3. A Photo of Viburnum species A Photo of Viburnum species

4. Abstract Bioassay-guided fractionation of the methanolic extract of Viburnum awabuki afforded two new lupane triterpene derivatives; 6b -hydroxyl-3,20-dioxo-30 norlupane-28-oic acid (1) and 3,4-secolup-4,20-dihydroxy-3,28-dioic acid-3-oic acid methyl ester (2), along with seven known lupane and oleanane- type triterpenes (3–9). The structure of the isolated compoundswas assigned using different spectroscopic techniques including 1D and 2D NMR. The 13 CNMR data of compounds 5 and 9 is reported for the first time. Triterpenes 1, 2, and 7–9 showed in vitro cytotoxic activity against several tumor cell line

5. Results Different chromatographic techniques utilizing normal and reversed phase silica gel have been used for fractionation of the methanol extract of the leaves and twigs of V. awabuki. The result of this separation yielded two new triterpenes derivatives, norlupane (1) and secolupane (2). In addition, seven known lupan- and oleanane-type triterpenes were also identified. The known oleanane-type triterpenes were idenitified as oleanolic acid (3  - hydroxyolean-12-en-28-oic acid) (3) and sumareesinolic acid (3 ,6  -dihydroxyolean-12-en-28-oic acid) (4), while the known lupane type were identified as 3  -hydroxylup- 12-en-28-oic acid (5), 3 ,20-dihydroxy lupane-28-oic acid (6), 6  -hydroxy-3-oxolup 20(29)-en-28 oic acid (7), 6 ,30- dihydroxy-3-oxolup 20(29)-en-28-oic acid (8), and 20 hydroxy-3-oxolup-28-oic acid (9). The 13 CNMR data of compounds 5 and 9 is reported for the first time. Triterpenes 1, 2, and 7–9 showed in vitro cytotoxic activity against several tumor cell line

6. Extraction, Separation and Purification Scheme of viburnum awabuki leaves and twigs

7. Triterpenes isolated from vibrnum awabuki

9. 1 HNMR of Compound 1 6 19 2 2’ 29 7 2426 25 23 27

10. DEPT Experiment of Compound 1 27 2526 11 24 23 29 12 21 15 16 2 22 13 7 1 19 99 9

11. HSQC of Compound 1 6192 29

12. HMBC of Compound 1 28 20 3 6 5 619 29 232724

13. 1 HNMR of Compound 2 OMe OH 26 2527 29 23 30 24

14. HSQC of Compound 2 OMe 18 195 OMe 149 8 1013725 2627 29233024 611 26 25 27 29 23 30 24 2 13 19 18

15. HMBC of Compound 2 283 4 29 30 23 24 26 25,27 20 OMe 2 18 22 17

16. Expanded HMBC of Compound 2 41720 5 19 26 25 27 29 30 24 23 18 14 9 8 10 13

17. NOESY of Compound 2

18. 1 HNMR of Compound 3 12 3 18

19. 13 C NMR of Compound 3 28 12 13 35

20. Expanded 13 C NMR of Compound 3 2524 26 6 16 11 30 27 2 15 2 15 24 x 20 22 21 297 10 1 4 8 18 14 19 17 9 5

21. HSQC of Compound 3 31812

22. 1 HNMR of Compound 4 12OH 6 3 18 x

23. 13 C NMR and DEPT Experiment of Compound 4 13 C NMR and DEPT Experiment of Compound 4 28 13365

24. HSQC of Compound 4 6123 18

25. 1 HNMR of Compound 5 in (CDCl 3 +CD 3 OD) 3 12 x

26. 13 C NMR of Compound 5 28 13 12 3518 9

27. DEPT Experiment of Compound 5 123 5 189 1920 23 30 26 2425 29 27 616 11 122 15 2

28. 1 HNMR of Compound 6

30. 13 C NMR and DEPT Experiment of Compound 6 28 3 2017 5 9 19 18 9 19 18 13 13 27242526 30 23 29

31. DEPT Experiment of Compound 6 27 2425 26 611 29 2330 2 1221 16 15 7 22 1 13 5 9 19 18

32. 1 HNMR of Compound 7 2929 6 30 2425 27 23 26

33. H HCOSY of Compound 7 29 6 19 2

34. DEPT Experiment of Compound 7 29 6 5 9 18 19 7 1 13 22 2 16 2115 12 23 24 11 30 2526 27

35. HSQC of Compound 7 29 6 20 28 3 296

36. 1 HNMR of Compound 8 29 6 30 26 23 27 2425

37. HSQC of Compound 8

38. 1 HNMR of Compound 9

39. 13 C NMR and DEPT Experiments of Compound 9

40. Expanded 13 C NMR of Compound 9 17 5 919 18 4 14 8 1 13 10 22 7 15 30 12 162 2329 11 246 26 25 27

41. DEPT Experiments of Compound 9

42. HSQC of Compound 9

43. HMBC of Compound 9