Supplementary Information

2344 2362 860 472 598 664 492 756 3412 926 3548 1072 992 1154 1174 1332 1248 1200 1358 1392 1466 1716 2848 2874 2930 Supplementary Information Zika Virus Activity of the Leaf and Branch Extracts of Tontelea micrantha and Its Hexane Extracts Phytochemical Study Fernanda L. Ferreira, a Marcela S. Hauck, b Lucienir P. Duarte, *,a José C. de Magalhães, b Louise S. M. da Silva, a Lúcia P. S. Pimenta, a Julio C. D. Lopes, a Maria O. Mercadante-Simões c and Sidney A. Vieira Filho d a Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais, 31270-901 Belo Horizonte-MG, Brazil b Departamento de Química, Biotecnologia e Engenharia de Bioprocessos, Universidade Federal de São João del-rei, Campus Alto Paraopeba, 36420-000 Ouro Branco-MG, Brazil c Departamento de Biologia Geral, Centro de Ciências Biológicas e da Saúde, Universidade Estadual de Montes Claros, 39401-089 Montes Claros-MG, Brazil d Departamento de Farmácia, Escola de Farmácia, Universidade Federal de Ouro, Campus Morro do Cruzeiro, 35400-000 Ouro Preto-MG, Brazil 96 88 80 72 64 %Transmittance 56 48 40 32 24 16 8 0 Figure S1. IR (KBr,) spectrum of 1. 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) *e-mail: lucienir@ufmg.br

Figure S2. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 1. Figure S3. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 1.

Figure S4. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 1. Figure S5. HSQC (400 MHz, CDCl 3 ) spectrum of 1.

Figure S6. Expanded HSQC (400 MHz, CDCl 3 ) spectrum of 1 in the region between H 2.4 to 0.6 ppm. Figure S7. HMBC spectrum of 1 (400 MHz, CDCl 3 ).

Figure S8. Expanded HMBC (400 MHz, CDCl 3 ) spectrum of 1 in the region between H 3.5 to 0.6 ppm. Figure S9. Expanded HMBC (400 MHz, CDCl 3 ) spectrum of 1 in the region between H 3.8 to 0.4 ppm.

Figure S10. COSY (400 MHz, CDCl 3 ) spectrum of 1. Figure S11. NOESY (400 MHz, CDCl 3 ) spectrum of 1.

Figure S12. Expanded NOESY (400 MHz, CDCl 3 ) spectrum of 1 in the region between H 2.7 to 1.1 ppm. Figure S13. HR-ESI-MS (positive-ion mode) spectrum of compound 1.

60 55 50 %Transmittance 45 40 35 30 25 1463 923 976 1002 1050 1073 1109 1220 1299 1362 20 15 1389 10 2869 5 2926 1715 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S14. IR (KBr) spectrum of 2. Figure S15. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 2.

Figure S16. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 2. Figure S17. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 2.

3.28 9.01 13.21 3.21 21.61 1.01 0.95 1.01 70 65 60 55 50 %Transmittance 45 40 35 30 25 20 15 10 5 0 4360 3588 3618 3474 3036 3006 2868 2932 2978 1458 1448 1362 1384 1258 1172 920 964 980 1000 1122 1020 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S18. IR (KBr) spectrum of 3. 8.61 7.66 7.33 7.27 3.75 3.74 1.92 1.90 1.75 1.72 1.58 1.56 1.48 1.42 1.37 1.37 1.31 1.31 1.27 1.17 1.01 1.00 0.99 0.96 0.95 0.94 0.87 0.00 9.00 ppm (t1) 8.50 8.00 7.50 7.00 6.50 6.00 5.50 5.00 4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00 Figure S19. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 3.

149.70 149.43 149.16 135.43 123.43 123.18 122.93 77.51 77.19 76.87 72.38 61.46 53.22 49.33 42.85 41.81 39.69 39.29 38.38 37.92 37.13 36.10 35.59 35.40 35.35 35.03 32.84 32.34 32.10 31.81 30.66 30.03 28.18 20.13 18.66 18.28 17.59 16.43 15.91 11.75 0.0000 150 ppm (t1) 125 100 75 50 25 0 Figure S20. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 3. 72.38 61.46 53.22 49.33 42.85 41.81 39.29 36.10 35.59 35.40 35.35 35.03 32.84 32.34 32.10 31.81 30.66 20.13 18.66 18.28 17.59 16.43 15.91 11.75 75.0 70.0 ppm (t1) 65.0 60.0 55.0 50.0 45.0 40.0 35.0 30.0 25.0 20.0 15.0 10.0 Figure S21. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 3.

%Transmittance 96 88 80 72 64 56 48 40 32 24 3422 2946 2928 2868 2260 2336 2366 1704 1214 1284 1388 1462 1060 914 734 628 470 16 8 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S22. IR (KBr) spectrum of 4. Figure S23. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 4.

Figure S24. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 4. Figure S25. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 4.

