In vitro Anti-Toxoplasma gondii and Antimicrobial Activity of Amides Derivated From Cinnamic Acid

Supplementary Materials In vitro Anti-Toxoplasma gondii and Antimicrobial Activity of Amides Derivated From Cinnamic Acid Graziela Rangel Silveira 1,*, Karoline Azerêdo Campelo 1, Gleice Rangel Silveira Lima 1, Lais Pessanha Carvalho 2, Solange Silva Samarão 3, Olney Vieira-da-Motta 3, Leda Mathias 1, Carlos Roberto Ribeiro Matos 1, Ivo José Curcino Vieira 1, Edesio José Tenório de Melo 2, Edmilson José Maria 1 1 Laboratório de Ciências Químicas, Centro de Ciências e Tecnologia, Universidade Estadual do Norte Fluminense-Darcy Ribeiro, Av. Alberto Lamego, 2000 - Parque Califórnia, Campos dos Goytacazes/RJ, 28013-602, Brasil; graziela_sil@hotmail.com (G. R. S.); karolcampelo16@yahoo.com.br (K. A. C.); gleice.sil@hotmail.com (G. R. S. L.); leddam8@gmail.com (L. M.); matos@uenf.br (C. R. R. M.); curcino@uenf.br (I. J. C. V.); edmilson_maria@yahoo.com.br (E. J. M.). 2 Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense-Darcy Ribeiro, Av. Alberto Lamego, 2000 - Parque Califórnia, Campos dos Goytacazes/RJ, 28013-602, Brasil; lais_pessanha@hotmail.com (L. P. C.); ejtm1202@gmail.com (E. J. T. M.). 3 Laboratório de Sanidade Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense-Darcy Ribeiro, Av. Alberto Lamego, 2000 - Parque Califórnia, Campos dos Goytacazes/RJ, 28013-602, Brasil; solangesamarao@gmail.com (S. S. S.); olney.motta@gmail.com (O. V.dM.). * Corresponding author: graziela_sil@hotmail.com; Tel.: +55-022-2728-6167 Molecules 2018, 23, 774; doi:10.3390/molecules23040774 www.mdpi.com/journal/molecules

Molecules 2018, 23, 774 2 of 49 Figure S1. 1 H NMR spectrum of compound 1 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 3 of 49 Figure S2. 13 C NMR spectrum of compound 1 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 4 of 49 Figure S3. HMBC spectrum of compound 1 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 5 of 49 001 HSQC.ESP 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 F2 Chemical Shift (ppm) -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Figure S4. HSQC spectrum of compound 1 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 6 of 49 Abundance 3200000 3000000 Scan 1852 (21.348 m in):001.d \data.m s (-1834)(-) 134.0 2800000 2600000 2400000 2200000 2000000 1800000 1600000 1400000 1200000 1000000 800000 600000 400000 77.0 103.0 200000 51.0 160.1 180.9 207.0 230.1 252.9 281.0 314.9 341.0 0 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 m /z--> Figure S5. Mass spectrum of compound 1 (CH2Cl2).

Molecules 2018, 23, 774 7 of 49 Figure S6. IR spectrum (KBr) of compound 1.

Molecules 2018, 23, 774 8 of 49 Figure S7. 1 H NMR spectrum of compound 2 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 9 of 49 Figure S8. 13 C NMR spectrum of compound 2 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 10 of 49 Figure S9. HMBC spectrum of compound 2 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 11 of 49 002 HSQC.ESP 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 F2 Chemical Shift (ppm) -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Figure S10. HSQC spectrum of compound 2 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 12 of 49 Abundance 6000000 Scan 1694 (19.854 m in):002.d\data.m s 131.1 5500000 5000000 4500000 4000000 3500000 3000000 103.1 2500000 2000000 1500000 77.0 265.1 1000000 500000 51.0 160.0 m/z--> 186.0 207.0 236.0 292.9 315.0 341.0 0 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 Figure S11. Mass spectrum of compound 2 (CH2Cl2).

Molecules 2018, 23, 774 13 of 49 Figure S12. IR spectrum (KBr) of compound 2.

Molecules 2018, 23, 774 14 of 49 Figure 13. 1 H NMR spectrum of compound 3 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 15 of 49 Figure S14. 13 C NMR spectrum of compound 3 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 16 of 49 Figure S15. HMBC spectrum of compound 3 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 17 of 49 003 HSQC.ESP 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 F2 Chemical Shift (ppm) Figure S16 HSQC spectrum of compound 3 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 18 of 49 Abundance 4500000 Scan 1817 (21.017 min):003.d\data.ms 131.1 4000000 3500000 3000000 2500000 2000000 103.0 1500000 77.0 1000000 160.0 285.1 500000 51.0 207.0 181.0 229.0 253.0 315.0 341.0 0 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 m/z-> Figure S17. Mass spectrum of compound 3 (CH2Cl2).

Molecules 2018, 23, 774 19 of 49 Figure S18. IR spectrum (KBr) of compound 3.

Molecules 2018, 23, 774 20 of 49 Figure S19. 1 H NMR spectrum of compound 4 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 21 of 49 Figure S20. 13 C NMR spectrum of compound 4 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 22 of 49 Figure S21. HMBC spectrum of compound 4 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 23 of 49 004 HSQCED.ESP 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 12 11 10 9 8 7 6 5 4 3 F2 Chemical Shift (ppm) Figure S22. HSQC spectrum of compound 4 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 24 of 49 Abundance 5000000 Scan 2212 (24.752 min):004.d\data.ms 131.1 4500000 4000000 3500000 3000000 2500000 103.0 2000000 1500000 1000000 77.0 160.0 500000 51.0 0 m/z--> 296.1 207.0 181.0 232.1 253.0 317.0 341.0 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 Figure S23. Mass spectrum of compound 4 (CH2Cl2).

