 (CH2CCH), 75.9 (CH2CCH), 124.1 (C-3), 125.8 (C-4a), 128.2 (C-7), 128.9 (C-5), 129.8 (C-8), 135.9 (C-6), 139.8 (C-4), 149.1 (C-8a), 153.7 (C-2). IR  νmax: 3211 (≡C-H), 2924 and 2852 , 2113 (C≡C), 1325 (S=O), 1141 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0121.4-Chloro-3-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (2c). Yield: 93%, m.p. 120–121 °C. 1H-NMR  δ: 2.12 , 3.07 , 4.23 , 7.77 , 7.93 , 8.20 , 8.45 , 9.39 . 13C-NMR  δ: 34.6 (CH3), 39.6 (CH2CCH), 74.1 (CH2CCH), 76.5 (CH2CCH), 125.4 (C-5), 126.1 (C-4a), 129.0 (C-6), 129.9 (C-3), 130.0 (C-8), 132.9 (C-7), 142.9 (C-4), 149.4 (C-2), 149.9 (C-8a). IR  νmax: 3265 (≡C-H), 2951 , 2118 (C≡C), 1334 (S=O), 1143 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0123.4-Chloro-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (2d). Yield: 91%, m.p. 124–125 °C. 1H-NMR  δ: 2.00 , 2.93 , 4.16 , 7.64 , 8.14 , 8.28 , 8.78 , 8.93 . 13C-NMR  δ: 34.4 (CH3), 39.9 (CH2), 74.5 (CH2CCH), 75.9 (CH2CCH), 122.6 (C-3), 125.7 (C-5), 125.9 (C-4a), 128.2 (C-7), 130.9 (C-8), 136.8 (C-6), 144.4 (C-4), 149.9 (C-8a), 152.2 (C-2). IR  νmax: 3267 (≡C-H), 3194 and 3086 , 2110 (C≡C), 1336 (S=O), 1155 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0125.4-Chloro-7-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (2e). Yield: 94%, m.p. 119–120 °C. 1H-NMR  δ: 2.01 , 2.93 , 4.15 , 7.64 , 8.02 , 8.39 , 8.64 , 9.92 . 13C-NMR  δ: 34.5 (CH3), 39.9 (CH2CCH), 74.5 (CH2CCH), 75.9 (CH2CCH), 123.4 (C-3), 125.1 (C-6), 125.7 (C-5), 128.6 (C-4a), 130.6 (C-8), 139.2 (C-7), 142.8 (C-4), 148.2 (C-8a), 151.6 (C-2). IR  νmax: 3282 (≡C-H), 3039 and 2935 , 2019 (C≡C), 1332 (S=O), 1161 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0128.4-Chloro-8-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (2f). Yield: 90%, m.p. 130–133 °C. 1H-NMR  δ: 2.03 , 3.00 , 4.39 , 7.62 , 7.74 , 8.49 , 8.56 , 8.93 . 13C-NMR  δ: 34.6 (CH3) 40.4 (CH2CCH), 72.7 (CH2CCH), 78.1 (CH2CCH), 122.3 (C-3), 126.5 (C-6), 127.3 (C-4a), 129.7 (C-5), 134.0 (C-7), 137.5 (C-8), 143.1 (C-4), 145.0 (C-8a), 150.6 (C-2). IR  νmax: 3254 (≡C-H), 2943 , 2015 (C≡C), 1319 (S=O), 1132 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0131.6-Chloro-3-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (14). Yield: 90%, m.p. 146–147 °C 1H-NMR  δ: 1.98 , 2.94 , 4.19 , 7.83 , 7.96 , 8.15 , 8.60 , 9.23 . 13C-NMR  δ: 34.5 (CH3), 39.9 (CH2CCH), 74.9 (CH2CCH), 75.7 (CH2CCH), 127.0 (C-6), 127.5 (C-5), 131.2 (C-8), 131.8 (C-3), 133.4 (C-7), 134.2 (C-4a), 136.3 (C-4), 147.5 (C-2), 147.6 (C-8a). IR  νmax: 3302 (≡C-H), 3041 and 2974 , 2129 (C≡C), 1340 (S=O), 1155 (S=O). HRMS (ESI) m/z: C13H11ClN2NaO2S [M + Na]+, Calcd. 317.0127; Found 317.0125.N-methyl-N-(3-propynyl)sulfamoylquinoline (5a). Yield: 86%, m.p. 100–101 °C. 1H-NMR  δ: 2.16–2.19 , 2.98 , 4.17 , 4.21 , 7.59 , 7.80–7.89 , 8.02 , 8.70 . 