2502 1
(PAs)PAs WHO PAs PAs PAs (PAs) (1) (Dictionary of Natural Products) SciFinder PAs PAs (2) PubMedEFSA MHRA PAs (3) (4) (1)(3) PAs (PAs) PAs PAs Senkirkine (Symphytum officinale) PAs Retronecine PAs 2
N- (PAs) (1) (2) PAs LC-MS/MS (3) PAs HPLC PAs LC-MS 9,000 8,850 3
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(PAs) (1) (Dictionary of Dictionary of Natural Products Natural Products) SciFinder SciFinder PAs PAs PAs PAs (Symphytum officinale) (Petasites japonicus) PA 1 Retronecine type 8 Heliotridine type 2 1 Otonesine type 3 Petasinesine type 2 (2) PubMedEFSA MHRA PAs (3) (4) (1)(3) PAs PAs PAs 5
1 (Symphytum officinale) (Petasites japonicus) 1) K. Araki et al., Chem. Pharm. Bull., 1968, 16, 2512-2516. 2) E. W. Monroe et al., J. Nat. Prod., 2001, 64, 251-253. 3) J. Brauchli et al., Experientia, 1982, 38, 1085-1087. 4) E. Roeder et al., Phytochemistry, 1992, 31, 4041-4042. 5) Y. G. Denisov et al., Rastitel nye Resursy, 1970, 6, 409-411. 6
1) K. Yamada et al., Chem. Lett., 1976, 461; K. Yamada et al., ibid., 1123. 2) T. Furuya et al., Chem. Pharm. Bull., 1976, 24, 1120. 3) K. Yamada et al., Phytochemistry, 1978, 17, 1667. 4) K. Yamada et al., Tetrahedron Lett., 1976, 17, 4543. 7
(PAs) MeOH 1.9 kgdry weight MeOH MeOH 219.0 g MeOH AcOEt 8.4 g2.4 g n-buoh 34.2 g LC-MS SiO2 amino-silica gel HPLC 7 PA PA2 NMR 7 PA 4 Retronesine type [Symphytine (1)Echimidine (2) Echimidine N-oxide (3)Lycopsamine (4)] 3 [SO-3 (5)SO-6 (6) SO-8 (7)] PA PA SO-4 (8) SOM-1 (9) 8
4 [SO-3 (5), SO-6 (6), SO-4 (8), SO-8 (7)] PA NMR SOM-1 NMR PAs 4.34 kgdry weight MeOH MeOH 424.6 gmeoh AcOEt 28.11 g2.77 gn-buoh 22.58 g SiO2 amino-silica gel HPLC 5 PA 6.8 kgdry weight 2 kg4 C 972 g) 2 kg (wet weight)1.2 kg (wet weight)2 kg (wet weight) MeOH 6.8 kg, dry weight MeOH MeOH MeOH AcOEt 20.6 g2.97 gn-buoh 90.9 g 97.8 mg 9
LC-MS 2.96 g SiO2 amino-silica gel HPLC 4 PA 3 MeOH 1920.27 g (wet weight) MeOH 8.0 L, rt 3 d; 7.0 L x 4 d; 6.0 L, reflux 8 h; 6.5 L, reflux 8 h MeOH 60.14 g4 C 191.56 g (dry weight) MeOH 3.0 L, rt 63 h; 2.3 L, rt 50.5 h; 2.5 L, reflux 8 h; 2.5 L, reflux 8 hmeoh 32.51 g4 C 429.39 g (dry weight) MeOH 4.3 L, rt 63 h; 3.5 L, rt 50.5 h; 3.8 L, reflux 8 h; 3.5 L, reflux 8 h MeOH 60.54 g4 C MeOH AcOEt 32.18 g1.85 gn-buoh 2.96 g SiO2 amino-silica gel HPLC 2 PA 10
921 g MeOH 9.5 L, rt 42 h; 8.9 L, rt 22.5 h; 6.3 L, reflux 8 h; 7.2 L, reflux 8 h MeOH 103.3 g MeOH AcOEt 24.6 g4.24 gn-buoh (10.9 g) 4.18 g SiO2 amino-silica gel HPLC 4 PA NMR 4 PA 3 Otonesine type [Neopetasitenine (13)Petasitenine (14) Senkirkine (15)] NMR 1 [PJ-4 (16)] Otonesine type Petasitenine (14) 5 25 11 1-2 (Petasites japonicus) 11
PAs Retronecine Retronecine (+)-Retronecine (1) Scheme 14 Retronecine Retronecine Loroquine PAs N- N- PAs PAs PAs Retrorsine CDCl3 NMR PA NMR 1,4- BTMSB-d4 Senkirkine 99.2% 12
LC-MS 13
