Virtual screening of ZINC database
We used the anchor motif as template to design and refine pharmacophore models to virtually screen more than 27 million compounds using the public server ZINCPharmer (Koes & Camacho, 2012). Based on A1R2 configuration we screened near to 27 million commercially available compounds using ZINCPharmer, resulting in the compounds studied here.
Synthesis of CBD3063
Step 1. 3-acetamidopiperidine-1-carbonyl chloride
In a cooled (0 °C) solution of N -(piperidin-3-yl)acetamide (500 mg, 3.52 mmol) in anhydrous dichloromethane (20 mL) was added NaHCO3 (1.10 g, 10.6 mmol) and triphosgene (696 mg, 2.34 mmol). The mixture was stirred at room temperature for 1 hour. After all starting material has been consumed, the mixture was filtered, and the collected filtrate was evaporated under reduced pressure. The resulting residue was then allowed to passed through a short silica plug (wash with 100% EtOAc) to yield crude 3-acetamidopiperidine-1-carbonyl chloride (467 mg, 65%) as sticky transparent liquid (HRMS calcd for C8H14ClN2O2+[M+H]+: 205.0748; found: 205.0738). The compound was used immediately for the next step without further purification.
Step 2. 3-acetamido-N -(3-(pyridin-2-ylamino)propyl)piperidine-1-carboxamide
3-acetamidopiperidine-1-carbonyl chloride (100 mg, 0.489 mmol) from the previous step was dissolved in anhydrous dichloromethane (5.0 mL). Into this solution was added Na2CO3 (104 mg, 0.977 mmol) andN 1-(pyridin-2-yl)propane-1,3-diamine (73.9 mg, 0.489 mmol). The mixture was stirred for 2 hours, upon which all starting material had reacted. The mixture was filtered, and the collected filtrate was evaporated under reduced pressure. The resulting residue was then purified by flash column chromatography (gradient elution of 0% to 10% MeOH in CH2Cl2) to yield3- acetamido-N -(3-(pyridin-2-ylamino)propyl)piperidine-1-carboxamide (137 mg, 88%) as white foam solid. 1H NMR (600 MHz, CDCl3) δ 7.99 (dd, J = 4.1, 0.8 Hz, 1H), 7.38 (ddd, J = 8.7, 7.1, 1.9 Hz, 1H), 6.53 (ddd, J = 7.0, 5.2, 0.8 Hz, 1H), 6.41 (d, J = 8.4 Hz, 1H), 6.12 (d, J = 6.5 Hz, 1H), 5.79 (t, J = 5.5 Hz, 1H), 5.10 – 4.95 (m, 1H), 3.99 – 3.86 (m, 1H), 3.45 – 3.36 (m, 6H), 3.32 (q, J = 6.1 Hz, 2H), 1.95 (s, 3H), 1.87 – 1.62 (m, 5H), 1.61 – 1.49 (m, 1H).13C NMR (151 MHz, CDCl3) δ 170.10, 158.60, 158.46, 147.02, 137.59, 112.50, 108.28, 48.66, 45.61, 44.60, 38.63, 37.69, 30.22, 29.40, 23.37, 22.23. HRMS calcd for C16H25N5O2Na [M+Na]+: 342.1900; found: 342.1908.