Supplementary Material

SUPPLEMENTARY MATERIAL

Two new isoquinoline alkaloids from the seeds of Nandina domestica

Juan Qin,a,1 Sheng-Yuan Zhang,b,1 Yu-Bo Zhang,a,c Li-Feng Chen,a Neng-Hua Chen,a Zhong-Nan Wu,a Ding Luo,a Guo-Cai Wang,*a and Yao-Lan Li*a a Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, Jinan University, Guangzhou 510632, China; b Medical College of Jiaying University, Meizhou 514031, P. R. China; c Department of Pharmacology, School of Medicine, Jinan University, Guangzhou, Guangdong 510632, China; e-mail: [email protected] (Y.L. Li); [email protected] (G.C. Wang)

Two new isoquinoline alkaloids, 6R,6aS-N-nantenine Nβ-oxide (1),

6S,6aS-N-nantenine Nα-oxide (2), along with nine known alkaloids, nantenine

(3), oxonantenine (4), protopine (5), nornantenine (6), N-methyl-laurotetanine

(7), isocorydine (8), O-methyflavinantine (9),

N-methyl-2,3,6-trimethoxymorphinan-dien-7-one Nβ-oxide (10) and

(+)-10-O-methylhernovine Nβ-oxide (11) were isolated from the seeds of

Nandina domestica. Their structures were elucidated by extensive analyses of

spectroscopic data (IR, UV, HRESIMS, 1D and 2D NMR), ECD calculation

and comparison with the related literatures. In addition, the cytotoxicity against

A549 cells of these alkaloids were determined by the MTT assay.

Keywords: Nandina domestica; isoquinoline alkaloid; cytotoxicity.

Contens: Pages:

Fig. S1. Key 1H-1H COSY and HMBC correlations of 1-2 ...... 3

Table S1. 1D NMR spectral data of 1 in CD3OD and 2 in DMSO (δ in ppm, J in Hz) ...... 4 Fig. S2. 1H NMR superimposed spectrum of 1-2 ...... 5 Fig. S3. HRESIMS of 1 ...... 5

Fig. S4. UV spectrum of 1 (CH3OH) ...... 6 Fig. S5. IR spectrum of 1 (KBr disc) ...... 6 1 Fig. S6. H NMR spectrum of 1 in CD3OD ...... 7 13 Fig. S7. C NMR spectrum of 1 in CD3OD ...... 7

Fig. S8. DEPT-135 spectrum of 1 in CD3OD ...... 8 1 1 Fig. S9. H- H COSY spectrum of 1 in CD3OD ...... 8

Fig. S10. HSQC spectrum of 1 in CD3OD ...... 9

Fig. S11. HMBC spectrum of 1 in CD3OD ...... 9

Fig. S12. NOESY spectrum of 1 in CD3OD ...... 10 Fig. S13. HRESIMS of 2 ...... 10

Fig. S14. UV spectrum of 2 (CH3OH) ...... 11 Fig. S15. IR spectrum of 2 (KBr disc) ...... 11 Fig. S16. 1H NMR spectrum of 2 in DMSO ...... 12 Fig. S17. 13C NMR spectrum of 2 in DMSO ...... 12 Fig. S18. DEPT-135 spectrum of 2 in DMSO ...... 13 Fig. S19. 1H-1H COSY spectrum of 2 in DMSO...... 13 Fig. S20. HSQC spectrum of 2 in DMSO ...... 14 Fig. S21. HMBC spectrum of 2 in DMSO ...... 14 Fig. S22. NOESY spectrum of 2 in DMSO ...... 15 Fig. S23. Experimental ECD and calculated ECD spectra of 1 and 2 ...... 15

Fig. S1. Key 1H-1H COSY and HMBC correlations of 1-2

Table S1. 1D NMR spectral data of 1 in CD3OD and 2 in DMSO (δ in ppm, J in Hz) 1a 2b

Position δH δC δH δC 1 - 146.4 - 144.6 1a - 129.8 - 126.2 1b - 122.4 - 122.9 2 - 154.4 - 152.7 3 6.76 (s) 111.8 6.87 (s) 111.1 3a - 128.0 - 125.8 4 2.73 (m) 25.5 2.98 (m) 27.0 5 α 3.53 (m), β 3.66 (m) 65.1 α 3.48, β 3.48 66.5 6 - - - - 6a 4.34 (d) 72.0 4.08 (d) 74.1 7 3.08 (m) 30.6 α 3.48 β 2.74 (t) 28.6 7a 6.83 (s) 128.6 - 124.5 8 - 109.5 7.03 (s) 108.6 9 - 148.4 - 146.4 10 7.76 (s) 148.3 - 146.5 11 - 109.5 7.72 (s) 107.8 11a 3.84 (s) 125.9 - 128.9

OCH3-1 3.57 (s) 56.4 3.61 (s) 59.9

OCH3-2 5.93 (s) 60.6 3.83 (s) 55.8

OCH2O-9,10 3.35 (s) 102.5 6.04 (s) 101.0

N-CH3 58.1 2.85 (s) 49.4 aMeasured at 400 (1H) and 100 (13C) MHz. bMeasured at 500 (1H) and 125 (13C) MHz. Overlapped signals were reported without designating multiplicity.

Fig. S2. 1H NMR superimposed spectrum of 1-2

Fig. S3. HRESIMS of 1

Fig. S4. UV spectrum of 1 (CH3OH)

Fig. S5. IR spectrum of 1 (KBr disc)

1 Fig. S6. H NMR spectrum of 1 in CD3OD

13 Fig. S7. C NMR spectrum of 1 in CD3OD

Fig. S8. DEPT-135 spectrum of 1 in CD3OD

1 1 Fig. S9. H- H COSY spectrum of 1 in CD3OD

Fig. S10. HSQC spectrum of 1 in CD3OD

Fig. S11. HMBC spectrum of 1 in CD3OD

Fig. S12. NOESY spectrum of 1 in CD3OD

Fig. S13. HRESIMS of 2

Fig. S14. UV spectrum of 2 (CH3OH)

Fig. S15. IR spectrum of 2 (KBr disc)

Fig. S16. 1H NMR spectrum of 2 in DMSO

13 Fig. S17. C NMR spectrum of 2 in DMSO

Fig. S18. DEPT-135 spectrum of 2 in DMSO

Fig. S19. 1H-1H COSY spectrum of 2 in DMSO

Fig. S20. HSQC spectrum of 2 in DMSO

Fig. S21. HMBC spectrum of 2 in DMSO

Fig. S22. NOESY spectrum of 2 in DMSO

30 calcd for (6aR)-1 30 calcd for (6aR)-2 calcd for (6aS)-1 calcd for (6aS)-2 20 exptl for 1 20 exptl for 2

10 10

0 0

Δε Δε

-10 -10

-20 -20

-30 -30

200 250 300 350 400 200 250 300 350 400 wavelength (nm) wavelength (nm)

Fig. S23. Experimental ECD and calculated ECD spectra of 1 and 2