1

SUPPLEMENTARY MATERIAL

Effect of indole alkaloids from roots of Rauvolfia ligustrina in

the noradrenergic neurotransmission

Herbert S. Magalhãesa, Alison B. da Silvaa, Nilberto R. F. Nascimentob, Luis Gustavo

F. de Sousab, Maria Júlia S. da Fonsecab, Maria Iracema B. Loiolac, Norberto K. V.

Monteirod, Francisco Wagner Q. Almeida Netod, Kirley M. Canutoe, Otilia Deusdênia

L. Pessoaa*

aDepartamento de Química Orgânica e Inoergânica – Universidade Federal do Ceará,

60971-270 Fortaleza, CE, Brazil

bInstituto Superior de Ciências Biomédicas – Universidade Estadual do Ceará, 60714-

903 Fortaleza, CE, Brazil

cHerbário Prisco Bezerra (EAC) – Universidade Federal do Ceará, 60440-900

Fortaleza, CE, Brazil

dDepartmento de Química Analítica e Físico-Química – Universidade Federal do Ceará,

60020-181 Fortaleza, CE, Brazil.

eEmbrapa Agroindústria Tropical, 60511-110 Fortaleza, CE, Brazil

*Corresponding author.

E-mail address: opessoa@ufc.br (O.D.L. Pessoa)

2

CONTENTS

Figure S1. ¹H NMR spectrum of compound 1 (500 MHz, MeOD). ............................................ 3

Figure S2. ¹³C NMR spectrum of compound 1 (125 MHz, MeOD). ........................................... 4

Figure S3. COSY NMR spectrum of compound 1. ...................................................................... 5

Figure S4. HSQC NMR spectrum of compound 1. ...................................................................... 6

Figure S5. HMBC NMR spectrum of compound 1. ..................................................................... 7

Figure S6. NOESY NMR spectrum of compound 1. ................................................................... 8

Figure S7. HRESIMS of compound 1. ......................................................................................... 9

Figure S8. ¹H NMR spectrum of compound 2 (300 MHz, CDCl3). ........................................... 10

Figure S9. ¹³C NMR spectrum of compound 2 (75 MHz, CDCl3). ............................................ 11

Figure S10. ¹H NMR spectrum of compound 3 (300 MHz, CDCl3). ......................................... 12

Figure S11. ¹³C NMR spectrum of compound 3 (75 MHz, CDCl3). .......................................... 13

Figure S12. ¹H NMR spectrum of compound 4 (300 MHz, CDCl3). ......................................... 14

Figure S13. ¹³C NMR spectrum of compound 4 (75 MHz, CDCl3). .......................................... 15

Figure S14. ¹H NMR spectrum of compound 5 (300 MHz, CDCl3). ......................................... 16

Figure S15. ¹³C NMR spectrum of compound 5 (75 MHz, CDCl3). .......................................... 17

Figure S16. ¹H NMR spectrum of compound 6 (300 MHz, MeOD). ........................................ 18

Figure S17. ¹³C NMR spectrum of compound 6 (75 MHz, MeOD). ......................................... 19

Figure S18. ¹H NMR spectrum of compound 7 (500 MHz, MeOD). ........................................ 20

Figure S19. ¹H NMR spectrum of compound 8 (500 MHz, MeOD). ........................................ 21

Figure S20. ¹³C NMR spectrum of compound 8 (125 MHz, MeOD). ....................................... 22

Figure S21. ¹H NMR spectrum of compound 9 (500 MHz, MeOD). ........................................ 23

Figure S22. ¹³C NMR spectrum of compound 9 (125 MHz, MeOD). ....................................... 24

Figure S23. ¹H NMR spectrum of compound 10 (300 MHz, MeOD). ...................................... 25

Figure S24. ¹³C NMR spectrum of compound 10 (75 MHz, MeOD). ....................................... 26

Figure S25. ¹H NMR spectrum of compound 11 (500 MHz, MeOD). ...................................... 27

Figure S26. ¹³C NMR spectrum of compound 11 (125 MHz, MeOD). ..................................... 28

Figure S27. ¹H NMR spectrum of compound 12 (300 MHz, MeOD). ...................................... 29

Figure S28. ¹³C NMR spectrum of compound 12 (75 MHz, MeOD). ....................................... 30

Figure S29. Optimized geometry of iso1 isomer using mPW1PW91/6-31G(d,p) level of theory.

