Supporting information Design, Synthesis, and Cancer Cell Growth Inhibitory Activity of Triphenylphosphonium Derivatives of the Triterpenoid Betulin Olga V. Tsepaeva, Andrey V. Nemtarev, Timur I. Abdullin, Leysan R. Grigor eva, Elena V. Kuznetsova, Rezeda A. Akhmadishina, Liliya E. Ziganshina, Hanh H. Cong, and Vladimir F. Mironov A. E. Arbuzov Institute of Organic and Physical Chemistry, Kazan Scientific Center of Russian Academy of Sciences, Arbuzov Street, 8, 420088, Kazan, Russian Federation Kazan (Volga Region) Federal University, Kremlevskaya Street, 18, 420008, Kazan, Tatarstan, Russian Federation *To whom correspondence should be addressed. Tel: +7 (843) 2727384. E-mail address: a.nemtarev@mail.ru S1
Table of Contents 1) Figure S1. Representative concentration-cell viability curves for cancer cell lines and human skin fibroblasts treated with the compound 9 (MTT assay). Page S5 2) Figure S2. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 2 Page S6 3) Figure S3. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 3 Page S7 4) Figure S4. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 4 Page S8 5) Figure S5. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 5 Page S9 6) Figure S6. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 6 Page S10 7) Figure S7. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 7 Page S11 8) Figure S8. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 8 Page S12 9) Figure S9. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 9 Page S13 10) Figure S10. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 10 Page S14 11) Figure S11. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 11 Page S15 12) Figure S12. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 12 Page S16 13) Figure S13. 1 H NMR (CDCl 3, 400 MHz) spectrum of compound 13 Page S17 14) Figure S14. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 3 Page S18 15) Figure S15. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 3 Page S19 16) Figure S16. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 4 Page S20 17) Figure S17. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 4 Page S21 S2
18) Figure S18. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 8 Page S22 19) Figure S19. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 8 Page S23 20) Figure S20. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 9 Page S24 21) Figure S21. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 9 Page S25 22) Figure S22. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 10 Page S26 23) Figure S23. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 10 Page S27 24) Figure S24. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 11 Page S28 25) Figure S25. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 11 Page S29 26) Figure S26. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 12 Page S30 27) Figure S27. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 12 Page S31 28) Figure S28. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of compound 13 Page S32 29) Figure S29. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of compound 13 Page S33 30) Figure S30. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 8 Page S34 31) Figure S31. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 9 Page S35 32) Figure S32. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 10 Page S36 33) Figure S33. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 11 Page S37 S3
34) Figure S34. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 12 Page S38 35) Figure S35. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of compound 13 Page S39 36) Figure S36. IR (KBr) spectrum of compound 2 Page S40 37) Figure S37. IR (KBr) spectrum of compound 3 Page S41 38) Figure S38. IR (KBr) spectrum of compound 4 Page S42 39) Figure S39. IR (KBr) spectrum of compound 5 Page S43 40) Figure S40. IR (KBr) spectrum of compound 6 Page S44 41) Figure S41. IR (KBr) spectrum of compound 7 Page S45 42) Figure S42. IR (KBr) spectrum of compound 8 Page S46 43) Figure S43. IR (KBr) spectrum of compound 9 Page S47 44) Figure S44. IR (KBr) spectrum of compound 10 Page S48 45) Figure S45. IR (KBr) spectrum of compound 11 Page S49 46) Figure S46. IR (KBr) spectrum of compound 12 Page S50 47) Figure S47. IR (KBr) spectrum of compound 13 Page S51 48) Table S48. Inhibitory Concentrations (IC 50, µм) of compounds 2-7 for Different Human Cells in vitro (MTT Assay) Page S52 S4