TLC TLC SiO2 10% MeOH/CHCl3 UV 254 nm Schlittler SiO2 flash 4.5 cm x h 8.0 cm, MeOH/CHCl3 gradient 14
Symphytine (1) Echimidine (2) Echimidine N-oxide (3) Lycopsamine (4) SO-3 (5) SO-6 (6) SO-8 (7) SO-4 (8) SOM-1 (9) 20.1 mg 47.0 mg 2.3 mg 1.6 mg 3.2 mg 7.0 mg 10.2 mg 1.6 mg 4.0 mg 1-4 NMR 1-3 ESI-MS PAs[SO-3 (5), SO-6 (6), SO-4 (8), SO-8 (7), SOM-1 (9)] SO-3 (5), SO-6 (6), SO-4 (8), SO-8 (7), SOM-1 (9) PAs 1 H-NMR, 13 C-NMR SOM-1 (9) 15
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Symphytine (1) Echimidine (2) Myoscorpine (10) Symlandine (11) 7-Acetyllycopsamine (12) 1.3 mg 37.6 mg 3.7 mg 0.4 mg 14.0 mg 10-12 NMR 10-12 ESI-MS 17
Petasites japonicus Whole plant, 6.8 Kg (dry weight) MeOH Extract 938.3 g extracted with MeOH (26.8 L, rt 4 d; 19.4 L, rt 1 d; 14 L, reflux 8 h; 17.8 L, reflux 8 h) dissolved in 1 N H 2 SO 4 aq. (2.9 L) extracted with AcOEt (2.0 L) AcOEt Layer extracted with 1 N H 2 SO 4 aq. (1.0 L) aq. Layer AcOEt Layer 20.6 g basified with conc. NH 3 (750 ml) (ph 9) extracted with 5% MeOH/CHCl 3 x 3-4, (total 7. 0 L) Crude Base 2.97 g aq. Layer extracted with n-buoh x 4 (total 8.0 L) n-buoh Layer 90.9 g aq. Layer LC-MS TLC TLC SiO2 20% MeOH/CHCl3 UV 254 nm Schlittler 18
SiO2 flash 4.6 cm x 9.0 cm, MeOH/CHCl3 gradient Neopetasitenine (13) Petasitenine (14) Senkirkine (15) PJ-4 (16) 172.2 mg 181.8 mg 24.7 mg 2.7 mg 13-16 NMR 13-16 ESI-MS 13-16 HPLC 13-16 LC-MS/MS 5 19
Parasenecio delphiniifolius Whole plant, 5.5 Kg (wet weight) MeOH Extract 153.2 g extracted with MeOH (rt, 2 times; reflux, 2 times) 152.3 g dissolved in 1 N H 2 SO 4 aq. (1.0 L) extracted with AcOEt (0.7 L) AcOEt Layer extracted with 1 N H 2 SO 4 aq. (0.8 L) aq. Layer AcOEt Layer 32.18 g basified with conc. NH 3 (ph 9) extracted with 5% MeOH/CHCl 3 (4 times, total 8.0 L) Crude Base 1.85 g aq. Layer extracted with n-buoh (3 times, total 2.8L) n-buoh Layer aq. Layer TLC TLC SiO2 UV 254 nm Anisaldehyde TLCSchlittler TLC MeOH (M) (C) SiO2 flash 3.0 cm x 8.0 cm, MeOH/CHCl3 gradient Fr. 1-4 10% MeOH/CHCl3 20
(C) Fr. 6-8 20% MeOH/CHCl3 HPLC Column InertSil ODS-3 (GL Science) 5 m (4.6 x 250 mm) A 0.2 % aq. HCOOH B 0.2 % HCOOH / MeOH Time (min) A (%) B (%) 0.5 ml / min 0 70 30 30 60 10 90 Inject0.21mg / 20mL MeOH 3L inject 61 0 100 90 stop 21
A, B, C1, 2, 3 A, B, C compounds isolated from Parasenecio delphiniifolius 22
Gynura bicolor Whole plant, 921 g (dry weight) MeOH Extract 103.3 g 101.9 g extracted with MeOH (9.5 L, rt, 42 h; 8.9 L, rt, 22.5 h; 6.3 L, reflux, 8 h; 7.2 L, reflux 8-9 h) dissolved in 1 N H 2 SO 4 aq. (810 ml) extracted with AcOEt (830 ml) AcOEt Layer aq. Layer AcOEt Layer 24.6 g basified with conc. NH 3 (ph 9) extracted with 5% MeOH/CHCl 3 (2.0 L x 3) extracted with 1 N H 2 SO 4 aq. (730 ml) Crude Base 4.24 g aq. Layer extracted with n-buoh (800 ml x 3) n-buoh Layer 10.9 g aq. Layer TLC TLC SiO2 UV 254 nm Hanessian 10% MeOH/CHCl3 23
