The structures were elucidated by the interpretation of HRESIMS, UV, and NMR data. The absolute configurations of Curtachalasins X1–X6 were determined by comparison of their experimental and calculated electronic circular dichroism (ECD) spectra. In bioassays, Curtachalasin X1 (1) and X5 (5) showed cytotoxic activity against the MCF-7 cell line with IC50 values of 2.03 μM and 0.85 μM, respectively.Six previously undescribed cytochalasins, Curtachalasins X1–X6 ( Cytochalasins are a class of cell membrane permeability mycotoxins, and they can lead to disruption of the filament mesh structure, preventing cell movement and changing cell morphology by binding to intracellular filaments and inhibiting actin polymerization at this point . MeanwhiXylaria was the largest genus of the family Xylariaceae. In the traditional view of this genus, they were saprotrophic fungi that usually appeared on deadwood, participated in the decomposition of organic waste, and even destructed the growth of plants +  in HRESIMS, indicating eleven degrees of unsaturation. Analysis of the 1H and 13C NMR data , 7.32 , and 7.24 ; four methyl groups at δH 2.19 , 1.48 , 1.01 , and 0.66 ; and ten methines including four oxygenated methines , two methenes , two quaternary carbons , a double bond , one amide group (δC 177.1), and one ketone group (δC 214.8). The established functional groups account for seven of eleven degrees of unsaturation, which indicated that compound 1 possessed a four-ring system. Compound /H-3 (δH 3.12)/H-4 (δH 2.91), H-7(δH 4.01)/H-8 (δH 1.59)/H-13 (δH 4.38)/H-14 (δH 1.52)/H-15 (δH 1.73)/H-16 (δH 1.77)/H-23 (δH 0.66), and H-14 (δH 1.52)/H-20(H-19)/H-21 (δH 4.03), allowing the connections as shown by bold lines in 1H-1H COSY correlations, established a six-membered ring D. Moreover, the HMBC correlations from H-21 to C-8, C-14, and C-19 and from H-13 to C-9, C-15 and C-20 established a six-membered ring C, which was fused to ring D via C-14 and C-20. According to the HMBC correlations from H-3 to C-1 and C-9, H-11 to C-4, C-5 and C-6, H-8 to C-4, and C-6 and C-9, the fusing pattern of rings A and B can be determined. In addition, the rings B and C of 1 were suggested to be fused through C-9 and C-8 based on the HMBC correlations between H-20 and C-9, between H-21 and C-4, and between H-13 and C-7. Locations of the acetyl group (δC 214.8 and 26.7) at C-17 (δC 84.6) and the phenyl group at C-10 in 1 were determined by HMBC correlations from CH3-22 to C-17 and C-18, and from H-10 to C-24 and C-25/29. The planar structure of J7,8 = 10.0 Hz), H-8/H-13 , H-19/H-20 , and H-20/H-21 , implied that H-7, H-13, H-20, and H-21 should be α-oriented, and H-8 and H-19 belong to the β-orientation. The NOE correlations of 13-OH/H-15a, H-19/H-14, and H-19/H-16 implied that they were cofacial and assigned as β-orientated. In contrast, the NOE correlations of H-15b/H-20 and H-15b/17-OH suggested that they were on the same side, with an α-orientation. Therefore, the relative configuration was determined as shown in Figure 4.Because the B, C, and D rings are typical cyclohexane structures, the coupling constants between vicinal protons are useful to determine their orientations. The NOESY correlation of H-7/H-13 and H-20/H-21, coupled with the coupling constant of H-7/H-8  as compound 1, according to HRESIMS. Their NMR  in 1, which indicated 2 as an epimer of 1. In addition, from 2D NMR correlations /6-311G*  level of theory, and the absolute configuration was deduced as 3S, 4R, 7R, 8S, 9R, 13S, 14R, 16S, 17S, 19R, 20R, and 21R-2 by comparison of the experimental and calculated ECD data . A comparison of the NMR data  and acetyl group (δC 172.5) was observed, which suggested that the acetyl group connects to the oxygen at position 21, as shown in 3 was assigned as 3S, 4R, 7R, 8S, 9R, 13S, 14R, 16S, 17S, 19R, 20S, and 21R by comparison of the calculated and experimental ECD data . Analysis of the 1D NMR data , 7.25 , and 7.20 ; five methyl groups ; and ten methines, including four oxygenated methines , two methenes (δC 44.2 and 33.4), and a double bond . The above data revealed that 4 was a tetracyclic cytochalasin bearing two acetyl groups, highly similar to those of 3. The only difference is that the hydroxyl group in 3 is replaced by a methoxyl group in 4, which was supported by HMBC correlations from -OCH3 (δH 3.29) to C-7 (δC 78.1) (Compound NMR data  showed cδC 78.1) .4 are similar to 3. Thus, the relative configuration of 4 can be deduced, as shown in 4 was determined to be 3S, 4R, 7R, 8S, 9R, 13S, 14R, 16S, 17S, 19R, 20S, and 21R by ECD calculation on the same level as for compound 1 . The NMR spectra of 5 highly matched with those of 4  to C-13 (δC 80.4). The similar experimental CD spectra  and the calculated ECD spectra of 5 . The 1H and 13C NMR spectra  with compound 1. For the difference, compound 6 showed one more double bond signal . The 1H-1H COSY correlation of H-8 (δH 2.20)/H-13 (δH 5.57) together with HMBC correlations from H-13 (δH 5.57) to C-7 (δC 68.1) and C-15 (δC 37.4) suggested that the double bond was located at C-13 (14) /6-311G*  level of theory (Compound  spectra  showed t-13 (14) . The simf theory .7), Curtachalasin O (8), Curtachalasin N (9), Curtachalasin I (10), Curtachalasin H (11), and Curtachalasin K (12) by the comparison of their spectral data with the data reported in the literature  + . Curtachalasin X2 (2): colorless powder;  + .Curtachalasin X3 (3): colorless powder; + .Curtachalasin X4 (4): colorless powder; + . Curtachalasin X5 (5): colorless powder; + . Curtachalasin X6 (6): colorless powder; + .1–6 were performed with the Gaussian 16 program package. Conformational analysis was initially performed using Spartan 14. The optimized conformation geometries, thermodynamic parameters, and populations of all conformations are provided in Theoretical calculation of ECD spectra for compounds 2 until 70–80% coverage. Cells were then transferred into 96-well culture plates with appropriate density. The plate was incubated for another 24 h at 37 °C, 5% CO2 for cell growth and adhesion. Then, the cells were treated with test samples prepared in culture media at different concentrations  for 48 h. The blank control  and the negative control  were performed simultaneously. After 48 h, 20 μL of MTS reagent was added into each well and optical density at 490 nm was read using a Spectra Max M5 microplate reader  after 1 h incubation at 37 °C, 5% CO2, in the dark. The percentage of cell inhibition was calculated based on the equation below.The cytotoxicity actions of the pure isolated compounds were tested on the breast cancer MCF-7 cell lines using the MTS assay. For this, cells were cultured in DMEM medium, supplemented with 10% fetal bovine serum FBS, 100 U/mL penicillin, and 100 U/mL streptomycin , and incubated at 37 °C, 5% CO1–6) were determined unambiguously by extensive spectroscopic analysis, with the absolute configuration being determined by quantum chemistry calculations. In the cytotoxicity assay, compounds 1 and 5 showed activity against MCF-7 cell lines with outstandingly low IC50 values (2.03 and 0.85 μM) compared to the positive anti-cancer drug cisplatin. Our studies have certain significances in exploring the structural diversity of cytochalasins.In summary, the structures of six undescribed cytochalasins with a tetracyclic skeleton (


Maerua siamensis (Capparaceae) roots are used for treating pain and inflammation in traditional Thai medicine. Eight new indole alkaloids, named maeruanitriles A and B, maeroximes A–C, and maeruabisindoles A–C, were isolated from them. Spectroscopic methods and computational analysis were applied to determine the structure of the isolated compounds. Maeroximes A–C possesses an unusual O-methyloxime moiety. The bisindole alkaloid maeruabisindoles A and B possess a rare azete ring, whereas maeruabisindole C is the first indolocarbazole derivative found in this plant family. Five compounds  displayed anti-inflammatory activity by inhibiting nitric oxide (NO) production in the lipopolysaccharide-induced RAW 264.7 cells. Maeruabisindole B was the most active inhibitor of NO production, with an IC50 of 31.1 ± 1.8 μM compared to indomethacin (IC50 = 150.0 ± 16.0 μM) as the positive control. Maerua is a genus in the family Capparaceae distributed in Africa, Arabia, South Asia, and Indo-China. Up to now, 69 accepted species have been discovered c 0.001, MeOH). Its molecular formula of C11H10N2O3, corresponding to an index hydrogen deficiency (IHD) of eight, was deduced from the sodium-adduct [M + Na]+ molecular ion peak at m/z 241.0585  in the HR-ESI-MS. Its IR spectrum displayed absorption bands due to the hydroxyl and amide NH , aromatic ring (1630 and 1463 cm−1), γ-lactam carbonyl (1722 cm−1), and nitrile (2256 cm−1) components. Its UV absorption maxima at 218, 268, and 322 nm indicated oxindole moiety . A methoxy group, at δH 3.81/ δc 56.5, was located at C-6 by the observed HMBC correlation between its proton resonance and C-6 (δC 163.4), and 1H-1H NOESY cross peaks of 6-OCH3 with both H-5 and H-7. The methylene resonances at δH 3.09 , and 2.89 , exhibited HMBC correlations to C-2 (δC 178.2), C-3 (δC 73.8), C-3a (δC 122.8) and C-9 (δC 117.7) signals, indicating the position of an acetonitrile group at C-3 of the oxindole structure. A hydroxy group (δH 5.44) was also located at C-3, as evidenced by its HMBC correlations to C-3 and C-3a. Selected HMBC and NOESY correlations are shown in 1 resembled that of plasiaticine