13C-NMR  δ: 19.2 (SCH2CCH), 34.5 (CH3), 39.8 (NCH2CCH), 70.8 (SCH2CCH), 73.9 (NCH2CCH), 76.9 (NCH2CCH), 79.5 (SCH2CCH), 124.2 (C-4a), 126.9 (C-6), 128.0 (C-8), 128.9 (C-5), 128.9 (C-5), 129.7 (C-3), 132.9 (C-7), 140.3 (C-4), 148.4 (c-8a), 154.3 (C-2). IR  νmax: 3271 (≡C-H), 3263 (≡C-H), 2951 and 2910 , 2115 (C≡C), 1323 (S=O), 1159 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0403.2-(3-Propynylthio)-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (5b). Yield: 81%, m.p. 164–165 °C. 1H-NMR  δ: 2.20 , 2.23 , 2.90  4.12 , 4.18 , 7.34 , 8.00–8.10 , 8.30 . 13C-NMR  δ: 18.2 (SCH2CCH), 34.4 (CH3), 39.8 (NCH2CCH), 70.9 (SCH2CCH), 74.3 (NCH2CCH), 76.1 (NCH2CCH), 79.4 (SCH2CCH), 121.9 (C-3), 125.0 (C-4a), 127.6 (C-7), 129.0 (C-5), 129.1 (C-8), 134.0 (C-6), 136.5 (C-4), 149.3 (C-8a), 161.2 (C-2). IR  νmax: 3286 (≡C-H), 3261 (≡C-H), 2974 and 2945 , 2113 (C≡C), 1325 (S=O), 1155 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0399.4-(3-Propynylthio)-3-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (5c). Yield: 81%, m.p. 101–102 °C. 1H-NMR  δ: 2.14–2.16 , 3.05 , 3.78 , 4.29 , 7.69 , 7.90 , 8.20 , 8.82 , 9.47 . 13C-NMR  δ: 25.9 (SCH2CCH), 34.7 (CH3), 39.9 (NCH2CCH), 73.6 (SCH2CCH), 74.1 (NCH2CCH), 77.2 (NCH2CCH), 78.4 (SCH2CCH), 127.8 (C-5), 128.7 (C-6), 130.0 (C-4a), 130.2 (C-6), 132.5 (C-7), 136.8 (C-3), 144.0 (C-4), 149.1 (C-2), 140.7 (C-8a). IR  νmax: 3294 (≡C-H), 3267 (≡C-H), 2968 and 2926 , 2013 (C≡C), 1330 (S=O), 1147 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0390.4-(3-Propynylthio)-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (5d). Yield: 88%, m.p. 139–140 °C. 1H-NMR  δ: 2.01 , 2.35 , 2.91 , 3.90 , 4.15 , 7.51 , 8.09 , 8.24 , 8.61 , 8.89 . 13C-NMR  δ: 20.0 (SCH2CCH), 34.5 (CH3) 39.9 (NCH2CCH), 73.0 (SCH2CCH), 74.6 (NCH2CCH), 75.9 (NCH2CCH), 77.3 (SCH2CCH), 117.8 (C-3), 124.8 (C-5), 125.5 (C-4a), 127.8 (C-7), 130.7 (C-8), 135.4 (C-6), 148.0 (C-4), 148.8 (C-8a), 151.0 (C-2). IR  νmax: 3292 (≡C-H), 3250 (≡C-H), 3012 and 2966 , 2121 (C≡C), 1334 (S=O), 1153 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0391.4-(3-Propynylthio)-7-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (5e). Yield: 90%, m.p. 113–114 °C. 1H-NMR  δ: 2.01 , 2.34 , 2.94 , 3.88 , 4.13 , 7.50 , 7.92 , 8.20 , 8.60 , 8.88 . 13C-NMR  δ: 19.8 (SCH2CCH), 34.5 (CH3), 39.9 (NCH2CCH), 72.8 (SCH2CCH), 74.4 (NCH2CCH), 75.9 (NCH2CCH), 76.6 (SCH2CCH), 118.4 (C-3), 124.0 (C-6), 124.7 (C-5), 128.4 (C-4a), 130.7 (C-8), 138.5 (C-7), 146.2 (C-4), 146.5 (C-8a), 150.9 (C-2). IR  νmax: 3255 (≡C-H), 2935 , 2014 (C≡C), 1348 (S=O), 1159 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0391.4-(3-Propynylthio)-8-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (5f). Yield: 93%, m.p. 119–120 °C. 1H-NMR  δ: 2.06 , 2.32 , 2.98 , 3.86 , 4.40 , 7.47 , 7.63 , 8.31 , 7.90 , 8.90 . 