462 480 570 714 594 746 820 2344 2368 892 914 960 1198 1316 3392 980 994 1006 1026 1124 1082 1054 1224 1362 1390 1468 2868 2930 3544 1706 96 88 80 72 64 %Transmittance 56 48 40 32 24 16 8 0 Figure S26. IR (KBr) spectrum of 5. 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S27. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 5.

Figure S28. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 5. Figure S29. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 5.

55 50 45 40 3402 2694 2332 2356 748 872 914 974 1006 1054 1072 1174 1192 1298 1364 %Transmittance 35 30 25 2866 1464 1388 20 15 2928 1714 10 5 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S30. IR (KBr) spectrum of 6. Figure S31. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 6.

Figure S32. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 6. Figure S33. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 6.

546 566 576 588 718 638 690 756 770 860 3408 914 1046 982 1080 1110 1146 1206 1358 1388 1466 1718 2846 2870 2930 2974 2998 96 88 80 72 64 %Transmittance 56 48 40 1282 1260 32 24 16 8 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S34. IR (KBr) spectrum of 7. Figure S35. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 7.

Figure S36. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 7. Figure S37. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 7.

60 55 820 50 45 40 2344 2366 1074 1110 1300 1204 1314 914 956 984 1004 588 734 %Transmittance 35 30 25 3534 1466 1450 1388 1040 1058 20 2868 15 10 5 2934 1716 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S38. IR (KBr) spectrum of 8. Figure S39. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 8.

Figure S40. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 8. Figure S41. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 8.

Figure S42. HSQC (400 MHz, CDCl 3 ) spectrum of 8. Figure S43. Expanded HSQC (400 MHz, CDCl 3 ) spectrum of 8 in the region between H 2.1 to 0.8 ppm.

Figure S44. HMBC (400 MHz, CDCl 3 ) spectrum of 8. Figure S45. Expanded HMBC (400 MHz, CDCl 3 ) spectrum of 8 in the region between H 1.6 to 1.0 ppm.

Figure S46. Expanded HMBC (400 MHz, CDCl 3 ) spectrum of 8 in the region between H 1.1 to 0.7 ppm. Figure S47. Expanded HMBC (400 MHz, CDCl 3 ) spectrum of 8 in the region between H 1.9 to 0.7 ppm. Figure S48. COSY (400 MHz, CDCl 3 ) spectrum of 8.

Figure S49. NOESY (400 MHz, CDCl 3 ) spectrum of 8. Figure S50. Expanded NOESY (400 MHz, CDCl 3 ) spectrum of 1 in the region between H 2.5 to 0.9 ppm.

Table S1. NMR spectral data of 30-hydroxyfriedelan-3-one (8) (400 MHz, CDCl 3 ) Position Type δ C δ H HMBC (C H) COSY NOESY 1 CH 2 22.29 1.70α 2β 1β 1.96β 2β, 10 1α, 2β, 11β 2 CH 2 41.52 2.30β 1β 2.40α 1α, 1β 3 C 213.21 4, 23 4 CH 58.24 2.24 23, 24 23 6α, 10, 23 5 C 42.15 4, 23, 24 6 CH 2 41.30 1.28α 24 7α 4, 6β 1.74β 7α 6α, 7β, 23, 24 7 CH 2 18.25 1.39β 6β 1.49α 6α, 6β, 8 8 CH 53.00 1.39 6, 10, 25, 26 7α 9 C 37.45 25 10 CH 59.51 1.52 1, 2, 24, 25 1β 4 11 CH 2 35.58 1.28α 8, 25 1.45β 1β 12 CH 2 30.53 1.35 27 13 C 39.81 19, 26, 27 14 C 38.38 12, 26, 27 15 CH 2 32.12 1.31 8, 26 1.50 16 16 CH 2 35.93 1.40 22, 28 15 1.57 17 C 30.00 18, 28 18 CH 42.74 1.51 19, 27, 28 19α 19 CH 2 29.34 1.24α 29, 30 18 1.48β 30 20 C 33.38 29, 30 21 CH 2 28.17 1.18α 22 22α, 22β 1.42β 22β 28 22 CH 2 38.11 1.00β 28 21α, 21β 28, 30 1.58α 21α 29 23 CH 3 6.83 0.88 4 4 4, 6β, 24 24 CH 3 14.67 0.72 4 6β, 23, 25 25 CH 3 18.02 0.87 24 26 CH 3 19.97 0.99 27 CH 3 18.59 1.07 18 28 CH 3 32.14 1.15 18, 22 21β, 22β, 30 29 CH 3 28.93 1.00 30 22a 30 CH 2 71.98 3.35 (d, H a ) 3.43 (d, H b ) (J a = 10.5/J b = 10.4 Hz) 19, 29 30 19β, 22β, 28 HMBC: heteronuclear multiple bond correlation; COSY: correlation spectroscopy; NOESY: nuclear Overhauser effect spectroscopy.

Figure S51. 1 H NMR (400 MHz, CDCl 3 ) spectrum of the mixture of 8 and 9. Figure S52. 13 C NMR (100 MHz, CDCl 3 ) spectrum of the mixture of 8 and 9.