C, isolated from Plantago asiatica (Plantaginaceae) + ion peak at m/z 271.0511 . The IR spectrum showed absorption bands that were attributable to amine NH , nitrile (2250 cm−1), the aromatic ring , and sulfoxide  functionalities. The 1H-NMR spectrum of 2 showed signals due to ABX coupling protons , one methylene [δH 4.18  and 4.13 ], one methoxy  and one methylsulfinyl group . Its 13C-NMR spectrum showed twelve carbon signals of eight unsaturated carbons , one aliphatic methylene carbon , one methoxy carbon , one nitrile carbon (δc 118.9) and one methylsulfinyl carbon   of the acetonitrile group appeared more downfield due to the anisotropic effect of a double bond between positions 2 and 3. This effect was also observed in indole-3-acetonitrile-2-S-β-glucopyranoside, which was isolated from the roots of I. indigotica + ion peak at m/z 265.0999 in the HR-ESI-MS . The IR spectrum showed absorption bands of amine NH , O-methyloxime, and the aromatic ring  + ion peak at m/z 279.0782 , was 14 mass units higher than compound 3, and thus, suggested the presence of a carbonyl instead of a methylene group. This was supported by a conjugated keto carbonyl signal (δC 182.5) in its 13C-NMR spectrum. The 3,6-disubstituted 1H-indole nucleus of 4 is identical to that of 3, as evidenced by their 1H- and 13C-NMR data in C 182.5), which displayed an HMBC correlation with an H-2 signal (δH 7.97)  and SCH3 groups  on the imine bond was proposed based on computational studies (4 was established as methyl (Z)-N-methoxy-2-(6-methoxy-1H-indol-3-yl)-2-oxoethanimidothioate and given the trivial name maeroxime B.Compound δH 7.97) . Similar studies . Thus, t5, which was obtained as a yellow amorphous solid, gave a protonated molecular ion [M + H]+ at m/z 279.0780 , indicating the same molecular formula as 4 (C13H14N2O3S). Its IR absorption bands at 3307 and 1729 cm−1 and UV maxima at 208, 272, and 316 nm were characteristic of the oxindole feature ethanimidothiolate and named maeroxime C.Compound  feature . The preNMR data , the difwith H-8 . The orianalysis . Therefo6 was isolated as a pale green amorphous solid. Its molecular formula of C22H19N3O2S (IHD 15) was deduced from an [M + H]+ ion peak observed in the HR-ESI-MS at m/z 390.1298 . The 1H-NMR data , 6.78 , and 6.99 ], one 1,2,3-trisubstituted benzene ring , two trisubstituted double bonds [δH 7.79  and 7.51 ], two NH [δH 10.71  and 11.52 ], two methoxys [δH 3.81  and 3.52 ] and one methylthio group , implying a bisindole alkaloid structure. Its 13C-NMR spectrum (C 144.8 (C-2), 139.5 (C-4), 133.4 (C-3)], and an SCH3 methyl signal (δC 14.0). The 6′-methoxy-1′H-indole-2′-yl substructure was confirmed by key HMBC correlations  and NOESY correlations . The 4″-methoxy-1″H-indole-3″-yl substructure was confirmed by HMBC correlations , 1H-1H COSY cross peak between NH-1″/H-2″ and NOESY correlations  (Isatis tinctoria roots -1H-indole. Compound 6 was named maeruabisindole A, and its biosynthetic pathway was proposed, as shown in Compound NMR data  displayespectrum  showed tCH3/H-5] . Both inia roots . The met7 was obtained as a pale green amorphous solid. Its molecular formula of C22H19N3O3S was confirmed by the pseudo-molecular [M + H]+ ion at m/z 406.1224  in the HR-ESI mass spectrum. Its IR spectrum was similar to 6 except for the presence of a sulfoxide absorption peak (1025 cm−1). In its 1H- and 13C-NMR spectra, resonances of a methylsulfinyl group were observed at δH 2.98 and δC 42.1, respectively, replacing the methylthio ones of compound 6. Otherwise, the NMR data representing both indole subunits of this bisindole alkaloid were similar to those of the previous compound (2 carbons of an azete ring resonated at δC 151.7 (C-2), δC 142.7 (C-4), and δC 137.8 (C-3) in its 13C-NMR spectrum. A methylsulfinyl substitution at C-2 and the linkage of the 6′-methoxy-1′H-indole-2′-yl moiety to C-3 of the azete ring were confirmed by HMBC correlations from the 2-SOCH3 signal to C-2 and from the H-3′ signal (δH 8.43) to C-3, respectively. The more downfield chemical shift of H-3′, compared to that of 6, could be due to the anisotropic effect of the S=O bond on the nearby methylsulfinyl group. NOESY correlations of 2-SOCH3/H-3′, 6′-OCH3/H-7′ and 4”-OCH3/H-5” supported the proposed structure of 7 as 4-methoxy-3-[3-(6-methoxy-1H-indol-2-yl)-2-(methylsulfinyl)azet-4-yl]-1H-indole (7 was named maeruabisindole B.Compound compound . Three sH-indole . Compoun8 was isolated as a dark green amorphous solid. Its molecular formula was established as C20H13N3O2 (IHD 16), based on the HR-ESI [M − H]− ion at m/z 326.0968 . The IR spectrum showed absorption bands due to hydroxyl and amine NH (3359 cm−1), nitrile (2212 cm−1), and the aromatic ring (1632 and 1468 cm−1). The 1H-NMR data , 7.40  and 7.24 ], one 1,2,4-trisubstituted benzene ring , an aromatic proton at δH 8.53 , two NH protons at δH 10.86  and 10.39 , a methoxy group at δH 4.12  and a hydroxyl group at δH 8.70 . The 13C NMR spectra exhibited eleven quaternary and seven methine carbon resonances, a methoxy signal at δC 56.0 (1-OCH3), and a nitrile carbon signal at δC 118.3 (6-CN). These data indicate that the structure of 8 comprised two indole rings connected into the core structure of indolocarbazole , and/or lipopolysaccharide (LPS). The inducible NOS gene is subsequently expressed, and NO is produced ,23. In c5) and maeruabisindoles B (7) and C (8) showed a nitric oxide inhibition activity stronger than indomethacin  and B (2) were as strong as indomethacin against nitric oxide production. Having an acetonitrile substituent at C-3, maeruanitriles A (1) and B (2) demonstrated an NO inhibitory activity similar to indole-3-acetonitrile compounds isolated from Isatis indigotica roots ethanimidothiolate. For bisindole alkaloids, maeruabisindole B (7) showed strong NO inhibitory activity at IC50 of 31.1 ± 1.8 μM, whereas maeruabisindole A (6) was too cytotoxic. Similar bisindoles with an azete ring, i.e., isatindigosides F and G, also exhibited NO inhibitory activity with IC50 of 70.3 ± 6.9 and 67.3 ± 5.5 μM, respectively carbazole derivative. Bisindole compounds from Isatis tinctoria roots  have bedigotica .TM iS50 FT-IR spectrometer  and Perkin Elmer FT-IR 1760X spectrometer . HR-ESI spectra were measured on a Bruker APEX II mass spectrometer  and Agilent 6540 UHD Accurate-Mass Q-TOF mass spectrometer . NMR was recorded on a Bruker Advance NEO 400 MHz NMR spectrometer  and a Varian VNMRS-600 spectrometer . Medium-performance liquid chromatography (MPLC) and flash column chromatography (Flash CC) were performed by using a PuriFlash® XS 420 , Sepacore® purification system , or a ceramic pump . Silica gel 60 , LiChroprep® RP-18 , and SephadexTM LH-20  were used as a stationary phase material for column chromatography (CC). Organic solvents  were redistilled prior to their use as a mobile phase composition.UV spectra were obtained by a Milton Roy Spectronic 3000 Array spectrophotometer . Optical rotations were measured on a JASCO P-2000 polarimeter . CD spectra were recorded using a Jasco J-815 CD spectrophotometer . IR spectra were recorded on a NicoletMaerua siamensis (Kurz) Pax. were collected in the Sikhio district, Nakhon Ratchasima province, and identified by one of the authors (C.C.) according to the Botanical Garden Organization (BGO) plant database, Thailand. A voucher specimen (CC-MS-0419) has been deposited at the herbarium of the Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Thailand.The roots of Dried roots were cut into small pieces and extracted with EtOAc (3 × 30 L) to yield the EtOAc extract (29.3 g). The marc was further extracted with MeOH (3 × 30 L) to obtain the MeOH extract (350 g). The MeOH extract was mixed with distilled water and partitioned with n-butanol (3 × 5 L) to give an n-butanol extract (50.8 g).n-hexane (C6H12)/acetone (Me2CO) from 15:1 to 6:1 to gain nine fractions (Fr.1-9). Fr.6 (274.50 mg) was loaded on MPLC  to yield ten fractions (Fr. 6-1 to 6-10). Fr. 6-5 (38.2 mg) was further separated by MPLC  to obtain ten fractions (Fr. 6-5-1 to 6-5-10). Fr. 6-5-2 (8 mg) was further purified by preparative RP-18 TLC, which was developed with DI water/MeCN (1:4) to afford compound 3 (2.4 mg). Fr. 8 (2.14 g) was subjected to MPLC  to yield twelve fractions (Fr. 8-1 to 8-12). Fr. 8-2 (66.6 mg) before being separated by MPLC  to gain twelve fractions (Fr. 8-2-1 to 8-2-12), and then thrice separated by the mobile phase ratio of 4:1:1 to 2:1:1 to give compound 1 (2.4 mg) and compound 2 (1.4 mg). Fr. 8-6 (31.9 mg) was subjected to MPLC  to yield sixteen fractions (Fr. 8-6-1 to 8-6-16). Fr. 8-6-13 was purified by MPLC  to gain five fractions (Fr. 8-6-13-1 to 8-6-13-5). Fr. 8-6-13-3 was separated by CC  to gain compound 4 (1.0 mg). Fr. 8-9 (158.00 mg) was purified by MPLC , and Sephadex LH-20 (MeOH) to give compound 5 (3.5 mg).The EtOAc extract was separated by FCC using silica gel (Si) as the stationary phase, and the mobile phase comprised 2Cl2/Me2CO (10:1 to 1:1)] to obtain sixteen fractions (Fr. C-1 to C-16). Fr. C1-7 (1.56 g), before it was separated by MPLC  to obtain 10 fractions (Fr. C-1-7-1 to C-1-7-10). Fr. C-1-7-5 (283.5 mg) was purified repeatedly by MPLC and Sephadex LH-20 (MeOH) to obtain compound 7 (1.1 mg). Fr. C-15 (3.58 g) was separated by MPLC  into five fractions (Fr. C15-1 to C15-5). Fr. C15-4 (8.6 mg) was purified by Sephadex LH-20 (MeOH) to obtain compound 6 (2.8 mg). Fr. E3 (2.31g) was loaded on MPLC  to yield compound 8 (3.5 mg).Butanol extract (50.8 