Euphorbia genus, with its remarkable diversity in species, is due to its variety of chemical compositions. Euphorbia turcomanica is one of the species of the spurge family growing wildly in Iran. This research aims to investigate the presence of secondary metabolites, specially terpenoid compounds, in terms of structural determination. Samples of E. tucomanica were finely powdered and macerated with acetone/dichloromethane 2:1. Repeated column chromatography using silica gel, MPLC, and HPLC methods followed by the analysis of data obtained from spectroscopic means was carried out to purify and identify the terpenoid constituents. The chemical structures of nine known terpenoids were determined for the first time from E. turcomanica during this study. Loliolide (1), a monoterpene, and eight steroids and triterpenes, including simiarenol (2), isomultiflorenol (3), cycloart-25-ene-3β,24-diol (4), β-sitosterol (5), cycloart-23-ene-3β,25-diol (6), 3α, 11α-dihydroxyurs-12-ene (7), 3β, 24β, 25-trihydroxycycloartane (8), and 7α-hydroxystigmasterol (9) were isolated and identified. E. tucomanica, with a rich terpenoid profile, can be one of the valuable and economic sources providing compounds for drug development.The attraction to the  Euphorbia or spurge, which is the largest genus of this family. These plants occur naturally in both tropical and temperate regions [Euphorbia, is one of the most diverse countries in southwest Asia, with the highest occurrence of endemics which are 21 species until 2020 [Euphorbia turcomanica Boiss., one of the species of the spurge family, is an annual herb growing wild in the lower fields and plains of Iran [The family Euphorbiaceae, with nearly around 7500 species and 300 genera, is one of the largest families of flowering plants on earth. More than 2000 species belong to the genus  regions ,2. Iran, of Iran .Euphorbia species, including bark, cortex, seeds, latex, lactiferous duct, stem, wood, stem bark, leaves, and flowers. The complex and diverse phytochemical properties of this genus include compounds from different chemical classes, especially the ones that consist of isoprene units known as terpenes and terpenoids, which are considered the largest and most diverse group of natural products. Between the major classes of terpenoids found in Euphorbia species, diterpenes and triterpenes are the most important constituents of the secondary metabolites, mainly in their ester forms [Euphorbia plants. Some of these diterpenoids are scarcely distributed in nature, like tigliane, which can be found in Euphorbia species [Secondary metabolites with a wide range of biological activities and broad structural diversity have been isolated from different parts of er forms ,6. Diter species ,8. The s species ,9.Euphorbia have a wide range of applications, but in particular, the latex of E. turcomanica is used as a laxative and diuretic. Also, it is externally applied to snake bites and scorpion stings and internally used for treating intestinal worms and purifying blood [Euphorbia genus in pharmacological research is due to the variety of chemical compositions in them. Part of these effects is associated with the presence of terpenoids, including diterpenes and triterpenes, especially cycloartanes. Plants-derived terpenoids have been identified to have anticancer and anti-inflammatory activities. The Euphorbia genus, with a rich terpenoid profile, can be one of the valuable and economically important sources providing compounds in drug development [In traditional medicine, species of ng blood . The attelopment ,11.E. turcomanica, but only a few phytochemical studies have been carried out to isolate and elucidate the chemical constituents present in this plant, including steroids, flavonoids, phenolics, aromatics, and especially terpenoid compounds, by preliminary screening [Most research has proven the biological and therapeutic properties of creening .E. turcomanica. The structures of these compounds were mainly established by spectroscopic methods.This study describes the isolation and structural determination of secondary metabolites from ® Gradient SystemC605 apparatus equipped with a fraction collector, using a glass column filled with silica gel , HPLC (High-Performance Liquid Chromatography) on a Waters® apparatus equipped with a 515 pump, a Waters®2487 Dual Wavelength Absorbance Detector and a Waters®2414 Refractive Index Detector, using Spherisorb®Sil (250 mm × 20 mm) column, 5 µm silica gel and hexane, with ethyl acetate as the mobile phase. The purity was checked using TLC (Thin-Layer Chromatography) with Merck® silica gel GF254 plates detected by spraying cerium sulfate. Determination of each structure was performed by NMR spectra recorded on a Bruker Avance® AV400 spectrometer.Medium pressure liquid chromatography (MPLC) was carried out on a BUCHIE. turcomanica was gathered in two places, Isfahan-Qom Road, Center of Iran, and Bandar Tourkaman, North of Iran, during the flowering time in July 2012. The plant was identified by Mr. Bahram Zehzad, Department of biology, University of Shahid Beheshti, Tehran, and a voucher specimen (No. 2410) is kept at the Herbarium of Pharmacognosy, Department of Pharmacognosy, Faculty of Pharmacy, Isfahan University of Medical Sciences.The whole plant of E. tucomanica were finely powdered and extracted with acetone/dichloromethane 2:1. The maceration was carried out at room temperature in 3 repeated procedures that each took 4 days and 10–15 liters of solvent. The extract was concentrated under a vacuum. The greasy extract was subjected to Celite and chromatographed using the VLC method with silica gel coated with paraffin as the stationary phase and MeOH–H2O (60:40) and MeOH–H2O (90:10) as a solvent system to eliminate chlorophyll and fats from the extract. Normal-phase (NP)-TLC analysis indicated that the fraction which was separated by MeOH–H2O (60:40) contained a series of terpenoids and fraction eluted with MeOH–H2O (90:10) affording steroid and triterpene-containing extract.Air-dried samples of 3, adsorbed onto Celite, and chromatographed using an open column and a linear gradient solvent system from n-hexane/ EtOAc 3–30%, which yielded 15 fractions. After comparing the TLC analysis of the obtained fractions, the ones with similar UV characterizations and Rf values were combined and concentrated by vacuum. Further purification of these fractions was carried out by MPLC using a linear gradient solvent system from MeOH/H2O (70:30) to MeOH/H2O (100:0), resulting in subfractions [1 was obtained from open column chromatography using a linear gradient solvent system from n-hexane/acetone 3% to n-hexane/acetone 30% of the first subfraction of fraction 10 in an n-hexane/acetone 20% mobile phase. After further vacuum concentration and analysis, it was identified as pure crystals of loliolide.The terpene-containing extract was dissolved in CHClractions . CompounThe steroid and triterpene-containing extract chromatographed by MPLC using a linear gradient solvent system from n-hexane 100% to EtOAc 100% . Fractio2 as simiarenol and [4 was isolated from fraction 5 as cycloartane. Compound 5 was purified from obtained crystals by recrystallization and identified as pure β-sitosterol. Fractions 7 and 8 were eluted with Hex/EtOAc (70:30) in MPLC and purified by HPLC using Hex/EtOAc (80:20) as a solvent and resulting in the isolation of compound 6 as a cycloartane. Fraction 8 contained pure crystals that were filtered and concentrated by a rotary evaporator, which were further purified by recrystallization and yielded compound 7. Fraction 12, eluted with Hex/EtOAc (50:50), was purified further on HPLC using Hex/EtOAc (37:63) as a solvent to yield compound 8 as a cycloartane-type triterpene. Fraction 14 was eluted with Hex/EtOAc (40:60) in MPLC and further purified by HPLC using Hex/EtOAc (50:50) as solvent. As a result, a significant amount of needle-shaped crystal was obtained and further purified by recrystallization and yielded compound 9 as a steroidal triterpene.Fraction 3 was eluted with Hex/EtOAc (90:10) in MPLC and further purified by HPLC using Hex/EtOAc (90:10) as the mobile phase and resulting in the isolation of compound Euphorbia turcomanica, as shown in 1H NMR and 13C NMR data of each compound are reported here.Nine compounds were extracted and purified from 1)Loliolide : δH 1.55 , 1.98 , 4.35 , 1.80 , 2.02 , 5.72 , 1.49 , 1.29 , 1.80 , 13C NMR data : 30.7 (C-1), 45.6 (C-2), 66.8 (C-3), 47.3 (C-4), 86.7 (C-5), 171.9 (C-6), 112.9 (C-7), 182.4 (C-8), 26.5 (C-9), 27.0 (C-10), 30.7 (C-11).White crystal, MW (g/mol): 196.1; 2.2)Simiarenol: 3β-hydroxy-E:B-friedo-hop-5-ene : δH 3.47 , 5.62 , 1.35 , 1.25 , 1.14 , 1.04 , 1.00 , 0.92 , 0.77 , 0.84 , 0.88 , 13C NMR data : 18.20 (C-1), 27.90 (C-2), 76.52 (C-3), 40.97 (C-4), 142.08 (C-5), 122.17 (C-6), 24.18 (C-7), 44.37 (C-8), 34.97 (C-9), 50.35 (C-10), 34.27 (C-11), 29.21 (C-12), 38.74 (C-13), 39.09 (C-14), 29.08 (C-15), 35.53 (C-16), 42.92 (C-17), 51.86 (C-18), 20.05 (C-19), 28.46 (C-20), 60.15 (C-21), 30.94 (C-22), 29.11 (C-23), 25.62 (C-24), 18.01 (C-25), 15.89 (C-26), 15.14 (C-27), 16.21 (C-28), 22.10 (C-29), 23.07 (C-30).Amorphous white powder, MW (g/mol): 426.70; 3.3)Isomultiflorenol : δH 3.23 , 0.80 , 0.95 , 0.95 , 1.05 , 0.96 , 1.06 , 0.98 , 1.00 , 13C NMR data : 34.33 (C-1), 26.34 (C-2), 79.17 (C-3), 38.94 (C-4), 50.81 (C-5), 19.36 (C-6), 28.04 (C-7), 135.19 (C-8), 133.63 (C-9), 37.72 (C-10), 20.95 (C-11), 30.80 (C-12), 37.39 (C-13), 41.13 (C-14), 27.63 (C-15), 36.89 (C-16), 30.93 (C-17), 44.23 (C-18), 34.48 (C-19), 28.47 (C-20), 35.15 (C-21), 36.84 (C-22), 28.18 (C-23), 15.76 (C-24), 19.96 (C-25), 19.14 (C-26), 24.78 (C-27), 31.65 (C-28), 34.74 (C-29), 33.09 (C-30).Amorphous white powder, MW (g/mol): 426.7; 4.4)Cycloart-25-ene-3β,24-diol : δH 3.28 , 0.96 , 0.33 , 0.55 , 0.88 , 4.02 , 1.72 , 0.88 , 0.80 , 0.96 , 13C NMR data : 32.04 (C-1), 30.49 (C-2), 78.99 (C-3), 40.61 (C-4), 47.21 (C-5), 21.25 (C-6), 28.22 (C-7), 48.14 (C-8), 20.09 (C-9), 26.48 (C-10), 26.15 (C-11), 35.67 (C-12), 45.38 (C-13), 48.93 (C-14), 32.08 (C-15), 26.56 (C-16), 52.27 (C-17), 18.19 (C-18), 29.85 (C-19), 36.05 (C-20), 18.44 (C-21), 32.10 (C-22), 28.28 (C-23), 76.52 (C-24), 147.87 (C-25), 111.10 (C-26), 17.74 (C-27), 19.45 (C-28), 25.56 (C-29), 14.15 (C-30).Amorphous white powder, MW (g/mol): 442.38; 5.5)β-sitosterol: Stigmast-5-en-3β-ol : δH 1.85 , 3.52 , 5.36 , 0.68 , 1.01 , 0.92 , 0.81 , 0.85 , 0.86 , 0.96 , 13C NMR data : 37.37 (C-1), 29.23 (C-2), 71.95 (C-3), 42.42 (C-4), 140.88 (C-5), 121.88 (C-6), 32.02 (C-7), 32.04 (C-8), 50.23 (C-9), 36.63 (C-10), 21.21 (C-11), 39.89 (C-12), 42.44 (C-13), 56.88 (C-14), 24.44 (C-15), 28.40 (C-16), 56.15 (C-17), 12.00 (C-18), 19.55 (C-19), 36.28 (C-20), 18.91 (C-21), 34.05 (C-22), 26.13 (C-23), 45.93 (C-24), 29.23 (C-25), 19.98 (C-26), 19.15 (C-27), 23.17 (C-28), 12.12 (C-29).White crystal, MW (g/mol): 414; 6.6)Cycloart-23-ene-3β,25-diol : δH 3.28 , 0.97 , 0.33 , 0.55 , 0.86 , 5.60 , 5.60 , 1.32 , 1.32 , 0.88 , 0.81 , 0.97 , 13C NMR data : 32.08 (C-1), 30.49 (C-2), 78.98 (C-3), 40.61 (C-4), 47.20 (C-5), 21.25 (C-6), 28.21 (C-7), 48.12 (C-8), 20.09 (C-9), 26.14 (C-10), 25.57 (C-11), 32.88 (C-12), 45.41 (C-13), 48.94 (C-14), 35.70 (C-15), 26.54 (C-16), 52.12 (C-17), 18.24 (C-18), 30.00 (C-19), 36.51 (C-20), 18.41 (C-21), 39.16 (C-22), 139.42 (C-23), 125.76 (C-24), 70.73 (C-25), 30.10 (C-26), 30.04 (C-27), 19.43 (C-28), 14.15 (C-29), 24.85 (C-30).White crystal, MW (g/mol): 442.38; 7.7)3α, 11α-dihydroxyurs-12-ene : 3.21 , 3.98 , 4.90 , 0.90 , 0.69 , 0.94 , 