SiO2 flash 4.5 cm x 8.0 cm, MeOH/CHCl3 gradient Fr. 1-5 5% MeOH/CHCl3 Fr. 6-10 10% MeOH/CHCl3 Fr. 11-16, MeOH (M) 20% MeOH/CHCl3 24
A, B, C1, 2, 3 A, B, C HPLC Fr. 7B7B Column Inertsil C18 (GL Science) 5 m (10 x 250 mm) A 0.2 % aq. HCOOH B acetonitrile 1.0 ml / min 30 Inject12.1 mg / 100 L (MeOH) 50 L Time (min) 0 60 A (%) 40 0 B (%) 60 100 25
(+)-Retronecine (RCM) (Scheme 1) ()- 3 Baylis -Hillman 4 4 RCM 6 Table 1 Baylis-Hillman 4 Entry reductant solvent condition result 1 DIBAL 70 complex THF 8.0 eq. 30 min mix. 2 BH3 70 to complex THF THF 0 mix. 6.0 eq. 30 min 3 LiAlH4 0 complex THF 15 eq. 30 min mix. 4 LiAlH4 0 complex THF 8.0 eq. 1 h mix. Table 1 26
(Scheme 2) (R)-3-Pyrrolizinol (8)N- 2 10 Table 2 11 6 Table 2 Entry Si source Base solvent condition 11 13 10 (S.M.) 1 TIPSOTf 2.5 eq. Et3N 2.0 eq. CH2Cl2 rt 3.5 h - 82% - 2 TIPSCl 2.0 eq. LDA 1.5 eq. THF 78 to 0 30 min - - trace 3 TIPSCl 3.0 eq. KHMDS 1.5 eq. toluene 78 to 0 30 min - 19% - 4 TIPSCl 3.0 eq. LDA 1.5 eq. THF/toluene 1 : 1 78 to 0 30 min decomp. Table 2 27
6 6 16 (Scheme 3) L-6 16 (Table 3) 17 Entry TESCl Base Solvent Condition 17 18 19 20 1 3.0 eq. LDA 1.4 eq. THF 78 to 0 30 min N.R. 2 3.0 eq. LHMDS 1.5 eq. THF 78 to 0 30 min N.R. 3 3.0 eq. KHMDS 1.5 eq. toluene 78 to 0 30 min - 25% 7% 17% 4 3.0 eq. KHMDS 1.5 eq. toluene 90 to 0 30 min - 24% 5% 21% Table 3 28
Pd (Scheme 4)(R)-3-Pyrrolizinol (8) 2 21 Pd Table 4 2324 23 TBS Loroquine (25) Entry Pd cat. Et3N additive solvent condition 22 23 24 1 Pd(PPh3)4 20 mol% 3.0 eq. - DMF 80 18 h - 24 % - 2 Pd(PPh3)4 20 mol% 3.0 eq. Na ascorbate 1.0 eq. DMF 80 18 h - 40 % - 3 Pd(PPh3)4 20 mol% - Na ascorbate 1.0 eq. DMF/Et3N (1 : 1) 80 18 h - 39% - 4 Pd(PPh3)4 20 mol% - - Et3N 80 18 h - 45% 18% Pd(OAc)2 5 - PPh3 DMF/Et3N 30 mol% 60 mol% (1 : 1) Table 4 80 9 h - 26% - 29
Retrorcine 1 H-NMR Senkirkine (15) NMR Senkirkine (15) NMR Senkirkine 7.78 mg 1,4-BTMSB-d4 0.66 mg NMR (CDCl3 )NMR 99.2 % 30
(PAs) (1) PA Mattocks Molyneux PA (2) PAs LC-MS/MS 5 PAs [Symphytine (1), Echimidine (2), Myoscorpine (10), Symlandine (11), 7- Acethyllycopsamine (12)] LC-MS/MS m/z 120 PA (3) PAs HPLC PAs LC-MSMS MeOH 31
PAs TLC Mattocks-Molyneux J. Chromatography, 1980, 195, 412-415 A 1% O-chloranil (tetrachloro-o-benzoquinone) / benzene B p-dimethylaminobenzaldehyde (2.0 g)(100 ml) Boron trifluoride etherate (2.0 ml) C 10% / benzene A N-oxide C B PAs Symlandine (11) TLC SiO2 10% MeOH / CHCl3 Hanessian Mattocks Molyneux 5mm 4 cm TLC 5 cm 6 mm Rf 0.15 Mattocks Molyneux PA 1, 2, 10-12 LC-MS/MS 32
PAs HPLC SampleEchimidine (2) main Echimidine (2) Column Inertsil ODS-3 (GL Science) 3 m (4.6 x 250 mm) A 0.2 % aq. HCOOH B MeOH 0.3 ml / min 30 Inject21.7 mg / 3mL (MeOH) 3L Time (min) 0 60 A (%) 40 0 B (%) 60 100 7.5 min Echimidine (2) 33
(PAs) 16 12 PAs LC-MS/MS HPLC 5 25 11 1-2 (Petasites japonicus) 108 26 12 3-6 25 6 14 25 26 2 19 25 26 6 4 26 27 3 5 26 34