# Simple Thalidomide Analogs in Melanoma: Synthesis and Biological Activity
## Materials and Methods  
### Chemistry  
All chemicals were supplied by Sigma-Aldrich or Alfa Aesar and were the highest-grade purity that were commercially available. Solvents were reagent grade unless otherwise indicated. Yields refer to purified products and were not optimized. Compound structures were confirmed by routine spectrometric analyses. Melting points were determined on a Gallenkamp melting point apparatus in open glass capillary tubes and are uncorrected. 1H NMR and 13C NMR spectra were recorded on a Varian VX Mercury spec  
Figure 1: Structure of thalidomide and correlated compounds (**2a-l**). trometer operating at 300 and 75 MHz for \({}^{1}\)H and \({}^{13}\)C, respectively, or an Agilent 500 MHz operating at 500 and 125 MHz for \({}^{1}\)H and \({}^{13}\)C, respectively. Chemical shifts are reported in parts per million (ppm) relative to solvent (CDCl\({}_{3}\)) resonance: \(\delta\) 7.26 (\({}^{1}\)H NMR) and \(\delta\) 77.0 (\({}^{13}\)C NMR). \(J\) values are given in Hz. The following abbreviations are used: s-singlet, d-doublet, t-triplet, and m-multiplet. Gas chromatography (GC)/mass spectroscopy (MS) was performed on a Hewlett-Packard 6890-5973 MSD at low resolution. Liquid chromatography (LC)/mass spectroscopy (MS) was performed on a spectrometer (Agilent 1100 series LC-MSD Trap System VL). Molecular ion was designed as \"M\({}^{+}\)\". Elemental analyses were carried out on a Eurovector Euro EA 3000 analyzer and the data for C, H, N were within \(\pm\)0.4 of theoretical values. _Synthesis of 2-phenyl-1H-isointol-1,3(2H)-dione_**(2a)**. Aniline (0.77 g, 8.3 mmol), phthal anhydride (1.23 g, 8.3 mmol) and triethylamine (0.12 mL, 0.9 mmol) were refluxed in toluene (10 mL) for 5 h. The solvent was removed in vacuo and the residue, taken up with CHCl\({}_{3}\), was washed 3 N HCl and water, and then dried over anhydrous Na\({}_{2}\)SO\({}_{4}\) to give 905 mg (49%) of **2a** as a white solid: mp 211-213 \({}^{\circ}\)C; lit. 200 \({}^{\circ}\)C [18]; 210-211 \({}^{\circ}\)C (acetone) [19]. GC/MS (70 eV) _m/z_ (%): 223 (M\({}^{+}\), 100); LC/MS: _m/z_ (%): 222 [M\({}^{+}\)-1]; \({}^{1}\)H NMR (300 MHz, CDCl\({}_{3}\)): \(\delta\) 7.36-7.54 (m, 5H, Ar), 7.54-7.82 (m, 2H, Ar), 7.90-7.98 (m, 2H, Ar); \({}^{13}\)C NMR (75 MHz, CDCl\({}_{3}\)): \(\delta\) 123.7 (2C), 126.6 (1C), 128.1 (2C), 129.1 (2C), 131.8 (2C), 134.4 (2C), 167.3 (2C). Anal C\({}_{13}\)H\({}_{9}\)NO\({}_{2}\) (223.06): calcd %: C 75.33; H 4.06; N 6.27. Found %: C 74.98; H 4.04; N 6.30. _Synthesis of 2-(2-fluorophenyl)-1H-isointol-1,3(2H)-dione_**(2b)**. Prepared as described for 2a starting from 2-fluoraniline. White solid. Yield: 65%: mp 190-191 \({}^{\circ}\)C (CHCl\({}_{3}\)/hexane); lit. 199 \({}^{\circ}\)C [19]; GC/MS (70 eV) _m/z_ (%): 241 (M\({}^{+}\), 92), 197 (100); \({}^{1}\)H NMR (500 MHz, CDCl\({}_{3}\)): \(\delta\) 7.23-7.30 (m, 2H, Ar), 7.35-7.38 (m, 1H, Ar), 7.42-7.47 (m, 1H, Ar), 7.79-7.82 (m, 2H, Ar), 7.95-7.98 (m, 2H, Ar); \({}^{13}\)C NMR (75 MHz, CDCl\({}_{3}\)): \(\delta\) 116.76 (d, \(J\) = 19.6 Hz, 1C), 119.4 (d, \(J_{\rm CF}\) = 13.7 Hz, 1C), 123.9 (1C), 124.6 (d, \(J_{\rm CF}\) = 4.6 Hz, 1C), 129.9 (2C), 130.7 (d, \(J_{\rm CF}\) = 8.0 Hz, 1C), 131.9 (2C), 134.4 (2C), 157.9 (d, \(J_{\rm CF}\) = 251.9 Hz, 1C), 166.5 (2C). Anal C\({}_{14}\)H\({}_{8}\)FNO\({}_{2}\) (241.05): calcd %: C 69.71; H 3.34; N 5.81. Found %: C 69.54; H 3.32; N 5.80. _Synthesis of 2-(4-(4-methylphenoxy)/phenyl)-1H-isointol-1,3(2H)-dione_**(2c)**. Prepared as described for **2a starting from 4-p-tolyloxyaniline. Grey solid.