0.76 , 1.00 , 0.82 , 13C NMR data : 33.5 (C-1), 25.4 (C-2), 76.0 (C-3), 37.5 (C-4), 48.8 (C-5), 18.2 (C-6), 35.2 (C-7), 43.5 (C-8), 55.8 (C-9), 38.2 (C-10), 68.4 (C-11), 128.7 (C-12), 142.9 (C-13), 42.2 (C-14), 27.9 (C-15), 27.7 (C-16), 33.6 (C-17), 58.1 (C-18), 39.4 (C-19), 39.3 (C-20), 31.1 (C-21), 41.3 (C-22), 28.7 (C-23), 22.4 (C-24), 16.6 (C-25), 18.0 (C-26), 23.3 (C-27), 28.6 (C-28), 17.5 (C-29), 21.3 (C-30).White crystal, MW (g/mol): 442.38; 8.8)3β, 24β, 25-trihydroxycycloartane : δH 3.27 , 0.96 , 0.33 , 0.55 , 0.88 , 3.31 , 1.16 , 1.22 , 0.89 , 0.96 , 0.80 , 13C NMR data : 32.10 (C-1), 30.49 (C-2), 78.99 (C-3), 40.62 (C-4), 47.23 (C-5), 21.26 (C-6), 28.54 (C-7), 48.14 (C-8), 20.10 (C-9), 26.58 (C-10), 26.15 (C-11), 35.68 (C-12), 45.42 (C-13), 50.59 (C-14), 33.03 (C-15), 26.68 (C-16), 52.44 (C-17), 18.22 (C-18), 30.05 (C-19), 36.52 (C-20), 18.57 (C-21), 33.66 (C-22), 28.68 (C-23), 79.77 (C-24), 73.36 (C-25), 23.36 (C-26), 26.72 (C-27), 19.44 (C-28), 14.15 (C-29), 25.57 (C-30).Amorphous white powder, MW (g/mol): 460.39; 9.9)7α-hydroxystigmasterol : δH 3.58 , 2.33 , 2.28 , 5.64 , 3.85 , 0.68 , 0.99 , 0.93 , 5.16 , 0.81 , 0.83 , 0.83 , 13C NMR data : 37.13 (C-1), 31.50 (C-2), 71.49 (C-3), 42.14 (C-4), 146.39 (C-5), 124.00 (C-6), 65.51 (C-7), 37.64 (C-8), 42.38 (C-9), 36.24 (C-10), 20.84 (C-11), 39.29 (C-12), 42.27 (C-13), 49.55 (C-14), 24.45 (C-15), 28.43 (C-16), 55.82 (C-17), 11.78 (C-18), 18.39 (C-19), 39.92 (C-20), 19.15 (C-21), 138.39 (C-22), 128.82 (C-23), 51.36 (C-24), 32.03 (C-25), 18.94 (C-26), 19.95 (C-27), 25.99 (C-28), 12.13 (C-29).Needle white crystal, MW (g/mol): 428.70; 1) was identified as C11H16O3. Based on 1H NMR, two methylic protons at δH 1.49 (H-9) and 1.29 (H-10) and one olefinic proton at δH 5.72 (H-7) were detected. The 13C NMR spectral data demonstrates the existence of a carbonyl group on the lactone ring (δc 182.4), a secondary hydroxyl group (δC 66.8), a trisubstituted olefinic bond (δC 112.9 and 171.9), and three singlet methyl groups  which has resemblance with the data from the literature. This structure was identified as loliolide [Compound , 0.95 (H-24), 1.05 (H-25), 0.96 (H-27), 1.06 (H-28), 0.98 (H-29), and 1.00 (H-30) revealed eight tertiary methyl groups, and the double doublet signal at δH 3.23 (H-3) refers to the axially oriented hydroxyl group. Based on the 13C NMR and DEPT data, eight CH3 carbons, eleven CH2 carbons, and eight quaternary carbons are revealed. The 13C NMR spectrum also indicates the presence of the double bond correlated to the olefinic carbons at δC 135.19 (C-8) and 133.63 (C-9).Compound terature . The 1H 4 was isolated with the molecular formula of C30H50O2 and identified as cycloart-25-en-3β,24-diol according to its 1H NMR and 13C NMR data, compared with published data [1H- NMR signals in the up-field area (δH 0.33 and 0.55) revealed a pair of doublets, which is characteristic of a cyclopropane ring. Also, it corroborated one secondary methyl group at 0.88 and five singlet methyls at δH 1.72 (H-27), 0.96 , 0.88 (H-28), and 0.80 (H-29). A double doublet carbinolic proton axially orientated at δH 3.28, assigned to the hydroxyl group as 3β-OH, is also determined from the 1H NMR data analysis. Compound hed data . The 1H-5 was found to be C29H50O, known as β-sitosterol. The 1H NMR data confirm the resemblance to Δ5-3β-hydroxy sterols by the presence of multiplets at δH 3.52, a characteristic peak for H-3α, and δH 5.36, determining the presence of the olefinic proton. The 1H NMR peaks at δH 1.01 (H-19) and 0.68 (H-18) reveal two singlet methyls, and the ones at δH 0.92 (H-21), 0.81 (H-26), and 0.92 (H-27) shows three secondary methyls and one methyl with a triplet signal at δH 0.86 (H-29). The 13C NMR spectrum shows two signals correlated with the olefinic carbons at δC 140.88 (C-5) and 121.88 (C-6), revealing the presence of the double bond. The signal at δC 71.95 suggests an oxymethine on C3, and the ones at δC 12.00 and 19.55 confirm the two methyl groups present at C18 and C19. The published spectroscopic data for β-sitosterol are all in agreement with these findings [Compound findings ,21,22.6 showed the molecular formula of C30H50O2 and was determined as cycloart-23-ene-3β,25-diol by comparison of the spectral data to those in the published literature [1H NMR spectra, one oxymethine proton at δH 3.28 (H-3), five singlet methyls at 1.72 (H-27), 0.88 (H-28), 0.80 (H-29), 0.96  and one further secondary doublet methyl at 0.88 (H-21) were detected. The pair of doublet signals at δH 0.55 and 0.33 in the up-field area are related to the cyclopropane ring in this structure.Compound terature ,25. Base7 assigned the molecular formula of C30H50O2 and, based on the comparison with those reported in the literature, was determined to be 3α, 11α- dihydroxyurs-12-ene [1H NMR spectrum singly supports the existence of two hydroxyl groups at δH 3.21 (H-3) and 3.98 (H-11). The peak at δH 3.21 (H-3) and the 13C NMR peak at δC 76.0 (C-3) revealed the presence of a 3α-hydroxyl group that was supported by the coupling constants. It also explains the strengthened α-effect at δC 76.0 (C-3) and the diminished β-effect at δC 25.4 (C-2) and 37.3 (C-4), comparing to the data from structures with axial orientation. Analyzing the 13C NMR spectra suggested the presence of a urs-12-ene by the olefinic carbons at δC 128.7 and 142.9. The olefinic signal at δH 4.90 from 1H NMR was assigned to the H-12. The urs-12-ene skeleton is determined from the 1H NMR signals of eight methyl groups, two of which are on a methine group and six are on quaternary carbons. Compound s-12-ene . The 1H 8 showed the molecular formula of C30H52O3, and, based on the data and comparison of the spectral values with the literature, it was elucidated as 3β, 24β, and 25-trihydroxycycloartane [1H NMR peaks at δH 1.16 (H-26), 1.22 (H-27), 0.89 (H-28), 0.96 , and 0.80 (H-30) revealed six tertiary methyl groups and the peak at δH 0.88 (H-21), a secondary methyl group. In the up-field area, the δH 0.33 and 0.55 peaks appeared as a pair of doublets, suggesting the existence of the cycloartane cyclopropane structure. Two signals at δH 3.27 (H-3) and 3.31 (H-24) reveal the presence of carbinolic protons related to the hydroxyl groups attached. The two carbinolic carbon shifts at δC 78.99 (C-3) and 79.77 (C-24) revealed the presence of two secondary hydroxyl groups and a tertiary hydroxyl group from a hydroxylated quaternary carbon at δC 73.36 (C-25).Compound loartane . The 1H 9 showed the molecular formula of C29H48O2. From the comparison of the spectroscopic data with other similar compounds published in the literature, it was determined to be 7α-hydroxystigmasterol [1H NMR showed three secondary methyls at δH 0.93 (H-21), 0.81 (H-26), and 0.93 (H-27), two singlet methyls at δH 0.68 (H-18), and 0.99 (H-19), and one triplet signal at δH 0.83 (H-29) indicating a tertiary methyl. The olefinic proton at δH 5.64 (H-6) in down-fields correlated to the double bond between C5 and C6. Another doublet peak at δH 5.16 (H-22) in the same area suggests another double bond between C22 and C23. The shifted signal at δC 146.39 (C-5) from the 13C NMR spectra is a reference to the α-orientation of the hydroxyl group on C27.Compound masterol ,29. The A great deal of ethnopharmacological and ethnomedicinal uses of natural terpenes from different plant resources are related to the presence of isoprenoid units. Recent scientific findings have demonstrated the important role of triterpenes in these therapeutic effects, especially anti-tumor, anti-inflammatory, and immunoregulatory aspects .Euphorbia species has motivated researchers to carry out further investigations on the biological and pharmacological potentials of these species. As a result, many therapeutic effects are introduced for them together with Euphorbia turcomanica, which is proven to have anti-cancer and immunomodulatory functions [The rich terpenoid profile of the Iranian unctions . On the E. turcomanica is not only remarkable, but also there is a vast variety of them with different subclasses, some of which are even considered to be scarce. The present study exhibits that the triterpenoid profile of Euphorbia species, being involved in the biosynthesis of sterols and, therefore, could be considered one of the distinctive chemotaxonomic biomarkers in this genus [4, 6, 8) are elucidated for the first time from E. turcomanica. Compounds 4 and 6 have been isolated from many members of Euphorbiaceae so far studied. They are also found simultaneously in species such as E. schimperi [E. altotibetic [E. segetalis and have shown antiviral activities against the African swine fever virus and Herpes simplex virus [4 has shown notable cancer chemopreventive effects [6, which has also been isolated from E. spinidens [Cycloartanes are triterpenoids identified by their tetracyclic skeletal framework, cyclopropane ring, and side chain. This class of triterpenoids is responsible for a range of biological activities and is considered one of the leading chemical compounds in is genus . Three cchimperi  and E. aotibetic . Also, bex virus . Compoun effects , and compinidens , is connpinidens ,36.8 is a less common cycloartane in comparison, but still, it has been reported from E. marschalliana [E.denticulata, which has shown potential cytotoxic activities [Compound halliana  and E.detivities .3, identified as isomultiflorenol, is another triterpene isolated from E. turcomanica during the present research, which has already been elucidated from other Euphorbia plants such as E. pubescens [E. supina [Compound ubescens  and E. s. supina . During . supina , which mEuphorbia species. Compound 7 is a known pentacyclic ursane-type triterpene that was previously isolated from other plant families [Although the biological effects from the triterpene-containing extracts are mostly associated with cycloartanes, there are also other types of triterpenes identified during this research that are noteworthy due to their scarcity in families , but to 2 is another compound isolated from E. turcomanica during the present research. It is a hopane-type triterpene which is considered one of the rare classes of triterpenes. The uncommon presence of a double bond between C-5 and C-6 causes the rare structural characterization of this compound. This compound has also been found in other Euphorbia species, including E. lathyris [E. aphylla [E. peplus [E. neriifolia and Cissampelos mucronate containing simiarenol as one of the present compounds in their extracts. Also, one recent research has shown this plant’s probable efficacy in treating a range of infectious diseases such as COVID-19, which might be related to this natural constituent but further investigations should be carried out to determine these claims and specifically relates them to simiarenol [Compound lathyris , and E.  aphylla . Moreove. peplus . Severalmiarenol ,44.5, 9) were also elucidated during the current research. Compound 5 is a commonly occurring phytosterol called β-sitosterol, which is a key component present in plants. Out of the many pharmacological and biological effects that have been reported for this compound over the past decades, anti-inflammatory, angiogenic, anti-adipogenic, anti-diabetic, anti-oxidant, anti-cancer, immunomodulatory activities and a role in cardiovascular diseases prevention can be mentioned, reported from recent findings that can be beneficial in drug developments and other therapeutic applications [E. hirta and its role in curing obesity have been linked to this natural compound [E. thymifolia [E. segetalis in the presence of other antimicrobial triterpenes [9) has not yet been reported from any of the Euphorbia species. Although this oxidized sterol has been isolated from other plant species [Two steroids  is also isolated during this study and identified as loliolide, which is a monoterpenoid lactone with 11 carbons and one of the degradation products of carotenoids that have been isolated from several higher plants and marine algae. Studies have revealed anti-bacterial and anti-fungal properties [Euphorbia genus, such as E. supina [E. micractina [E. cooperi [E. alatavica [E. turcomanica before.One monoterpene (operties  as well operties ,55, antioperties ,57, poteoperties , and neuoperties ,60. Manyoperties ,65,66,67. supina , E. micrcractina , E. coop cooperi , and E. latavica  but has E. turcomanica for the first time, including an iridoid monoterpene, as well as steroids and triterpenes. The results suggest that this plant comprises a variety of compounds that can be considered a potential source of natural bioactive constituents with applications in medical and pharmacological sciences. Further investigation should be done for the isolation and identification of other phytochemical compounds in the Euphorbia genus, and it is worth exploring their prospective applications.In the present study, nine known terpenoids were isolated from