g) was separated by Sephadex LH-20 (MeOH) to obtain five fractions (Fr. A-E). Fr. C (5.7 g) was subjected to MPLC max (MeOH) nm (log ε): 218 (5.49), 268 (4.66), 322 (4.04); CD  nm (mdeg): 240 (+8.20), 265.5 (−9.64), 283.0 (0.07); Rh2(OCOCF3)4-induced CD (CH2Cl2) nm (Mol CD): 350.0 (+1.51); IR (ATR) νmax: 3291, 2256, 1789, 1629, 1462, 1342, 1722 cm−1; 1H and 13C-NMR data (acetone-d6): see m/z 241.0585 .Compound 2 (maeruanitrile B): reddish brown amorphous; UV λmax (MeOH) nm (log ε): 228 (4.85), 300 (4.39), 342 (3.77); IR (ATR) νmax: 3163, 2924, 2850, 2360, 2249, 1626, 1451, 1298, 1208, 1160, 1022 cm−1; 1H and 13C-NMR data (CD3OD): see m/z 271.05112 .Compound 3 (maeroxime A): reddish brown amorphous; UV λmax (MeOH) nm (log ε): 212 (5.12), 217 (5.26), 225 (4.80), 269 (4.15); IR (ATR) νmax: 3369, 2923, 2852, 1714, 1627,1501, 1457, 1337, 1198, 1093 cm−1; 1H and 13C-NMR data (DMSO-d6): see m/z 265.0999 .Compound 4 (maeroxime B): orange-brown amorphous; UV λmax (MeOH) nm (log ε): 212 (4.69), 280 (3.25), 314 (4.18); IR (ATR) νmax: 3283, 2924, 2851, 1718, 1617, 1521, 1421, 1241, 1197, 1074, 1032 cm−1; 1H and 13C-NMR data (DMSO-d6): see m/z 279.0780 .Compound 5 (maeroxime C): yellow amorphous; UV λmax (MeOH) nm (log ε): 208 (4.80), 272 (4.47), 316 (4.15); IR (ATR) νmax: 3306, 2956, 2924, 2854, 1729, 1618, 1461, 1378, 1283, 1074, 1037 cm−1; 1H and 13C-NMR data : see m/z 279.0782 .Compound 6 (maeruabisindole A): pale green amorphous; UV λmax (MeOH) nm (log ε): 210 (3.72), 270 (4.03), 315 (3.95), 355 (3.49), 365 (3.44); IR (ATR) νmax: 3384, 2919, 2850, 1625, 1559, 1508, 1458, 1420, 1325, 1286, 1246, 1228, 1196, 1162, 1089, 1029 cm−1; 1H and 13C-NMR data (DMSO-d6): see m/z 390.1298 .Compound 7 (maeruabisindole B): pale green amorphous; UV λmax (MeOH) nm (log ε): 210 (4.47), 230 (4.15), 310 (3.93), 340 (3.31), 355 (3.31); IR (ATR) νmax: 3396, 2921, 2851, 1602, 1465, 1377, 1258, 1172, 1117, 1025 cm−1; 1H and 13C-NMR data (CD3OD): see m/z 406.1224 .Compound 8 (maeruabisindole C): dark green amorphous; UV λmax (MeOH) nm (log ε): 210 (4.07), 285(2.93), 355(2.21), 365(2.36); IR (ATR) νmax: 3397, 2921, 2851, 2212, 1595, 1464, 1395, 1260, 1121, 1041 cm−1; 1H and 13C-NMR data (acetone-d6): see m/z 326.0968 .Compound 1), the possible configurations were computed at a B3LYP/6-31G level. The ECD spectra were calculated using the time-dependent density functional theory (TD-DFT) method with a B3LYP functional and 6-311++G basis set. The geometry optimization and TD-DFT calculations were both performed with a polarizable continuum model (PCM) solvation model using methanol (MeOH). The rotary strengths of 70 excited states were calculated. All calculations were performed using the Gaussian16 program package  were used to treat cells.4 cells/well in a 96-well plate. The cells were pre-treated with various concentrations of samples for 24 h. Cells were induced with 100 ng/mL LPS for 24 h. The culture supernatant was collected for NO production analysis, and cells were further examined for their viability.Raw264.7 cells were seeded at a density of 5 × 10The cell viability was measured by the MTT assay . An MTT The NO production was measured using a Griess reagent kit  . Ninety A: The concentration of the nitric oxide in the cells induced by LPS and with sample pre-treatment .B: The concentration of nitric oxide in the cells induced by LPS without sample pre-treatment .50) of the inhibitory concentration calculated using GraphPad Prism 9.The NO inhibitory activity was expressed as half the maximum  from at least three independent experiments. The mean differences of the compounds vs. indomethacin (the positive control) were evaluated by one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test . Statistical significance was defined as p < 0.05, p < 0.005, p < 0.001, and p < 0.0001.The ICMaerua siamensis (Capparaceae). Their structures were elucidated based on spectroscopic methods and computational analysis. Among them, maeruanitriles A (1) and B (2), maeroxime C (5), and maeruabisindoles B (7) and C (8) inhibited nitric oxide production in LPS-induced RAW 264.7 cells. This finding supports the traditional use of M. siamensis roots for analgesic and anti-inflammatory purposes in traditional Thai medicine.Eight new indole alkaloids were isolated from the roots of


The Murcia Region (south-eastern Iberian Peninsula) has a great diversity of Lepidopteran fauna, as a zoogeographical crossroads and biodiversity hotspot with more than 850 butterflies and moth species recorded.Pyralidae moths  is synthesised. In total, three subfamilies, 67 genera and 142 species have been recorded and these are listed, along with their collection, literature references and biological data, including chorotype, voltinism and the flight period in the study area. The subfamilies are Galleriinae, Phycitinae and Pyralinae. Seventy-three species are newly recorded, sixty-two species are confirmed from literature and only seven species have not been observed for the Murcia Region.In the present paper, based on an examination of museum specimens, published records and new samples, a comprehensive and critical species list of  Pyralidae, belonging to the superfamily Pyraloidea, are mainly nocturnal micromoths (Microlepidoptera) with an estimated 6,000 named species worldwide, of which the European fauna is represented by ca. 470 species , Acrobasisobliqua (Zeller), Amphithrixsublineatella (Staudinger), Epischniaillotella Zeller, Homoeosomanebulella (Denis & Schiffermüller), Homoeosomasinuella (Fabricius) and Pterothrixidiarufella (Duponchel) and Aphomiamurciella from Sierra Espuña and recorded Alophiacombustella (Herrich-Schäffer), Asalebriaflorella (Mann), Assaraconicolella (Constant), Dioryctriasylvestrella (Ratzeburg), Ephestiawelseriella (Zeller), Euzopheralunulella (O. Costa), Psorosamediterranella Amsel, Aglossabrabanti Ragonot, Lorymaegregialis (Herrich-Schaffer) and Stemmatophoravulpecalis Ragonot. Subsequently, Aphomiamurciella Zerny and recorded Eurhodopecruentella (Duponchel), Stemmatophoragadesialis Ragonot and Synaphediffidalis (Guenée). Asalebriapseudoflorella from Sierra Espuña, considered as a subspecies of A.ferruginella (Zerny) at the present time.The tiamiegi  and, latraphalis  was alsoEpischniaperoni, currently synonymised with Epischniaasteris (Staudinger) and E.prodromella (Hübner) and E.illotella (Zeller) were recorded for the first time, while Cryptoblabesgnidiella (Millière) and Pempeliapalumbella (Denis & Schiffermüller) amongst others, although Coenochroaablutella (Zeller) and Homoeosomanimbella (Duponchel)  have not been collected during the present study. Subsequently, Archiephestiamurciella considered as a synonym with Archiephestiaadpiscinella (Chrétien) and Ancylosisuncinatella (Ragonot) and Hypotiamiegi (Ragonot).Later, Merulempistaturturella (Zeller) (cited as M.numidella (Ragonot)), Subsequent contributions are those of PseudoinsalebriaibericaGymnancylahillneriellaCeutolophaisidis (Zeller) has been recorded by Gymnancylahornigii (Lederer) by Psorosaferrugatella (Turati) and Epischniaampliatella (Heinemann) by Recently, The Region of Murcia has a great diversity of Lepidopteran fauna, as a zoogeographical crossroads and biodiversity hotspot, with more than 850 butterfly and moth species (Considering various bioclimatic approaches relative to temperature (thermotypes) and rainfall (ombrotypes), four different bioclimatic areas can be recognised according to Pyralidae family from the Murcia Region (south-eastern Iberian Peninsula) for taxonomists providing data about distribution, chorology, phenology and voltinism.This present checklist is intended to update the recorded species and to facilitate access to the most recent data on the H)-furanone). Adult specimens were examined externally and the genitalia structures were dissected using standard procedures  with minPyralidae collected by the authors until the end of 2021, along with the material deposited in the private collections of J.A. de la Calle, F. Lencina, F. Albert and F. Arcas. It also includes all of those records previously referenced in the bibliography.The list contains all species of Black and actinic (6 and 15 W) Heath traps, 125 W Robinson traps, 125 W mercury vapour traps and 4 W LED light traps were used for nocturnal sampling. Catches taken during daytime and in the urban environment (street lighting) are also included. All these sampling points are located within the study area and, especially, in the natural protected areas like the mountainous Parks of Sierra Espuña, Sierra de la Pila, El Valle and Carrascoy etc. and the coastal Parks of Calblanque, Monte de las Cenizas and Peña del Águila, Salinas and Arenales de San Pedro del Pinatar, etc.Pyralidae by The subfamilies are systematically ordered and identified, based on the most recent classification of B1365F49-907C-52FF-BAA3-F17166C467DF5AE84485-CB19-5C81-B70F-C6364D0CCDFA0A6192CD-1A75-51D2-8019-C5557A4D4004CosmopolitanBiological data: Polyvoltine. Flight period: VI, X. First record in Murcia Region.FAE03D7A-7152-5811-AA48-B745BA7C5155CosmopolitanBiological data: Polyvoltine. Flight period: VII-X. First record in Murcia Region.4B46AEEC-5E64-54CC-8086-65B991E76888Atlanto-MediterraneanBiological data: Polyvoltine. Flight period: I, VII-IX. First record in Murcia Region.35FEB8CA-0719-5C9E-AD36-4447AB07C1F4HolarcticReferences: Zerny, 1914F478453F-F3FF-594E-AF22-D763E6F797B9EndemicReferences: A0ECD27D-0594-517C-8C0D-BC2F7BCB6C26Mediterranean-AsiaticBiological data: Polyvoltine. Flight period: VII-VIII. First record in Murcia Region.F0FBDD9A-7EB6-524E-A063-D49BAC5FD1E4EurasiaticReferences: 2272FDA4-F131-5F10-8849-4F557F45B70FCosmopolitanBiological data: Bivoltine. Flight period: IV-XI. First record in Murcia Region.17B0D194-E067-5F05-B5E7-6D810A60FFC40E2AD0D4-FCB2-5ADE-AC67-4C414D6ADA8DTropicalReferences: 7DB78095-D32B-5AFE-B9DA-0C6EFE0C89CAMediterranean-AsiaticBiological data: Bivoltine. Flight period: IX. First record in Murcia Region.789DBD4B-BF8A-56C0-931D-B039BA177DE1Atlanto-MediterraneanBiological data: Univoltine. Flight period: IX-X. First record in Murcia Region.37480A67-6B70-5298-8FAB-FD6749D6665EMediterranean-AsiaticReferences: Chrétien, 1911BB1A3E32-4C4E-5BF1-B827-6123A110CF23Atlanto-MediterraneanBiological data: Univoltine. Flight period: VI. First record in Murcia Region.B7CFDD97-DB5F-5284-84EF-17BD094BA727Mediterranean-AsiaticReferences: B8B718BA-BECE-52C0-9A33-3F0DB59E18A9Atlanto-MediterraneanReferences: A8CD58EE-AEEC-5D4E-899F-590124D2AD11Atlanto-MediterraneanBiological data: Bivoltine. Flight period: IX. First record in Murcia Region.BA4E0A35-5AB3-5E78-9D23-2F6BEF8F163CMediterranean-AsiaticReferences: 639A12E0-94E7-5DAE-AB2A-47F9D42E0C91Mediterranean-AsiaticBiological data: Bivoltine. Flight period: II-VIII. First record in Murcia Region.Slamka, Ylla & Macià, 2018ED634723-CBA6-5519-80D2-3DFBC22A0BECEndemicReferences: B8DB76BF-9753-509C-8D68-11DBBAB26348EurasiaticReferences: 01EB8731-84CA-5E98-9785-220A7855C6B1Mediterranean-AsiaticReferences: DCFB5389-24DD-59BF-B94A-DDDB530F1AD0Mediterranean-AsiaticReferences: Turati, 192445B87397-2A29-5DAE-BA94-DE6EE2B868EDMediterranean-AsiaticReferences: Amsel, 1953A23EF6F8-C1FF-585E-B163-8B570D4C8432Mediterranean-AsiaticReferences: DEF67F81-160C-5154-B943-B1BABB95439EMediterranean-AsiaticBiological data: Bivoltine. Flight period: III-X. First record in Murcia Region.EC48BB17-AF3D-5F45-88F1-56BD78046182EurasiaticBiological data: Bivoltine. Flight period: V. First record in Murcia Region.8B0E1772-8621-5647-84F1-C97DD6E03FD2Atlanto-MediterraneanBiological data: Polyvoltine. Flight period: V-IX. First record in Murcia Region.03B91912-4290-5897-9D35-3D4D7218ED7ATropicalReferences: 30CF38D9-89D7-59BF-AC87-37BFED1870B4Mediterranean-AsiaticBiological data: Bivoltine. Flight period: II, VI, IX-X. First record in Murcia Region.F5F78982-1E11-51BF-9A49-223B90362ACBMediterranean-AsiaticReferences: DAE30773-A43D-5939-9BDB-A64D799C84A0Mediterranean-AsiaticReferences: 444EA283-B578-5C1A-A3B1-677052C800A6TropicalReferences: 4D024112-5A2C-53E6-BA85-CDD8EB70D86EPalaearcticBiological data: Polyvoltine. Flight period: V, VIII-IX. First record in Murcia Region.97D1F5DB-69DE-5CFA-AACE-A87B61EFB115CosmopolitanReferences: 57B71507-332C-544C-89AC-F48FCFA032A2HolarcticBiological data: Bivoltine. Flight period: II-V. First record in Murcia Region.9F7C46A4-B250-562D-9E42-F3DEA4D2A74EMediterranean-AsiaticReferences: BAB965E5-0DD5-5061-AD01-F7D13ACC4A94Atlanto-MediterraneanBiological data: Bivoltine. Flight period: II-IV, VII. First record in Murcia Region.Ragonot, 1893C2EC597B-E2DC-5140-BAB6-251FCA1C3F0BMediterranean-AsiaticReferences: FAAFAC44-3054-5AAA-998D-B5B3322C2AFEEurasiaticReferences: Zeller, 1839D6B7DA59-09FC-52EE-9D21-C75CF6CD79CCMediterranean-AsiaticBiological data: Bivoltine. Flight period: V-VII. First record in Murcia Region.621D0868-B6DD-5885-8399-CF0F5D50CD70Atlanto-MediterraneanBiological data: Univoltine. Flight period: VIII. First record in Murcia Region.37157B27-C710-5D2C-917E-193FCB97E507Mediterranean-AsiaticReferences: 7DD749AF-7E8E-53CF-820D-3CA0C426D770EurasiaticBiological data: Bivoltine. Flight period: III-VIII. First record in Murcia Region.071A0E06-D24C-539E-BCDA-01FF970A46D2EurasiaticBiological data: Univoltine. Flight period: IX. First record in Murcia Region.8356E7C7-E831-5565-8639-BE4CAF14E960TropicalBiological data: Polyvoltine. Flight period: IV, IX-X. First record in Murcia Region.553B882A-068F-533E-A049-5B974C370973TropicalReferences: 121A236C-9DA7-5673-8D78-F2914C7DA382EurasiaticBiological data: Bivoltine. Flight period: V, IX. First record in Murcia Region.E8274CFF-3F96-57EC-B048-73715FB8B2C3EurasiaticReferences: EAB0DF15-3E87-5BC2-B24A-DAC1AF8FC30CMediterranean-AsiaticReferences: FD038B84-20BF-5677-9FE2-8AB88C2D383BMediterranean-AsiaticReferences: A6C3182F-AAEA-5857-AEBF-9A0B9EDDEDE3Mediterranean-AsiaticReferences: Zeller, 18390665BCA0-0F7C-5054-B029-1D369B519051Mediterranean-AsiaticReferences: Staudinger, 187117ABCAFD-A12A-5E16-8FBD-80056A8584BFAtlanto-MediterraneanReferences: 12381DC0-DDF0-5769-BE22-1EEBB0AE5BD3EurasiaticReferences: 22553273-51F0-5FD8-9AD4-1DEE10388E0DEurasiaticBiological data: Bivoltine. Flight period: VIII. First record in Murcia Region.C18D44B3-36B6-598D-B2D7-234C33E8651CMediterranean-AsiaticReferences: Zeller, 18486154404B-13EA-5737-AC83-82CF1BBAB1D7Mediterranean-AsiaticBiological data: Univoltine. Flight period: VII-X. First record in Murcia Region.1119A135-1FAF-585D-81EE-AC5596566C31Mediterranean-AsiaticReferences: D4119D31-DBBC-5118-94AF-6599758A5BB5Atlanto-MediterraneanBiological data: Univoltine. Flight period: VIII-X. First record in Murcia Region.Zeller, 1848E927AFD2-E62A-55BD-BB03-F0EB56E7FB4DEurasiaticBiological data: Univoltine. Flight period: VIII. First record in Murcia Region.7DC9A89A-9D8A-50CD-B26F-FF9D5785F071Mediterranean-AsiaticBiological data: Univoltine. Flight period: VI-VIII. First record in Murcia Region.ED605580-82FA-5C63-9DAE-153ECBB7BCE6Mediterranean-AsiaticReferences: B2819911-F4FD-5AA1-B837-13ED1E1A67ADEurasiaticBiological data: Univoltine. Flight period: VI. First record in Murcia Region.C7D99C8C-09DB-5913-BBE3-C724F32F0BCFHolarcticBiological data: Univoltine. Flight period: VIII. First record in Murcia Region.6C783D22-AAB1-5DA4-8219-4EAD690FB458CosmopolitanBiological data: Polyvoltine. Flight period: IV-V, VII-X. First record in Murcia Region.A2E0146B-9D8C-5F52-9DC1-660E6E94454DEurasiaticBiological data: Univoltine. Flight period: VI-VII. First record in Murcia Region.3AFE583D-0F31-5685-9125-FC0897C3BD1AMediterranean-AsiaticReferences: FCF630A5-56BF-544C-93AE-ED53D6A90766EurasiaticBiological data: Univoltine. Flight period: IV. First record in Murcia Region.Mann, 1861C6C2217F-E663-5DF3-A76D-060B33D8E2BDMediterranean-AsiaticBiological data: Univoltine. Flight period: III-V. First record in Murcia Region.E8828C6A-2B62-522E-B6E4-F1F80046BE6BAtlanto-MediterraneanReferences: B1F701B8-7A72-5A82-ABFE-F91976981FD6Mediterranean-AsiaticReferences: Ragonot, 1887899F4A61-55B0-5E9A-8B01-1BA709B82D7EEurasiaticBiological data: Univoltine. Flight period: IV, VIII. First record in Murcia Region.42F6CA84-5A08-5643-AA28-995718573BD4Mediterranean-AsiaticBiological data: Univoltine. Flight period: VIII-IX. First record in Murcia Region.E1969B6F-A5BB-51D8-8298-B892D50F05AEAtlanto-MediterraneanReferences: Gastón & Vives, 20182C69656B-1D31-590A-88A9-BEABA63AF15FEndemicReferences: 0C044E3F-1B8E-5ACC-BEAD-C447BADDF7E8Mediterranean-AsiaticReferences: C38D623B-B870-500B-9BD0-0F472571C307Mediterranean-AsiaticBiological data: Bivoltine. Flight period: IX. First record in Murcia Region.2FA61D57-6DC3-559A-9B3E-7E96FE849C80Mediterranean-AsiaticReferences: AC1E1019-CD19-5DF7-B299-A1DD0B34908CEurasiaticBiological data: Bivoltine. Flight period: V, VII-X. First record in Murcia Region.49E66EA3-8D08-5F6D-87B7-30CF57572EF2Mediterranean-AsiaticReferences: 612758CD-8ED3-5E95-93F1-F3AC38D005FAMediterranean-AsiaticBiological data: Polyvoltine. Flight period: V, X. First record in Murcia Region.9435B0DE-F5F2-5346-84D3-E6AA1CA64A9DMediterranean-AsiaticBiological data: Polyvoltine. Flight period: II-V, X-XI. First record in Murcia Region.D08B34FF-622C-5841-804B-0A5FC857910EMediterranean-AsiaticReferences: 5E998E23-CC38-54DE-851A-F71CCEE43C11EurasiaticReferences: B237244F-7F60-504F-9B95-6B017E2AFC32Atlanto-MediterraneanReferences: Staudinger, 1870223DFB8E-A4CD-5880-93E0-F0E23A250BE3EurasiaticBiological data: Bivoltine. Flight period: V. First record in Murcia Region.F1FCC22E-0E6F-599A-9BD4-79F9FEFD817DMediterranean-AsiaticBiological data: Bivoltine. Flight period: VII. First record in Murcia Region.451A1832-1CE3-5FB3-A624-8162F6297028EurasiaticReferences: AE09AE31-4E07-53C9-9729-8A94853A5C7DMediterranean-AsiaticBiological data: Polyvoltine. Flight period: IV-V, IX. First record in Murcia Region.A52C6515-2E0B-593D-AB41-DFC9E2F95847Mediterranean-AsiaticBiological data: Bivoltine. Flight period: IV-V, X. First record in Murcia Region.65D5628B-8ED5-57F7-8A4A-6CD3C00CE394Mediterranean-AsiaticBiological data: Bivoltine. Flight period: V, VIII. First record in Murcia Region.E525C474-85BE-598E-9B64-B6039E2CA510EurasiaticBiological data: Bivoltine. Flight period: IV, VII-XI. First record in Murcia Region.Ragonot, 1901BC9CF625-1B14-5D15-9AAE-A02C058D4952Atlanto-MediterraneanBiological data: Bivoltine. Flight period: IV-V, VII-X. First record in Murcia Region.83E72189-63C0-597B-BBEB-8688189CED0DMediterranean-AsiaticReferences: 66B75B88-CF88-5CA9-86F4-FF96DEB567C1EurasiaticReferences: 08652440-6142-5989-B1BB-E725FA0FC88CEurasiaticReferences: 401CA697-754D-5C6C-B4D2-5613D03BFF2FEurasiaticReferences: CCADBE18-5A66-5639-BC06-103F21114234Atlanto-MediterraneanBiological data: Univoltine. Flight period: V. First record in Murcia Region.F19A08EC-DD99-530A-9386-9593213B9CFCEurasiaticBiological data: Univoltine. Flight period: III-V. First record in Murcia Region.3D9DC5D5-3029-524E-908A-8D144AD85B78EurasiaticReferences: 21DAB015-1641-5DFC-BABD-DAB70E2865EAEurasiaticBiological