hERG Channels

A successive column chromatography approach was used to separate the major compounds of the active fraction, including bakkenolide B (1), bakkenolide D (2), 1,5-di-could be an excellent source of bacterial NA inhibitors for food and medicinal uses

A successive column chromatography approach was used to separate the major compounds of the active fraction, including bakkenolide B (1), bakkenolide D (2), 1,5-di-could be an excellent source of bacterial NA inhibitors for food and medicinal uses. Supplementary Materials The following are available online at https://www.mdpi.com/2218-273X/10/6/888/s1, Figures S1CS20: 1D and 2D NMR spectra of compounds 1?4, Figures S21 and S22: Isolation of bioactive compounds from the aerial portion of with preparative HPLC, Figure S23: HPLC profiles of extract and compounds present K145 in the aerial portion of em P. of wild or cultivated species are consumed fresh as a vegetable, and are processed into different food products. Previous phytochemical investigations have shown that species contain many bioactive components, such as triterpenoids, sterols, fatty acids, phenolic compounds, sesquiterpenoids (e.g., bakkenolide), and other trace minerals [15,16,17,18,19]. Some of these compounds are responsible for the antioxidant, anti-allergenic, anti-inflammation, anti-carcinogenic, anti-mutagenic, antimicrobial, and neuroprotective effects of products [20,21,22,23,24]. To date, K145 no report exists on bacterial NA inhibition by chemical constituents of on bacterial NA and isolated active compounds from the extract based on the enzyme activity assay. Enzyme kinetic analyses and molecular-modeling studies have been performed using the most active compound to provide insights into the interactions between the active compounds and bacterial NA. 2. Material and Methods 2.1. General Experimental Procedures CC was performed using Diaion HP-20 (Mitsubishi-Chemical, Tokyo, Japan), silica gel (230C400 mesh; Merck Co., Darmstadt, Germany) and Sephadex LH-20 (GE Healthcare Bio-Science AB, Uppsala, Sweden). Recycling Preparative HPLC was performed by using a LaboACE LC-5060 (JAI Co., Ltd., Tokyo, Japan). A JAIGEL-ODS column (20 500 mm, 15 m, JAI) was used for preparative HPLC. Enzymatic assays were carried out on a Spectra Max M3 Multi-Mode Microplate Reader (Molecular Device, Sunnyvale, CA, USA). The NMR spectra of 1C4 were recorded on Bruker K145 700 and 900 MHz spectrometer (Bruker, Karlsruhe, Germany), using standard Bruker pulse programs. Chemical shifts are given as whole plants were collected in Baekdudaegan National Arboretum in April 2019 (voucher BDNA-2019-3004). Voucher specimen is deposited in the herbarium of Baekdudaegan National Arboretum for future reference, and the plant species were identified by the taxonomist Lee D.H. This plant was dried at room temperature in an airtight place under dark conditions. 2.3. Extraction and Isolation The dried and powdered leaves and stems of (1.6 kg) were successively extracted using methanol, at room temperature (2 times at 3-day intervals, totaling 6 days). After the solvent was removed under reduced pressure at 45 C, a residue (33.9 g) was obtained. After the removal of methanol under reduced pressure, the MeOH extracts was passed thought a Diaion HP-20 column and partitioned between 5.91 (1H, dd, = 7.2, 15 Hz, H-3), 5.72 (1H, d, = 11.2 Hz, H-9), 5.17 (1H, s, H-13a), 5.14 (1H, s, H-13b), 5.10 (1H, m, H-1), 4.63 (2H, m, H-12), 2.78 (1H, dd, = 11.2, 5.0 Hz, H-10), 2.21 (1H, d, = 14.3 Hz, H-6), 1.91 (1H, d, = 14.3 Hz, H-6), 1.91 (3H, s, H-2), 1.85 (3H, dd, = 7.2, 1.6 Hz, H-4), 1.78 K145 (2H, m, H-2), 1.75 (3H, s, H-5), 1.66 (1H, m, H-3), 1.55 (1H, m, H-4), 1.34 (1H, m, H-3), 1.09 (3H, s, H-15), 0.87 (3H, d, = 6.8 Hz, H-14). 13C-NMR (175 MHz, Chloroform-177.5 (C-8), 169.9 (C-1), 167.3 (C-1), 147.7 (C-11), 136.7 (C-3), 128.2 (C-2), 108.3 (C-13), 80.8 (C-9), 70.6 (C-12), 70.5 (C-1), 54.9 (C-7), 51.4 (C-10), 45.8 (C-6), 43.4 (C-5), 35.2 (C-4), 29.5 (C-3), 26.8 (C-2), 20.9 (C-2), 20.3 (C-5), 19.5 (C-15), 15.5 (C-14), 15.5 (C-4). 2.3.2. Bakkenolide D (2) 1H-NMR (900 MHz, Chloroform-7.04 (1H, d, = 10.1 Hz, H-3), 5.76 (1H, d, = 11.2 Hz, H-9), 5.62 (1H, d, = 10.14 Hz, H-2), 5.21 (1H, s, H-13), 5.17 (1H, s, H-13), 5.15 (1H, m, H-1), 4.67 (2H, m, H-12), 2.75 (1H, dd, = 11.2, 5.0 Hz, H-10), 2.39 (3H, s, H-4), 2.24 (1H, d, = 14.3 Hz, H-6), 2.02 (3H, s, H-2), 1.95 (1H, d, = 14.3 Hz, H-6), 1.84 (1H, m, H-2), 1.76 (1H, m, H-2), 1.67 (1H, dd, = 14.1, 3.6 Hz, H-3), 1.57 (1H, m, H-4), 1.37 (1H, dd, = 12.9, 3.7 Hz), H-3), 1.11 (3H, s, H-15), 0.90 (3H, d, H-14). 13C-NMR (225 MHz, Rabbit Polyclonal to HSL (phospho-Ser855/554) Chloroform-177.5 (C-8), 169.9 (C-1), 165.6 (C-1), 152.8 (C-3),.