13C-NMR  δ: 19.9 (SCH2CCH), 34.6 (CH3) 40.6 (NCH2CCH), 72.6 (SCH2CCH), 72.7 (NCH2CCH), 77.6 (SCH2CCH), 78.3 (NCH2CCH), 117.5 (C-3), 125.3 (C-6), 127.0 (C-4a), 128.9 (C-5), 133.4 (C-7), 137.4 (C-8), 143.7 (C-8a), 146.4 (C-4), 149.9 (C-2). IR  νmax: 3261 (≡C-H), 3253 (≡C-H), 2957 and 2922 , 2016 (C≡C), 1325 (S=O), 1132 (S=O). HRMS (ESI) m/z: C16H14N2NaO2S2 [M + Na]+, Calcd. 353.0394; Found 353.0394.2-(3-Propynylseleno)-3-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6a). Yield: 89%, m.p. 109–110 °C. 1H-NMR  δ: 2.13 , 2.19 , 2.96 , 4.05 , 4.21 , 7.59 , 7.81–7.90 , 8.04 , 8.64 . 13C-NMR  δ: 11.7 (SeCH2CCH), 34.6 (CH3), 39.9 (NCH2CCH), 70.8 (SeCH2CCH), 74.2 (NCH2CCH), 77.2 (NCH2CCH), 80.8 (SeCH2CCH), 124.4 (C-4a), 127.1 (C-6), 128.3 (C-8), 128.9 (C-5), 131.0 (C-3), 132.9 (C-7), 139.6 (C-4), 149.0 (C-8a), 152.5 (C-2). IR  νmax: 3275 (≡C-H), 2918 and 2852 , 2121 (C≡C), 1321 (S=O), 1155 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS [M + Na]+, Calcd. 400.9839; Found 400.9841.2-(3-Propynylseleno)-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6b). Yield: 89%, m.p. 179–180 °C. 1H-NMR  δ: 1.99 , 2.25 , 2.90 , 4.09 , 4.12 , 7.47 , 7.99–8.04 , 8.09 , 8.31 . 13C-NMR  δ: 10.7 (SeCH2CCH), 34.4 (CH3), 39.9 (NCH2CCH), 71.2 (SeCH2CCH), 74.3 (NCH2CCH), 76.0 (NCH2CCH), 80.6 (SeCH2CCH), 124.2 (C-3), 125.5 (C-4a), 127.6 (C-7), 129.2 (C-5 and C-8), 134.3 (C-6), 136.2 (C-4), 149.9 (C-8a), 159.5 (C-2). IR  νmax: 3284 (≡C-H), 3261 (≡C-H), 2960 and 2904 , 2113 (C≡C), 1325 (S=O), 1157 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS [M + Na]+, Calcd. 400.9839; Found 400.9836.4-(3-Propynylseleno)-3-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6c). Yield: 85%, m.p. 122–123 °C. 1H-NMR  δ: 2.15 , 2.22 , 3.04 , 3.61 , 4.31 , 7.73 , 7.88 , 8.18 , 8.68 , 9.42 . 13C-NMR  δ: 16.0 (SeCH2CCH), 34.6 (CH3), 40.0 (NCH2CCH), 73.5 (SeCH2CCH), 74.2 (NCH2CCH), 77.4 (NCH2CCH), 79.5 (SeCH2CCH), 128.8 (C-6), 129.7 (C-4a), 130.0 (C-5), 130.4 (C-8), 132.4 (C-7), 137.0 (C-3), 142.0 (C-4), 148.5 (C-2), 149.2 (C-8a). IR  νmax: 3273 (≡C-H), 3055 and 2978 , 2123 (C≡C), 1342 (S=O), 1155 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS [M + Na]+, Calcd. 400.9839; Found 400.9838.4-(3-Propynylseleno)-6-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6d). Yield: 91%, m.p. 139–140 °C. 1H-NMR  δ: 2.02 , 2.34 , 2.92 , 3.78 , 4.15 , 7.75 , 8.11 , 8.32 , 8.57 , 8.85 . 13C-NMR  δ: 11.9 (SeCH2CCH), 34.5 (CH3), 39.9 (NCH2CCH), 73.3 (SeCH2CCH), 74.6 (NCH2CCH), 75.9 (NCH2CCH), 78.6 (SeCH2CCH), 123.3 (C-3), 126.9 (C-5), 127.7 (C-4a), 128.0 (C-7), 130.6 (C-8), 136.0 (C-6), 146.6 (C-4), 147.9 (C-8a), 150.6 (C-2). IR  νmax: 3290 (≡C-H), 3248 (≡C-H), 3012 and 2968 , 2010 (C≡C), 1332 (S=O), 1153 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS [M + Na]+, Calcd. 