data: Univoltine. Flight period: II, VI, IX-X. First record in Murcia Region.835D6C61-9107-5B04-8EC4-F21F370B2F38Atlanto-MediterraneanReferences: 9A62225D-E44F-507C-929C-DBCE17D4E424HolarcticReferences: 18579E9D-E272-57FD-8EA4-C1A6AF7E7E44Mediterranean-AsiaticBiological data: Bivoltine. Flight period: V, VII-VIII. First record in Murcia Region.F53CC5F6-082D-5C8D-B4AD-CF41FB2127ACMediterranean-AsiaticReferences: 93C34B57-5F2E-56B0-BE47-0D17525A711FCosmopolitanBiological data: Polyvoltine. Flight period: V-IX. First record in Murcia Region.Ragonot, 190117EDE3B5-6E48-554A-A6AB-E6008D26B734Mediterranean-AsiaticReferences: Staudinger, 185910B29580-86BE-5922-81B3-A0869A9A23ADAtlanto-MediterraneanBiological data: Polyvoltine. Flight period: V-VI, IX. First record in Murcia Region.Richards & Thomson, 1832782FE72B-F955-52CD-AE43-FE2B3CD0A86FEurasiaticBiological data: Polyvoltine. Flight period: IX. First record in Murcia Region.2E44A65E-ADA2-5BC7-ACA8-AB7E74B05679CosmopolitanBiological data: Polyvoltine. Flight period: VI, XII. First record in Murcia Region.06C726D2-5E4D-543E-8B87-CCE1A805690DMediterranean-AsiaticReferences: 54C0C01A-9F13-5843-8ECF-EEAB62485D8CMediterranean-AsiaticBiological data: Bivoltine. Flight period: VI. First record in Murcia Region.7FB071BC-ACBC-597D-A028-3C86FCA41171CosmopolitanBiological data: Polyvoltine. Flight period: V-X. First record in Murcia Region.EC08A98A-E792-5427-A1FD-F9D8D71178D6CosmopolitanBiological data: Polyvoltine. Flight period: VIII. First record in Murcia Region.0F2C5CF0-B754-54C4-AB24-75DE4ABD3E0AMediterranean-AsiaticBiological data: Polyvoltine. Flight period: IX. First record in Murcia Region.59A5EF68-B6C1-5580-B840-64EDCE2CA7455C010146-13D2-54D8-95D0-5C48F8B4B3FEEurasiaticBiological data: Bivoltine. Flight period: VI-IX. First record in Murcia Region.1D92F0A5-F734-516D-92A8-F38EB84909BDMediterranean-AsiaticReferences: Lederer, 1858B7152D2B-36AE-5CDD-82EB-381448CAE0F2Mediterranean-AsiaticReferences: 526F0322-E337-54A1-936E-9A1832A29231Mediterranean-AsiaticBiological data: Bivoltine. Flight period: IV-V, VII-VIII. First record in Murcia Region.7BAEEF1D-E0E5-5EDF-BE07-1DC0A51B4751EndemicReferences: 188E575D-F81C-5C06-9531-3FFA7009B628EndemicReferences: 1A268E1F-3F16-5DCF-B825-32DA85C1F013Mediterranean-AsiaticBiological data: Univoltine. Flight period: V-VI. First record in Murcia Region.722AE73C-61C0-5E1A-BE7C-2DF11AA2829AAtlanto-MediterraneanReferences: 1C57E773-BE72-5AEF-9449-29329762EB88Atlanto-MediterraneanBiological data: Univoltine. Flight period: VI-VIII. First record in Murcia Region.943D861D-FFCD-5B5A-A556-E285BB799642EurasiaticReferences: D0AE5B16-E21E-5621-9F72-0FE6A2F0C16ECosmopolitanBiological data: Polyvoltine. Flight period: III-IV, VI-XI. First record in Murcia Region.AF4C1B6F-0186-5536-B584-3453255CC7E0CosmopolitanBiological data: Univoltine. Flight period: IV, IX-X. First record in Murcia Region.EED07796-5AC4-5E5B-87AD-06504E2800FECosmopolitanBiological data: Univoltine. Flight period: VI. First record in Murcia Region.Ragonot, 18846E7BDC6B-1D6A-5E92-8793-8D9F124379D2Atlanto-MediterraneanReferences: Ylla, Šumpich, Gastón, Huertas & Macià, 20177F3D22C5-799C-5056-BB12-FDABC58D4AABEndemicBiological data: Bivoltine. Flight period: IV-V. First record in Murcia Region.05EA0707-93D0-5356-B4FE-0761625F4A65Mediterranean-AsiaticReferences: Ragonot, 1882F1455122-10BF-533E-ABDB-3FC8AFE2E856Atlanto-MediterraneanReferences: Ragonot, 18918FCB2901-4EC8-5DF6-B908-7BD6729F5CDEAtlanto-MediterraneanReferences: D88A5953-3505-572C-A7CF-06917243F870Mediterranean-AsiaticBiological data: Univoltine. Flight period: VI-VIII. First record in Murcia Region.9F32FB2F-4E60-519E-9E8E-A63A7E6FB9C1Atlanto-MediterraneanBiological data: Univoltine. Flight period: IX. First record in Murcia Region.45609B3D-DD3D-5DEF-82E1-84B62A4FAAE6Mediterranean-AsiaticBiological data: Univoltine. Flight period: VIII-X. First record in Murcia Region.88F2FAE2-BB33-5D57-86F0-D285135C4095Atlanto-MediterraneanBiological data: Univoltine. Flight period: VII-X. First record in Murcia Region.14B8F838-54EA-5C38-95C2-416D1650A393Atlanto-MediterraneanBiological data: Polyvoltine. Flight period: IV-XI. First record in Murcia Region.A01BE3E4-1D6E-56C4-8E5F-4E6E50A47885Mediterranean-AsiaticReferences: BD40F19E-7A3C-5C7A-AAEE-158609647A9CMediterranean-AsiaticReferences: 249A0195-DFBA-54D0-ADA5-5BE4B614308CHolarcticBiological data: Bivoltine. Flight period: VI-X. First record in Murcia Region.3A541C97-25A4-5D39-BBF9-F4A55BE7BBF8Mediterranean-AsiaticBiological data: Univoltine. Flight period: IX-X. First record in Murcia Region.Galleriinae (8 species), Phycitinae (107 species) and Pyralinae (27 species). Seventy-three new records (51%) from the Murcia Region are added to its Lepidopteran fauna.The list includes 142 species in 67 genera and three subfamilies: Phycitinae, comprises 77.6% of all genera and 75.3% of all species, while Pyralinae comprise 14.9% and 19.1% and Galleriinae with 7.5% and 5.6%, respectively , whilst the Iberian Pyralidae fauna comprise 262 extant species  showed a low dominance value of 0.96  and a Chao1 estimate of total species richness amongst 140 species  to 165.5 species  which is close to 142 species studied and foresees the addition of new species in the future.Pyralidae diversity in the Murcia Region seem relatively rich when compared to those in other Iberian Regions and with the whole of the Iberian Peninsula, as for instance, similar Iberian Regions extensively surveyed like Catalonia , Acrobasis , Phycitoides and Stemmatophora , Pempelia, Ephestia and Hypotia  and Aphomia, Cadra, Dioryctria, Epischinia, Aglossa and Synaphe . The majority of genera (12) are species-poor (2-3 species) or known in the Murcia Region from a single species (42 genera).The most species-rich Pyralidae fauna with 116 species recorded, followed by the Mesomediterranean area with 73 species, while the Supra- and Oromediteranean areas appear to be less diverse with 39 species  are more frequent than the Atlanto-Mediterranean elements (17.6%). On the other hand, the elements of wide distribution, such as the Eurasiatic, Holarctic and Palaearctic (24.6%), are the most common in the mountainous biotopes of the centre and north of the study area, while the tropical and cosmopolitan species (12.0%) have their origin mainly in Africa. The presence of opportunistic species is due to the agricultural crop fields that dominate part of the Murcian territory. The Iberian endemisms are represented with six species (4.2%).Chorological analysis for the family Phyctinae recorded species feed on plant species belonging to the Asteraceae, Cistaceae, Fabaceae, Pinaceae, Fagaceae, Oleaceae, Chenopodiaceae and Lamiaceae families, amongst others. The most particular cases are those related to the genera Cadra, Ephestia and Plodia which are pests on stored products. Some species, such as Apomyeolisceratoniae (Zeller), Cryptoblabesgnidiella (Millière), Etiellazinckeniella (Treitschke) and Euzopherapinguis (Haworth) must be controlled since they are agricultural crop pests. Many of the species of the subfamily Galleriinae live in bee, bumblebee or wasp nests as well as on plant detritus. Others, such as Aphomiasabella (Hampson) and Cathayiainsularum (Speidel & Schmitz), are parasites of palm trees (Phoenix spp.). In relation to the subfamily Pyralinae, most of the species feed on plant and animal detritus with Pyralisfarinalis (Linnaeus) being also a particular pest on cereal flour. Finally, the food source and diet of 27.4% of species are unknown, so it will be necessary to carry out complementary studies for further biological understanding.Regarding the biology of the species, the environmental conditions of the study area, which affect the availability of trophic resources for reproduction, suggest that most of the species are bivoltins (40.8%) and univoltins (38.0%), while the rest are polyvoltins (21.1%). Most of the Epischniamuscidella Ragonot, cited in Hypotiasyrtalis (Ragonot) also cited in Psorosanucleolella (Möschler) cited in Tephriscyriella (Erschoff), cited from Sierra Espuña mountains in T.ochreella Ragonot in Laristaniaalbipunctella (Chrétien), cited in Pseudoinsalebriaiberica Slamka, Ylla & Macià (Gymnancylahornigii (Lederer) was cited in Epischidiafulvostrigella (Eversmann) according to morphological and barcode data (unpublished data).Some taxa cited in the references have been removed from the checklist as  & Macià . FinallyPyralidae moth species in the Murcia Region was 69. Our study increases this number to a total of 142, based on an examination of museum specimens, published records and sampled individuals, accounting for 54.1% of all of the Iberian species known. This study presents an updated checklist of current Pyralidae moth species with their distribution and biological information for the Murcia Region in the south-eastern Iberian Peninsula.Prior to our investigation, the number of known Pyralidae taxa and localities where conservation is an important priority for policy-makers, conservation planners and for the management of insect diversity in Spain.This study serves as both a guide for collection in the poorly sampled south-western European continent and a comprehensive reference list with the We encourage lepidopterists holding additional data on systematically collected pyralids to produce an updated dataset. Additionally, new intensive surveys in adjacent regions are being conducted and unknown specimens are continuously identified to species level.