Phaseolus vulgaris L. cvs. ICA-Cerinza and Uribe Rosado, tolerant and susceptible to anthracnose, respectively). Indanoyl derivatives  were obtained from 1-oxo-indane-4-carboxylic acid. In general, the accumulation of isoflavonoid-type phytoalexins, such as isoflavones , isoflavanones , isoflavan (phaseollinisoflavan), coumestrol, and pterocarpans (phaseollidin and phaseollin), was dependent on the common bean cultivar, the post-induction time, and the elicitor structure. Isoflavones, dalbergioidin, and coumestrol reached their highest amounts during the first 48 to 72 h, whereas kievitone, phaseollinisoflavano, and the pterocarpans reached maximum levels between 72 and 96 h. The 1-oxo-indanoyl-L-isoleucine methyl ester elicited the highest levels of phytoalexins (similar to those elicited by the methyl jasmonate) and showed no significant phytotoxic effects on common bean seedlings. The indanoyl-type synthetic elicitor, 1-oxo-indanoyl-L-isoleucine methyl ester, may represent a promising agronomic alternative for disease control in common bean by enhancing the accumulation of antimicrobial isoflavonoid phytoalexins.Eleven indanoyl derivatives were synthesized and, along with methyl jasmonate, evaluated as isoflavonoid-phytoalexin elicitors in two cultivars of common bean ( Plants often respond to microbial infection by producing antimicrobial, low-molecular-weight secondary metabolites known as phytoalexins. These compounds also accumulate in plants in response to various agents called elicitors, including substances of biotic (from pathogens or the same plant) and abiotic origin, such as synthetic compounds. The exogenous application of elicitors allows for an increase the phytoalexin concentration and, consequently, enhances the resistance of plants to fungal infections . TherefoPseudomonas syringae that mimics the plant hormone jasmonoyl-L-isoleucine, together with some structurally-related compounds (such as indanoyl-amino acid conjugates) elicits secondary metabolite accumulation in both plant tissues and cell cultures N,N′-dicyP. vulgaris L.), cultivars ICA Cerinza and Uribe Rosado, were purchased from Semicol Ltd.a.  and Semillas & Semillas Ltd.a. , respectively. ICA Cerinza is tolerant to anthracnose (C. lindemuthianum Sacc.), rust (Uromyces phaseoli), oidium (Erysiphe potygoni DC), and Fusarium root rot (Fusarium sp.)  were dissolved in DCM at room temperature and prepared according to the process described in +: 304.1543), 304.1539  and 290.1385 , respectively. Spectroscopic data for (7), (8), and indanoyl amides are presented as follows: 1-oxo-indanoyl-glycyl methyl ester (7) (Yield: 19%). M. p. = 105 °C. 1H NMR : 2.73 , 3.45 , 3.84 , 4.3 , 6.79 (NH), 7.48 , 7.89 . 13C NMR : 26.1 (C3), 36.2 (C2), 41.6 (C2′), 52.65 (–OCH3), 126.6 (C7), 127.7 (C6), 132.5 (C5), 132.7 (C4), 138.3 (C7a), 154.4 (C3a), 167.1 (–CONH), 170.4 (–COO–), 206.5 (C1). MS (ESI) m/z: calcd for C13H13NO4 [M + H]+: 248.0917. Found: 248.0916. 6-bromo-1-oxo-indanoyl-L-isoleucyl methyl ester (8) (Yield: 59%). 1H NMR : 1.05 , 1.32 , 1.55 , 2.08 , 2.78 , 3.39 , 3.85 , 4.86 , 6.65 (NH), 7.98 , 8.02 . 13C NMR : 11.7 (C5′), 15.6 (C6′), 25.4 (C3), 25.8 (C4′), 36.3 (C2), 38.2 (C3′), 52.5 (–OCH3), 56.9 (C2′), 121.7 (C6), 129.3 (C7), 134.6 (C4), 135.5 (C5), 140.1 (C7a), 152.4 (C3a), 165.3 (–CONH), 172.4 (–COO), 204.81 (C1). N-(3-(dimethylamine)propyl)-1-oxo-indane-4-carboxamide (9) (Yield: 22%). Amorphous solid. M. p. = 218 °C. TLC: Rf ; 0.29 (red spot with Dragendorff’s reagent). 1H NMR : 1.93 , 2.42 , 2.67 , 2.75 , 3.52 , 3.62 , 7.48 , 7.88 , 8.39 (–NH–). 13C NMR : 26.3 (C2), 34.0 (C2′), 36.3 (C3), 39.8 (C1′), 45.0 , 58.6 (C3′), 126.1 (C7), 127.6 (C6), 132.4 (C5), 133.3 (C4), 138.3 (C7a), 154.7 (C3a), 167.0 (–NHCO–), 207.0 (C1). MS (ESI) m/z: calcd for C15H20N2O2 [M + H]+: 261.1597. Found: 261.1594. N-(3-(diethylamine)propyl)-1-oxo-indane-4-carboxamide (10) (Yield: 35%). Colorless oil. TLC: Rf ; 0.37 (red spot with Dragendorff’s reagent). 1H NMR : 1.09 , 1.84 , 2.62–2.66 , 2.69 , 3.32 , 3.37 , 3.44 , 7.52 , 7.86 . 13C NMR : 9.7 , 25.2 , 25.5 (C3), 37.9 (C1′), 46.3 , 49.9 (C2), 125.3 (C7), 127.4 (C6), 132.7 (C5), 133.9 (C4), 137.7 (C7a), 154.3 (C3a), 168.6 (–NHCO–), 207.7 (C1). MS (ESI) m/z: calcd for C17H24N2O2 [M + H]+: 289.1910. Found: 289.19081-oxo-indane-4-carboxylic acid (ribed in . The res2) according to the process described in +: 306.1699. Found: 306.1696.1-oxo-indanoyl-L-isoleucyl methyl ester (rding to . 1-hydro4); the mixture was stirred manually for 1 min, after which 1.5 equivalents of 3-bromobenzaldehyde were added. The reaction was stirred periodically at room temperature for 12 h. The product was obtained as an insoluble solid; it was recrystallized with n-hex-DCM-methanol mixture, 2:1:1, v/v and filtered. 2-(3′-bromobenzylidene)-1-oxo-indanoyl-L-isoleucyl methyl ester (13) (Yield: 33%). Light green solid. M.p. = 198 °C. TLC: Rf ; 0.46 (yellow spot with oleum). 1H NMR : 1.05 , 1.10 , 1.98 , 3.51 , 3.89 , 4.36 , 4.92 , 6.73 , 7.41 , 7.56 , 7.61 , 7.64 , 7.71 , 7.84 , 7.95 , 8.07 . 13C NMR : 11.7 (C5′), 15.7 (C6′), 33.9 (C3), 35.6 (C4′), 49.2 (C3′), 53.0 (–OCH3), 56.9 (C2′), 123.1 (C3″), 127.3 (C6), 128.2 (C6″), 128.9 (C5″), 130.5 (C7), 132.3 (C4), 132.5 (C4″), 132.7 (C2″), 133.1 (Ca), 133.8 (C5), 135.4 (C4), 137.2 (C1″), 148.9 (C7a), 156.8 (C3a), 166.6 (–NHCO–), 172.5 (–COO–), 193.5 (C1). MS (ESI) m/z: calcd for C24H24BrNO4 [M + H]+: 470.0961. Found: 470.0964.A few drops of piperidine were added to (4) dissolved in ethanol. The reaction was stirred at reflux for 24 h. The resulting product was subjected to two stages of purification by column chromatography, first using a Sephadex LH-20  and then using silica gel (mobile phase: chloroform). 1-benzylhydrazono-indanoyl-L-isoleucyl methyl ester (14) (Yield: 37%). White solid. M. p. = 65 °C. TLC: Rf ; 0.39 (yellow spot with oleum). 1H NMR : 1.05 , 1.31–1.39 , 1.5–1.61 , 2.00–2.05 , 3.03–3.07 , 3.32 , 3.36 , 3.78 (–OCH3), 4.58 , 7.43 , 7.48–7.56 , 7.60 , 7.68 , 7.91 , 8.14 . 13C NMR : 10.2 (C5′), 14.7 (C6′), 25.3 (C4′), 27.0 (C2), 28.1 (C3), 36.7 (C3′), 48.5 (Ca), 51.2 (–OCH3), 57.4 (C2′), 124.8 (C7), 127.2 (C6), 127.5 , 128.3 , 129.7 (C5), 131.8 (C4″), 132.5 (C4), 133.1 (C3a), 138.6 (C7a), 147.9 (C1″), 164.7 (C1), 169.6 (–NHCO–), 172.4 (–COO–).Benzylhydrazine  was added to (v/v) was used to improve the solubility of each compound in water. As a negative control, common bean seedlings were submerged in a hydroalcoholic solution (0.05%). Plant material was then transferred to sterilized plastic containers and sown on moist cellucotton in the dark at 25 °C for 72 h. Subsequently, cotyledons, epicotyls, and hypocotyls-roots were carefully separated. The cotyledons and epicotyls were discarded, and the hypocotyls-roots (5.0 g) were used for elicitation analysis. All experiments were tested three times.Common bean seedlings (average height of 4 to 6 cm) cvs. ICA Cerinza and Uribe Rosado were immersed for 3 h in 1.0 mM of the 1-oxo-indanoyl derivatives (4 to 14) or 1.0 mM MeJA (positive control). A low concentration of ethanol  cvs. ICA Cerinza and Uribe Rosado were immersed for 3 h in hydroalcoholic solution at 1.00 mM of the elicitor that generated the highest phytoalexin accumulation. In plastic containers, the plant material was sown in moist cellucotton and stored for 24, 48, 72, and 96 h (post-induction times) at room temperature in the dark. Hypocotyls-roots (5 g) were separated and analyzed after extraction [Hypocotyl-root samples were processed according to . A chromv/v) were used as positive and negative control, respectively, for comparison purposes. Tests were carried out on the cultivar Uribe Rosado, analyzing seed germination, cotyledon hardness, and seedling and root growth after regular and periodic spraying with the elicitor or ethanol . Eight seeds were used in each plate, and tests were performed in quintuplicate.The phytotoxic effect on common bean of the elicitor that generated the highest phytoalexin accumulation was evaluated at a concentration of 1.0 mM. In addition, MeJA (1.0 mM) and ethanol  with 5-day growth were generously sprayed with the treatments . This process was repeated every two days for one week. On the non-treatment days, seedlings were sprayed with water to avoid wilting. On the seventh day, seedlings were removed from the vermiculite; root, seedling, stem, and leaf size, along with the number of secondary roots and leaf coloration, were measured. Seedlings were exposed to diffused light at room temperature and 80% relative humidity. Chlorophyll content: fresh leaves (200 mg) were extracted with an aqueous solution of ethanol (80%) for 20 min in a water bath at 80 °C. The mixture was then filtered, and the filtrate was adjusted to a volume of 5 mL. The absorbance of the solution was measured at 654 nm in a spectrophotometer, and the chlorophyll content was calculated by taking into account the extinction coefficient of chlorophyll. The results were expressed as mg/g fresh weight .p ≤ 0.05.Values are expressed as mean ± standard deviation (s.d.). Data within the groups were analyzed using one-way analysis of variance (ANOVA), followed by Fischer’s least significant differences (LSD) test at The accumulation of isoflavonoid phytoalexins in common bean tissues was found to depend on the time after induction and the structure of the elicitor. During the first 48 h, the biosynthetic precursors, isoflavones and isoflavanones, were the main phytoalexins; then, coumestrol and pterocarpan-type phytoalexins increased, reaching maximum levels. A metabolic blockage in the biosynthetic pathway in the 5-hydroxyisoflavonoid biosynthetic pathway could be suggested. Analysis of a series of eleven indanoyl derivatives revealed that some structural requirements are necessary for the elicitor effect. Variations in the C-4 or C-6 position of the indanoyl system and the carbonyl group of the oxo-cyclopentyl system resulted in significant changes in activity. The compound 1-oxo-indanoyl-L-isoleucine methyl ester is a promising elicitor, stimulating the synthesis and accumulation of phytoalexins in common bean tissues without significant phytotoxic effects on seeds and seedlings.


