# _applied sciences_
## 2 Materials and Methods
### Chemistry
#### 2.1.2 General Procedure for the Synthesis of Compounds **4** and **5**  
To a solution of compound **2** (1 mmol; 0.44 g) in dry pyridine (15 mL), 3-(3-methoxyphenyl) acryloyl chloride (2 mmol; 0.39 g) or nicotinoyl chloride (2 mmol; 0.28 g) and DMAP (cat) was added. The reaction mixture was refluxed for 4 h, poured into H\({}_{2}\)O/H\({}^{+}\), filtered, washed with water until neutral, and dried. The residue was purified via Al\({}_{2}\)O\({}_{3}\) column chromatography (eluent petroleum ether-ethyl acetate 20:1\(\rightarrow\)5:1). 3,28-Di-O-[3-(3-methoxyyphenyl)acrylate]-olean-12(13)-en **4**  
Beige solid; mp: 114 \({}^{\circ}\)C; yield 78%; [\(\alpha\)]\({}_{D}\)\({}^{20}\) +17\({}^{\circ}\) (\(c\) 0.1, CHCl\({}_{3}\)). \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)) 8 7.64 (m, 2H, H3\({}^{\prime\prime}\), H8\({}^{\prime\prime}\)), 7.62 (m, 2H, H3\({}^{\prime}\), H8\({}^{\prime}\)), 7.48 (d, \(J\) = 8.7 Hz, 2H, H9\({}^{\prime}\), H9\({}^{\prime\prime}\)), 6.92 (d, \(J\) = 1.5 Hz, 2H, H5\({}^{\prime}\), H5\({}^{\prime\prime}\)), 6.86 (d, \(J\) = 1.7 Hz, 2H, H7\({}^{\prime}\), H7\({}^{\prime\prime}\)), 6.33 (d, \(J\) = 2.7 Hz, 1H, H2\({}^{\prime\prime}\)), 6.29 (d, \(J\) = 2.5 Hz, 1H, H2\({}^{\prime}\)), 5.23 (s, 1H, H12), 4.64 (dd, \(J\) = 9.2 Hz, 1H, H3), 4.17 (d, \(J\) = 11.0 Hz, 1H, H28), 3.83 (2s, 6H), 3.80 (m, 1H, H28), 2.16-1.00 (m, 23H, CH, CH\({}_{2}\)), 1.18, 0.99, 0.98, 0.94, 0.91, 0.90, 0.88 (7s, 2H, 7 CH\({}_{3}\)). 13C NMR (125.5 MHz, CDCl\({}_{3}\)) 8 167.50 (C1\({}^{\prime\prime}\)), 161.32 (C6\({}^{\prime\prime}\)), 161.27 (C6\({}^{\prime}\)), 144.14, 143.96, 143.67 (C13), 131.97, 129.71, 129.68, 127.30, 127.25, 122.86 (C12), 116.35, 115.87, 114.30, 113.47, 113.40, 80.74 (C3), 70.68 (C28), 55.37 (OCH\({}_{3}\)), 55.32 (C5), 47.55 (C18), 46.27 (C19), 42.60, 41.69, 39.83, 38.34, 37.98, 36.86 (C10), 36.08, 34.05, 33.18, 32.50, 31.55, 30.95 (C20), 28.10, 25.99, 25.65, 23.71, 23.60, 22.37, 18.25, 16.89, 16.82, 16.74, 15.60. MS (APCI) _m_/z 763.6 [M+H]\({}^{+}\) (calcd for C\({}_{50}\)H\({}_{66}\)O\({}_{6}\), 763.07). 3,28-Di-O-[isonicotinate]-olean-12(13)-en **5**  
Beige solid; mp: 123\({}^{\circ}\)C; yield 72%; [\(\alpha\)]\({}_{D}\)\({}^{20}\) +5\({}^{\circ}\) (\(c\) 0.1, CHCl\({}_{3}\)). \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)) 8 8.80-8.73 (m, 4H, H4\({}^{\prime\prime}\), H6\({}^{\prime\prime}\), H4\({}^{\prime}\), H6\({}^{\prime}\)), 7.85-7.80 (m, 4H, H3\({}^{\prime\prime}\), H7\({}^{\prime\prime}\), H3\({}^{\prime}\), H7\({}^{\prime\prime}\)), 5.25 (s, 1H, H12), 4.79-4.72 (m, 1H, H3), 4.33 (d, \(J\) = 11.0 Hz, 1H, H28), 3.97 (d, \(J\) = 11.1 Hz, 1H, H28), 1.02-2.18 (m, 23H, CH, CH\({}_{2}\)), 1.19, 1.01, 0.99, 0.93, 0.91 (7s, 21H, 7 CH\({}_{3}\)). \({}^{13}\)C NMR (125.5 MHz, CDCl\({}_{3}\)) 8 165.06 (C1\({}^{\prime\prime}\)), 164.75 (C1\({}^{\prime}\)), 150.67 (C6\({}^{\prime\prime}\), C4\({}^{\prime\prime}\)), 150.56 (C6\({}^{\prime}\)), 150.51 (C4\({}^{\prime}\)), 143.37 (C13), 138.14 (C2\({}^{\prime}\)), 137.76 (C2\({}^{\prime}\)), 123.07 (C12), 122.86 (C3\({}^{\prime}\), C7\({}^{\prime\prime}\), C3\({}^{\prime}\), C7\({}^{\prime\prime}\)), 82.72 (C3), 71.98 (C28), 55.29 (C5), 47.51 (C18), 46.15 (C19), 42.59, 41.69, 40.13, 39.83, 38.25, 38.10, 37.45, 36.85 (C10), 36.24, 34.15, 33.94, 33.13, 32.43, 31.62, 30.91 (C20), 28.20, 26.20, 26.02, 25.62, 23.57, 23.52, 22.45, 21.50, 18.20, 16.94, 16.73, 15.57. MS (APCI) _m_/z 653.5 [M+ H]\({}^{+}\) (calcd for C\({}_{42}\)H\({}_{56}\)N\({}_{2}\)O\({}_{4}\), 652.92). 2.1.3. Synthesis of Compound **17**  
\(N\)-Methylpiperazine (1.6 mmol; 0.16 mL), paraformaldehyde(0.3 g), NaOAc (5 mmol; 0.41 g), and CuI (0.05 mmol; 9.5 mg) were added to a solution of compound **11** (1 mmol; 0.56 g) in dry dioxane (10 mL). The reaction mixture was stirred under argon for 10 h at 60 \({}^{\circ}\)C. After the reaction, the mixture was diluted with water and extracted with CHCl\({}_{3}\) (3 \(\times\) 20 mL). The combined organic layer was washed with water (3 \(\times\) 50 mL), dried over CaCl\({}_{2}\) and evaporated under reduced pressure.