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

Molecules 2018, 23, 774; doi:10.3390/molecules23040774 www.mdpi.com/journal/molecules Supplementary Materials

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

Graziela Rangel Silveira^1,*, Karoline Azerêdo Campelo¹, Gleice Rangel Silveira Lima¹, Lais Pessanha Carvalho², Solange Silva Samarão³, Olney Vieira-da-Motta³, Leda Mathias¹, Carlos

Roberto Ribeiro Matos¹, Ivo José Curcino Vieira¹, Edesio José Tenório de Melo², Edmilson José Maria¹

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; [email protected] (G. R. S.); [email protected] (K. A. C.);

[email protected] (G. R. S. L.); [email protected] (L. M.); [email protected] (C. R. R. M.);

[email protected] (I. J. C. V.); [email protected] (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; [email protected] (L. P. C.); [email protected] (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; [email protected] (S. S. S.); [email protected] (O.

V.dM.).

* Corresponding author: [email protected]; Tel.: +55-022-2728-6167

Figure S1. ¹H NMR spectrum of compound 1 (CDCl3, 500 MHz).

Figure S2. ¹³C NMR spectrum of compound 1 (CDCl3, 125 MHz).

Figure S3. HMBC spectrum of compound 1 (CDCl3, 500 MHz).

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 Shif t (ppm)

-10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210

F1 Chemical Shift (ppm)

Figure S4. HSQC spectrum of compound 1 (CDCl3, 500 MHz).

40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 0

200000 400000 600000 800000 1000000 1200000 1400000 1600000 1800000 2000000 2200000 2400000 2600000 2800000 3000000 3200000

m /z-->

Abundance

Scan 1852 (21.348 m in): 001.D \data.m s (-1834) (-) 134.0

103.0 77.0

51.0 160.1180.9 207.0230.1252.9 281.0 314.9 341.0

Figure S5. Mass spectrum of compound 1 (CH²Cl²).

Figure S6. IR spectrum (KBr) of compound 1.

Figure S7. ¹H NMR spectrum of compound 2 (CDCl3, 500 MHz).

Figure S8. ¹³C NMR spectrum of compound 2 (CDCl3, 125 MHz).

Figure S9. HMBC spectrum of compound 2 (CDCl3, 500 MHz).

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 Shif t (ppm)

-10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210

F1 Chemical Shift (ppm)

Figure S10. HSQC spectrum of compound 2 (CDCl3, 500 MHz).

40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 0

500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000 5000000 5500000 6000000

m /z-->

Abundance

Scan 1694 (19.854 m in): 002.D \data.m s 131.1

103.1

77.0

265.1

51.0 160.0

207.0 236.0 315.0 341.0

186.0 292.9

Figure S11. Mass spectrum of compound 2 (CH2Cl2).

Figure S12. IR spectrum (KBr) of compound 2.

Figure 13. ¹H NMR spectrum of compound 3 (CDCl3, 500 MHz).

Figure S14. ¹³C NMR spectrum of compound 3 (CDCl3, 125 MHz).

Figure S15. HMBC spectrum of compound 3 (CDCl3, 500 MHz).

003 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 2.5

F2 Chemical Shif t (ppm)

0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180

F1 Chemical Shift (ppm)

Figure S16 HSQC spectrum of compound 3 (CDCl3, 500 MHz).

40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 0

500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000

m/z-->

Abundance

Scan 1817 (21.017 min):003.D\data.ms 131.1

103.0

77.0

160.0

285.1 51.0

207.0 253.0 341.0

181.0 229.0 315.0

Figure S17. Mass spectrum of compound 3 (CH2Cl2).

Figure S18. IR spectrum (KBr) of compound 3.

Figure S19. ¹H NMR spectrum of compound 4 (CDCl3, 500 MHz).

Figure S20. ¹³C NMR spectrum of compound 4 (CDCl3, 125 MHz).

Figure S21. HMBC spectrum of compound 4 (CDCl3, 500 MHz).

004 HSQCED.ESP

12 11 10 9 8 7 6 5 4 3

F2 Chemical Shif t (ppm)

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)

Figure S22. HSQC spectrum of compound 4 (CDCl3, 500 MHz).

