# Design, Synthesis, and Cell Lines Studies of Oleanolic Acid--Hydrogen Sulfide Donor Hybrids
## 2 Materials and Methods
### General Procedure for the Synthesis of Targeted Compounds
#### 2.1.1 General Procedure for the Preparation of **2a-c**  
Oleanolic acid, white needles (CH\({}_{3}\)OH): \({}^{1}\)H NMR (CDCl\({}_{3}\), 400 MHz) \(\delta\) ppm 5.27 (t, 1H, \(J\) = 3.2 Hz, 12-H), 3.22 (dd, 1H, \(J\) = 11.2, 4.0 Hz, 3-H), 2.81 (dd, 1H, \(J\) = 14.4, 4.0 Hz, 18-H), 1.13-2.20 (m, 22H, methylene and methine protons), 1.12 (s, 3H, 29-H), 0.98 (s, 3H, 30-H), 0.92 (s, 3H, 26-H), 0.90 (s, 3H, 23-H), 0.90 (s, 3H, 25-H), 0.77 (s, 3H, 27-H), 0.74 (s, 3H, 24-H), OH and COOH show no signal; \({}^{13}\)C NMR (CDCl\({}_{3}\), 100 MHz) \(\delta\) ppm 183.1, 143.6,122.6, 79.1, 55.2, 47.6, 46.5, 45.9, 41.6, 41.0, 39.3, 38.8, 38.4, 37.1, 33.8, 33.1, 32.6, 32.4, 30.7, 28.1, 27.7, 27.2, 25.9, 23.6, 23.4, 22.9, 18.3, 17.1, 15.5, 15.3. EDCI (2.4 mmol), DMAP (0.1 mmol) were added to OA (1.0 mmol) in anhydrous CH\({}_{2}\)Cl\({}_{2}\) (20 mL) solution. The mixture was stirred for 20 min under room temperature [36,37]. Then, ADT-OH or 4-OH-TBZ or N-benzyl-4-hydroxybenzothamide (2.0 mmol, dissolved in 10 mL dry CH\({}_{2}\)Cl\({}_{2}\)) was added dropwise to the mixture and the reaction solution was whisked until the end of the reaction. Then, we washed the whole solution using water (20 mL \(\times\) 3), saturated NaHCO\({}_{3}\) aqueous solution (20 mL \(\times\) 3) and saturated brine (20 mL \(\times\) 3), dried and filtered it with anhydrous Na\({}_{2}\)SO\({}_{4}\), inspissated it under reduced pressure to obtain a remainder, and purified it by flash column chromatography to acquire pure product. _Oleanolic acid 28-(4-thiocarbamoyl) benzoate_ (**2a**). Yield: 41%. White solid, m.p. : 200.3-202.0 \({}^{\circ}\)C: \({}^{1}\)H NMR (CDCl\({}_{3}\), 300 MHz) \(\delta\) ppm 7.65 (d, 2H, \(J\) = 8.2 Hz, Ar-H), 7.13 (d, 2H, \(J\) = 8.2 Hz, Ar-H), 5.34 (brs, 1H, 12-H), 3.20 (m, 1H, 3-H), 2.93 (br d, 1H, \(J\) = 11.3 Hz, 18-H), 1.18-2.20 (m, 22H, methylene and methine protons), 1.17 (s, 3H, 29-H), 0.98 (s, 3H, 30-H), 0.95 (s, 3H, 26-H), 0.93 (s, 3H, 23-H), 0.90 (s, 3H, 25-H), 0.80 (s, 3H, 27-H), 0.76 (s, 3H, 24-H), OH and NH\({}_{2}\) show no signal; \({}^{13}\)C NMR (CDCl\({}_{3}\), 75 MHz) \(\delta\) ppm 175.5, 154.5, 143.0, 133.6, 122.8, 123.1, 109.4, 78.9, 55.2, 47.5, 47.4, 45.7, 41.8, 41.4, 39.5, 38.8, 38.5, 37.0, 33.8, 33.1, 32.8, 32.3, 30.7, 28.1, 27.8, 27.2, 25.8, 23.6, 23.4, 23.0, 18.3, 17.4, 15.6, 15.4; HRMS (ESI) Calcd for C\({}_{3}\)T\({}_{54}\)NO\({}_{5}\)S [M + H]\({}^{+}\) 592.3824, found: 592.3813. _Oleanolic acid 28-[4-(5-thioxo-5H-1,2-dithiol-3-yl)] benzoate_ (**2b**). Yield: 57%. Yellow solid, m.p. : 154.5-156.1 \({}^{\circ}\)C: \({}^{1}\)H NMR (CDCl\({}_{3}\), 300 MHz) \(\delta\) ppm 7.64 (d, 2H, \(J\) = 8.2 Hz, Ar-H), 7.39 (s, 1H, 8\({}^{\prime}\)H), 7.15 (d, 2H, \(J\) = 8.2 Hz, Ar-H), 5.35 (br s, 1H, 12-H), 3.21 (m, 1H, 3-H), 2.80 (br d, 1H, \(J\) = 11.7 Hz, 18-H), 1.19-2.20 (m, 22H, methylene and methine protons), 1.18 (s, 3H, 29-H), 0.96 (s, 3H, 30-H), 0.96 (s, 3H, 26-H), 0.93 (s, 3H, 23-H), 0.90 (s, 3H, 25-H), 0.82 (s, 3H, 27-H), 0.77 (s, 3H, 24-H), OH shows no no signal, \({}^{13}\)C NMR (CDCl\({}_{3}\), 75 MHz) \(\delta\) ppm 215.3, 175.7, 172.0, 147.1, 143.1, 128.9, 128.2, 124.0, 123.1, 122.8, 79.0, 55.2, 47.6, 47.4, 45.7, 41.8, 41.5, 39.5, 38.8, 38.5, 37.0, 33.8, 33.1, 32.8, 32.4, 30.7, 28.1, 27.8, 27.1, 25.9, 23.6, 23.5, 23.1, 18.3, 17.5, 15.7, 15.4; HRMS (ESI) Calcd for C39H53O3S [M + H]\({}^{+}\) 665.3157, found: 665.3153. _Oleanolic acid 28-[4-(N-benzyl-thioxo-4)thiocarbamoyl] benzoate_ (**2c**). Yield: 40%. Yellow solid, m.p.