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1
Supporting Information
Anti-Acanthamoeba activity of brominated sesquiterpenes from Laurencia johnstonii
Sara García-Davis1,2, Ines Sifaoui3, María Reyes-Batlle3, Ezequiel Viveros-Valdez2, José E. Piñero3, Jacob Lorenzo-Morales3, José J. Fernández1,4* and Ana R. Díaz-Marrero1*
1 Instituto Universitario de Bio-Orgánica Antonio González (IUBO AG), Centro de Investigaciones Biomédicas de Canarias (CIBICAN), Universidad de La Laguna (ULL), Avda. Astrofísico F. Sánchez, 2, 38206 La Laguna, Tenerife, España; [email protected] (S.G.D.); [email protected] (A.R.D.M); [email protected] (J.J.F.)
2 Universidad Autónoma de Nuevo León (UANL), Facultad de Ciencias Biológicas. Av. Pedro de Alba s/n, 66450 San Nicolás de los Garza, Nuevo León, México; [email protected] (E.V.V.)
3 Instituto Universitario de Enfermedades Tropicales y Salud Pública de Islas Canarias, Universidad de La Laguna. Av. Astrofísico Francisco Sánchez s/n, 38206 La Laguna, Tenerife, España.; [email protected] (I.S.); [email protected] (J.L.M.)
4 Departamento de Química Orgánica, Universidad de La Laguna (ULL), Avda. Astrofísico F. Sánchez, 2, 38206 La Laguna, Tenerife, España
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TABLE OF CONTENTS
Page Scheme S1. Isolation process of sesquiterpenes 1-5 from Laurencia johnstonii 3 Figure S1. 1H-NMR spectrum for laurinterol (1) (500 MHz, CDCl3) 4 Figure S2. 1H-NMR spectrum for isolaurinterol (2) (500 MHz, CDCl3) 5 Figure S3. 1H-NMR spectrum for aplysin (3) (500 MHz, CDCl3) 6 Figure S4. 1H-NMR spectrum for α-bromocuparane (4) (500 MHz, CDCl3) 7 Figure S5. 1H-NMR spectrum for α-isobromocuparane (5) (500 MHz, CDCl3) 8 Figure S6. 1H-NMR spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3) 9 Figure S7. COSY spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3) 10 Figure S8. HSQC-ed spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3) 11 Figure S9. HMBC spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3) 12 Figure S10. 13C NMR spectrum of 8-bromoaplysin (6) (150 MHz, CDCl3) 13 Figure S11. HREIMS spectrum of 8-bromoaplysin (6) 14 Figure S12. 1H NMR spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3) 15 Figure S13. COSY spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3) 16 Figure S14. HSQC-ed spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3) 17 Figure S15. HMBC spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3) 18 Figure S16. 13C NMR spectrum of 3a-bromojohnstane (7) (150 MHz, CDCl3) 19 Figure S17. 1D-NOE experiments of 3a-bromojohnstane (7) (600 MHz, CDCl3) 20 Figure S18. HREIMS spectrum of 3a-bromojohnstane (7) 21 Figure S19. 1H NMR spectrum of 8,10-dibromoisoaplysin (8) (500 MHz, CDCl3) 22 Figure S20. 1H NMR spectrum of 8,10-dibromoaplysinol (9) (500 MHz, CDCl3) 23
3
Scheme S1. Isolation process of sesquiterpenes 1-5 from Laurencia johnstonii
Br
OH
laurinterol (1)
Br
OH
isolaurinterol (2)
Br
O
aplysin (3)
α-bromocuparane (4)
Br
α-isobromocuparane (5)
Br
4
Figure S1. 1H-NMR spectrum for laurinterol (1) (500 MHz, CDCl3).
012345678
G9
5
Figure S2. 1H-NMR spectrum for isolaurinterol (2) (500 MHz, CDCl3).
6
Figure S3. 1H-NMR spectrum for aplysin (3) (500 MHz, CDCl3).
7
Figure S4. 1H-NMR spectrum for α-bromocuparane (4) (500 MHz, CDCl3).
8
Figure S5. 1H-NMR spectrum of α-isobromocuparane (5) (500 MHz, CDCl3).
9
Figure S6. 1H-NMR spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3).
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
10
Figure S7. COSY spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3).
0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
f2(ppm)
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
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
11
Figure S8. HSQC-ed spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.07.5f2(ppm)
10
20
30
40
50
60
70
80
90
100
110
120
130
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
12
Figure S9. HMBC spectrum of 8-bromoaplysin (6) (600 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.07.5
f2(ppm)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
13
Figure S10. 13C NMR spectrum of 8-bromoaplysin (6) (150 MHz, CDCl3).
102030405060708090100110120130140150160f1(ppm)
14
Figure S11. HREIMS spectrum of 8-bromoaplysin (6).
15
Figure S12. 1H NMR spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3).
0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
16
Figure S13. COSY spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.07.5
f2(ppm)
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
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
17
Figure S14. HSQC-ed spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.0
f2(ppm)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
18
Figure S15. HMBC spectrum of 3a-bromojohnstane (7) (600 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.0
f2(ppm)
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
19
Figure S16. 13C NMR spectrum of 3a-bromojohnstane (7) (150 MHz, CDCl3).
102030405060708090100110120130140150160
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
20
Figure S17. 1D-NOE experiments of 3a-bromojohnstane (7) (600 MHz, CDCl3).
0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5f1(ppm)
H4 – SELECTIVE NOESY PULSE AT dH 3.84 ppm
H14 – SELECTIVE NOESY PULSE AT dH 1.04 ppm
H13/H5b – SELECTIVE NOESY PULSE AT dH 1.54 ppm
H3-13
H3-13
H3-14H-6a/H-5b
H-12
H-4
H-3b
H-6a
H-3b
Br
OBr
Br14
COSY
14
1513
2
11
9
7
H
HMBC
H4
Me13
Me14
H3β
(a) (b)
H6β
21
Figure S18. HREIMS spectrum of 3a-bromojohnstane (7).
22
Figure S19. 1H NMR spectrum of 8,10-dibromoisoaplysin (8) (500 MHz, CDCl3).
1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14
23
Figure S20. 1H NMR spectrum of 8,10-dibromoaplysinol (9) (500 MHz, CDCl3).
0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
f1(ppm)
Br
OH
laurinterol (1)
Br
OBr
Br
OBr
Br
8
6 (15%) 3α-bromojohnstane (7) (30%)
Br
OH
isolaurinterol (2)
Br
OBr
6 (24%)
Br
OBr Br
Br
OBr OH
8 (6%) 9 (27%)
13
5
6
1312
14
15
24
17
14
1315 9
11
Br2 (1.95 mmol)
Et2O, rt, 15 min
Br2 (1.95 mmol)
Et2O, rt, 15 min
HBr (10 µL, 48%)
Et2O, rt, 15 min
Br
O8
13
5
6
1312
14