Determination of 5-LO Activity in a Cell-Free Assay To determine 5-LO activity, aliquots of purified 5-LO (0

Determination of 5-LO Activity in a Cell-Free Assay To determine 5-LO activity, aliquots of purified 5-LO (0.5 g 5-LO in 1 mL PBS plus 1 mM EDTA) were pre-incubated with the test compounds or vehicle (0.1% dimethyl sulfoxide, DMSO) on ice for 15 min, pre-warmed for 30 s at 37 C in a water bath, and then stimulated with 20 M AA and CaCl2 2 mM for 10 min at 37 C. neutrophils. These triterpene acids were also investigated for their direct interference with 5-LO and cathepsin G in cell-free assays. Furthermore, our studies were expanded to 10 semi-synthetic BA derivatives. Our data reveal that besides BAs, several tetra- and pentacyclic triterpene acids are effective or even superior inhibitors of 5-LO product formation in human neutrophils, and in parallel, inhibit cathepsin G. Their beneficial target profile may qualify triterpene acids as anti-inflammatory natural products and pharmacological prospects for intervention with diseases related to aging. species, termed frankincense, is usually a traditional Ayurvedic medicine that has experienced increasing popularity also in Western countries during the past decades [6]. Frankincense is usually a natural and rich source for a variety of triterpene acids, including boswellic acids (BAs), tirucallic acids, roburic acids, and lupeolic acids [7,8,9]. In folk medicine, lipophilic frankincense extracts are used as alternative to anti-inflammatory steroidal drugs (i.e., glucocorticoids) or NSAIDs for treatment of inflammatory diseases, such as rheumatoid arthritis, osteoarthritis, asthma, atopic dermatitis, and inflammatory bowel diseases [6]. The 3-species by preparative high-performance liquid chromatography (HPLC), as described previously [9,22]. The < 0.05, (**) < 0.01, (***) < 0.001; student 2.1 Hz, 1H, H-12), 4.21 (d, 9.1 Hz, 1H, H-11), 3.98 (bs, 1H, H-3), 2.22C2.06 (m, 3H, H-1, H-2, H-16), 1.94C1.86 (m, 1H, H-6), 1.84C1.75 (m, 1H, H-15), 1.75C1.68 (m, 2H, H-6, H-9), 1.68C1.60 (m, 2H, H-1, H-5), 1.60C1.50 (m, 1H, H-7), 1.50C1.30 (m, 8H, H-2, H-7, H-18, H-19, H-21, H-21, H-22, H-22), 1.29 (s, 3H, H-23), 1.22 (bs, gamma-secretase modulator 1 4H, H-15, H-27), 1.14 (s, 3H, H-26), 1.09 (s, 3H, H-25), 0.93 (bs, 5H, H-16, H-20, H-30), 0.90 (d, 6.2 Hz, 3H, H-29), 0.83 (s, 3H, H-28). 13C-NMR ((CD3)2CO, 125.76 MHz): [ppm] = 180.3 (C-24, COOH), 142.2 (C-13, H>C=C<), 132.8 (C-12, H>C=C<), 71.7 (C-3, HO>CH-), 69.6 (C-11, HO>CH-), gamma-secretase modulator 1 60.3 (C-18, >CH-), 55.4 (C-9, >CH-), 50.7 (C-5, gamma-secretase modulator 1 >CHC), 49.3 (C-4, >C<), 45.0 (C-8, >C<), 44.1 (C-14, >C<), 43.2 (C-22, CCH2C), 41.4 (C-19, >CHC), 41.4 (C-20, >CHC), 40.5 (C-10, >C<), 37.8 (C-1, CCH2C), 36.1 (C-7, CCH2C), 35.5 (C-17, >C<), 32.9 (C-21, CCH2C), 30.2 (C-28, CCH3), 29.8 (C-16, CCH2C), 28.4 (C-15, CCH2C), 28.3 (C-2, CCH2C), 26.2 (C-23, CCH3), 24.3 (C-27, CCH3), 22.6 (C-30, CCH3), 21.6 (C-6, CCH2C), 19.7 (C-26, CCH3), 18.9 (C-29, CCH3), 15.8 (C-25, CCH3). MS (EI, gamma-secretase modulator 1 70 eV): (%) = 454 (100) gamma-secretase modulator 1 [M-H2O]+, 439 (8), 421 (12), 325 (4), 301 (12), 269 (8), 255 (45), 253 (6), 237 (4), 215 (5). HRMS (EI, 70 eV): calculated: 472.3587 for C30H48O4; found: 472.3570. []D: +51.7 (c = 1.61, acetone). m.p. 166C169 C Rabbit Polyclonal to UGDH (dec). 11–Hydroxy–boswellic acid 19: 1H-NMR ((CD3)2CO, 500.13 MHz): [ppm] = 5.30 (d, 4.3 Hz, 1H, H-12), 4.45 (t, 4.7 Hz, 1H, H-11), 4.00 (t, 2.4 Hz, 1H, H-3), 2.33C2.25 (m, 1H, H-2), 2.12C2.05 (m, 1H, H-16), 2.00C1.88 (m, 2H, H-6, H-15), 1.84C1.78 (m, 1H, H-1), 1.76C1.70 (m, 1H, H-6), 1.70C1.60 (m, 2H, H-1, H-7), 1.58C1.50 (m, 3H, H-2, H-5, H-9), 1.47C1.43 (m, 1H, H-22), 1.43 (s, 3H, H-25), 1.43C1.36 (m, 3H, H-18, H-19, H-21), 1.36C1.33 (m, 3H, H-7, H-21, H-22), 1.33 (s, 3H, H-26), 1.28 (s, 3H, H-23), 1.15C1.10 (m, 1H, H-15), 1.09 (s, 3H, H-27), 1.09C1.06 (m, 1H, H-16), 0.93 (s, 3H, H-30), 0.93C0.87 (m, 1H, H-20), 0.87 (s, 3H, H-28), 0.84 (d, 5.9 Hz, 3H, H-29). 13C-NMR ((CD3)2CO, 125.76 MHz): [ppm] = 180.2 (C-24, COOH), 142.1 (C-13, H>C=C<), 131.7 (C-12, H>C=C<), 71.8 (C-3, HO>CHC), 66.8 (C-11, HO>CHC), 60.8 (C-18, >CHC), 53.9 (C-9, >CHC), 51.7 (C-5, >CHC), 48.7 (C-4, >C<), 44.4 (C-14, >C<), 43.3 (C-22, CCH2C), 41.6 (C-8, >C<), 41.4 (C-19, >CHC), 41.4 (C-20, >CHC), 40.6 (C-10,.