Stewartia sinensis (Chinese Stewartia), a ‘vulnerable’ species endemic to China. The new structures were elucidated on the basis of spectroscopic data, single crystal X-ray diffraction, and electronic circular dichroism (ECD) analyses. Stewartiacids A (1) and B (2) are isoursenol derivatives. Stewartiacid C (3) is a 12-oxo-γ-amyrin analogue. Both isoursenol and γ-amyrin derivatives are quite rare in nature. Stewartiacids D (4) and E (5) are 13,27-cycloursane-type compounds. Stewartiacids K (11) and L (12) are ursane-type triterpene and phenylpropanol adducts built through a 1,4-dioxane ring, which are also seldom reported in the literature. The rest are common C-23 carboxylated ursane-type (6–10) and oleanane-type  pentacyclic triterpenoids. Stewartiacids G (7), K (11), and L (12) showed moderate inhibitory effects against ATP-citrate lyase (ACL), with IC50 values of 12.5, 2.8, and 10.6 μM, respectively. Stewartiacid K (11) also exhibited moderate inhibition (IC50: 16.8 μM) of NF-κB.Fourteen previously undescribed naturally occurring C-23 carboxylated triterpenoids, stewartiacids A–N (1–14), were isolated and characterized from the twigs and leaves of the ornamental and medicinal plant  Stewartia sinensis. 11 and 12 showed inhibitory effects against ACL.Fourteen new C-23 carboxylated triterpenoids  were obtained from the ‘vulnerable’ plant  Stewartia, one member of the family Theaceae, comprises about twenty species worldwide and three quarters are distributed in China.1 Several species of Stewartia are not only grown as ornamental plants but also utilized as folk medicines.2 There are three well-known Stewartia species, which are mainly distributed in East Asia, i.e., Chinese Stewartia (S. sinensis Rehd. et Wils.), Korean Stewartia (S. koreana Nakai), and Japanese Stewartia (S. pseudocamellia Maxim.3). Phenolic compounds  and sterols have been reported from the leaves/twigs4–6 and stems7 of Korean Stewartia with anti-inflammatory,4,7 anti-photoaging,5 and antioxidative6 effects. Phenolic derivatives have also been obtained from the leaves/twigs,8 flowers,9 and stems10 of Japanese Stewartia with anti-melanogenic,8 antioxidative,8 and anti-allergic effects.9 It is uncertain that both S. pseudocamellia and S. koreana have been regarded as the same plant species by some Korean researchers.8 In addition, a few triterpenoids have been encountered from some Stewartia species.8The small plant genus 1,12 The population of S. sinensis is dwindling due to the destruction of vegetation and the poor ability of natural regeneration. This ornamental plant has been recorded as a ‘vulnerable’ species in the China Plant Red Data Book (CPRDB) published in 1992.13 It is also on the International Union for Conservation of Nature (IUCN) Red List.14 Its stems and roots have been used as a folk medicine to treat rheumatic and traumatic injury.13,15 However, S. sinensis has never been phytochemically and pharmacologically investigated. Recently, several statistical surveys unveiled that plant-originated natural products (NPs) and their intricate molecular frameworks still offer medicinal chemists a range of uncharted chemotypes for drug discovery,16 among which the rare and endangered plants (REPs) could serve as better sources than other botanic sources.17 A pioneering phylogenetic study of the terrestrial plants showed that NPs-derived drugs are mainly produced by specific drug-productive plant families, and most REPs species are distinctly in drug-producing families.17 Therefore, there is a tremendous need to prioritize protection and utilization of these REPs species at extinction risk. Since 2013, a special program has been launched to systematically identify bioactive/novel NPs from REPs. For examples, a number of structurally diverse sesquiterpenoids, triterpenoids, and diterpenoids with protein tyrosine phosphatase 1B (PTP1B) inhibitory and cytotoxic effects were isolated and characterized from the leaves and twigs of the endangered ornamental plants Michelia shiluensis,18Camellia crapnelliana ,19 and the shed trunk barks of the endangered plant Pinus dabeshanensis,20 respectively. In the course of our continuous interest in identifying bioactive compounds from REPs endemic to China,21 the chemical constituents of the EtOAc-soluble fraction of the 90% MeOH extract of the twigs and leaves of S. sinensis have been investigated. It is worth mentioning that the Chinese Stewartia is generally a 6–16 m tall tree,1,12 making the plant samples (the renewable leaves and twigs) easier to harvest. As a result, a total of 18 triterpenoids were isolated and characterized, including 14 previously undescribed C-23 carboxylated ones . Reported herein are their isolation, structure determination, and inhibitory effects against ATP-citrate lyase (ACL) and NF-κB.Chinese Stewartia is a flowering camellia plant endemic to central and eastern China. This relict species is highly regarded horticulturally for the combination of its beautiful, buff/tan colour bark—carried high up into the crown—and absolutely smooth with little or no flaking, cup-shaped/fragrant white single flowers in midsummer, elliptical papery leaves, and intensely red fall foliage.S. sinensis  was suspended in H2O and then partitioned successively with petroleum ether, EtOAc, and n-BuOH. The entire EtOAc-soluble fraction was subjected repeatedly to column chromatography (CC) over silica gel, MCI gel, Sephadex LH-20, and semi-preparative HPLC to afford eighteen triterpenoids (1–18). By comparing the observed and reported spectroscopic data and physicochemical properties, the known compounds 15–18 were identified as urs-12-en-3β-ol ,22 ursolic acid (16),23 olean-12-en-3β-ol ,22 and oleanic acid (18),23 respectively.The 90% MeOH extract of the twigs and leaves of 30H46O6, was determined by the HRESIMS at m/z 525.3182 [M + Na]+  and 13C NMR data , 0.86 , 1.03 , 1.09 , and 1.46 , and two secondary methyl groups at δH 0.92  and 0.98  were observed. In addition, signals for three oxymethine resonances at δH 3.51 , 3.96 , and 4.05 , and an olefinic proton at δH 5.68  were also readily distinguished. Its 13C NMR spectrum resolved 30 carbon resonances (δC 214.2 (C-12)], one carboxyl group [δC 181.3 (C-23)], and a double bond [δC 119.9 (C-15) and 157.1 (C-14)] corresponded to three IHDs, thus requiring 1 to feature a pentacyclic skeleton. The NMR data of 1 were found to closely related to those of 3β,6α-dihydroxy-urs-14-en-12-one,24 an unusual isoursenol derivative previously isolated from the roots of Rubia schumanniana. Unlike common ursane-type triterpenoids /H-20 (H3-30)/H-21/H-22, and the HMBC correlations of H-9/C-11 and H3-28/C-22  was obtained as colorless crystals from MeOH. Its molecular formula, CNMR data  with eigspectrum , resonansonances , which we.g., 15  and 16 , the Me-28/C-22 . In addiδC 181.3 .JH-2β/H-3 (10.1 Hz), JH-5/H-6β (11.0 Hz), JH-9/H-11 (11.0 Hz), and JH-21α/H-22 (10.6 Hz) indicated that H-3, H-5, H-9, H-11, and H-22 were in axial positions. In the ROESY spectrum, the correlations between H-3/H-5, H-5/H-9, H-9/H3-27, and H3-27/H3-30 revealed their cofacial relationship and were arbitrarily assigned as α-oriented. In turn, the ROE interactions of H3-24/H3-25, H3-25/H-11, H-11/H3-26, H-18/H-20, H-18/H-22, H-18/H3-28, and H-18/H3-29 indicated their β-orientation. Finally, a single-crystal X-ray diffraction experiment with Ga Kα radiation for 1 defined its  absolute configuration [absolute structure parameter: 0.19 (17)]. Based on the above findings, the structure of compound 1 was characterized as -3β,11α,22α-trihydroxy-12-oxo-isours-14-en-23-oic acid.The relative configuration of 1 was assigned by analysis of the key proton–proton coupling constants and ROESY interactions . The mag30H46O5, was deduced from the deprotonated ion at m/z 485.3665 [M − H]− in its HRESIMS and the 13C NMR data , δC 78.6 (C-22)] when compared with those of 1, suggested the disappearance of 22-OH group in 2. This was confirmed by the HMBC cross-peak from H3-28 to C-22 -3β,11α-dihydroxy-12-oxo-isours-14-en-23-oic acid.Stewartiacid B (2) was obtained as a white powder, and its molecular formula, CNMR data . The clo30H44O6 as deduced from the deprotonated ion peak at m/z 499.3069 [M − H]− in its HRESIMS and the 13C NMR data , 1.13 , 1.18 , 1.24 , and 1.33 ] two secondary methyl groups [δH 1.03  and 1.12 ], and two oxymethine resonances at δH 3.95  and 4.38 . A total of 30 carbon signals, including two ketone carbonyls at δC 209.8 (C-12) and 216.5 (C-22), a carboxyl carbon at δC 181.5 (C-23), two olefinic carbons at δC 141.9 (C-13) and 147.9 (C-18), and two oxymethine carbons at δC 76.0 (C-3) and 78.4 (C-11), were displayed in its 13C NMR spectrum. Further HMBC correlations from H3-27 to C-8/C-13/C-14/C-15, and from H3-28/H3-29 to C-18 revealed that compound 3 is a γ-amyrin analogue featuring an uncommon Δ13(18) double bond. The two oxymethine groups were anchored at C-3 and C-11 based on the HMBC correlations from H3-24 to C-3 and from H-9 to C-11 . The relative configuration of 3 was determined via analysing the coupling constants and ROESY data. The large J values between H-2β and H-3 (11.5 Hz) and between H-9 and H-11 (11.5 Hz) were indicative of their trans-diaxial relationship. The ROE correlations -en-23-oic acid. Finally, the absolute configuration  of 3 was unequivocally established by a single-crystal X-ray diffraction analysis [Flack parameter = −0.01 (19)] using Ga Kα radiation (Stewartiacid C (3) was obtained as colorless crystals from MeOH, with a molecular formula of CNMR data . Inspectpic data  indicate to C-11 . Similarelations  of H-3/Hadiation .30H46O6, as 1 based on the HRESIMS and 13C NMR data , δC 18.4 (C-27)] in a cyclopropyl ring appeared. This implied that 4 possesses a 13,27-cycloursane-type hexacyclic triterpenoid framework. This deduction was confirmed by the HMBC