SUPPLEMENTARY MATERIAL

New Dammarane and Ursane type Triterpenoids from the Flower of Ixora

coccinea Linn.

Ambreen Ikram,a Muhammad Ali Versiani,

*,a Amna Khatoon,

a Salman Khalid

Ahmed,a Samina Abdul Sattar

a and Shaheen Faizi

b

aDepartment of Chemistry, Federal Urdu University of Arts, Sciences and Technology, Gulshan-

e- Iqbal, Science Campus, Karachi-75300, Pakistan

bHEJ Research Institute of Chemistry, International Center for Chemical and Biological

Sciences, University of Karachi, Karachi-75270, Pakistan

Two new esters of dammarane triterpenoids ixorene isovalerate (1), ixorene 3´,8´-

dimethyloctanoate (2) and a new ursane type triterpenoids Ixoroid acid (3) were

isolated from the methanolic extract of flowers of Ixora coccinea Linn., along

with the three known constituents. The structures of compound 1 and 3 were

elucidated on the basis of extensive 1D-, 2D-NMR studies and mass spectrometry

as 17β-dammara-12, 20-diene-3β-isovelarate and 3β-Hydroxy-18β-urs-12ene-

29β-oic acid respectively, while 2 was identified as 17β-dammara-12,20-diene-

3β-3´,8´-dimethyloctanoate through 1H-NMR and mass spectral data. Compounds

1, 2, 4 and 5 were evaluated for their in vitro cytotoxic activity, which exhibited

weak activity against the 3T3, PC3 and Hela cell lines with the IC50 value ˃30

µM. Antioxidant results of 1-5 revealed that only compound 5 showed antioxidant

activity in DPPH radical scavenging inhibition with the IC50 0.0467±0.0018 mg

mL-1

. Both activities are the first records of these isolated compounds from the

flowers of Ixora coccinea Linn.

Keywords: Dammarane triterpenoid; ixorene isovalerate; ixorene 3´, 8´-

dimethyloctanoate; cytotoxic activity; antioxidant activity

Table S1. NMR spectral dataa 17β-dammara-12,20-diene-3β-isovalerate (1)

Position δH δC COSY-45° HMBC

1 1.25 (m),1.23 (m) 35.57 2 2

2 1.59 (m),1.22 (m) 29.36 1,3 1,1΄,3

3 4.47 (dd J 10.5 and

5.5 Hz)

80.62 2 1΄,2,29

4 - 40.85 - -

5 0.75 (m) 55.38 6 4,7

6 1.23 (m),1.23 (m) 22.68 5,7 7

7 1.36( m),1.23 (m) 34.21 6 5,6,9

8 - 42.83 - -

9 1.26 (m) 50.34 11 7

10 - 37.08 - -

11 1.59 (m), 1.23 (m) 24.96 9,12 12,17

12 5.35 (brs) 122.57 11 11,17

13 - 139.71 - -

14 - 42.93 - -

15 1.65 (m), 1.35 (m) 39.99 16 17,30

16 1.25 (m), 1.36 (m) 34.12 15,17 17

17 2.35 (m) 48.01 16 12,13,20,21

18 0.99 (s) 15.97 - 9,8,14

19 0.81 (s) 16.16 - 5,9

20 - 150.92 - -

21 4.66 (brs), 4.54

(brs)

109.35 - 17,20

22 1.27 (m),1.23 (m) 29.59 23 20,23,24

23 1.26 (m),1.24 (m) 29.46 22 22, 25

24 1.23 (m),1.23 (m) 25.16 23,25 22,25

25 1.23 (m) 32.01 24 23,24,26,27

26 0.85 (d J 6.5 Hz) 14.51 25 25

27 0.82 (d J 6.5 Hz) 14.10 25 25

28 0.81(s) 27.96 - 3,5,10

29 0.76 (s) 17.99 - 4,7,10

30

1΄

2΄

3΄

4΄

5΄

1.01(s)

-

2.27 (dd J 7.5 and

15 Hz)

1.63 (m)

0.91(d J 6.5 Hz)

0.92(d J 6.5 Hz)

16.65

173.72

34.85

38.05

14.51

14.10

-

-

3΄

2΄,4΄,5΄

3΄

3΄

7,8,14

-

1΄,3΄,3

2΄,4΄,5΄

3΄,2΄

3΄,2΄

aCDCl3, 500 MHz, δ in ppm, J in Hz

Table S2. NMR spectral dataa 3β-Hydroxy-18β-urs-12ene-29β-oic acid (3)