400.9839; Found 400.9838.4-(3-Propynylseleno)-7-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6e). Yield: 82%, m.p. 108–109 °C. 1H-NMR  δ: 2.02 , 2.32 , 2.92 , 3.76 , 4.13 , 7.73 , 7.95 , 8.15 , 8.63 , 8.84 . 13C-NMR  δ: 11.6 (SeCH2CCH), 34.5 (CH3), 39.9 (NCH2CCH), 73.1 (SeCH2CCH), 74.5 (NCH2CCH), 76.0 (NCH2CCH), 78.7 (SeCH2CCH), 123.6 (C-3), 124.7 (C-6), 126.8 (C-5), 129.8 (C-8), 130.3 (C-4a), 139.3 (C-7), 145.3 (C-4), 145.5 (C-8a), 149.8 (C-2). IR  νmax: 3296 (≡C-H), 3275 (≡C-H), 3074 and 2980 , 2015 (C≡C), 1342 (S=O), 1161 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS [M + Na]+, Calcd. 400.9839; Found 400.9834.4-(3-Propynylseleno)-8-[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (6f). Yield: 89%, m.p. 111–115 °C. 1H-NMR  δ: 2.05 , 2.31 , 2.99 , 3.72 , 4.40 , 7.63–7.69 , 8.27 , 8.52 , 8.86 . 13C-NMR  δ: 19.9 (SCH2CCH), 34.6 (CH3) 40.6 (NCH2CCH), 72.6 (NCH2CCH), 72.9 (SeCH2CCH), 78.3 (NCH2CCH), 78.9 (SeCH2CCH), 123.0 (C-3), 125.6 (C-6), 129.0 (C-4a), 131.1 (C-5), 133.5 (C-7), 137.5 (C-8), 143.3 (C-4), 143.9 (C-8a), 149.9 (C-2). IR  νmax: 3257 (≡C-H), 2957 and 2925 , 2015 (C≡C), 1325 (S=O), 1132 (S=O). HRMS (ESI) m/z: C16H1N2NaO2SeS M + Na]+, Calcd. 400.9839; Found 400.9833.11).A mixture of 6-chloro-3-bromoquinoline , sodium methanethiolate , and dry DMF (10 mL) was boiled with stirring under argon atmosphere for 6 h. The mixture was then cooled to room temperature and poured into cold 8% aqueous NaOH (30 mL), and prpargyl bromide  was added dropwise with stirring. The stirring cotinued at room temperature for 1 h. The solid was filtered off, washed with water, and air-dried. The crude product was purified by column chromatography (ethyl acetate) to give pure 3,6-di(3-propynylsulfanyl)quinoline quinoline (11). Yield: 91%, m.p. 97–98 °C. 1H-NMR  δ: 2.27–2.30 , 3.70 , 3.74 , 7.70 , 7.78 , 8.05 , 8.18 , 8.88 . 13C-NMR  δ: 21.9 (SCH2CCH), 22.7 (SCH2CCH), 72.1 (SCH2CCH), 72.8 (SCH2CCH), 78.8 (SCH2CCH), 79.1 (SCH2CCH), 125.8 (C-5), 128.5 (C-4a), 129.2 (C-8), 129.7 (C-3), 131.0 (C-7), 135.4 (C-6), 136.6 (C-4), 144.6 (C-8a), 150.9 (C-2). IR  νmax: 3278 (≡C-H), 2926 and 2885 (CH2), 2110 (C≡C). HRMS (ESI) m/z: C15H12NS2 [M + H]+, Calcd. 270.0411; Found 270.0415.N-methylpropargylamine  in 20 mL of chloroform was cooled to 0 °C and then, with stirring, 3,6-dichlorosulfonylquinoline 9  was added. The stirring was continued at 0–5 °C for 5 h. Then, the reaction mixture was washed with cold water (2 × 6 mL), dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (ethyl acetate) to give pure product 12.A solution of triethylamine  and 3,6-Di[N-methyl-N-(3-propynyl)sulfamoyl]quinoline (12). Yield: 96%, m.p. 138–139 °C. 1H-NMR  δ: 1.97–1.99 , 2.95 , 2.97 , 4.18 , 4.22 , 8.23 , 8.36 , 8.55 , 8.82 , 9.38 . 