The structure of one complex was established by X-ray diffraction analysis. The complexation led to a slight distortion of the cyclophane conformations.In this study, 28-membered macrocyclic 1,5-di-2,4,6,8-tetrabenzenacyclooctaphane were synthesized by condensation of pyridinephosphine, paraformaldehyde, and primary diamines (bis(4-aminophenyl)methane or -sulfide. The first representatives of binuclear copper(I) complexes of P,N-containing cyclophanes with two 1,5-diaza-3,7-diphosphacyclooctane rings incorporated into a macrocyclic core and intracavity location of unusual, developed angle Cu A distinctive feature of phosphorus-containing cyclophanes is the combination of two binding sites with different features. The first site is a phosphorus atom that is able to form rather stable coordination bonds with various soft transition metal ions in a low oxidation state. The second site is a hydrophobic intramolecular cavity bounded by phenylene fragments which is suitable for host–guest interactions with small organic molecules or for encapsulation of a metal-containing fragment connected with a phosphorus atom. This explains why phosphorus-containing cyclophanes could be regarded to be a promising scaffold for tailor-made transition metal complexes which may be of interest both for homogeneous catalysis and for the creation of novel recognition systems and molecular materials ,7,8,9,102-pincer macrocyclic ligands were useful synthons for the effective synthesis of dihydrogen, ethylene, and carbonyl derivatives of rhodium(I) . . 1 and 21 and 2 were moderate  and noticeably lower than the yields of analogous P-aryl substituted macrocycles (18–60%) + along with additional peaks [LCu2 + Na]+ (for both 3 and 4) and [LCu2 + 2Na]+ and [LCu2 + O + (CH3)2SO]+ (for 4). The composition of the main peaks was additionally confirmed by isotope distribution analysis  and −27.7 (4) ppm. The signals of both complexes in the 31P NMR spectra were shifted to the low field by ca. 16 ppm according to the signals of free ligands. The chemical shifts of products and the Δδ values were in the typical range for P,P-chelate copper(I) complexes of 1,5-diaza-3,7-diphosphacyclooctanes +, 959 [1 + Na]+, 982 [1 + 2Na]+, 998 [1 + Na + K]+. Anal. Calc. for C54H52N8P4 (936.96): C 69.23, H 5.55, N 11.97, P 13.25%. Found: C 69.78; H 5.81; N 11.61; P 12.80%.3,17,53,57-Tetra(pyridine-2-yl)-3,7-dithia-1,5-di-2,4,6,8-tetrabenzenacyclooctaphane1 (2). Cyclophane 2 was obtained similar to 1 from (pyridine-2-yl)phosphine , paraformaldehyde , and bis(4-aminophenyl)sulfide . The yield of 2 was 0.35 g (15%). M.p. 222 °C. 1H NMR (DMSO-d6): δ 4.40 ax), 4.56 eq), 6.73 ), 7.34 ), 7.44 ), 7.74 ), 7.80 ), 8.74 ). 31P{1H} NMR (DMSO-d6): δP −43.96 (s). ESI MS: m/z 973 [2 + H]+, 995 [2 + Na]+, 1018 [2 + 2Na]+. Anal. Calc. for C52H48N8P4S2 (973.03): C 64.20, H 4.94, N 11.52, P 12.76, S 6.58%. Found: C 64.67; H 5.17; N 11.38; P 12.30, S 6.28%.3,17,53,57-Tetra(pyridine-2-yl)-1,5-di-2,4,6,8-tetrabenzenacyclooctaphane]-μ-iododicopper(I)}iodide{+, 1191 [3–I]+. Anal. Calc. for C54H52N8P4Cu2I2 (1317.84): C 49.22, H 3.98, N 8.50, P 9.40%. Found: C 48.77; H 3.49; N 8.38; P 9.30%.1H NMR (DMSO-d6): δ 3.58 ), 4.68 A), 4.81 B), 6.73 ), 7.13 ), 7.50–7.55 ), 7.64–7.71 ), 7.90 ), 8.84 ). 31P{1H} NMR (DMSO-d6): δP −29.42 (s).3,17,53,57-Tetra(pyridine-2-yl)-3,7-dithia-1,5-di-2,4,6,8-tetrabenzenacyclooctaphane}bis(copper(I)iodide){1 (4). Complex 4 was obtained similar to 3 from 2  and copper(I) iodide . The yield of 4 was 0.034 g (48%). ESI MS: m/z 1126 [4–2I + Na + H]+, 1147 [4–2I + 2Na + H]+, 1191 [4–2I + O+(CH3)2SO]+, 1227 [4–I]+. Anal. Calc. for C52H48N8P4S2Cu2I2 (1353.92): C 46.13; H 3.57; N 8.28; P 9.15; S 4.74%. Found: C 46.57; H 3.19; N 8.39; P 9.28, S 4.28%. 1H NMR (DMSO-d6): δ 4.63 A), 4.74 B), 6.84 ), 7.48 , partially overlapped by H(9)), 7.49 ), 7.80–7.85 ), 7.89 ), 8.80 ). 31P{1H} NMR (DMSO-d6): δP −27.70 (s). The row of 28-membered cyclophanes containing two 1,5-diaza-3,7-diphosphacyclooctane fragments was extended by the synthesis of two new representatives with pyridyl substitutents at the phosphorus atoms. Condensation of pyridine-2-yl-phosphine with paraformaldehyde and (bis(4-aminophenyl)methane or -sulfide resulted in macrocyclic 1,5-di-2,4,6,8-tetrabenzenacyclooctaphane in lower yields of 15–23% than its P-aryl substitiuted analogues. For the first time, it is shown that the reaction of 28-membered cage cyclophanes containing two 1,5-diaza-3,7-diphosphacyclooctane fragments with two molecules of CuI leads to binuclear P,P-chelate complexes with retention, in general, of the cyclophane geometry. An unusual, charged, and almost linear Cu-I-Cu moiety with trigonal copper atoms is situated within the intramolecular cavity of the cyclophane ligands.


Preliminary mechanistic studies suggest that trityl cation dominates the catalysis, while the silyl cation plays a minor role.(Ph This success led us to extend trityl cation catalysis to allylsilane-mediated reactions for which metal-based Lewis acids do not work well. We focused on β,γ-unsaturated α-ketoesters as electrophiles. Typical enones undergo Michael-type allylation ]  is well-Ph(F)4]] , the usePh(F)4]] ,14,15,16Ph(F)4]] ,18, MichPh(F)4]] , Diels–APh(F)4]] ,29,30,31Ph(F)4]] , epoxidePh(F)4]] , ene reaPh(F)4]] ,34 and oPh(F)4]] ,36,37,38Ph(F)4]] ,40,41,42ldehydes . In thislylation ,45,46 orlylation ,49,50,51products a52]. Su. Su3C)[Bproducts b 53]. A. A3C)[BP3C)[BPh(F)4]-catalyzed Hosomi-Sakurai allylation of allylsilanes with β,γ-unsaturated α-ketoesters -4a was obtained in 97% yield; no desired Hosomi-Sakurai allylation product 3a was detected (entry 1). SnCl4, AlCl3 and FeCl3 provided a mixture of 3a and (±)-4a, in which 3a was the minor isomer and the 3a:(±)-4a ratio ranged from 23:77 to 45:55 (entries 2–4). BF3•Et2O and TMSOTf proved to be ineffective catalysts, leading to less than 30% conversion even after 4 days (entries 5 and 6). We also tested lanthanide-based Lewis acids such as Sc(OTf)3 and Yb(OTf)3 (entries 7 and 8). Sc(OTf)3 afforded undesired (±)-4a as the sole detectable product; no reaction occurred using Yb(OTf)3. In sharp contrast to metal-based Lewis acids, the trityl salt (Ph3C)[BPh(F)4] displayed excellent catalytic ability, generating 3a as a single chemoisomer in 97% yield (entry 9). In fact, 1 mol % of (Ph3C)[BPh(F)4] was efficient enough to provide 3a in comparably high yield and selectivity (entry 10).We initially screened the metal-based Lewis acid catalysts using β,γ-unsaturated α-ketoester at 25 °C . In the 3b–3j in excellent yields. The reaction tolerated substrates containing functionalized phenyl rings, naphthyl rings or heterocycles. An electron-donating substitution on the phenyl ring slightly decreased chemoselectivity, as shown in 3e (H-S/D-A = 90:10) and 3f (H-S/D-A = 95:5). The reaction generated 3k and 3l from the corresponding alkyl-substituted ketoesters. Allylation of dienyl ketoester with 2a gave 3m in 80% yield, with 1,4-regioselectivity dominating over 1,6-regioselectivity. β,γ-unsaturated α-ketimine ester performed well in the reaction, giving enamido ester 3n in 75% yield. Interestingly, propargyl α-keto ester underwent 1,2-allylation exclusively, leading to α-tertiary hydroxy ester 3o in 93% yield. Switching the ester group from OMe to a bulkier i-Pr group decreased chemoselectivity .With the optimal reaction conditions in hand, we examined the scope of β,γ-unsaturated α-ketoesters . Reactio2b–2e bearing alkyl or aryl substituents at the 2-position reacted well with 1a, giving 3q–3t in 85–96% yields (3C)[BPh(F)4] also allowed facile anti-SE’ allylation of the bulky 3,3-dimethyl-1-trimethylallylsilanes 2f with 1a (entry 5). However, this catalyst did not efficiently control the diastereoselectivity of allylation: the reaction of Z-crotyltrimethylsilane Z2g- afforded 3v in 96% yield but as a 3:2 (1.0 mol %) at 25 °C. After stirring for 10 min, the reaction was quenched with p-TsOH . The mixture was directly concentrated under reduced pressure. Purification of the crude residue via silica gel flash column chromatography (gradient eluent: 0–2.0% of EtOAc/petroleum ether) afforded γ,γ-disubstituted α-ketoester 3. The characterization data for all synthetic compounds are provided in the To a solution of β,γ-unsaturated α-ketoester 3a): 1H NMR  δ 2.37 , 2.45 , 3.19 , 3.34 , 3.80 , 5.00 , 5.03 , 5.66 , 7.20 , 7.28 , 7.31 ; 13C NMR  δ 40.2, 40.8, 44.9, 52.9, 117.3, 126.7, 127.5, 128.5, 135.8, 143.2, 161.3, 192.9; IR (neat) cm−1 3029, 2954, 2923, 1731, 1442, 1277, 1253, 1089, 919; HRMS  calcd for C14H16O3Na (M+Na)+: 255.0992, found 255.0989.Methyl 2-oxo-4-phenylhept-6-enoate (1 (0.22 mmol) and allylsilane 2 (0.26 mmol) and in anhyd. CH2Cl2 (4 mL) under argon atmosphere was added (Ph3C)[BPh(F)4] (1.0 mol %) at 25 °C. After stirring for 10 min, the reaction was quenched with NEt3 (1.32 mmol). The mixture was directly concentrated under reduced pressure. Purification of the crude residue via silica gel flash column chromatography (gradient eluent: 0–2.0% of EtOAc/petroleum ether) afforded silyl enol ether 5. The characterization data for all synthetic compounds are provided in the To a solution of β,γ-unsaturated α-ketoester Z)-4-phenyl-2-((triethylsilyl)oxy)hepta-2,6-dienoate (5a): 1H NMR  δ 0.79 , 1.05 , 2.56 , 3.82 , 3.99 , 5.05 , 5.11 , 5.78 , 7.30 , 7.38 ; 13C NMR  δ 5.5, 6.8, 40.6, 41.8, 51.9, 116.4, 124.6, 126.4, 127.5, 128.5, 135.0, 140.1, 143.3, 165.3; IR (neat) cm−1 2955, 2878, 1727, 1642, 1439, 1372, 1266, 1232, 1143, 1010, 913; HRMS  calcd for C20H30O3SiNa (M+Na)+: 369.1856, found 369.1856.Methyl (3C)[BPh(F)4]-catalyzed Hosomi-Sakurai allylation of allylsilanes with β,γ-unsaturated α-ketoesters. Various γ,γ-disubstituted α-ketoesters α-ketoesters were synthesized in high yields with excellent chemoselectivity. Mechanistic studies suggest that the trityl cation dominates catalysis, while the silyl cation plays only a minor role. Verification of this mechanism also makes the trityl cation-catalyzed asymmetric reaction possible, which is a challenging task and little progress has been achieved. The related work is ongoing in our group.In summary, we have developed a (Ph