Cell viability (%) 100 80 60 40 20 PC-3 Cell viability (%) 140 120 100 80 60 40 20 MCF-7 0 1E-4 1E-3 0.01 0.1 Concentration (µm) 0 1E-4 1E-3 0.01 0.1 Concentration (µm) Cell viability (%) 140 120 100 80 60 40 20 0 MCF-7/Vinb 1E-6 1E-5 1E-4 1E-3 0.01 Concentration (µm) Cell viability (%) 100 80 60 40 20 0 HSF 1E-4 1E-3 0.01 0.1 Concentration (µm) Figure S1. Representative concentration-cell viability curves for cancer cell lines and human skin fibroblasts treated with the compound 9 (MTT assay). S5
Figure S2. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 2. S6
Figure S3. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 3. S7
Figure S4. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 4. S8
Figure S5. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 5. S9
Figure S6. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 6. S10
Figure S7. 1 H NMR (CDCl 3, 400 MHz) spectrum of bromide 7. S11
Figure S8. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 8. S12
Figure S9. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 9. S13
Figure S10. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 10. S14
Figure S11. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 11. S15
Figure S12. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 12. S16
Figure S13. 1 H NMR (CDCl 3, 400 MHz) spectrum of phosphonium salt 13. S17
Figure S14. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of bromide 3. S18
Figure S15. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of bromide 3. S19
Figure S16. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of bromide 4. S20
Figure S17. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of bromide 4. S21
Figure S18. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 8. S22
Figure S19. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 8. S23
Figure S20. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 9. S24
Figure S21. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 9. S25
Figure S22. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 10. S26
Figure S23. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 10. S27
Figure S24. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 11. S28
Figure S25. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 11. S29
Figure S26. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 12. S30
Figure S27. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 12. S31
Figure S28. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum of phosphonium salt 13. S32
Figure S29. 13 C-{ 1 H} NMR (CDCl 3, 100.6 MHz) spectrum fragment of phosphonium salt 13. S33
Figure S30. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 8. S34
Figure S31. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 9. S35
Figure S32. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 10. S36
Figure S33. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 11. S37
Figure S34. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 12. S38
Figure S35. 31 P-{ 1 H} NMR (CDCl 3, 162 MHz) spectrum of phosphonium salt 13. S39
Figure S36. IR (KBr) spectrum of bromide 2. S40
Figure S37. IR (KBr) spectrum of bromide 3. S41
Figure S38. IR (KBr) spectrum of bromide 4. S42
Figure S39. IR (KBr) spectrum of bromide 5. S43
Figure S40. IR (KBr) spectrum of bromide 6. S44
Figure S41. IR (KBr) spectrum of bromide 7. S45
Figure S42. IR (KBr) spectrum of phosphonium salt 8. S46
Figure S43. IR (KBr) spectrum of phosphonium salt 9. S47
Figure S44. IR (KBr) spectrum of phosphonium salt 10. S48
Figure S45. IR (KBr) spectrum of phosphonium salt 11. S49
Figure S46. IR (KBr) spectrum of phosphonium salt 12. S50
Figure S47. IR (KBr) spectrum of phosphonium salt 13. S51
IC 50, µм compound PC-3 MCF-7 МСF-7/Vinb HSF 2 22.34 ± 2.92 22.3 ± 2.0 20.24 ± 2.0 9.67 ± 0.98 3 53.31 ± 5.65 33.37 ± 3.27 44.89 ± 6.83 51.48 ± 2.97 4 22.86 ± 6.38 23.27 ± 2.73 52.86 ± 9.36 > 100 5 64.2 ± 2.4 82.57 ± 10.63 11.88 ± 4.0 11.09 ± 1.46 6 71.68 ± 8.66 105.36 ± 15.38 115.0 ± 0.66 191.35 ± 16.84 7 21.82 ± 1.89 92.72 ± 8.22 137.07 ± 27.32 29.93 ± 8.26 betuline 148.65 ± 13.09 227.04 ± 19.42 233.42 ± 13.29 164.4 ± 12.26 Table S48. Inhibitory Concentrations (IC 50, µм) of compounds 2-7 for Different Human Cells in vitro (MTT Assay) S52