13C-NMR  δ: 34.5 (2 × CH3), 39.9 (2 × CH2), 74.6 (CH2CCH), 75.0 (CH2CCH), 75.7 (CH2CCH), 75.8 (CH2CCH), 125.6 (C-4a), 129.8 (C-7), 130.2 (C-5), 130.7 (C-8), 132.7 (C-3), 137.5 (C-6), 138.4 (C-4), 149.7 (C-2), 150.0 (C-8a). IR  νmax: 3300 (≡C-H), 3282 (≡C-H), 3059 and 2980 , 2129 (C≡C), 1365, 1348, and 1328 (S=O), 1170 and 1143 (S=O). HRMS (ESI) m/z: C17H16N3NaO4S2 [M + Na]+, Calcd. 413.0480; Found 413.0482.7)  was added, and heating and stirring continued for 6 h. The mixture was cooled to room temperature and poured into ice water (80 mL). The solid was filtered off, washed with water, and air-dried. 6-Chloro-3-phenylmethyl-sulfanylquinoline was recrystallized from a mixture of hexane–CH2Cl2  to give pure compound (10) (89%). m.p. 77–78 °C , NaOH , and dry DMF (10 mL) was heated with stirring at 80 °C under an argon atmosphere for 1 h. Then, 6-chloro-3-bromoquinoline (77–78 °C ).10  was cooled to 5 °C and then dropped over 60 min to a well-stirred mixture of 6% aqueous solution of sodium hypochlorite  at such a rate that the temperature remained below 10 °C. The mixture was poured onto 15 g of ice. The organic layer was separated, and the aqueous layer was extracted with chlorform (3 × 3 mL). The chloroform extracts were combined, washed with water, and dried over anhdrous sodium sulfate. CHCl3 was evaporated to leave a solid residue. The residue was recrystallized from benzene to give 6-chloro-3-chlorosulfonylquinoline (13) (91%). m.p. 109–110 °C (lit. m.p. 109–110 °C quinoline (5b) and 2-(3-propynyl-seleno)-6-[N-methyl-N-(3-propynyl)sulfamoyl]- quinoline (6b), were ranked highly by the PLANTSPLP scoring function for CYP1A1 and CYP1B1. The quinoline moiety of acetylenic quinolinesulfonamides form a number of hydrophobic interactions with phenylalanines of CYP1A1  and CYP1B1 . The key to understanding the differences between the binding mechanisms of selected compounds is the orientation of the sulfamoyl and/or thiopropargyl groups inside CYP catalytic sites, very close to the heme–iron center. The superposition of acetylenic quinolinesulfonamide functional groups is stabilized by essential hydrophobic interactions with Ile115, Leu312, Ala317, Val382, Leu386, and Leu490 (CYP1A1) and Val 126, Ala133, Leu264, Ala330, Val395, Ile399, and Leu509 (CYP1B1). The orientation of sufur-containing substituents (sulfamoyl and/or thiopropargyl groups) inside active sites can be additionally enhanced by π–sulfur miscellaneous interactions, mainly with Phe123 or Phe224 (CYP1A1) and Phe134 or Phe268 (CYP1B1) residues. Docking data remain in good correlation with cytotoxic activity results.In conclusion, the design, synthesis, structure, and in vitro antiproliferative activity of new quinolinesulfonamide derivatives were presented in this paper. Compounds were prepared using a straightforward and efficient method of synthesis. The molecular structures of the title compounds were confirmed by 5b and 6b may be attributed to the inhibition of the CYP routes. These findings can be exploited in favor of the development and testing of novel quinoline-based molecules for better anticancer activity.This observation suggests that, although the WST-1 screening protocol did not decide on a mechanism for the observed cytotoxic activity of the tested compounds, the activity of


S*,8R*,7’R*,8’S*)-icariol A2-9-O-β-xylopyranoside (1), and a new indole alkaloid, hygarine (2), were isolated from the extracts of Hygroryza aristata (Gramineae). The structures of all compounds were elucidated on the basis of NMR spectral analysis. The compounds (-)-epigallocatechin-3-O-gallate (4) and (-)-epicatechin-3-O-gallate (5) possess free radical scavenging activities and compound 1 could inhibit superoxide anion generation and elastase release by fMLP/CB-induced human neutrophils with IC50 values of 19.33 ± 0.86 and 24.14 ± 1.59 μM, respectively. Twenty-six known compounds and two new compounds, including a new lignan, (7 Hygroryza aristata (Gramineae) is a stoloniferous perennial plant widely distributed in tropical Asia +m/z 323.3734). The IR spectrum of 2 displayed absorption bands from a hydroxyl group  and an amide group . In the UV spectrum of 2, absorption maxima were observed at 224 (sh), 288, and 312 (sh) nm, revealing that the compound has an indole skeleton D +0.63° . UV λmaxnm (logε) : 237 (2.89), 279 (2.70). IRνmax cm−1: 3,375, 1,613, 1,517, 1,113. FAB-MS m/z (rel. int.%): 591 , 175 (7), 161 (9), 153 (6), 149 (20), 133 (9). 1H- and 13C-NMR: see Hygarine (2). Brown syrup. UV λmaxnm (logε): 224 (sh) (3.52), 288 (3.27), 312 (sh) (3.22). IRνmax cm−1: 3,404, 1,643, 1,048. FAB-MS m/z (rel. int.%): 323 , 147 (30), 55 (100), 94 (30). HRFAB-MS: Anal. Calcd. for C19H18N2O3 323.3726, found 323.3734. 1H- and 13C-NMR: see S*,8R*,7’R*,8’S*)-icariol A2-9-O-β-xylopyranoside (1) and hygarine (2), together with twenty-six known compounds, were isolated from the extracts of Hygroryza aristata. (-)-Epigallocatechin-3-O-gallate (4) and (-)-epicatechin-3-O-gallate (5) possess free radical scavenging activities. -Icariol A2-9-O-β-xylopyranoside (1) could inhibit superoxide anion generation and elastase release by fMLP/CB-induced human neutrophils with IC50 values of 19.33 ± 0.86 and 24.14 ± 1.59 μM, respectively. Two new compounds, (7