Molecules 2018, 23, 774 25 of 49 Figure S24. IR spectrum (KBr) of compound 4.

Molecules 2018, 23, 774 26 of 49 Figure S25. 1 H NMR spectrum of compound 5 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 27 of 49 Figure S26. 13 C NMR spectrum of compound 5 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 28 of 49 Figure S27. HMBC spectrum of compound 5 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 29 of 49 005 HSQC.ESP 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 F1 Chemical Shift (ppm) 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 F2 Chemical Shift (ppm) Figure S28. HSQC spectrum of compound 5 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 30 of 49 Abundance 6000000 164.1 Scan 2047 (23.192 m in):006.d \data.m s 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 103.0 1500000 1000000 500000 51.0 311.1 207.0 253.0 355.0 429.1 503.1 553.1 627.1 0 50 100 150 200 250 300 350 400 450 500 550 600 m /z--> Figure S29. Mass spectrum of compound 5 (CH2Cl2).

Molecules 2018, 23, 774 31 of 49 Figure S30. IR spectrum (KBr) of compound 5.

Molecules 2018, 23, 774 32 of 49 Figure S31. 1 H NMR spectrum of compound 6 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 33 of 49 Figure S32. 13 C NMR spectrum of compound 6 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 34 of 49 Figure S33. HMBC spectrum of compound 6 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 35 of 49 006 HSQC.ESP 10 9 8 7 6 5 4 3 F2 Chemical Shift (ppm) -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Figure S34. HSQC spectrum of compound 6 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 36 of 49 A b u n d a n c e 5 0 0 0 0 0 S c a n 2 9 3 6 (3 1.5 9 8 m in ): 0 0 9.D \d a ta.m s (-2 8 3 7 ) (-) 1 7 5.9 4 5 0 0 0 0 4 0 0 0 0 0 3 5 0 0 0 0 3 0 0 0 0 0 2 5 0 0 0 0 2 0 0 0 0 0 1 0 2.0 1 5 0 0 0 0 1 3 0.1 1 0 0 0 0 0 m /z--> 5 0 0 0 0 6 3.0 2 0 5.0 3 7 4.0 2 5 3.0 2 8 1.0 3 3 1.1 4 0 6.1 4 5 2.1 0 40 60 80 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0 2 6 0 2 8 0 3 0 0 3 2 0 3 4 0 3 6 0 3 8 0 4 0 0 4 2 0 4 4 0 Figure S35. Mass spectrum of compound 6 (CH2Cl2).

Molecules 2018, 23, 774 37 of 49 Figure S36. IR spectrum (KBr) of compound 6.

Molecules 2018, 23, 774 38 of 49 Figure S37. 1 H NMR spectrum of compound 7 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 39 of 49 Figure S38. 13 C NMR spectrum of compound 7 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 40 of 49 Figure S39. HMBC spectrum of compound 7 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 41 of 49 007 HSQC.ESP 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 F2 Chemical Shift (ppm) -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Figure S40. HSQC spectrum of compound 7 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 42 of 49 A b u n d a n c e 4 0 0 0 0 0 S c a n 3 8 6 2 (4 0.3 5 3 m in ): 0 1 0.D \d a ta.m s (-3 9 4 9 ) (-) 1 7 6.0 3 5 0 0 0 0 3 0 0 0 0 0 2 5 0 0 0 0 2 0 0 0 0 0 1 5 0 0 0 0 1 0 0 0 0 0 1 0 2.0 1 2 9.9 2 0 5.1 5 0 0 0 0 5 1.0 3 4 1.1 2 5 4.0 2 8 3.1 3 1 1.1 3 7 3.0 4 0 5.1 4 5 1.0 0 40 60 80 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0 2 6 0 2 8 0 3 0 0 3 2 0 3 4 0 3 6 0 3 8 0 4 0 0 4 2 0 4 4 0 m /z--> Figure S41. Mass spectrum of compound 7 (CH2Cl2).

Molecules 2018, 23, 774 43 of 49 Figure S42. IR spectrum (KBr) of compound 7.

Molecules 2018, 23, 774 44 of 49 Figure S43. 1 H NMR spectrum of compound 8 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 45 of 49 Figure S44. 13 C NMR spectrum of compound 8 (CDCl3, 125 MHz).

Molecules 2018, 23, 774 46 of 49 Figure S45. HMBC spectrum of compound 8 (CDCl3, 500 MHz).

F1 Chemical Shift (ppm) Molecules 2018, 23, 774 47 of 49 117.200.001.2rr.esp 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 F2 Chemical Shift (ppm) 8 16 24 32 40 48 56 64 72 80 88 96 104 112 120 128 136 144 152 160 168 176 Figure S46. HSQC spectrum of compound 8 (CDCl3, 500 MHz).

Molecules 2018, 23, 774 48 of 49 Abundance 5000000 Scan 1971 (22.473 m in): 011.D \data.m s (-1947) (-) 138.0 4500000 4000000 3500000 3000000 2500000 2000000 1500000 m /z--> 1000000 500000 0 77.0 183.0 240.0 367.0 287.1 330.0 405.1 467.1 503.1 554.2 50 100 150 200 250 300 350 400 450 500 550 Figure S47. Mass spectrum of compound 8 (CH2Cl2).

Molecules 2018, 23, 774 49 of 49 Figure S48. IR spectrum (KBr) of compound 8.