540 568 634 670 756 912 2344 2366 980 1016 1054 1080 1198 1618 1362 1458 3496 1388 2870 1710 2932 Figure S53. DEPT-135 (100 MHz, CDCl 3 ) spectrum of the mixture of 8 and 9. 65 60 55 50 45 40 %Transmittance 35 30 25 20 15 10 5 4500 4000 3500 3000 2500 2000 1500 1000 500 Wavenumber (cm-1) Figure S54. IR (KBr) spectrum of 10.

Figure S55. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 10. Figure S56. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 10.

3.23 2.06 2.02 1.00 Figure S57. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 10. 7.26 5.13 5.12 5.10 2.08 2.06 2.04 2.00 1.98 1.96 1.60 0.00 7.00 ppm (t1) 6.50 6.00 5.50 5.00 4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00 Figure S58. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 11.

134.93 124.25 77.30 76.98 76.67 39.74 26.74 16.01 ppm (t1) 125 100 75 50 25 0 Figure S59. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 11. 124.24 39.74 26.73 16.01 125 ppm (t1) 100 75 50 25 0 Figure S60. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 11.

Figure S61. 1 H NMR (400 MHz, CDCl 3 ) spectrum of 12. Figure S62. 13 C NMR (100 MHz, CDCl 3 ) spectrum of 12.

Figure S63. DEPT-135 (100 MHz, CDCl 3 ) spectrum of 12. Figure S64. 1 H NMR (400 MHz, CDCl 3 ) spectrum of mixture of 13.

Figure S65. 13 C NMR (100 MHz, CDCl 3 ) spectrum of mixture of 13, with expansion between C 30.2 to 28.3 ppm. Figure S66. DEPT-135 (100 MHz, CDCl 3 ) spectrum of mixture of 13. 260 Name Back Signal HISS.dat C16:0 260 240 240 220 220 200 200 180 180 160 160 pa 140 140 pa 120 120 100 100 80 C18:0 80 60 C18:1 60 40 C14:0 C16:1 C17:0 C18:2 C18:3 C20:0 40 20 20 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 Minutes Figure S67. GC spectrum of mixture of 13.

Table S2. 13 C NMR data for compounds 2-7, 9 and 10 (100 MHz, CDCl 3 ) Position 2 3 a 4 5 6 7 9 10 1 22.30 15.91 21.08 22.27 22.3 22.28 22.29 22.40 2 41.54 36.10 37.41 41.51 41.5 41.49 41.50 41.64 3 213.20 72.38 179.05 213.99 212.9 212.85 213.1 213.17 4 58.25 49.33 36.08 58.25 58.2 58.23 58.20 58.37 5 42.16 37.92 37.85 42.11 42.0 42.10 42.08 42.37 6 41.32 41.81 38.97 41.29 41.1 41.22 41.28 41.42 7 18.25 17.59 18.12 18.26 18.1 18.28 18.27 18.39 8 53.13 53.22 53.01 52.53 52.9 53.37 51.01 53.36 9 37.47 37.13 39.08 37.50 37.2 37.49 37.53 37.56 10 59.51 61.46 59.05 59.54 59.3 59.46 59.54 59.64 11 35.65 35.40 35.21 35.47 35.48 35.55 35.33 35.78 12 30.52 30.66 30.22 30.12 30.6 32.79 30.21 30.83 13 39.72 38.38 39.65 39.40 38.8 38.20 39.06 39.82 14 38.32 39.69 38.34 38.18 37.7 39.93 38.83 38.38 15 32.44 32.34 32.31 31.31 32.4 30.63 30.51 32.53 16 36.03 35.59 36.11 29.15 35.0 37.03 36.12 36.84 17 30.02 30.03 30.03 35.19 47.8 33.22 32.50 30.01 18 42.82 42.85 42.87 39.48 36.5 41.97 44.33 42.84 19 35.37 35.35 35.33 34.53 35.5 35.11 35.96 36.01 20 28.18 28.18 28.18 28.16 28.4 42.74 34.37 33.94 21 32.80 32.84 32.88 31.44 32.5 218.74 74.32 75.82 22 39.27 39.29 39.31 33.41 28.1 54.99 47.07 48.64 23 6.83 11.75 7.60 6.82 6.8 6.82 6.81 6.94 24 14.67 16.43 19.41 14.68 14.6 14.66 14.68 14.79 25 17.96 18.28 17.94 18.08 17.2 17.74 17.80 18.05 26 20.27 20.13 20.16 19.09 20.0 18.46 18.21 20.84 27 18.67 18.66 18.75 19.20 18.8 21.20 19.28 18.88 28 32.11 32.10 32.13 68.08 209.0 33.22 33.18 34.54 29 35.03 35.03 34.98 32.85 34.5 28.82 31.86 34.88 30 31.79 31.81 31.86 34.27 29.4 24.99 24.94 24.10 a CDCl 3 /C 5 D 5 N.