cross-peaks from H2-27 to C-12, C-13, and C-14 (b-27 (δH 1.16) with H-9 positioned the cyclopropyl ring in the α-face of the molecule. The absolute configuration of 4 was established to be  by a single-crystal X-ray diffraction experiment using Ga Kα radiation -3β,11α,22α-trihydroxy-12-oxo-13α,27-cycloursan-23-oic acid.Stewartiacid D (4) was obtained as colorless crystals (in MeOH), with the same molecular formula, CNMR data . Its 1H data  and 13C NMR data  and C-13 (S) in 5 to be the same as 4. Accordingly, the structure of 5 was defined as -3β,11α-dihydroxy-12-oxo-13α,27-cycloursan-23-oic acid.Stewartiacid E (5) was obtained as a white powder, and the close similarity of its UV, NMR data . The difNMR data . The relm/z 517.3665 [M − H]−, establishing the molecular formula C31H50O6 for 6. Its 1H and 13C NMR data  and a carboxyl group at C-23 (δC 180.5) based on the key HMBC correlations from H3-28 to C-22 and from H3-24 to C-23 in 6  and ROE correlations  of 6 was established by a single-crystal X-ray diffraction experiment using Ga Kα radiation [Flack parameter = 0.10 (8)] (Stewartiacid F (6) was obtained as colorless crystals from MeOH. The HRESIMS spectrum displayed a deprotonated ion at NMR data  and 3 weNMR data  and 16,2-23 in 6 . The α-oelations . Hence, .10 (8)] .m/z 501.3577 [M − H]−, the molecular formula of stewartiacid G (7) was determined to be C31H50O5, with one O-atom less than that of 6. Inspection of its NMR data  and the HMBC cross-peak from H3-28 to C-22. The relative configuration of 7 was identical to that of 6, as evidenced by the ROESY experiment , C-14 (S), and C-18 (R) in 7 to be the same as 6. Therefore, the structure of compound 7 was defined as -11α-methoxy-3β,12-dihydroxy-urs-12-en-23-oic acid.On the basis of an HRESIMS ion at NMR data  and 3 im− ion peak at m/z 501.3222 established its molecular formula to be C30H46O6. The 1H and 13C NMR spectroscopic data (δC 196.2) in 8 along with the absence of methoxy signal at δH 3.42 . Indeed, the presence of such a Δ12,13-11-one moiety in 8 was in accordance with the UV absorption at 287 nm. Further HMBC correlations of H-9/C-11, H-18/C-12, and H3-27/C-13 confirmed the above deduction .In the HRESIMS spectrum of stewartiacid H (8), the [M − H]pic data  and 3 cleduction . The releduction , and comeduction . Accordi. 300 nm , well ma. 300 nm . Thus, t30H44O6, as indicated by the 13C NMR data  from the overlaid experimental and calculated ECD curves -3β,12-dihydroxy-11,22-dioxo-urs-12-en-23-oic acid.Stewartiacid I (9) possesses a molecular formula of CNMR data  and the D curves . Therefo30H42O6 by analysis of HRESIMS and 13C NMR data (18(19) double bond . This was corroborated by the HMBC correlations from H3-29/H3-30 to C-19 and from H3-28 to C-18  was determined as CNMR data , with tw42H60O9, was determined by HRESIMS  and 13C NMR data , 7.28 , 7.23 ]. The aforementioned data were closely related to those of 11α,12--urs-12-en-3β,15α-diol (11a), which was previously isolated from Siphonodon celastrineus.26 As with 11a, compound 11 is a hybrid of ursane-type triterpenoid and phenylpropanol by forming a unique 1,4-dioxane ring. The oxygen bridge from C-11 to C-8′ was verified by the HMBC cross-peak between H-11 and C-8′ (δH 5.75) in 11 and the HMBC correlation from H-3 to the acetyl carbonyl carbon suggested that an acetoxy moiety was located at C-3 in 11. Differing from 11a the 15-OH was absent in 11, which was corroborated by the HMBC correlation from H3-27 to C-15. In addition, as with compounds 1–10, the carboxyl group at C-23 was confirmed by the distinct HMBC correlations from H-3/H3-24 to C-23. The relative configuration of 11 was determined by analysing the vicinal coupling constants of key protons , between H-9 and H-11 (8.8 Hz), and between H-18 and H-19 (10.7 Hz) were indicative of their trans-diaxial relationship. The ROE correlations of H-3/H-5, H-5/H-9, and H-9/H3-27 indicated that H-3, H-5, H-9, and H3-27 assumed α-axial orientations. In turn, the ROE correlations of H3-24/H3-25, H3-25/H-11, H3-26/H-11, H3-28/H-18, and H3-29/H-18 suggested these protons to be β-oriented in 11. Meanwhile, the magnitude of JH-7′/H-8′ (10.4 Hz) indicated a trans configuration of the dioxane ring,27 and the ROE correlation between H-8′ and H-11 supported their β-orientation. The ROE correlation between H-2′ and the methoxy group also confirmed the latter to be located at C-3′. Thus, the structure of 11 was defined as 11α,12--3β-acetoxy-urs-12-en-23-oic acid.Stewartiacid K (11) was isolated as a white powder and its molecular formula, CNMR data , correspand C-8′ . Another protons  and ROE  protons . The lar42H60O9) as that of 11 based on the HRESIMS ion at m/z 731.4125 . Its 1H and 13C NMR data resembled those of 11, with noticeable difference being observed around the 1,4-dioxane ring (JH-7′/H-8′ = 3.7 Hz) was indicative of a 7′,8′-cis relative configuration. H-7′ and H-8′ were then assigned as α-orientation from the ROE correlations of H-2′/H-6′ with H-11, and of H′-8 with H3-29, as well as the absence of correlation between H-11 and H-7′ . Accordingly, the structure of 12 was established as -11α,12--3β-acetoxy-urs-12-en-23-oic acid. Interestingly, the formation of the 1,4-dioxane rings in compounds 11 and 12 has been considered to be resulted via a free radical coupling reaction.26Stewartiacid L (12) was found to possess the same molecular formula  as compound 9 based on the HRESIMS and 13C NMR data. Comparsion of the 1D NMR data of 13  in 13 rather than two secondary methyl groups in 9. This implied that, unlike compounds 1–12 with an ursane-type skeleton, 13 is an oleanane-type triterpenoid, as with the co-occurring compounds 17 , H-5 and H-6β (11.2 Hz), and between H-18 and H-19α (12.2 Hz) were indicative of their axial orientations. The ROE correlations  has the same molecular formula . This implied that they should be a pair of C-4 epimers. As for 14, a clear ROE correlation between H3-24 (δH 1.75) and H3-25 (δH 1.39) undoubtedly allowed Me-24 to be β-axially oriented (28 the 12-OH was concluded to adopt the β-orientation as evident from the ROE correlations of 12-OH (δH 7.13) with H-18 (δH 2.68) and H3-26 (δH 1.67). Thus, the configuration at C-4 in camellisin C28 seems to be wrongly assigned, and its structure should be revised as 3β,11α,13β-trihydroxy-11,22-dioxo-olean-24-oic acid. Herein, the 1H and 13C NMR data for 3β,11α,13β-trihydroxy-11,22-dioxo-olean-23-oic acid (14) are accurately assigned in this study.Stewartiacid N (14) has a molecular formula of CNMR data . The NMRoriented . Consist29 since it affects nutrient catabolism and cholesterol and fatty acid biosynthesis. Recently, a series of diterpenoids and norditerpenoids from the endangered conifers Pseudotsuga sinensis21 and Picea brachytyla21 were found to have significant ACL inhibitory effects. In the present study, all the isolated compounds (1–18) were evaluated for their ACL inhibitory activities. Among them, compound 11 displayed the most potent inhibitory effect, with an IC50 value of 2.8 μM. Compounds 7 and 12 showed moderate inhibition against ACL, with IC50 values of 12.5 and 10.6 μM, respectively (50: 0.4 ± 0.1 μM).30 To our knowledge, this is the first report on the naturally occurring ursane-type triterpenoids as ACL inhibitors. In addition, compounds 1–18 were also tested for their inhibitory activities against NF-κB, and only 11 showed moderate inhibition (IC50: 16.8 μM). Bortezomib (PS-341) was used as the positive control (IC50: 0.06 ± 0.01 μM) (31The ATP-citrate lyase (ACL) is a promising target for the treatment of metabolic disorders and cancer,ectively . The res0.01 μM) .31Stewartia sinensis. This is the first phytochemical and pharmacological investigation on this plant. Unlike 28-carboxylated ursane-/oleanane-type triterpenoids  that are widely distributed in the plant kingdom, the ursane-/oleanane-type triterpenoids featuring a 23-COOH group are quite rare in nature. To our knowledge, only a dozen C-23 monocarboxylated derivatives have been so far reported . From a chemical point of view, stewartiacids A (1) and B (2) are rare isoursenol derivatives featuring the 27-methyl group at C-13,24,25 whereas stewartiacid C (3) is a rare 12-oxo-γ-amyrin analogue.25 Stewartiacids D (4) and E (5) are two 13,27-cycloursane-type triterpenoids. Stewartiacids K (11) and L (12) are uncommon ursane-type triterpene and phenylpropanol hybrids by forming a unique 1,4-dioxane bridge.26,32 The absolute configurations of compounds 1–9 and 12 were well established either by single crystal X-ray diffraction or by ECD analyses. The absolute configurations of the rest new stewartiacids  are still open; however, they could be assigned as shown in both e.g., -10] based on biogenetic considerations.In the present study, 14 new C-23 carboxylated triterpenoid derivatives, stewartiacids A–N, were isolated from the twigs and leaves of the endangered ornamental plant Stewartiacids G (7), K (11), and L (12) showed inhibitory effects against ACL. Compound 11 also exhibited inhibition on NF-κB. The above findings may provide useful clues for discovery and development of new therapeutic or preventive agents for treatment of metabolic disorders and other ACL or NF-κB related diseases. Moreover, the identification of new molecules from endangered plants reveals the importance in conservation efforts to prevent species diversity loss in the control of emerging druggable targets.−1) and a X-bridge C18 column  were utilized. Thin-layer chromatography (TLC) was performed on pre-coated plates . TLC spots were visualized under UV light (254 or 365 nm) and by spraying with 5% H2SO4/vanillin followed by heating to 120 °C.Melting points were measured with a WRS-1B capillary melting point apparatus. Optical rotations were obtained with a Rudolf Autopol IV at 22 °C. UV and IR spectra were recorded on a Hitachi U-2900E UV spectrophotometer and a Thermo Scientific Nicolet Is5 FT-IR spectrometer, respectively. ECD spectra were recorded on a JASCO-810 spectropolarimeter. ESI-MS and HRESIMS were acquired on an Agilent 1100 LC/MSD mass spectrometer and an AB Sciex Triple TOF 5600 spectrometer, respectively. X-ray crystallographic data were measured on a Bruker Apex Duo Diffractometer (Ga Kα). 