Their structures were elucidated mainly through 1D- and 2D-NMR spectra analysis and ECD comparison. Compound 1 has a 6/5/5/6/6/8 hexacyclic ring system bearing 2,2-dimethyl-1,3-dioxocane, which is rare in natural products. Compound 2 has an unusual open F-ring structure. The cytotoxic activities against 10 cancer cell lines and antimicrobial activities against model bacteria and fungi of all compounds were assayed. No compound showed antimicrobial activity, but at a concentration of 1 μM, compounds 1 and 6 exhibited the highest inhibition rates of 71.2% and 83.4% against JeKo-1 cells and U2OS cells, respectively.Five new indole diterpenoids named paspaline C–D ( The 1H-NMR spectrum , one acetal carbon (C-31), one oxygenated 3sp carbon (C-27), three non-oxygenated 3sp carbons , and eight methine groups, including four olefinic methines (C-20–C-23) and four 3sp methine carbons .Paspaline C  search resulted in many hits, most of which were indole diterpenoids, mainly the derivatives of paspaline (6), which was co-isolated with 1. Detailed comparison of the NMR data between 1 and 6 showed that compound 1 had an additional 2-oxypropyl unit. The observed HMBC correlations from H-9 to C-31 and from H-32 to C-7 established a 2,2-dimethyl-1,3-dioxocane ring (ring F), so the planar structure of compound 1 was solved. It had a 6/5/5/6/6/8 hexacyclic system. The H-H COSY from H-20 to H-23, as well as the HMBC correlations  from H-11 was determined mainly by analysis of the NOESY spectrum  of compound 1 was 3S*, 4S*, 7S*, 9S*, 12S*, 13R*, and 16S*.The relative configuration of spectrum . The NOE(2) had the molecular formula of C30H43NO4 established by the HR-ESI-MS analysis which exhibited the [M + H]+ ion at m/z 504.3081 . Comparison of the 1H- and 13C- NMR spectra of 1 and 2 showed two similar sets of data, the main differences being C-7 and C-31. For compound 1, C-31 (δ106.5 ppm) was an aldehyde acetal carbon, and H-7 was a typical oxygenous chemical shift (δ3.59 ppm), while in compound 2, C-31 (δ170.7 ppm) was a carbonyl carbon, and the chemical shift of H-7 (δ4.80 ppm) shifted downfield obviously. Detailed analysis of 2D-NMR spectra  of compounds 1 and 2 . A negatole ring . Compount 230 nm  and posimpound 6 .3) had a molecular formula of C34H43NO5 based on the HR-ESI-MS analysis which exhibited an [M + H]+ ion at m/z 546.3217 . The 1H- and 13C-NMR spectra of 3  /7.03 /7.01  suggested a 1, 2, 3-trisubstitued phenyl nucleus, indicating an isopentenyl group located at potion 20 or 23 of indole deterpenoid skeleton. The HMBC correlations of H-21 to C-32, CH3-35 and CH3-36 to C-33 and C-34 confirmed a 3-methyl-2-butenyl unit at position 20, so the planar structure of 3 was 20-prenylated -27-O-acetyl paxilline.Paxilline B (tra of 3  are very4) had the same molecular formula C34H43NO5 as compound 3 based on (+)-HR-ESI-MS [M + H] + value 546.3220. The two compounds had highly similar 1H-NMR and 13C-NMR spectra  was obtained as a colorless solid, and its molecular formula was determined as C28H35NO4 based on (+)-HR-ESI-MS data [M + H]+ 450.2638 . It had highly similar 1H-NMR and 13C-NMR spectra with 13-desoxypaxilline (10) , which was consistent with their molecular formulas. HMBC correlations from H3-30 to C-7, as well as the typical aldehyde acetal chemical shift of C-7 (δC 96.5) established the planar structure of 5 as 7-methoxy-13-desoxypaxilline.Paxilline D  (O-acetyl paxilline (7) , 1’, 1’1–5, line (7) , 20-pren (7) 25D: 0 , HR-ESI-MS at m/z 480.3474 [M + H]+, calculated for C31H45NO3, 480.3472. UV (MeOH), λmax (log ε): 284 (1.4), 225 (5.1). IR (KBr) νmax 3430, 2939, 2253, 1651, 1455, 1086, 1049, 1025, 1004, 824, 762, 615. For 1H-NMR and 13C-NMR data, see Paspaline C (2): colorless solid, [α]25D: +34 , HR-ESI-MS at m/z 504.3081 [M + Na]+, calculated for C35H45NO4Na, 504.3084. UV (MeOH), λmax (log ε): 280 (1.2). IR (KBr) νmax 3440, 2974, 2114, 1634, 1456, 1364, 1276, 1160, 1048, 1032, 911, 471. For 1H-NMR  and 13C-NMR  data, see Paspaline D (3): colorless solid, [α]25D: +24 , HR-ESI-MS at m/z 546.3217 [M + H]+, calculated for C34H44NO5, 546.3214. UV. UV (MeOH), λmax (log ε): 285 (1.0), 225 (3.8). IR (KBr) νmax 3430, 2948, 2843, 1658, 1453, 1377, 1179, 1113, 1020, 635. 1H-NMR  and 13C-NMR  data, see Paxilline B (4): colorless solid, [α]25D: +60 , HR-ESI-MS at m/z 546.3220 [M + H]+, calculated for C34H44NO5, 546.3214. UV (MeOH), λmax (log ε): 290 (1.1), 240 (4.3). IR (KBr) νmax 3403, 2982, 2937, 2849, 1709, 1676, 1624, 1449, 1368, 1259, 1124, 1025, 960, 928, 742, 709, 606, 583. For 1H-NMR  and 13C-NMR  data, see Paxilline C (5): colorless solid, [α]25D: 0 , HR-ESI-MS at m/z 450.2638 [M + H]+, calculated for C28H36NO4, 450.2639. UV (MeOH), λmax (log ε): 230 (1.5). IR (KBr) νmax 3400, 2928, 1658, 1448, 1373, 1303, 1258, 1127, 1032, 742. For 1H-NMR  and 13C-NMR  data, see Paxilline D  were used. The spores at 1×106/mL were poured onto culture media on a Petri dish (φ = 9 cm): PDA medium for fungal indicators, and Luria–Bertain (LB) medium for bacterial indicators. The sterilized filter paper (φ = 5 cm) with 5 µL of compound solution at 10 mg/mL was put on the surface of the culture medium; 24–48 h later, the inhibition diameters were measured. Amphotricin B and gentamicin at 1 mg/mL were used as positive controls for the fungal and bacterial indicators, respectively. The antimicrobial activity was evaluated with the disk diffusion methodology . Four in® AQueous Assay Reagents according to the instruction manual at 490 nm. Data were normalized to the control group (DMSO).A375, MDA-MB-231, HepG2, BGC823, U2OS and MCF-7 cell lines were grown in DMEM medium. SKGT4, Kyse450, JeKo-1 and HL60 cells were grown in RPMI1640 medium. Two media were supplemented with 10% FBS and 1% penicillin or streptomycin. Then, 198 µL of the medium with about 5000 cancer cells was plated in each well of a 96-well plate. Then, 2 µL of the test compound at 10 mM in DMSO was added in triplicate. Forty-eight hours later, the cell viability was determined using CellTiter 961–5) and 11 known congeners (6–16) were identified from sea-mud-derived fungal Penicillium Brefeldianum WZW-F-69. Even though the main structural elements resemble the reported indole diterpenes, paspaline C (1) has a rare 2,2-dimethyl-1,3-dioxocane ring F, while paspaline D (2) has an unusual open ring F structure. Paxilline B (3) and paxilline C (4) are isopentenyl ring A derivatives of paxilline, while paxilline D (5) has an aldehyde acetyl carbon (C-7), which makes them chemically unique. Although the AC of new compounds was established mainly by the comparison of ECD spectra with known compounds 6 and 7, the consistence with known compounds, such as 6-hydroxylpaspslinine [1 and 6 exhibited cytotoxic activity against several cancer cell lines. In summary, the identification of diverse structural compounds from WZW-F-69 illustrates the huge potential of finding more new bioactive lead compounds from marine fungi for future research.In summary, 5 new indole diterpenoids (carbon C-, which m


According to the results of an X-ray diffraction study, some of the anions and solvate molecules were disordered. The cations (n-Bu4N)+ and anions [2-B10Br9SR2]− were associated via C-H…Br and H…H contacts. In addition, Br…Br interactions between anions were revealed. The role of these contacts was analysed in terms of Hirshfeld surface analysis, QTAIM theory and the NCI method using quantum chemical calculations. An increase in the size of the alkyl R moiety led to significant strengthening of the total energy of H…H interactions. In the case of R = -n-C18H37, a parallel mutual orientation of alkyl moieties was established that was similar to the packing of salts of fatty acids. The nature of C-H…Br and Br…Br interionic interactions was found to be attractive, in contrast to the repulsive nature of intermolecular Br…Br interactions.A new series of compounds based on perbrominated disubstituted sulfonium derivatives of the  Non-covalent interactions continue to receive significant attention, due to their crucial role in various fields of chemistry, such as the stabilisation of metal complexes with various types of ligands, or the interaction of halogens with Lewis acids, which have applications in crystal engineering ,2, organAn example of a context where non-covalent interactions play an important role is provided by compounds that are based on boron cluster anions. In this case, various types of weak interactions are formed, depending on the type of compound used ,6,7,8. Dcloso-borate anions, and only a few to perbrominated anions. The most extensive studies of the properties of this class of compounds are devoted to the study of perhalogenated alkoxy-   is most conveniently carried out using elemental bromine in acetonitrile . After complete pouring of the reagent solution  to the mixture, the solution is slowly warmed to room temperature and after 24 h the only final reaction product is a completely substituted derivative.onitrile . Before 11B NMR spectra of the final compounds practically did not differ in their patterns from their perchlorinated counterparts [2-B10Br9SR2] , melting points were examined and found to be virtually identical to their perchlorinated analogues. Compounds with short hydrocarbon chains (up to n-Bu) had a melting point above 200 °C, while for a compound with –n-C8H17 substituents it was 127–128 °C; for –n-C12H25 it was 98–99 °C and for –n-C18H37 it was 60–62 °C.X-ray diffraction analysis and Hirschfeld surface analysis. Structure of anions (n-Bu4N)[2–B10Br9S(n-Pr)2] (1), (n-Bu4N)[2–B10Br9S(i-Pr)2] (2), (n-Bu4N)[2–B10Br9S(n-Bu)2] (3), (n-Bu4N)[2–B10Br9S(n-C12H25)2] (5), and (n-Bu4N)[2–B10Br9S(n-C18H37)2] (6) are shown in 1/c) salt 5 and triclinic cells (P-1) of salts 1, 2, 3, and 6 contain one cation and one anion each. The cell of the last salt also includes a highly disordered DMSO molecule that cannot be modelled. Hence, an integral contribution of the strongly disordered DMSO molecule was excluded from total electron density; it was removed from the electron density map using the solvent masking implemented in the OLEX2 program. Exopolyhedral iso-propyl groups of the [2–B10Br9S(i-Pr)2]− anion were disordered and rotated relative to one another, so that the C1C2C3 and C4C5C6 planes formed angles of 61.2 and 66.0°.The crystallographically independent parts of the monoclinic unit cell . The B-te anion ,28,29. Tte anion ,28. The 1 (n-Bu4N)[2–B10Cl9S(n-Pr)2]   and compPr)2]  to (n-Bu4N)[2–B10Br9S(n-C18H37)2].The energies of separate interatomic interactions estimated from the QTAIM study can be used to calculate the interactions between charged molecular fragments (cations and anions). These calculated energy values are presented in n-Bu4N)[2–B10Br9S(n-C12H25)2] was 9.8 kJ/mol. In addition to intermolecular interactions, some intramolecular H…H and Br…H interactions were revealed. The energy of the strongest interactions of this type was up to 2.4 kcal/mol, which exceeded the values for analogous intermolecular interactions.In addition to H…H interactions, Br…H and Br…Br interactions were responsible for anion–anion and cation–anion bonding. Most Br…H interactions are weak (less than 5 kJ/mol), and they are mostly responsible for cation–anion bonding. Conversely, Br…Br interactions play a noticeable role in the association between anions. In most cases, the energy of Br…Br does not exceed 5 kJ/mol, but in salts studied in the current work there were several interactions that were considerably stronger. For example, the energy of the Br6…Br6 interaction in 2] was 80 kJ/mol.The presence of sufficiently bulky alkyl substituents near sulphur atoms led to the formation of numerous intermolecular interactions that were both intermolecular and intramolecular. Most of these were of the Br…H type. In addition, in the case of -C2) ρ(r) function. The latter is the product of multiplications of the values of λ2 eigenvalues of the Hessian matrix (it can be positive or negative) to the values of ρ(r) at a given point. With the NCI method, any non-covalent interaction is defined as the region between atoms, in contrast to the QTAIM definition, where bcp is indicative of interaction. Attractive interactions are characterised by negative values of λ2, while repulsive interactions are described by positive λ2.To visualise and describe the nature of the interactions in a comprehensive manner, the features of electron density distribution were analysed using the NCI method . Similar2 < 0. The isosurfaces of the RDG function related to intermolecular Br…H interactions can be described as being ‘attractive’. In addition, the NCI study revealed intermolecular Br…Br interactions between adjacent bromine atoms that can be described as being mostly repulsive (λ2 > 0).Thus, the analysis of interatomic interactions using the NCI method indicated the attractive character of Br…Br interactions between borane moieties. One of the strongest Br…Br interactions is shown in . It can nHn]2-  with elemental chlorine, which requires a significant excess of the reagent and a temperature above 100 °C [closo-decaborate anions [2-B10H9SR2]− at a threefold excess of bromine at room temperature for 24 h. If a smaller amount of elemental