Anti-Osteoclastogenic Effect of Isoquinoline Alkaloids from the Rhizome Extract of Sinomenium

Anti-osteoclastogenic effect of isoquinoline alkaloids from the rhizome extract of Sinomenium acutum

Ji Young Lee1,2, Kwang-Jin Kim3, Jinhee Kim1, Sang Un Choi1, Seong Hwan Kim4, Shi Yong Ryu1,2,*

Affiliation 1 Research Center for Medicinal Chemistry, Korea Research Institute of Chemical Technology, Daejeon 305-600, Korea 2 Graduate School of New Drug Discovery and Development, Chungnam National University, Daejeon 305-764, Korea 3 Research Institute of Basic Science, Sunchon National University, Suncheon 540-742, Korea 4 Laboratory of Translational Therapeutics, Pharmacology Research Center, Division of Drug Discovery Research, Korea Research Institute of Chemical Technology, Daejeon 305-600, Korea

Address for correspondence Dr. Shi Yong Ryu Korea Research Institute of Chemical Technology 141 Gajeongro, Yuseong, Daejeon 305-600, Korea Tel. + 82-42-860-7163 Fax + 82-42-861-7160 E-Mail: [email protected]

S1 List of Supplementary Materials

Page Figure S1. Effect of S. acutum extract in osteoclast differentiation S3 Figure S2. 1H NMR spectrum of 1 S4 Figure S3. 13C NMR spectrum of 1 S5 Figure S4. 1H-1H COSY spectrum of 1 S6 Figure S5. 1H-1H COSY spectrum of 1_expansion S7 Figure S6. HMQC spectrum of 1 S8 Figure S7. HMBC spectrum of 1 S9 Figure S8. NOESY spectrum of 1 S10 Figure S9. 1H NMR spectrum of 2 S11 Figure S10. 13C NMR spectrum of 2 S12 Figure S11. 1H-1H COSY spectrum of 2 S13 Figure S12. HSQC spectrum of 2 S14 Figure S13. HMBC spectrum of 2 S15 Figure S14. NOESY spectrum of 2 S16 Figure S15. NOESY spectrum of 2_expansion S17 Figure S16. 1H NMR spectrum of 7S-dihydrosinomenine S18

Figure S1. S. acutum MeOH extract suppresses ostoclastogenesis. (A) BMMs were cultured with RANKL (10 ng/ml) and M-CSF (30 ng/ml) in the presence of the indicated concentrations of S. Acutum MeOH extract. After 4 days, cells were fixed in

3.7% formalin for 5 min, permeabilized in 0.1% Triton X-100 for 10 min, and stained for TRAP, a marker enzyme of osteoclasts. (B) TRAP-positive multinuclear cells (nuclei

≥ 10) were counted as osteoclasts. *** P < 0.001.

S3 Figure S2. 1H NMR spectrum of 1 Figure S3. 13C NMR spectrum of 1

S5 Figure S4. 1H-1H COSY spectrum of 1 Figure S5. 1H-1H COSY spectrum of 1_expansion

S7 Figure S6. HMQC spectrum of 1 Figure S7. HMBC spectrum of 1

S9 Figure S8. NOESY spectrum of 1 Figure S9. 1H NMR spectrum of 2

S11 Figure S10. 13C NMR spectrum of 2 Figure S11. 1H-1H COSY spectrum of 2

S13 Figure S12. HSQC spectrum of 2 Figure S13. HMBC spectrum of 2

S15 Figure S14. NOESY spectrum of 2 Figure S15. NOESY spectrum of 2_expansion

S17 Figure S16. 1H NMR spectrum of 7S-dihydrosinomenine