Position δH δC COSY-

45°

HMBC

1 1.60 (m)

1.70 (m)

36.68 2 C-2,3

2 1.58 (m) 27.16 1,3 C-1, 2,10

3 3.204 (dd J

11.5 and 5 Hz)

79.05 2 C-1,2

4 - 38.97 - -

5 0.70 (m) 55.19 6 C-4, 24

6 1.52(m)

1.35 (m)

18.28 5,7 C-5,8

7 0.88 (m) 32.92 6 C-4,5

8 - 41.96 - -

9 1.5 (m) 47.52 11 C-10

10 - 37.05 - -

11 1.61(m)

1.90 (m)

24.73 9,12 C-9,12,13

12 5.22 (brs) 125.83 11 C-9,11,14,18

13 - 137.90 - -

14 - 41.63 - -

15 1.84 (m) 27.98 16 C-14

16 1.58 (m) 27.16 15 C-14,17

17 - 30.67 - -

18 2.15 (d J 8.5

Hz)

52.61 19 C-19,12,13,29

19 2.80 (m) 41.03 18,20 C-18

20 1.61(m) 24.12 19,21 C-18, 22

21 1.46 (m) 33.74 20, 22 C-20

22 1.49 (m) 30.59 21 -

23 0.97 (s) 28.12 - C-3,5,24

24 0.75 (s) 15.55 - C-3,5,24

25 0.90 (s) 15.47 - C-5,9

26 0.75 (s) 15.32 - C-8,9,14

27 0.92 (s) 21.17 - C-8,14

28 0.83 (s) 17.02 - C-16,18

29 8.08 (s) 182.32 - -

30 0.76 (d J 7.5

Hz)

17.01 20 C-17,20,21

aCDCl3, 500 MHz, δ in ppm, J in Hz

O

1

2

3

4 5

6

7

8

9

10

11

12

13

14

15

16

17

1819

20

21 2223

24

25

26

29

30

28

H

5

O

123

4

5

H

H

H

H

HMBC NOSEY

Figure S2. Important HMBCs and NOSEYs correlation of ixorene isovelarate

HO

HO2C

H

H

3

29

1817

12

9

1920

30

5

H

H

(1)

Figure S3. Important HMBCs correlation of ixoroid acid (3)

SUPPLEMENTARY MATERIALS

New Dammarane and Ursane type Triterpenoids from the Flower of Ixora

coccinea Linn.

Ambreen Ikram,a Muhammad Ali Versiani,

*,a Amna Khatoon,

a Salman Khalid

Ahmed,a Samina Abdul Sattar

a and Shaheen Faizi

b

aDepartment of Chemistry, Federal Urdu University of Arts, Sciences and Technology, Gulshan-

e- Iqbal, Science Campus, Karachi-75300, Pakistan

bHEJ Research Institute of Chemistry, International Center for Chemical and Biological

Sciences, University of Karachi, Karachi-75270, Pakistan

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

Figure S 2. 1H NMR spectrum of compound 1 (region 3ppm to 6ppm, CDCl3, 500 MHz).

Figure S 3. 13

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

Figure S 4. Subspectrum DEPT135 of compound 1 (CDCl3, 125MHZ).

Figure S 5. Subspectrum DEPT 90 of compound 1 (CDCl3, 125MHZ).

Figure S 6. HSQC correlation of compound 1 (CDCl3, 500MHz).

Figure S 7. HSQC expended spectrum of compound 1 (CDCl3, 500MHz).

Figure S 8. HMBC correlation of compound 1 (CDCl3, 500MHz).

Figure S 9. HMBC expended spectrum of compound 1 (CDCl3, 500MHz).

Figure S 10. COSY correlation of compound 1 (CDCl3, 500MHz).

Figure S 11. NOESY correlation of compound 1 (CDCl3, 500MHz).

Figure S 12. IR spectrum of compound 1.

Figure S 13. EIMS spectrum of compound 1.

Figure S 14. HR-EIMS spectrum of compound 1.

Figure S 15. 1H NMR spectrum of compound 2 (CDCl3, 500 MHz).

Figure S 16. 1H NMR spectrum of compound 2 (region 3ppm to 6ppm, CDCl3, 500 MHz).