bromine is used, a mixture of fully and partially halogenated substituted derivatives is observed in the reaction mixture over the same time period. With an elemental bromine:boron cluster ratio of 2:1, the halogenation process can also be completed, but it is necessary to increase the reaction time to 3 days or more.In contrast to chlorination of boron cluster anions − [closo-dodecaborate anion [B12Cl11OR]− and [B12Br11OR]− [Investigation of the melting points of perchlorinated  and perbH11NR3]− ,36 and sBr11OR]− . This maAt first glance, NCI analysis of intermolecular interactions in the obtained compounds shows that the values of the structural properties of anions  increase, but from a thermodynamic point of view, it seems that the melting temperature values are determined by the molecular flexibility of alkyl substituents in anions, which can be described as conformational entropy.n-Bu4N)[2-B10H9SR2]  in the range 4000–400 cm–1 at a resolution of 1 cm–1. Samples were prepared as suspensions in CCl4.111B and 13C NMR spectraH,  of samples dissolved in DMF-d7 were recorded on a Bruker Avance II 300 spectrometer operating at a frequency of 300.3, 96.32 and 75.49 MHz, using an internal deuterium lock. Tetramethylsilane and boron trifluoride etherate were used as external references. The spectra of the compounds obtained are presented in Melting point was measured on a REACH Devices RD-MP in the temperature range of 25−250 °C with a heating interval of 1.5, 3, 6 and 12 °C/min.The X-ray diffraction. Single crystals were obtained by crystallisation from dimethylsulfoxide solution. X-ray diffraction data were collected with a Bruker SMART APEX II diffractometer using graphite monochromated Mo Kα radiation with multilayer optics. The structures were solved using the direct method and refined using the full-matrix least squares method against F2 of all data, using SHELXL-2014 [LXL-2014  and OLEXLXL-2014  softwaren-Bu4N)[2–B10Br9S(n-C18H37)2] was found to be disordered over multiple positions and could therefore not be modelled satisfactorily. It was removed from the electron density map using the OLEX solvent mask command. One molecule of DMSO in the asymmetric unit of  was done using equipment from the NRC “Kurchatov Institute”—IREA Shared Knowledge Centre. Using a Bruker D8 ADVANCE X-ray diffractometer  in the angle range 2Theta = 5–50 deg., with a step of 0.01125 deg. and accumulation time 0.1–0.25 s. in cuvettes with a substrate of oriented single-crystal silicon. Immediately before recording, the test samples were ground in a synthetic jasper mortar and placed in cuvettes, levelling the sample level flush. Diffractograms of the compounds obtained are presented in Hirshfeld surface analysis. The Crystal Explorer 17.5 program  (2), (n-Bu4N)[2–B10Br9S(n-Bu)2] (3), (n-Bu4N)[2–B10Br9S(n-C12H25)2] (5) and (n-Bu4N)[2–B10Br9S(n-C18H37)2] (6), which have alkyl substituents of different lengths. Between (n-Bu4N)[2–B10Br9S(i-Pr)2] and (n-Bu4N)[2–B10Br9S(n-Pr)2], it was decided to calculate only the first one to save computational time, because i-Pr and borane form intramolecular interactions that can be characterised through a study of the electron density function. All quantum chemical calculations were performed at 0 K.The structures studied with quantum chemical calculations were characterised by substantial uncertainties relating to atomic coordinates, due to thermal motion and because it was impossible to precisely refine the position of hydrogen atoms. Some structures were disordered. To overcome these problems, the disorder of some molecular fragments was removed and the atomic coordinates were optimised. After that, all atomic coordinates and cell vectors were relaxed. The values of optimised atomic coordinates, cell vectors and total energies can be found in −7 eV). DFT (PW-PBE-D3) periodic calculations were performed using the VASP package .General method for the synthesis of compounds [2-B10Br9S(n-Pr)2] (1).( From (n-Bu4N)[2-B10H9S(n-Pr)2]  and Br2 , (n-Bu4N)[2-B10Br9S(n-Pr)2]  was obtained. Yield: 85%. Anal. calc. of C22H50B10Br9NS: C, 22.2; H, 4.2; N, 1.2; S, 2.7; Found: C, 22.0; H, 4.1; N, 1.0; S, 2.5. 11B NMR  ppm: −0.5 , −4.9 , −10.6 , −13.2 , −17.0 . 1H NMR  ppm: 3.56 , 3.56 , 3.40 , 1.94, 1.87 , 1.77 , 1.39 , 1.04 , 0.97 . 13C NMR  ppm: 59.3 (Bu4N+), 41.0 (SCH2), 24.6 (Bu4N+), 21.7 (SCH2CH2), 20.6 (Bu4N+), 14.2 (Bu4N+), 13.4 (CH3). IR (CCl4) cm−1: 2961, 2931, 2873, 1470, 1378, 1308, 1242, 1155, 1123, 1067, 970, 880, 795, 759, 433.n-Bu4N)[2-B10Br9S(i-Pr)2] (2).( From (n-Bu4N)[2-B10H9S(i-Pr)2]  and Br2 , (n-Bu4N)[2-B10Br9S(i-Pr)2]  was obtained. Yield: 89%. Anal. calc. of C22H50B10Br9NS: C, 22.2; H, 4.2; N, 1.2; S, 2.7; Found: C, 22.0; H, 4.0; N, 1.0; S, 2.5. 11B NMR  ppm: −0.5 , −5.9 , −10.2 , −13.2 , −17.2 . 1H NMR  ppm: 4.32 , 3.40 , 1.81 , 1.78 , 1.39 , 0.97 . 13C NMR  ppm: 59.3 (Bu4N+), 45.1 (SCH), 24.6 (Bu4N+), 23.2 (CH3), 20.6 (Bu4N+), 14.2 (Bu4N+). IR (CCl4) cm−1: 2961, 2932, 2873, 1469, 1378, 1307, 1240, 1156, 1120, 1066, 969, 896, 880, 797, 760, 432.n-Bu4N)[2-B10Br9S(n-Bu)2] (3).( From (n-Bu4N)[2-B10H9S(n-Bu)2]  and Br2 , (n-Bu4N)[2-B10Br9S(n-Bu)2]  was obtained. Yield: 87%. Anal. calc. of C24H54B10Br9NS: C, 23.7; H, 4.5; N, 1.1; S, 2.6; Found: C, 23.6; H, 4.2; N, 1.0; S, 2.4. 11B NMR  ppm: −0.5 , −4.9 , −10.6 , −13.2 , −17.0 . 1H NMR  ppm: 3.58 , 3.39 , 1.92 ), 1.86 ), 1.77 , 1.47 , 1.42 , 0.97 , 0.95 ,. 13C NMR  ppm: 59.4 (Bu4N+), 38.9 (SCH2), 30.0 (SCH2CH2), 24.6 (Bu4N+), 22.4 (CH2CH3), 20.6 (Bu4N+), 14.2 (Bu4N+), 13.9 (CH3). IR (CCl4) cm−1: 2959, 2932, 2872, 1470, 1417, 1380, 1340, 1309, 1242, 1229, 1150, 1122, 1032, 969, 896, 882, 795, 760, 435.n-Bu4N)[2-B10Br9S(n-C8H17)2] (4).( From (n-Bu4N)[2-B10H9S(n-C8H17)2]  and Br2 , (n-Bu4N)[2-B10Br9S(n-C8H17)2]  was obtained. Yield: 86%. Anal. calc. of C32H70B10Br9NS: C, 28.9; H, 5.3; N, 1.0; S, 2.4; Found: C, 28.7; H, 5.1; N, 0.9; S, 2.2. 11B NMR  ppm: −0.4 , −4.9 , −13.2 . 1H NMR  ppm: 3.59 , 3.39 , 1.94 ), 1.87 ), 1.77 , 1.42 , 1.28 , 0.98 , 0.88 . 13C NMR  ppm: 59.4 (Bu4N+), 39.2 (SCH2), 32.6 (SCH2CH2), 29.8, 29.5, 29.1, 28.0 (C3-C6), 24.7 (Bu4N+), 23.5 (CH2CH3), 20.6 (Bu4N+), 14.7 (CH3), 14.2 (Bu4N+). IR (CCl4) cm−1: 2958, 2932, 2874, 1470, 1417, 1380, 1362, 1340, 1309, 1242, 1122, 969, 896, 882, 795, 762, 435.n-Bu4N)[2-B10Br9S(n-C12H25)2] (5).( From (n-Bu4N)[2-B10H9S(n-C12H25)2]  and Br2 , (n-Bu4N)[2-B10Br9S(n-C12H25)2]  was obtained. Yield: 82%. Anal. calc. of C40H86B10Br9NS: C, 33.3; H, 6.0; N, 1.0; S, 2.2; Found: C, 33.1; H, 5.8; N, 0.8; S, 2.0. 11B NMR  ppm: −0.4 , −4.9 , −13.2 . 1H NMR  ppm: 3.59 , 3.39 , 1.95 ), 1.88 ), 1.77 , 1.39 , 1.28 , 0.97 , 0.88 . 13C NMR  ppm: 59.4 (Bu4N+), 39.2 (SCH2), 32.8 (SCH2CH2), 30.6, 30.4, 30.3, 30.2, 29.6, 29.1, 28.0 (C3-C10), 24.7 (Bu4N+), 23.6 (CH2CH3), 20.6 (Bu4N+), 14.8 (CH3), 14.3 (Bu4N+). IR (CCl4) cm−1: 2962, 2926, 2874, 2854, 1469, 1418, 1379, 1308, 1242, 1150, 1121, 968, 895, 839, 787, 759, 432.n-Bu4N)[2-B10Br9S(n-C18H37)2] (6).( From (n-Bu4N)[2-B10H9S(n-C18H37)2]  and Br2 , (n-Bu4N)[2-B10Br9S(n-C18H37)2]  was obtained. Yield: 84%. Anal. calc. of C52H110B10Br9NS: C, 38.8; H, 6.9; N, 0.9; S, 2.0; Found: C, 38.7; H, 6.7; N, 0.7; S, 1.7. 11B NMR  ppm: −0.4 , −4.9 , −13.2 . 1H NMR  ppm: 3.60 , 3.39 , 1.95 ), 1.88 ), 1.77 , 1.39 , 1.29 , 0.97 , 0.88 . 13C NMR  ppm: 59.4 (Bu4N+), 39.2 (SCH2), 32.9 (SCH2CH2), 30.6, 30.5, 30.3, 30.2, 29.6, 29.0, 28.0 (C3-C16), 24.6 (Bu4N+), 23.6 (CH2CH3), 20.6 (Bu4N+), 14.8 (CH3), 14.2 (Bu4N+). IR (CCl4) cm−1: 2961, 2925, 2874, 2854, 1468, 1406, 1378, 1308, 1243, 1150, 1121, 968, 895, 838, 786, 762, 433.closo-decaborate anion with alkyl substituents of various hydrocarbon chain lengths can be easily prepared with elemental bromine from their non-halogenated compounds in about a day. The melting points of the compounds obtained are practically the same as their perchlorinated analogues. It can be argued that they depend little on the type of halogen atom, but significantly decrease with the growth of the chain of the alkyl substituent. According to quantum chemical calculations and Hirschfeld surface analysis, the crystal structures are stabilised mainly by cation–anion and anion–anion interactions. The nature of Br…Br interactions between brominated borane fragments was found to be attractive, in contrast to intramolecular Br…Br interactions between adjacent bromine atoms.Perbrominated sulfonium derivatives of the


H-chromen-3-yl-1,3,5-triazines 2a-h, 3-(benzoxazol-2-yl)-2H-chromen-2-imines 4a-b, and 3-(benzothiazol-2-yl)-2H-chromen-2-imines 6a-c were obtained by reacting of appropriate 2-iminocoumarin ligands L1a-h, L3a-b, and L5a-c with 3-fold molar excess of copper(II) chloride. The structure of these compounds was confirmed by IR spectroscopy, elemental analysis, and single-crystal X-ray diffraction data . All the synthesized complexes were screened for their activity against five human cancer cell lines: DAN-G, A-427, LCLC-103H, SISO, and RT-4 by using a crystal violet microtiter plate assay and relationships between structure and in vitro cytotoxic activity are discussed. The coordination of 2-iminocoumarins with copper(II) ions resulted in complexes 2a-h, 4a-b, and 6a-c with significant inhibitory properties toward tested tumor cell lines with IC50 values ranging from 0.04 μM to 15.66 μM. In comparison to the free ligands L1a-h, L3a-b, and L5a-c, the newly prepared Cu(II) complexes often displayed increased activity. In the series of copper(II) complexes of 2-imino-2H-chromen-3-yl-1,3,5-triazines 2a-h the most potent compound 2g contained a 4-phenylpiperazine moiety at position 6 of the 1,3,5-triazine ring and an electron-donating diethylamino group at position 7′ of the 2-iminocoumarin scaffold. Among the Cu(II) complexes of 3-(benzoxazol-2-yl)-2H-chromen-2-imines 4a-b and 3-(benzothiazol-2-yl)-2H-chromen-2-imines 6a-c the most active was benzoxazole-2-iminocoumarin 4b that also possessed a diethylamino group at position 7′ of the 2-iminocoumarin moiety. Moreover, compound 4b was found to be the most prominent agent and displayed the higher potency than cisplatin against tested cell lines.A series of copper(II) complexes of 2-imino-2 Cancer—a multifactorial disease—constitutes the second leading cause of worldwide mortality, being a major public health issue. Despite the fact that there are several various approaches in cancer treatment, chemotherapy still remains one of the most important strategies . Among cAmong transition metals, copper occupies an important position with respect to its presence in living organisms and biological role in physiological processes, such as cell metabolism or mitochondrial respiration ,6,7,8. MThe antiproliferative effects of the copper(II) complexes may result from various mechanisms. For example, copper(II) complexes with a well-defined antitumor mechanism of action include intercalators of DNA ,21,22,23H-chromene)—less known coumarin analogs—are studied for many interesting potential applications due to their optical or fluorescent properties  or binuclear [(LCuCl)2(μ-Cl)2] complexes with CuCl2. In complexes 2f, 2g, and 2h, the ligand on binding to the metal ion adopts the B conformation. In mononuclear complexes 2h, 2g, and 6c, the Cu (II) atom is tetracoordinated with the coordination environment intermediate between tetrahedral and square planar as indicated by the analysis with the SHAPE program (version 2.1). In the dinuclear complex, 2f, the pentacoordinated metal center is in a slightly distorted tetragonal pyramidal environment. In this compound, one of the Cl ligands is acting in a bridging mode binding asymmetrically the metal centers in the Cu2Cl2 coordination ring [Cu-Cl distances are 2.3330(6) and 2.6959(7) Å]. The bond formed