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 0

500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000 5000000

m/z-->

Abundance

Scan 2212 (24.752 min): 004.D\data.ms 131.1

103.0

77.0

160.0

296.1 51.0

207.0 253.0 341.0

181.0 232.1 317.0

Figure S23. Mass spectrum of compound 4 (CH2Cl2).

Figure S24. IR spectrum (KBr) of compound 4.

Figure S25. ¹H NMR spectrum of compound 5 (CDCl3, 500 MHz).

Figure S26. ¹³C NMR spectrum of compound 5 (CDCl3, 125 MHz).

Figure S27. HMBC spectrum of compound 5 (CDCl3, 500 MHz).

005 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

F2 Chemical Shif t (ppm)

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)

Figure S28. HSQC spectrum of compound 5 (CDCl³, 500 MHz).

50 100 150 200 250 300 350 400 450 500 550 600 0

500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000 5000000 5500000 6000000

m /z-->

Abundance

Scan 2047 (23.192 m in): 006.D \data.m s 164.1

103.0

311.1

51.0 207.0 253.0 355.0 429.1 503.1 553.1 627.1

Figure S29. Mass spectrum of compound 5 (CH2Cl2).

Figure S30. IR spectrum (KBr) of compound 5.

Figure S31. ¹H NMR spectrum of compound 6 (CDCl3, 500 MHz).

Figure S32. ¹³C NMR spectrum of compound 6 (CDCl3, 125 MHz).

Figure S33. HMBC spectrum of compound 6 (CDCl3, 500 MHz).

006 HSQC.ESP

10 9 8 7 6 5 4 3

F2 Chemical Shif t (ppm)

-10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210

F1 Chemical Shift (ppm)

Figure S34. HSQC spectrum of compound 6 (CDCl3, 500 MHz).

4 0 6 0 8 0 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 0

5 0 0 0 0 1 0 0 0 0 0 1 5 0 0 0 0 2 0 0 0 0 0 2 5 0 0 0 0 3 0 0 0 0 0 3 5 0 0 0 0 4 0 0 0 0 0 4 5 0 0 0 0 5 0 0 0 0 0

m / z -->

A b u n d a n c e

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

1 0 2 .0

1 3 0 .1

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

Figure S35. Mass spectrum of compound 6 (CH2Cl2).

Figure S36. IR spectrum (KBr) of compound 6.

Figure S37. ¹H NMR spectrum of compound 7 (CDCl3, 500 MHz).

Figure S38. ¹³C NMR spectrum of compound 7 (CDCl3, 125 MHz).

Figure S39. HMBC spectrum of compound 7 (CDCl3, 500 MHz).

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 Shif t (ppm)

-10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210

F1 Chemical Shift (ppm)

Figure S40. HSQC spectrum of compound 7 (CDCl3, 500 MHz).

4 0 6 0 8 0 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 0

5 0 0 0 0 1 0 0 0 0 0 1 5 0 0 0 0 2 0 0 0 0 0 2 5 0 0 0 0 3 0 0 0 0 0 3 5 0 0 0 0 4 0 0 0 0 0

m /z-->

A b u n d a n c e

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

2 0 5 .1 1 0 2 .0

1 2 9 .9

3 4 1 .1 5 1 .0

3 1 1 .1

2 8 3 .1 4 0 5 .1

2 5 4 .0 3 7 3 .0 4 5 1 .0

Figure S41. Mass spectrum of compound 7 (CH2Cl2).

Figure S42. IR spectrum (KBr) of compound 7.

Figure S43. ¹H NMR spectrum of compound 8 (CDCl3, 500 MHz).

Figure S44. ¹³C NMR spectrum of compound 8 (CDCl3, 125 MHz).

Figure S45. HMBC spectrum of compound 8 (CDCl3, 500 MHz).

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 Shif t (ppm)

8 16 24 32 40 48 56 64 72 80 88 96 104 112 120 128 136 144 152 160 168 176

F1 Chemical Shift (ppm)

Figure S46. HSQC spectrum of compound 8 (CDCl3, 500 MHz).

50 100 150 200 250 300 350 400 450 500 550 0

500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000 5000000

m /z-->

A bundance

S can 1971 (22.473 m in): 011.D \data.m s (-1947) (-) 138.0

77.0

367.0 183.0

240.0

330.0

287.1 405.1 467.1503.1 554.2

Figure S47. Mass spectrum of compound 8 (CH2Cl2).

Figure S48. IR spectrum (KBr) of compound 8.