# 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
#### 2.1.2 General Procedure for the Preparation of **4a-d**
: 164.9-165.9 \({}^{\circ}\)C: \({}^{1}\)H NMR (CDC1\({}_{3}\), 300 MHz) \(\delta\) ppm 7.61 (d, 2H, \(J\) = 8.4 Hz, Ar-H), 7.39 (s, 1H, 8\({}^{\prime\prime}\)-H), 6.95 (d, 2H, \(J\) = 8.4 Hz, Ar-H), 5.27 (brs, 1H, 12-H), 4.05 (t, 2H, \(J\) = 6.8 Hz, 1\({}^{\prime}\)-H), 4.03 (t, 2H, \(J\) = 6.8 Hz, 5\({}^{\prime}\)-H), 3.19 (m, 1H, 3-H), 2.86 (brd, 1H, \(J\) = 10.9 Hz, 18-H), 1.13-2.20 (m, 28H, methylene and methine protons), 1.12 (s, 3H, 29-H), 0.97 (s, 3H, 30-H), 0.92 (s, 3H, 26-H), 0.90 (s, 3H, 23-H), 0.87 (s, 3H, 25-H), 0.76 (s, 3H, 27-H), 0.72 (s, 3H, 24-H), OH shows no signal; \({}^{13}\)C NMR (CDC1\({}_{3}\), 75 MHz) \(\delta\) ppm 215.3, 177.8, 162.4, 143.8, 134.6, 128.6, 122.3, 115.4, 79.0, 68.2, 63.9, 55.2, 47.6, 46.7, 45.9, 41.7, 41.3, 39.3, 38.7, 38.4, 37.0, 33.8, 33.1, 32.6, 32.5, 31.6, 30.7, 28.7, 27.81, 27.  
or 7. The intermediate product 5 or 7 (1.0 mmol) was dissolved in 20 mL of anhydrous dichloromethane solvent, dehydrating agent EDCI (2.4 mmol) was added, and a certain amount of dimethylaminopyridine catalyst was present in the flask. This was stirred for 20 min at room temperature. Then, N-benzyl-4-hydroxybenzothioamide or 4-OH-TBZ or ADT-OH 2.0 mmol was dissolved in 10 mL of anhydrous dichloromethane solvent, dropwise added to the flask, and continued to react until the reaction was completed. Then, the whole solution was washed 3 times with water (20 mL \(\times\) 3), saturated aqueous sodium bicarbonate solution (20 mL \(\times\) 3), and saturated brine (20 mL \(\times\) 3), in that order. The organic layer was collected, dried over anhydrous sodium sulfate, filtered, evaporated in vacuo, and passed through the column; the residues obtained after separation and purification were the corresponding products **6a**, **6b** and **8a-c**. _Oleanolic acid 3-[4-(4-carbamoylomethyl)] succinate_ (**6a**). Yield: 29%. Yellow solid, m.p. : 140.4-142.0 \({}^{\circ}\)C: \({}^{1}\)H NMR (CDCl\({}_{3}\), 400 MHz) \(\delta\) ppm 7.14 (d, 2H, \(J\) = 7.6 Hz, Ar-H), 7.06 (d, 2H, \(J\) = 7.6 Hz, Ar-H), 5.35 (brs, 1H, 12-H), 4.55 (t, 1H, \(J\) = 7.2 Hz, 3-H), 2.95 (dd, 1H, \(J\) = 13.6, 2.4 Hz, 18-H), 2.90 (t, 2H, \(J\) = 6.0 Hz, 3\({}^{\prime}\)-H), 2.75 (t, 2H, \(J\) = 6.0 Hz, 2\({}^{\prime}\)-H), 1.19-2.20 (m, 22H, methylene and methine protons), 1.18 (s, 3H, 29-H), 0.97 (s, 3H, 30-H), 0.94 (s, 3H, 26-H), 0.93 (s, 3H, 23-H), 0.87 (s, 3H, 25-H), 0.87 (s, 3H, 27-H), 0.81 (s, 3H, 24-H), COOH and NH2 show no signal; \({}^{13}\)C NMR (CDCl\({}_{3}\), 100 MHz) \(\delta\) ppm 182.5, 173.8, 172.8, 154.5, 143.6, 133.6, 122.8, 122.6, 109.4, 80.5, 55.2, 47.6, 46.5, 45.9, 41.6, 41.0, 39.3, 38.4, 38.0, 37.1, 33.8, 33.1, 32.6, 32.4, 30.7, 30.1, 29.8, 28.1, 27.7, 25.9, 24.7, 23.6, 23.4, 22.9, 18.3, 17.1, 15.5, 15.3; HRMS (ESI) Calcd for C\({}_{41}\)H\({}_{88}\)NO\({}_{6}\)S [M + H]\({}^{+}\) 692.3979, found: 692.3973. _Oleanolic acid 3-[4-(5-thixo-5H-1,2-dithiol-3-yl)phenyl] succinate_ (**6b**). Yield: 30%. Red brown solid, m.p. : 126.6-127.6 \({}^{\circ}\)C: \({}^{1}\)H NMR (CDCl\({}_{3}\), 300 MHz) \(\delta\) ppm 7.40 (s, 1H, \(\delta^{\prime}\)-H), 7.25 (d, 2H, \(J\) = 8.8 Hz, Ar-H), 7.17 (d, 2H, \(J\) = 8.8 Hz, Ar-H), 5.36 (t, 1H, \(J\) = 3.2 Hz, 12-H), 4.57 (t, 1H, \(J\) = 7.6 Hz, 3-H), 2.97 (dd, 1H, \(J\) = 10.2, 2.4 Hz, 18-H), 2.91 (t, 2H, \(J\) = 6.4 Hz, 2\({}^{\prime}\)-H), 2.76 (t, 2H, \(J\) = 6.4 Hz, 3\({}^{\prime}\)-H), 1.27-2.0 (m, 22H, methylene and methine protons), 1.26 (s, 6H, 29-H and 30-H), 1.19 (s, 3H, 26-H), 0.98 (s, 3H, 23-H), 0.95 (s, 3H, 25-H), 0.88 (s, 3H, 27-H), 0.83 (s, 3H, 27-H), 0.83 (s, 3H, 24-H), COOH shows no signal; \({}^{13}\)C NMR (CDCl\({}_{3}\), 75 MHz) \(\delta\) ppm 125.3, 182.5, 173.8, 172.8,172.0, 147.1, 143.6, 128.9, 128.2, 124.0, 123.1, 122.6, 80.5, 55.2, 47.6, 46.5, 45.9, 41.6, 41.0, 39.3, 38.4, 38.0, 37.1, 33.8, 33.1, 32.6, 32.4, 30.7, 30.1, 29.8, 28.1, 27.7, 25.9, 24.7, 23.6, 23.4, 22.9, 18.3, 17.1, 15.5, 15.3; HRMS (ESI) Calcd for C\({}_{43}\)H\({}_{57}\)O\({}_{6}\)S\({}_{3}\) [M + H]\({}^{+}\) 765.3312, found: 765.3317.