by the metal center with the primary imine group N atom of the 2-iminocoumarin fragment, which is in the range 1.930(2)–1.957(2) Å, is always shorter than the bond to the secondary imine group N atom in the chelate ring (1.994(2)–2.053(2) Å).In all studied compounds, the organic ligands 2f, 2h, and 2g primary NH2 amino groups are involved in intramolecular N-H···Cl hydrogen bonds and intermolecular N-H···N or N-H···Cl interactions, whereas the imino =N-H group does not take part in any hydrogen bonding. In the case of 6c, where the primary NH2 group is missing, the imino C=N-H proton takes part in N-H···Cl interaction assembling the coordination molecules into hydrogen-bonded centrosymmetric dimer, where the distance between the two-metal center is 3.9916(5) Å. In Some additional geometrical details can be found in the 2a-h, 4a-b, and 6a-c in phosphate-buffered saline solution  at 37 °C was investigated using UV-Vis spectroscopy. The 2b and 2d over 6 h when they were incubated at 37 °C in PBS. In general, all copper(II) complexes tested proved to be relatively stable in the PBS solution, as exemplified by the complexes 2b and 2d, since no new spectra with the formation of isosbestic points were observed. The complex 2d  complexes mplex 2d A showed x 2d -2H-chromen-2-imines 4a-b and 3-(benzothiazol-2-yl)-2H-chromen-2-imines 6a-c were investigated for antiproliferative activity in a panel of five human cancer cell lines of different origin including: human pancreas adenocarcinoma DAN-G, human lung carcinoma A-427, human non-small cell lung cancer LCLC-103H, human cervix cancer SISO, and human urinary bladder carcinoma RT-4.All copper(II) complexes of 2-imino-21-Cl2) of copper(II) complex 2f is 2.696 Å (2g-h (2.24–2.33 Å), the dimer 2f under physiological conditions can easily dissociate to form a mononuclear copper(II) complex with the structure analogous to the discussed complexes 2a-e and 2g-h. Thus, it can be assumed that in the solution, the tested complex 2f possesses a similar structure to that of the complexes 2a-e and 2g-h.In regards to the cytotoxicity studies, it should be mentioned that the bond length of the halide bridge  complexes, 2a-h, 4a-b, and 6a-c, together with the data obtained for previously reported free ligands, 2-imino-2H-chromen-3-yl-1,3,5-triazines L1a-h copper(II) (2a). Yield 13% as orange colored powder; m.p. 261–262 °C; IR (KBr) νmax (cm−1): 3468, 2934, 2855, 1647, 1550, 1482, 1240, 803, 776. Anal. calcd for C17H18Cl2CuN6O (456.82): C, 44.70; H, 3.97; N, 18.40. Found: C, 44.62; H, 3.78; N, 18.32.Dichlorocopper(II) (2d). Yield 23% as green crystals; m.p. 211–212 °C; IR (KBr) νmax (cm−1): 3298, 2923, 1643, 1544, 1493, 1233, 804, 756. Anal. calcd for C18H18BrCl2CuN7O (562.74): C, 38.42; H, 3.22; N, 17.42. Found: C, 38.36; H, 3.12; N, 17.23.An appropriate 2-iminocoumarin-1,3,5-triazine derivative L1b, L1c, or L1f (1 mmol) was dissolved in 98% ethanol (4–10 mL) at a temperature of 40 °C. After cooling to room temperature (20–22 °C), copper(II) chloride dihydrate (3 mmol) dissolved in 1 mL of 98% ethanol and was added gradually. If precipitate of 1,3,5-triazine derivative was formed after cooling to room temperature the second portion of 98% ethanol was added. Upon slow evaporation of the solvent over 1–4 days, the copper(II) complex was formed. Precipitate was filtered off, washed with cold 98% ethanol (2 × 0.5 mL) and dried in a desiccator. The following complexes were obtained to above procedure:Dichloro{4-[7-(diethylamino)-2-imino-2H-chromen-3-yl]-6-morpholino-1,3,5-triazin-2-amine}copper(II) (2b). Yield 26% as violet crystals; m.p. 189–190 °C; IR (KBr) νmax (cm−1): 3307, 1653, 1598, 1498, 1351, 1237, 1173, 800. Anal. Calcd for C20H25Cl2CuN7O2 (529.91): C, 45.33; H, 4.76; N, 18.50. Found: C, 45.14; H, 4.68; N, 18.42.Dichloro{4-[7-(diethylamino)-2-imino-2H-chromen-3-yl]-6--1,3,5-triazin-2-amine}copper(II) (2c). Yield 18% as dark brown crystals; m.p. 189 °C (dec.); IR (KBr) νmax (cm−1): 3110, 1654, 1595, 1500, 1466, 1340, 1247, 1183, 1015, 800, 693. Anal. calcd for C22H28Cl2CuN8O (554.96): C, 47.61; H, 5.09; N, 20.19. Found: C, 47.58; H, 5.01; N, 20.08.Tetrachloro-bis-{copper(II)} (2f). Yield 23% as dark green crystals; m.p. 169–170 °C; IR (KBr) νmax (cm−1): 3489, 3336, 1649, 1546, 1494, 1231, 804, 757.2f ([(C22H21N7O)2Cl4Cu2]·C2H5OH, M = 1113.86 g/mol): triclinic, space group P-1 (no. 2), a = 10.4108(4) Å, b = 11.3726(4) Å, c = 11.4595(5) Å, α = 83.833(3)°, β = 64.171(4)°, γ = 77.430(3)°, V = 1191.84(9) Å3, Z = 1, T = 293(2) K, μ(Cu Kα) = 3.641 mm−1, Dcalc = 1.552 g/cm3, 24,433 reflections measured (8.574° ≤ 2Θ ≤ 151.718°), 4866 unique  which were used in all calculations. The final R1 was 0.0393 (I > 2σ(I)) and wR2 was 0.1187 . The crystals were unstable in the air. The solvent ethanol molecule is disordered around an inversion center. The methylene and methyl group C atoms from the ethanol molecules occupying different sites were found in overlapping positions. The molecule of 2f is shown in Crystal data for An appropriate 2-iminocoumarin-1,3,5-triazine derivative L1e, L1g or L1h (1 mmol) was dissolved in methanol (6 mL) at a temperature of 40 °C. After cooling to room temperature (20–22 °C) copper(II) chloride dihydrate (3 mmol) dissolved in 1 mL of methanol and was added gradually. Upon slow evaporation of the solvent over 1–4 days, copper(II) complex was formed. Precipitate was filtered off, washed with cold methanol (2 × 0.5 mL) and dried in a desiccator. The following complexes were obtained to above procedure:Dichlorocopper(II) (2e). Yield 21% as green crystals; m.p. 171–172 °C; IR (KBr) νmax (cm−1): 3227, 2923, 1654, 1582, 1508, 1467, 1380, 1340, 1250, 803. Anal. calcd for C19H21Cl2CuN7O (497.87): C, 45.84; H, 4.25; N, 19.69. Found: C, 45.68; H, 4.12; N, 19.54.Dichloro{4-[7-(diethylamino)-2-imino-2H-chromen-3-yl]-6-(4-phenylpiperazin-1-yl)-1,3,5-triazin-2-amine}copper(II) (2g). Yield 27% as violet crystals; m.p. 194–195 °C; IR (KBr) νmax (cm−1): 3352, 2923, 1654, 1586, 1501, 1232, 804, 748.2g : triclinic, space group P-1 (no. 2), a = 8.8366(7) Å, b = 11.1171(8) Å, c = 15.0104(9) Å, α = 100.138(6)°, β = 93.417(6)°, γ = 105.259(7)°, V = 1391.65(18) Å3, Z = 2, T = 293(2) K, μ(Cu Kα) = 3.161 mm−1, Dcalc = 1.444 g/cm3, 14,878 reflections measured (6.018° ≤ 2Θ ≤ 133.202°), 4927 unique  which were used in all calculations. The final R1 was 0.0423 (I > 2σ(I)) and wR2 was 0.1293 . One of the ethyl groups of the diethylamino substituent is disordered over two sites. The molecule of 2g is shown in Crystal data for Dichlorocopper(II) (2h). Yield 22% as greenish crystals; m.p. 191 °C (dec.); IR (KBr) νmax (cm−1): 3295, 1643, 1570, 1545, 1496, 1233, 803, 761.2h ([(C22H20ClN7O]Cl2Cu]·0.5(H2O), M = 577.35g/mol): monoclinic, space group P21/n (no. 14), a = 16.9813(4) Å, b = 8.8458(2) Å, c = 17.1929(4) Å, β = 109.043(3)°, V = 2441.26(11) Å3, Z = 4, T = 294 K, μ(Cu Kα) = 4.563 mm−1, Dcalc = 1.571 g/cm3, 13,722 reflections measured (6.352° ≤ 2Θ ≤ 133.198°), 4315 unique  which were used in all calculations. The final R1 was 0.0453 (I > 2σ(I)) and wR2 was 0.1335 . The water molecule is disordered over two sites related by an inversion center. Hydrogen atoms of the water molecule were not located. The molecule of 2h is shown in Crystal data for H-chromen-2-imine derivative L3a or L3b (1 mmol) was dissolved in 98% ethanol (4–10 mL) at a temperature of 40 °C. To the solution was added dropwise at ambient temperature (20–22 °C) copper(II) chloride dihydrate (3 mmol) dissolved in ethanol (1 mL). The solution was left at room temperature (20–22 °C) and the solvent was slowly evaporated. The resulting precipitate (24 h to 4 days) was filtered, washed with cold ethanol (2 × 0.5 mL), and dried in a desiccator. The following complexes were prepared according to above given procedure.An appropriate 3-(benzoxazol-2-yl)-2Dichloro[3-(benzo[d]oxazol-2-yl)-6-fluoro-2H-chromen-2-imine]copper(II) (4a). Yield 20% as violet-brown crystals; m.p. > 350 °C; IR (KBr) νmax (cm−1): 3119, 3067, 2927, 1635, 1586, 1560, 1471, 1430, 1374, 1269, 1205, 1155, 1054, 876, 794, 722. Anal. calcd for C16H9Cl2CuFN2O2 (414.71): C, 46.34; H, 2.19; N, 6.76. Found: C, 46.17; H, 2.35; N, 6.85.Dichloro[3-(benzo[d]oxazol-2-yl)-N,N-diethyl-2-imino-2H-chromen-7-amine]copper(II) (4b). Yield 24% as violet crystals; m.p. 229–230 °C; IR (KBr) νmax (cm−1): 3292, 2976, 1654, 1601, 1574, 1527, 1497, 1399, 1343, 1240, 1184, 1123, 1077, 973, 773, 757. Anal. calcd for C20H19Cl2CuN3O2 (467.84): C, 51.35; H, 4.09; N, 8.98. Found: C, 51.43; H, 3.89; N, 9.05.H-chromen-2-imine derivative L5a, L5b, or L5c (1 mmol) was dissolved in dimethylformamide (2–4 mL) containing 1–2% water at a temperature of 80 °C. To the resulting solution, copper(II) chloride dihydrate (3 mmol) was added dropwise at ambient temperature (20–22 °C) and dissolved in dimethylformamide (1 mL). The solution was left at room temperature (20–22 °C) and the solvent was slowly evaporated. The resulting precipitate (24 h to 4 days) was filtered, washed with cold dimethylformamide (2 × 0.5 mL) and dried in a desiccator. The following complexes were prepared according to above given procedure.An appropriate 3-(benzothiazol-2-yl)-2Dichloro[3-(benzo[d]thiazol-2-yl)-6-fluoro-2H-chromen-2-imine]copper(II) (6a). Yield 29% as violet-dark blue crystals; m.p. 231–232 °C; IR (KBr) νmax (cm−1): 3178, 3062, 3017, 1664, 1642, 1596, 1566, 1491, 1413, 1272, 1248, 957, 880, 784, 763, 753. Anal. calcd for C16H9Cl2CuFN2OS (430.77): C, 44.61; H, 2.11; N, 6.50. Found: C, 44.49; H, 2.31; N, 5.87.Dichloro[3-(benzo[d]thiazol-2-yl)-N,N-diethyl-2-imino-2H-chromen-7-amine]copper(II) (6b). Yield 29% as violet-dark blue crystals; m.p. 231–232 °C; IR (KBr) νmax (cm−1): 3338, 2963, 2922, 1646, 1592, 1573, 1510, 1380, 1350, 1271, 1185, 1115, 756, 745. Anal. calcd for C20H19Cl2CuN3OS (483.90): C, 49.64; H, 3.96; N, 8.63. Found: C, 49.55; H, 3.83; N, 8.57.Dichloro[3-(benzo[d]thiazol-2-yl)-6-methyl-2H-chromen-2-imine]copper(II) (6c). Yield 24% as dark green crystals; m.p. 248–249 °C; IR (KBr) νmax (cm−1): 3190, 3007, 2922, 1636, 1613, 1588, 1568, 1457, 1412, 1327, 1247, 1209, 1127, 956, 861, 825, 752.6c ([(C17H12N2OS)Cl2Cu]·C3H7NO, M = 499.88 g/mol): triclinic, space group P-1 (no. 2), a = 8.8247(3) Å, b = 9.9302(4) Å, c = 12.6944(5) Å, α = 104.339(3)°, β = 90.641(3)°, γ = 102.895(3)°, V = 1047.99(7) Å3, Z = 2, T = 293(2) K, μ(Cu Kα) = 4.935 mm−1, Dcalc = 1.584 g/cm3, 8178 reflections measured (7.206° ≤ 2Θ ≤ 136.502°), 3828 unique  which were used in all calculations. The final R1 was 0.0266 (I > 2σ(I)) and wR2 was 0.0770 . The asymmetric unit of 6c is shown in Crystal data for 2a-h, 4a-b, or 6a-c, resulting in a final concentration of 40 µM. The solution was then transferred to a 1.0 cm quartz cuvette and placed in a heated cuvette holder maintained at 37 °C. UV-Vis spectra were recorded at 60 min intervals between wavelengths of 200 nm and 600 nm by means of UV-Vis spectrophotometer  connected to a personal computer running the Vision 32 software.To 5.0 mL of phosphate-buffered saline at pH 7.4 and pre-warmed at 37 °C was added 10 µL of a 20 mM DMF solution of the copper(II) complex 3, and 10% sterile calf serum, suitable for cell culture. Cells were incubated in a humid atmosphere of 5% CO2 at 37 °C in 75 cm2 plastic culture flasks  and were passaged shortly before becoming confluent. For the cytotoxicity studies, 100 μL of a cell suspension was seeded into 96-well microtiter plates (Sarstedt) at a density of 1000 cells per well except for the LCLC-103H cell line, which was plated out at 250 cells per well. One day after plating, the cells were treated with a test substance at five concentrations per compound. The 1000-fold concentrated