SUPPLEMENTARY MATERIAL New phenolic acids from Salvia ... · SUPPLEMENTARY MATERIAL New phenolic acids from Salvia yunnanensis C.H.Wright Yun Yu a, Yin-ru Wang a, Zhen-huan Dong a,

SUPPLEMENTARY MATERIAL

New phenolic acids from Salvia yunnanensis C.H.Wright

Yun Yu a, Yin-ru Wang

a, Zhen-huan Dong

a, Wei Li

b, Shu-Ming Li

b, Xue-feng

Huang a*

aDepartment of Natural Medicinal Chemistry, China Pharmaceutical University,

Nanjing, 210009, P. R. China; bTasly R&D Institute, Tasly Pharmaceutical Co., Ltd.,

Tianjin 300410, P. R. China

* Corresponding author: Xue-feng Huang

Tel.: +86-25-52716876; Fax: +86-25-85301528.

E-mail address: [email protected].

Abstract

Two new phenolic acids, ethyl pro-lithospermate (1), n-butyl pro-lithospermate (2)

were isolated from Salvia yunnanensis C.H.Wright, along with nineteen known

compounds (3-21). The structures of the isolated compounds were elucidated on the

basis of extensive spectrometry and by comparing their physical and spectroscopic

data to literature. Among them, compounds 11, 12 and 14-16 were firstly isolated

from Salvia yunnanensis C.H.Wright. Some of the isolated compounds were

evaluated for their neuroprotection. Compound 10-12 showed significant

neuroprotective effects in PC12 cells and compounds 1, 4-7 displayed moderate

neuroprotective effects.

Keywords:

Salvia yunnanensis; phenolic acids; phenylpropanoids; neuroprotection

Figure S1. Key 1H-

1H COSY and HMBC correlations (H→C) in compound 1 and

compound 2

Figure S2. The CD spectrum of 1 (black in MeOH) and salvianolic acid B

(green in MeOH)

Figure S3. The CD spectrum of 2 (black in MeOH) and salvianolic acid B

(green in MeOH)

Figure S4. 1H-NMR spectrum of 1

Figure S5. 13

C-NMR spectrum of 1

Figure S6. 1H-

1H COSY spectrum of 1

Figure S7. HSQC spectrum of 1

Figure S8. HMBC spectrum of 1

Figure S9. NOESY spectrum of 1

Figure S10. 1H-NMR spectrum of 2

Figure S11. 13

C-NMR spectrum of 2

Figure S12. 1H-

1H COSY spectrum of 2

Figure S13. HSQC spectrum of 2

Figure S14. HMBC spectrum of 2

Figure S15. NOESY spectrum of 2

Figure S16. HR-ESI-MS spectrum of 1

Figure S17. HR-ESI-MS spectrum of 2

Figure S18. HPLC chromatogram of compound 1 (rose) and the DMSO extract

of S. yunnanensis (black)

2.5 5.0 7.5 10.0 12.5 15.0 min

0

25000

50000

75000

100000

125000

150000

175000

200000

225000

uV

2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0min

0

25000

50000

75000

100000

125000

150000

175000

uV

Figure S19. HPLC chromatogram of compound 2 (rose) and the DMSO extract

of S. yunnanensis (black)

Table S1. 1H- NMR (300 MHz) and

13C- NMR (75 MHz) of compound 1 and

compound 2 in DMSO-d6 (δ in ppm, J in Hz)

Position 1 2

δH δC δH δC

1 - 122.5 122.6

2 - 125.6 125.4

3 - 147.0 146.9

4 - 143.6 143.6

5 6.81 (1H, d, J = 8.4) 117.3 6.79 (1H, d, J = 8.4) 117.3

6 7.21 (1H, d, J = 8.4) 120.4 7.21 (1H, d, J = 8.4) 120.3

7 7.55 (1H, d, J = 15.9) 140.7 7.54 (1H, d, J = 15.9) 140.7

8 6.25 (1H, d, J = 15.9) 117.2 6.24 (1H, d, J = 15.9) 117.3

9 - 167.6 167.7

1' - 131.0 130.9

2' 6.73 (1H, br.s) 112.9 6.72 (1H, br.s) 112.9

3′ - 145.6 145.4

4' - 145.6 145.6

5'′ 6.71 (1H, d, J = 7.8) 115.6 6.71 (1H, d, J = 7.8) 115.4

6' 6.65 (1H, d, J = 7.8) 117.0 6.64 (1H, d, J = 7.8) 116.9

7' 5.79 (1H, d, J = 5.0) 86.3 5.79 (1H, d, J = 4.7) 86.2

8' 4.43 (1H, d, J = 5.0) 55.2 4.43 (1H, d, J = 4.7) 55.2

9' - 171.2 171.2

10' 4.14 (2H, q, J = 7.0) 61.3 4.08 (2H, m) 64.9

11' 1.19 (3H, t, J = 7.0) 13.7 1.53 (2H, m) 29.9

12' 1.23(2H, m) 18.5

13' 0.82 (3H, t, J = 7.2) 13.4

Assignments were based on 1H-1H COSY, HSQC, HMBC and NOESY.

Table S2. Composition and gradient of the mobile phase for HPLC analysis

Time/min A (H2O/CF3COOH

, 100 : 0.02 , v/v)% B (CH3CN)%

0 85 15

5 77 23

10 70 30

15 67 33

30 67 33