..................................................................................................................................................... 31

Figure S30. Optimized geometry of iso2 isomer using mPW1PW91/6-31G(d,p) level of theory.

..................................................................................................................................................... 31

Table S1. ¹³C chemical shifts of compounds 2-6 and 8-12. ........................................................ 32

Table S2. Efficiency and potency of the Rauwolfia ligustrina constituents in the inhibition of

the noradrenergic neurotransmission. ......................................................................................... 33

3

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

4

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

5

Figure S3. COSY NMR spectrum of compound 1.

6

Figure S4. HSQC NMR spectrum of compound 1.

7

Figure S5. HMBC NMR spectrum of compound 1.

8

Figure S6. NOESY NMR spectrum of compound 1.

9

Figure S7. HRESIMS of compound 1.

10

Figure S8. ¹H NMR spectrum of compound 2 (300 MHz, CDCl3).

11

Figure S9. ¹³C NMR spectrum of compound 2 (75 MHz, CDCl3).

12

Figure S10. ¹H NMR spectrum of compound 3 (300 MHz, CDCl3).

13

Figure S11. ¹³C NMR spectrum of compound 3 (75 MHz, CDCl3).

14

Figure S12. ¹H NMR spectrum of compound 4 (300 MHz, CDCl3).

15

Figure S13. ¹³C NMR spectrum of compound 4 (75 MHz, CDCl3).

16

Figure S14. ¹H NMR spectrum of compound 5 (300 MHz, CDCl3).

17

Figure S15. ¹³C NMR spectrum of compound 5 (75 MHz, CDCl3).

18

Figure S16. ¹H NMR spectrum of compound 6 (300 MHz, MeOD).

19

Figure S17. ¹³C NMR spectrum of compound 6 (75 MHz, MeOD).

20

Figure S18. ¹H NMR spectrum of compound 7 (500 MHz, MeOD).

21

Figure S19. ¹H NMR spectrum of compound 8 (500 MHz, MeOD).

22

Figure S20. ¹³C NMR spectrum of compound 8 (125 MHz, MeOD).

23

Figure S21. ¹H NMR spectrum of compound 9 (500 MHz, MeOD).

24

Figure S22. ¹³C NMR spectrum of compound 9 (125 MHz, MeOD).

25

Figure S23. ¹H NMR spectrum of compound 10 (300 MHz, MeOD).

26

Figure S24. ¹³C NMR spectrum of compound 10 (75 MHz, MeOD).

27

Figure S25. ¹H NMR spectrum of compound 11 (500 MHz, MeOD).

28

Figure S26. ¹³C NMR spectrum of compound 11 (125 MHz, MeOD).

29

Figure S27. ¹H NMR spectrum of compound 12 (300 MHz, MeOD).

30

Figure S28. ¹³C NMR spectrum of compound 12 (75 MHz, MeOD).

31

Figure S29. Optimized geometry of iso1 isomer using mPW1PW91/6-31G(d,p) level of

theory.

Figure S30. Optimized geometry of iso2 isomer using mPW1PW91/6-31G(d,p) level of

theory.

32

Table S1. ¹³C chemical shifts of alkaloids 2-6 and 8-12.