stock solutions in dimethylformamide (DMF) or dimethylsulfoxide (DMSO) were serially diluted by 50% in DMF or DMSO to give the feed solutions, which were diluted 500-fold into the culture medium. The controls received DMF or DMSO at a final concentration of 0.1%. Each concentration was tested in eight wells, with each well receiving 100 μL of the medium containing the substance. The concentration ranges were chosen to bracket the expected IC50 values as best as possible. Cells were then incubated for 96 h, after which time the medium was removed and replaced with 1% glutaraldehyde/phosphate-buffered saline for 30 min. The glutaraldehyde solution was discarded, and the plates washed with water before staining cells for 30 min with an aqueous solution of 0.02% Crystal Violet. After washing out excess Crystal Violet from the plates, optical density (OD) was measured at λ = 570 nm by use of an Anthos 2010 plate reader . Corrected T/C values were calculated according to the Equation (1): 50 values were estimated by linear least-squares regression of the (T/C)corr values versus the logarithm of the substance concentration by using MS-Excel; only concentrations that yielded (T/C)corr values between 10% and 90% were used in the calculation of the IC50 values. The reported IC50 values are the averages of three independent experiments [The antiproliferative activities of the compounds on human cancer cells were determined by the Crystal Violet assay as previously described . All celeriments .50 values ranging from 0.04 to 15.66 μM. Furthermore, it was observed that newly prepared Cu(II) complexes often possess higher activity than the related ligands. As expected, in the series of Cu(II) complexes of 2-imino-2H-chromen-3-yl-1,3,5-triazine derivatives, the highest potency was shown for compound 2g bearing 4-phenylpiperazine moiety at position 6 of the 1,3,5-triazine ring and an electron-donating diethylamino group at position 7’ of the 2-iminocoumarin scaffold. On the other hand, the SAR studies of this series indicate that 2-iminocoumarin scaffold is open for variation such that halogen, diethylamino, and methyl substituents are allowed. In turn, in the category of Cu(II) complexes of benzoxazole/benzothiazole-2H-chromen-2-imines the best antiproliferative activity was found for 7’-diethylamino-substituted benzoxazole derivative 4b. Additionally, compound 4b was found to be the most promising agent with higher potency than cisplatin against the LCLC-103H, SISO, and RT-4 cell lines. Based on the results obtained, one may conclude that the most active compounds in either triazine or benzazole series possess a diethylamino group in the 2-iminocoumarin skeleton as evidenced by compounds 2g and 4b, suggesting that this group is somehow involved in the mechanism of action. In this context, it is worth noting that coumarin-based agents [In this paper we have described the synthesis, structure, and cytotoxic activity of two series of novel Cu(II) complexes of 2-iminocoumarins bearing a 1,3,5-triazine or benzoxazole/benzothiazole moiety. The coordination of 2-iminocoumarin ligands with copper(II) results in metal complexes with significant growth inhibitory properties toward tested human tumor cell lines with ICd agents ,74,75,76


Achillea mellifolium belongs to a highly reputed family of medicinal plants, with plant extract being used as medicine in indigenous system. However, limited data is available regarding the exploitation of the medicinal potential of isolated pure compounds from this family; (2) Methods: A whole plant extract was partitioned into fractions and on the basis of biological activity, an ethyl acetate fraction was selected for isolation of pure compounds. Isolated compounds were characterized using different spectroscopic techniques. The compounds isolated from this study were tested for their medicinal potential using in-vitro enzyme assay, coupled with in-silico studies; (3) Results: Three new acrylic acid derivatives (1–3) have been isolated from the ethyl acetate fraction of Achillea mellifolium. The characterization of these compounds (1–3) was carried out using UV/Vis, FT-IR, 1D and 2D-NMR spectroscopy  and mass spectrometry. These acrylic acid derivatives were further evaluated for their enzyme inhibition potential against urease from jack bean and α glucosidase from Saccharomyces cerevisiae, using both in-silico and in-vitro approaches. In-vitro studies showed that compound 3 has the highest inhibition against urease enzyme (IC50 =10.46 ± 0.03 μΜ), followed by compound 1 and compound 2 with percent inhibition and IC50 value of 16.87 ± 0.02 c and 13.71 ± 0.07 μΜ, respectively, compared to the standard (thiourea-IC50 = 21.5 ± 0.01 μΜ). The investigated IC50 value of compound 3 against the urease enzyme is two times lower compared to thiourea, suggesting that this compound is twice as active compared to the standard drug. On the other hand, all three compounds (1–3) revealed mild inhibition potential against α-glucosidase. In-silico molecular docking studies, in combination with MD simulations and free energy, calculations were also performed to rationalize their time evolved mode of interaction inside the active pocket. Binding energies were computed using a MMPBSA approach, and the role of individual residues to overall binding of the inhibitors inside the active pockets were also computed; (4) Conclusions: Together, these studies confirm the inhibitory potential of isolated acrylic acid derivatives against both urease and α-glucosidase enzymes; however, their inhibition potential is better for urease enzyme even when compared to the standard.(1) Background:  Achillea millefolium, locally named as Kangi Jari or Kala Chahu, is a perennial herb belonging to the Asteraceae family, also referred to as a composite family. Asteraceae is the largest angiosperm family, comprised of over 1620 genera and 23,000 species. Achillea is a genus of 115 species that are mostly perennial herbs. Achillea millefolium is native to temperate and sub-tropical regions of the world and grows well in meadows + 250, 222, 208, 180, 172, 150, 124, 70 and 58; HR-EI-MS: m/z [M+] Calcd. 266.1518 for Mol. formula C15H22O4; Observed 266.1510; 3); 1H-NMR  δ (ppm): 3.80 , 5.80 , 2.81 , 6.30 , 5.98 , 3.78 , 1.35 , 0.98 , 1.02 , 3.42 , 1.95 ; 13C-NMR  δ (ppm): 39.6 (C-1), 72.1 (C-2), 194.8 (C-3), 125.1 (C-4), 164.4 (C-5), 59.2 (C-6), 132.4 (C-7), 137.6 (C-8), 170.1 (C-9), 70.6 (C-10), 18.1 (C-11), 19.6 (C-12), 23.6 (C-13), 55.7 (C-14), 22.6 (C-15)  λ6 (C-15) .max 244 (4.01), 220 (3.8) nm; IR (KBr) υmax 1680, 1740 and 2980 cm−1; EI-MS m/z: 252 [M]+ 220, 202, 236, 196, 174, 152, 122, 70 and 58; HR-EI-MS: m/z [M+] Calcd. 252.1362 for Mol. formula C14H20O4; Observed 252.1356; 3); 1H-NMR  δ (ppm): 3.74 , 5.82 , 2.78 , 6.28 , 5.92 , 3.85 , 0.92 , 1.08 , 3.35 , 1.90 ; 13C-NMR  δ (ppm): 38.2 (C-1), 70.8 (C-2), 192.1 (C-3), 126.7 (C-4), 167.1 (C-5), 58.1 (C-6), 134.8 (C-7), 141.1 (C-8), 172.1 (C-9), 56.1 (C-10), 19.8 (C-11), 23.4 (C-12), 57.9 (C-13), 22.6 (C-14)  λ6 (C-14) .max 238 (2.8) and 216 (3.7) nm; IR (KBr) υmax 2920, 1720 and 1875 cm−1; EI-MS m/z: 236 [M]+ 218, 204, 174, 168, 150, 122, 70 and 56; HR-EI-MS: m/z [M+] Calcd. 236.1412 for Mol. formula C14H20O3; Observed 236.1406; 3); 1H-NMR  δ (ppm): 3.69 , 5.85 , 2.80 , 6.27 , 5.96 , 2.10 , 0.97 , 1.02 , 3.40 , 1.88 ; 13C-NMR  δ (ppm): 40.2 (C-1), 72.3 (C-2), 193.6 (C-3), 128.7 (C-4), 165.6 (C-5), 59.1 (C-6), 131.8 (C-7), 136.5 (C-8), 184.4 (C-9), 24.1 (C-10), 20.8 (C-11), 23.6 (C-12), 22.2 (C-13), 57.2 (C-14)  λ2 (C-14) .The column chromatography of the ethyl acetate fraction resulted in isolation and purification of three new derivatives of acrylic acid, and their characterization was carried out using various spectroscopic techniques.1 was isolated as a light-yellow colored gummy solid, having highest molecular ion peak at m/z 266.1510 in HR-EI-MS, suggesting the molecular formula of C15H22O4. The fragment peaks obtained were 250, 222, 208, 180, 172, 150, 124, 70 and 58. The absorption band in the UV spectrum were obtained at λmax 224 (4.01), 220 (3.8), while IR spectrum confirmed presence of hydroxyl group, ester group and conjugated carbonyl moiety, as suggested by absorption band at υmax 1672, 1730 and 2960 cm−1.Compound 1H-NMR spectrum showed chemical shift values for three methyl groups at δH 0.98 , 1.02  and 1.95  while two other singlets were also obtained at δH 3.80  and 5.80 . Similarly, doublet was obtained at chemical shift value of 2.81  and a doublet of doublet was obtained at δH 6.30  while proton at position 8 also showed doublet at δH 5.98 .The J = 6.8 Hz, H-10) and triplet at δH 1.35  was also obtained, as given in In addition, a quartet for methylene protons at position 10 was obtained at δH 3.78  carbon atoms comprising of four methyl, one methylene, five methine and four quaternary and one methoxy carbon atom, as given in 13C-NMR spectrum revealed the presence of five methine carbon at δC 72.1 (C-2), 125.1 (C-4), 59.2 (C-6), 132.4 (C-7) and 137.6 (C-8). Similarly, four quaternary carbons resonated at δC 39.6 (C-1), 194.8 (C-3) and 170.1 (C-9) and signal for methoxy carbon was obtained at δC 55.7 (C-14).The 1 was done on the basis of HMBC and NOE spectra, as shown in The placement of substituents for compound 1 was confirmed to be ethyl (E)-3--5-methoxy-2,6,6-trimethyl-4-oxocyclohex-2-en-1-yl) acrylate, as suggested by all spectral data, as well as literature comparison  were utilized for energy minimization of proteins. The active site of both urease and α-glucosidase was specified within 10 Å of the co-crystallized ligand. The lowest energy minimized docked pose was used for further analysis. The MOE’s ligand-interaction module was used for two-dimensional ligand-enzyme interactions analysis. Docked complexes were visualized and further analyzed using MOE and the Discovery Studio Visualizer. All the structures of test compounds  namely ethyl (E)-3--5-methoxy-2,6,6-trimethyl-4-oxocyclohex-2-en-1-yl)acrylate (1), methyl (E)-3--5-methoxy-2,6,6-trimethyl-4-oxocyclohex-2-en-1-yl)acrylate (2) and -6-methoxy-3,5,5-trimethyl-4-((E)-3-oxobut-1-en-1-yl)cyclohex-2-en-1-one (3). The characterization and structure elucidation of these acrylic acid derivatives was done through various spectroscopic techniques such as UV, IR, 1D and 2D-NMR spectroscopy  and mass spectrometry. Enzyme inhibition studies revealed these test compounds (1–3) as potential inhibitors for jack bean urease and mild inhibition potential against α-glucosidase enzyme. All three compounds showed promising inhibitory potential against urease, with compound 3 having an IC50 of 10.46 ± 0.033 µM and compound 1 and 2 with IC50s of 16.87 ± 0.02 µM and 13.71 ± 0.07 µM, respectively. Moderate to low inhibition potential was observed for all test compounds in case of α-glucosidase. All three compounds showed IC50 value higher than standard i.e., acarbose (IC50 = 287.1 ± 0.03 µM).The urease and 1 and 2 showed results comparable to the standard in case of urease enzyme. Compound 1 had the best results in MD simulations and MMPBSA calculations, indicating that these compounds could have better inhibitory performance in experimental tests than standard, which is currently the compound with the best IC50 value against urease.According to a molecular docking study, the most active compounds demonstrated a good protein ligand interaction profile against the target proteins more pronounced in case of urease enzyme, with important interactions such as hydrogen bonding, van der Waal and anion interactions. MD simulations rationalize the docking and in-vitro studies since all compounds were retained inside the active pocket. Evidently, compound