1D and 2D NMR spectra were performed on a Bruker Avance III 400 MHz or a Bruker Avance III 600 MHz spectrometers using the residual solvent signals as the internal standard. All chemical shifts were expressed in ppm. Semi-preparative HPLC was performed on a Waters e2695 system coupled with a 2998 photodiode array (PDA) detector and a 2424 evaporative light-scattering detector (ELSD). A SunFire C18 column  were collected by one of the authors (Mr Dao-An Xiao) from Mingyue Mountain in Yichun, Jiangxi Province of China, in October 2017. The plant was also identified by Mr Xiao . A voucher specimen (no. 20171007) was deposited at the Herbarium of the School of Pharmacy at Fudan University.The twigs and leaves of S. sinensis were extracted with 90% MeOH  at room temperature. The resultant dark green residue  was suspended in H2O (1.5 L) and extracted successively with petroleum ether , EtOAc (3 × 1.5 L), and n-BuOH (3 × 1.5 L). The EtOAc-soluble extract (52.2 g) was subjected to a silica gel column with a stepwise gradient-elution, employing a mixture of PE–EtOAc (30 : 1 → 20 : 1 → 10 : 1 → 5 : 1 → 1 : 1 → 1 : 5 → neat EtOAc) as solvents, to afford nine fractions (Fr.1–Fr.9), according to TLC analysis. Fr.2 (910 mg) was separated by gel permeation chromatography (GPC) on Sephadex LH-20 , followed by semi-preparative HPLC  to afford compounds 15  and 17 . Fr.3 (1.05 g) was fractioned by Sephadex LH-20 (MeOH), and five fractions (Fr.3A-Fr.3E) were obtained. Fr.3B (127 mg) was purified by semi-preparative HPLC  to afford compounds 16  and 18 . Fr.4 (410 mg) was fractioned on a MCI column with a step gradient elution of MeOH–H2O (50 : 50 → 70 : 30 → 85 : 15 → 100 : 0) and six fractions (Fr.4A–Fr.4F) were collected. Separation of Fr.4C (63 mg) over Sephadex LH-20 (MeOH) and semi-preparative HPLC  afforded compounds 12  and 11 . Fr.5 (1.9 g) was fractioned on an MCI column with a step gradient elution of MeOH–H2O (50 : 50 → 70 : 30 → 85 : 15 → 100 : 0) and seven fractions (Fr.5A–Fr.5G) were obtained. Fr.5B (74.0 mg) was purified by semi-preparative HPLC to furnish compound 14 . Fr.5C (197.0 mg) was further separated on Sephadex LH-20 (MeOH) to give subfractions Fr.5C-1–Fr.5C-6. Fr.5C-3 (22 mg) was purified by semi-preparative HPLC  to afford compound 4 . By employing the same HPLC  system, compounds 9  and 13  were purified from Fr.5C-4 (17.4 mg), whereas compound 10  was purified from Fr.5C-5 (12.6 mg). Compound 3 (4.0 mg) was obtained from Fr.5D (130 mg) by Sephadex LH-20 (MeOH) followed by HPLC purification . Purification of fraction Fr.5E (218 mg) by semi-preparative HPLC  afforded compounds 2  and 7 . Fr.7 (3.5 g) was chromatographed over a silica gel column  to give six subfractions, Fr.7A–Fr.7F. Fr.7D (650 mg) was rechromatographed by Sephadex LH-20 (MeOH), and seven subfractions (Fr.7D-1–Fr.7D-7) were obtained. Purification of subfraction Fr.7D-4 (233 mg) by semi-preparative HPLC  yielded compounds 6  and 8 . Compound 1 (5.0 mg) was purified from Fr.7F by Sephadex LH-20 (MeOH) followed by semi-preparative HPLC purification . Fr.8 (1.1 g) was fractioned by Sephadex LH-20 (MeOH), and six subfractions (Fr.8A–Fr.8F) were generated. Fr.8C (488 mg) was purified by semi-preparative HPLC  to afford compound 5 .The air-dried, powdered twigs and leaves (3.5 kg) of α]22D −7.8 ; UV (MeOH) λmax (log ε) 203 (4.62) nm; ECD  λmax (Δε): 233 (−1.7), 316 (−1.4); IR (KBr) vmax 3476, 2945, 2920, 2868, 1706, 1674, 1464, 1384 1210, 1132, 1013, 801 cm−1; 1H and 13C NMR data, see m/z 525 [M + Na]+, 501 [M − H]−; HRESIMS m/z 525.3182 [M + Na]+ .Colorless crystals (MeOH), mp 322.4–323.0; [α]22D −4.6 ; UV (MeOH) λmax (log ε) 207 (1.54) nm; ECD  λmax (Δε): 233 (−0.5), 316 (−0.4); IR (KBr) vmax 3451, 2967, 2918, 2863, 1705, 1661, 1621, 1459, 1382, 1207, 1153, 1008, 868 cm−1; 1H and 13C NMR data, see m/z 485 [M − H]−; HRESIMS m/z 485.3268 [M − H]− .White powder; [α]22D −5.2 ; UV (MeOH) λmax (log ε) 210 (1.24), 257 (1.15) nm; ECD  λmax (Δε): 220 (+3.9), 257 (−3.5), 289 (−3.5); IR (KBr) vmax 3441, 2972, 2932, 2860, 1696 (br.), 1644, 1457, 1379, 1339, 1205, 1060, 1015, 798, 726 cm−1; 1H and 13C NMR data, see m/z 499 [M − H]−; HRESIMS m/z 499.3069 [M − H]− .Colorless crystals (MeOH); [α]22D +59.6 ; UV (MeOH) λmax (log ε) 215 (3.43) nm; ECD  λmax (Δε): 214 (+3.2), 291 (+3.6); IR (KBr) vmax 3661, 2972, 2940, 2825, 1703, 1663, 1454, 1344, 1205, 1054, 1012 cm−1; 1H and 13C NMR data, see m/z 501 [M − H]−, 1003 [2M − H]−; HRESIMS m/z 501.3277 [M − H]− .Colorless crystals (MeOH), mp 272.0–273.1; [α]22D +34.7 ; UV (MeOH) λmax (log ε) 217 (2.49) nm; ECD  λmax (Δε): 233 (−2.4), 291 (+3.8); IR (KBr) vmax 3436, 2985, 2915, 2860, 1694, 1659, 1619, 1452, 1384, 1212, 1058, 1013, 866 cm−1; 1H and 13C NMR data, see m/z 509 [M + Na]+, 485 [M − H]−, 971 [2M − H]−; HRESIMS m/z 509.3238 [M + Na]+ .White powder; [α]22D +23.3 ; UV (MeOH) λmax (log ε) 209 (2.02) nm; ECD  λmax (Δε): 207 (+7.2); IR (KBr) vmax 3405, 2993, 2925, 2860, 2823, 1701, 1681, 1454, 1344, 1314, 1202, 1054, 1032, 1014, 721 cm−1; 1H and 13C NMR data, see m/z 517 [M − H]−; HRESIMS m/z 517.3539 [M − H]− .Colorless crystals (MeOH), mp 220.9–221.5; [α]22D +17.3 ; UV (MeOH) λmax (log ε) 203 (1.10) nm; ECD  λmax (Δε): 206 (+17.4); IR (KBr) vmax 3681, 2920, 2841, 1701, 1681, 1642, 1522, 1457, 1053, 1036, 874, 809, 724 cm−1; 1H and 13C NMR data, see m/z 517 [M − H]−; HRESIMS m/z 501.3577 [M − H]− .White powder; [α]22D +10.6 ; UV (MeOH) λmax (log ε) 287 (2.67) nm; ECD  λmax (Δε): 209 (+2.6), 234 (−0.5), 285 (+4.2); IR (KBr) vmax 3663, 2970, 2868, 1702, 1671, 1452, 1347, 1205, 1058, 1005, 729 cm−1; 1H and 13C NMR data, see m/z 501 [M − H]−; HRESIMS m/z 501.3225 [M − H]− .White powder; [α]22D +5.8 ; UV (MeOH) λmax (log ε) 285 (1.97) nm; ECD  λmax (Δε): 214 (+3.0), 236 (−0.7), 266 (+2.3), 300 (+10.1); IR (KBr) vmax 3463, 2970, 2910, 2830, 1703, 1661, 1449, 1357, 1182, 1063, 1010, 871 cm−1; 1H and 13C NMR data, see m/z 499 [M − H]−; HRESIMS m/z 499.3065 [M − H]− .White powder; [α]22D +18.0 ; UV (MeOH) λmax (log ε) 299 (2.70) nm; IR (KBr) vmax 3449, 2975, 2915, 2866, 1700, 1656, 1474, 1384, 1210, 1055, 1030, 803 cm−1; 1H and 13C NMR data, see m/z 497 [M − H]−; HRESIMS m/z 497.2933 [M − H]− .White powder; [α]22D +25.0 ; UV (MeOH) λmax (log ε) 203 (7.94), 279 (0.65) nm; ECD  λmax (Δε): 228 (+0.4), 278 (+0.2); IR (KBr) vmax 3439, 2980, 2925, 2860, 1715, 1671, 1619, 1454, 1384, 1267, 1055, 1010, 918 cm−1; 1H and 13C NMR data, see m/z 731 [M + Na]+; HRESIMS m/z 731.4111 [M + Na]+ .White powder; [α]22D +16.7 ; UV (MeOH) λmax (log ε) 203 (7.30), 279 (0.72) nm; ECD  λmax (Δε): 214 (+8.6), 279 (+0.3); IR (KBr) vmax 3441, 2927, 2853, 1713, 1661, 1619, 1439, 1380, 1207, 1080, 1020, 941 cm−1; 1H and 13C NMR data, see m/z 731 [M + Na]+; HRESIMS m/z 731.4125 [M + Na]+ .White powder; [α]22D +4.2 ; UV (MeOH) λmax (log ε) 285 (2.02) nm; IR (KBr) vmax 3446, 2977, 2922, 2873, 1704, 1661, 1454, 1377, 1207, 1140, 1060, 1033, 1008, 948, 843, 721 cm−1; 1H and 13C NMR data, see m/z 499 [M − H]−; HRESIMS m/z 499.3061 [M − H]− .White powder; [α]22D +22.0  {camellisin C:28 [α]24D +10.0 }; UV (MeOH) λmax (log ε) 210 (0.66) nm; IR (KBr) vmax 3446, 2977, 2922, 2873, 1703, 1661, 1454, 1377, 1207, 1060, 1033, 843 cm−1; 1H and 13C NMR data, see m/z 517 [M − H]−, 541 [M + Na]+; HRESIMS m/z 541.3140 [M + Na]+ .White powder; [ε) 210 0.6 nm; IR 33 structure solution program using Intrinsic Phasing and refined with the SheIXT34 refinement using Least Squares minimization. Crystallographic data of compounds 1, 3, 4, and 6 have been deposited in the Cambridge Crystallographic Data Centre as CCDC-1959096, CCDC-1972126, CCDC-1959098, and CCDC-1959097, respectively.Detailed data are included in the ESI.35 for all conformers of stewartiacids H (8), I (9), and L (12). Rotatory strengths for a total of 30 excited states were calculated. ECD spectra were generated using the program SpecDis 1.6  and GraphPad Prism 5  from dipole-length rotational strengths by applying Gaussian band shapes with sigma = 0.3 eV.The Monte Carlo conformational searches were carried out by Spartan's 10 software  using Merck Molecular Force Field (MMFF). The conformers with Boltzmann-population of over 7% were chosen for ECD calculations, and then the conformers were initially optimized at B3LYP/6-31g  level in MeOH using the conductor-like polarizable calculation model (CPCM). The theoretical calculation of ECD was conducted in MeOH using time-dependent density functional theory (TDDFT) at the CAM-B3LYP/def2-TZVP level30 The kinase assay was carried out in a 384-well plate  in a volume of 5 μL reaction mixture containing 2.0 μL of ACL, 2.0 μL of ATP, and 1.0 μL of the tested compound with different concentrations. Reactions in each well were kept going for 30 min under 37 °C. After the enzymatic reaction, 2.5 μL of ADP-Glo™ reagent was added to each well to terminate the kinase reaction and deplete the unconsumed ATP within 60 min at room temperature. In the end, 5.0 μL of kinase detection reagent (reagent 2) was added to each well and incubated for 1 h to simultaneously convert ADP to ATP. The luminescence signal was measured using a PerkinElmer EnVision reader. The known inhibitor BMS 303141  based on a sigmoidal dose–response curve (variable slope) and computed using Graphpad Prism 5 (Graphpad Software). Bortezomib  was used as the positive control.31The HEK293 with stable NF-κB expression cell line was used for the luciferase assay.There are no conflicts to declare.RA-010-C9RA09542J-s001RA-010-C9RA09542J-s002