Position 2 3 4 5 6 8 9 10 11 12

δC

2 130.2 181.7 126.2 130.0 178.3 127.2 132.3 129.2 129.0 132.5

3 54.1 71.3 55.6 69.8 54.6 58.8 62.7 63.5 63.4 55.5

5 51.3 54.1 48.4 67.4 60.9 54.3 54.0 55.0 55.0 52.4

6 16.8 34.6 15.9 19.6 35.7 20.3 21.2 20.1 20.0 17.6

7 108.0 57.5 105.8 105.8 65.3 107.4 107.3 107.3 107.3 107.9

8 122.2 124.7 125.1 120.4 136.3 120.4 127.8 127.5 127.5 128.8

9 118.8 109.5 118.3 119.1 125.8 102.0 119.0 119.2 119.3 118.7

10 109.4 145.1 120.6 110.5 128.4 146.5 120.4 120.9 120.9 120.0

11 156.6 149.3 123.4 157.4 130.7 149.0 123.0 123.7 123.7 122.2

12 95.4 95.7 112.2 95.3 122.6 96.6 112.2 112.7 112.7 112.2

13 136.7 133.9 137.1 138.3 156.6 133.3 138.4 138.7 138.7 137.9

14 24.3 30.2 23.4 21.2 25.9 29.6 33.3 34.9 26.3 25.3

15 32.2 30.6 30.4 30.8 27.7 25.7 36.9 37.5 37.5 33.9

16 51.8 110.1 51.2 51.4 48.4 104.4 51.9 55.3 55.5 53.9

17 78.0 155.2 77.1 77.8 77.5 156.8 68.0 66.8 66.7 72.5

18 77.8 - 76.9 77.4 13.3 - 29.3 26.3 35.0 75.7

19 29.8 72.3 29.3 31.2 130.5 74.7 24.3 24.9 24.8 33.5

20 34.0 38.1 32.7 33.1 131.8 36.6 35.0 36.2 35.9 35.8

21 49.1 53.6 51.1 59.9 92.3 53.6 61.5 59.7 59.5 50.5

22 172.9 167.8 171.9 171.9 171.6 168.7 174.0 176.3 174.5 174.5

23 165.6 18.6 165.7 165.6 - 18.0 - - - -

24 56.0 56.9 - 56.0 - 57.1 - - - -

25 60.9 56.4 61.0 61.1 - 56.8 - - - -

26 52.1 51.1 52.7 52.7 - 52.0 52.0 - 52.5 52.4

27 56.4 - 56.5 56.5 - - - - - -

28 61.1 - 61.1 61.1 - - - - - -

29 56.4 - 56.5 56.5 - - - - - -

1’ 125.5 - 125.3 125.2 105.2 - - - - -

2’ 107.0 - 107.2 107.1 74.8 - - - - -

3’ 153.1 - 153.1 153.2 77.8 - - - - -

4’ 142.5 - 142.6 142.6 71.5 - - - - -

5’ 153.1 - 153.1 153.2 78.7 - - - - -

6’ 107.0 - 107.2 107.1 62.7 - - - - -

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Table S2. Efficiency and potency of the Rauvolfia ligustrina alkaloids in the inhibition

of the noradrenergic neurotransmission.

*Alkaloids presented neuroexcitatory effect.

Alkaloids Maximal inhibitory response in the

noradrenergic neurotransmission

(% Control)

Concentration necessary to

induce half maximal inhibition

(IC50 mol/L [CI 95%])

1 87.9 ± 2.0 1.2x10-7 [7.7x10-9 - 2.0x10-6]

2 86.3 ± 4.4 3.9x10-9 [3.0x10-12 - 5.0x10-6]

3* - -

4 43.5 ± 4.1 2.0x10-8 [4.5x10-9 - 9.0x10-8]

5 18.6 ± 3.1 2.2x10-7 [3.2x10-8 - 1.5x10-6]

6 16.0 ± 1.9 1.6x10-7 [2.0x10-8 - 1.2x10-6]

7 44.6 ± 4.8 7.2x10-8 [1.3x10-8 - 4.0x10-7]

8 52.2 ± 4.2 7.1x10-7 [2.1x10-7 - 2.1x10-6]

9 81.2 ± 3.7 7.1x10-7 [2.1x10-7 - 2.1x10-6]

10 76.0 ± 4.3 4.4x10-7 [1.8x10-7 - 1.1x10-6]

11* - -

12 23.0 ± 1.6 2.5x10-7[1.6x10-7 - 3.8x10-7]