# _applied sciences_
## 2 Materials and Methods
### Chemistry
#### 2.1.2 General Procedure for the Synthesis of Compounds **4** and **5**
The residue was purified by SiO\({}_{2}\) column chromatography (eluent CHCl\({}_{3}\)-MeOH 100:0\(\rightarrow\)90:10), obtaining compound **17**. [3,2b]Indolo-N-(4-(piperazin-1-yl))butt-2 yn-1-yl)-olean-12(13)-en-28-amide **17**  
Yellow solid; mp: 139\({}^{\circ}\)C; yield 81%; [\(\alpha\)]\({}_{D}\)\({}^{20}\) -21\({}^{\circ}\) (\(c\) 0.1, CHCl\({}_{3}\)). \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)) 8 7.45-7.00 (m, 4H), 6.12 (br, s, NH), 5.50 (s, 1H, H12), 3.90 and 4.10 (d, 2H, \(J\) = 17.2 Hz, NHCH\({}_{2}\)), 3.30 (d, \(J\) = 9.1 Hz, NCH\({}_{2}\)), 2.80-2.50 (m, 8H, 4 CH\({}_{2}\)), 2.30\(-\)1.30 (m, 23H, CH and CH\({}_{2}\)), 1.32, 1.30, 1.22, 1.20, 0.90, 0.89, 0.81 (7s, 21H, 7 CH\({}_{3}\)). \({}^{13}\)C NMR (125.5 MHz, CDCl\({}_{3}\)) 8 178.03 (CONH), 144.62 (C13), 140.90, 136.16, 128.10, 123.30 (C12), 120.92, 118.81, 117.85, 110.47, 106.50, 81.10 (C\(\equiv\)C), 79.11 (C=C), 53.07 (2C), 51.32 (2C), 46.91, 46.66, 46.39, 46.24, 42.20, 40.94, 39.50, 38.01, 37.45, 36.82, 34.11, 34.01, 33.01, 32.26, 31.78, 30.97, 30.76,with CH\({}_{2}\)Cl\({}_{2}\) (2 \(\times\) 15 mL), the combined extracts were washed with 5% HCl (3 \(\times\) 15 mL), H\({}_{2}\)O (2 \(\times\) 15 mL), dried over CaCl\({}_{2}\), and the solvent was evaporated in a water jet vacuum. Purification of the crude product was performed using column chromatography on Al\({}_{2}\)O\({}_{3}\) by elution with a mixture of petroleum ether-chloroform (1:0 \(\rightarrow\) 1:1). [3,2b]Indolo-N-piperazin-olean-12(13)-en-28-amide **14**  
Yellow solid; mp: 117 \({}^{\circ}\)C; yield 85%; [\(\alpha\)]\({}_{D}\)\({}^{20}\) +4\({}^{\circ}\) (\(c\) 0.1, CHCl\({}_{3}\)). \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)) \(\delta\) 7.45-7.00 (m, 4H), 5.38 (s, 1H, H12), 3.15-2.70 (m, 8H, 4 CH\({}_{2}\)), 3.10-1.20 (m, 22H, CH and CH\({}_{2}\)), 1.30, 1.21, 1.19, 0.95, 0.93, 0.91, 0.81 (7s, 21H, 7 CH\({}_{3}\)). \({}^{13}\)C NMR (125.5 MHz, CDCl\({}_{3}\)) \(\delta\) 175.34 (C28), 144.17 (C13), 140.91 (C3), 136.12 (C6\({}^{\prime}\)), 128.22 (C1\({}^{\prime}\)), 122.09 (C12), 120.89 (C4\({}^{\prime}\)), 118.82 (C3\({}^{\prime}\)), 117.98 (C2\({}^{\prime}\)), 110.36 (C5\({}^{\prime}\)), 106.81 (C2), 53.27 (2C), 51.63 (2C), 47.59, 46.47, 46.32, 44.60, 43.70, 42.05, 39.26, 38.15, 36.73, 34.01, 33.04, 32.36, 31.01, 30.41, 29.99, 27.99, 25.82, 24.03, 23.44, 23.32, 22.83, 19.31, 16.74, 16.68, 15.59. MS (APCI) _m/z_ 596.5 [M+ H]\({}^{+}\) (calcd for C\({}_{40}\)H\({}_{57}\)N\({}_{3}\)O, 595.92). 2-[(E)-pyridine-4-ylmethylene]-3-oxo-olean-12(13)-en-28-carboxylate **26**  
White solid; mp: 166 \({}^{\circ}\)C; yield 72%; [\(\alpha\)]\({}_{D}\)\({}^{20}\)\({}^{49}\)\({}^{\circ}\) (\(c\) 0.1, CHCl\({}_{3}\)). \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)) \(\delta\) 8.71-8.65 (d, \(J\) = 6 Hz, 2H), 7.40-7.30 (d, \(J\) = 6.2 Hz, 2H), 6.95 (s, 1H), 5.85 (brs, NH), 5.40 (d, \(J\) = 3.4 Hz, 1H, H12), 2.95-1.25 (m, 22H, CH, CH\({}_{2}\)), 1.24, 11.18, 1.15, 0.90, 0.95, 0.88, 0.87 (7s, 21H, 7 CH\({}_{3}\)). \({}^{13}\)C NMR (125.5 MHz, CDCl\({}_{3}\)) \(\delta\) 207.09 (C3), 181.01 (C28), 147.92, 145.54, 145.04 (C13), 142.22, 139.08, 133.21, 126.21, 124.69, 122.19 (C12), 53.00, 46.62, 46.49, 45.42, 44.03, 42.69, 42.37, 39.36, 39.13, 36.35, 34.07, 32.97, 32.51, 31.56, 30.73, 29.50, 27.28, 25.72, 25.49, 23.50, 22.68, 20.24, 19.05, 16.59, 15.39. MS (APCI) _m/z_ 543.5 [M+ H]\({}^{+}\) (calcd for C\({}_{36}\)H\({}_{50}\)N\({}_{2}\)O\({}_{2}\), 542.81).