Figure S 17. EIMS spectrum of compound 2.

Figure S 18. HR-EIMS spectrum of compound 2.

Figure S 19. 1

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

Figure S 20. 1H NMR spectrum of compound 3 (region 0 ppm to 1.4 ppm, CDCl3, 500 MHz).

Figure S 21. 1H NMR spectrum of compound 3 (region 2.5 ppm to 8 ppm, CDCl3, 500 MHz).

Figure S 22. 13

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

Figure S 23. 13

C NMR expended spectrum of compound 3 (CDCl3, 125 MHz).

Figure S 24. Subspectrum DEPT135 of compound 3 (CDCl3, 125MHZ).

Figure S 25. Subspectrum DEPT 90 of compound 3 (CDCl3, 125MHZ).

Figure S 26. HSQC correlation of compound 3 (CDCl3, 500MHz).

Figure S 27. HSQC expended spectrum of compound 3 (CDCl3, 500MHz).

Figure S 28. HMBC correlation of compound 3 (CDCl3, 500MHz).

Figure S 29. HMBC expended spectrum of compound 3 (CDCl3, 500MHz).

Figure S 30. COSY correlation of compound 3 (CDCl3, 500MHz).

Figure S 31. NOESY correlation of compound 3 (CDCl3, 500MHz).

Figure S 32. IR spectrum of compound 3.

Figure S 33. EIMS spectrum of compound 3.

Figure S 34. HR-EIMS spectrum of compound 3.

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

Figure S2. 1H NMR spectrum of compound 1 (CDCl3, 500 MHz)

Figure S 2. 1H NMR spectrum of compound 1 (region 3ppm to 6ppm, CDCl3, 500 MHz).

Figure S 3. 13

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

Figure S 4. Subspectrum DEPT135 of compound 1 (CDCl3, 125MHZ).

Figure S 5. Subspectrum DEPT 90 of compound 1 (CDCl3, 125MHZ).

Figure S 6. HSQC correlation of compound 1 (CDCl3, 500MHz).

Figure S 7. HSQC expended spectrum of compound 1 (CDCl3, 500MHz).

Figure S 8. HMBC correlation of compound 1 (CDCl3, 500MHz).

Figure S 9. HMBC expended spectrum of compound 1 (CDCl3, 500MHz).

Figure S 10. COSY correlation of compound 1 (CDCl3, 500MHz).

Figure S 11. NOESY correlation of compound 1 (CDCl3, 500MHz).

Figure S 12. IR spectrum (KBr)of compound 1.

Figure S 13. EIMS spectrum of compound 1.

Figure S 14. HR-EIMS spectrum of compound 1.

Figure S 15. 1H NMR spectrum of compound 2 (CDCl3, 500 MHz).

Figure S 16. 1H NMR spectrum of compound 2 (region 3ppm to 6ppm, CDCl3, 500 MHz).

Figure S 17. EIMS spectrum of compound 2.

Figure S 18. HR-EIMS spectrum of compound 2.

Figure S 19. 1

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

Figure S2. 1H NMR spectrum of compound 1 (CDCl3, 500 MHz)

Figure S 20. 1H NMR spectrum of compound 3 (region 0 ppm to 1.4 ppm, CDCl3, 500

MHz).

Figure S 21. 1H NMR spectrum of compound 3 (region 2.5 ppm to 8 ppm, CDCl3, 500 MHz).

Figure S 22. 13

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

Figure S 23. 13

C NMR expended spectrum of compound 3 ( CDCl3, 125 MHz).

Figure S 24. Subspectrum DEPT135 of compound 3 (CDCl3, 125MHZ).

Figure S 25. Subspectrum DEPT 90 of compound 3 (CDCl3, 125MHZ).

Figure S 26. HSQC correlation of compound 3 (CDCl3, 500MHz).

Figure S 27. HSQC expended spectrum of compound 3 (CDCl3, 500MHz).

Figure S 28. HMBC correlation of compound 3 (CDCl3, 500MHz).

Figure S 29. HMBC expended spectrum of compound 3 (CDCl3, 500MHz).

Figure S 30. COSY correlation of compound 3 (CDCl3, 500MHz).

Figure S 31. COSY correlation of compound 3 (CDCl3, 500MHz).

Figure S 32. IR spectrum of compound 3

Figure S 33. EIMS spectrum of compound 3.

Figure S 34. HR-EIMS spectrum of compound 3.