Electronic Supplementary Information: towards cialis ...

Electronic Supplementary Information:

Highly stereoselective one-pot construction of trisubstituted tetrahydro--carboline-fused diketopiperazines: A synthetic route towards cialis analogues

Mouhamad Jida,* Dirk Tourwé and Steven Ballet*

Research Group of Organic Chemistry, Departments of Chemistry and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, [email protected] and [email protected]

Contents

1. General information..................................................................................................................12. General procedure for the synthesis of THBC-based DKPs (8a-n) .........................................23. Characterization of compounds 8a-o........................................................................................34. NOESY studies of 8a and 8l ....................................................................................................85. NMR Spectra.......................................................................................................................9-265. 2D NMR Spectra (COSY, NOESY, HSQC) and HPLC of 8a .........................................27-30

NH

HN

OOH

R3 NC

R1 NH2R2 CHO

Boc

NH

N

R4

NR1O

O

R2

( R3 = Cy, Bn, tBu)

1) Ugi-4CR2) Boc-deprotection

3) Pictet-Spengler

4) Cyclisationup to > 98% de

15 examples up to 81% yield

one-pot

*

*

R4 CHO

Electronic Supplementary Material (ESI) for RSC Advances.This journal is © The Royal Society of Chemistry 2014

mailto:[email protected]

mailto:[email protected]

S

1. General information1H and 13C NMR spectra (25°C) were recorded at 250 MHz and 63 MHz on a Bruker Avance

DRX 250 spectrometer or at 500 MHz and 126 MHz respectively on a Bruker Avance II 500

spectrometer. Chemical shifts are in parts per million (ppm). The assignments were made using

one dimensional (1D) 1H and 13C spectra or two-dimensional (2D) HSQC and COSY spectra.

HRMS data was recorded with a Micromass QTOF-micro system. Mass spectra were recorded

with a LCMS-MS triple-quadrupole system. Analytical HPLC was performed on an Agilent 1100

Series system with a Supelco Discovery BIO Wide Pore RP column (25 cm × 4.6 mm, 5 μm).

Flow rates of 1 ml/min were used and detection was done at 215 nm. The solvent system

consisted of 0.1% TFA in water (A) and 0.1% TFA in acetonitrile (B). The gradient consisted of

a 26 min. run from 3% B to 100% B. Flash chromatography was performed with silica gel 60

(Davisil, 0.040-0.063 mm) from Merck. Glass plates with silica gel 60 F254 (Merck) were used

for thin layer chromatography. Visualization of the products on TLC plates was realized using

UV light (254 nm), KMnO4 spray. Melting points were determined on a Büchi B-540 apparatus

and are uncorrected. Reactions were performed using a Biotage® Initiator+ Microwave

Synthesizer. All commercial reagents and solvents were used without further purification.

Specific rotations were measured on a polarimeter polartronic M Schmidt-Haenschusing a 5 cm

cell in standard conditions (20°C / 589.3 nmair / 589.44 nmvac).

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2. General procedure for one-pot tandem Ugi-4CR/Boc-deprotection/Pictet-Spengler/Cyclisation sequence (8a-n).

In a capped 5 mL microwave-vessel containing a stirring bar, the N-Boc protected (L/D)-

tryptophan (0.5 mmol), the aldehyde (0.5 mmol) and the amine (0.5 mmol) were dissolved and

mixed in methanol (3 mL). The solution was stirred at room temperature for 30 min. Isocyanide

(0.5 mmol) was then added, and the resulting mixture was stirred at room temperature. After 18h,

total conversion was determined from LC-MS analysis and the solvent was removed in vacuo.

The crude Ugi-4CR product was dissolved in 30% TFA in DCM (3 mL) and the resulting

mixture was stirred at room temperature for 3 hours. After completion of the N-Boc deprotection

step (monitored by HPLC and LC-MS), the solvent was evaporated in vacuo. Next, the crude

deprotected Ugi product was dissolved in a mixture of TFA (1.5 mmol) in DCM (3 mL) and the

corresponding aldehyde (0.55 mmol) was then added, and the resulting mixture was stirred

overnight at room temperature. Total conversion was observed from HPLC and LC-MS analysis

and the solvent was removed under reduced pressure.

The crude Ugi Pictet-Spengler product was dissolved in glacial acetic acid (3 mL) and the

reaction vessel was capped. The reaction mixtures was heated at 180°C under microwave

irradiation for 80 minutes using the Biotage initiator+ microwave reactor (average effective ramp

time =1 min), the power was set at 400 W and the pressure was set at 30 bar (average effective

pressure = 5 bar). After completion of the reaction as indicated by LC-MS, the solvent was

evaporated under reduced pressure and the residue was dissolved in EtOAc (20 mL). The organic

layer was washed with sat. NaHCO3 (3 X 10 mL), 1N HCl (3 X 10 mL), sat. NaCl (3 X 10 mL),

dried over MgSO4 and the solvent was removed under reduced pressure.The crude products were

purified by flash chromatography. The resulting products (8a-n) were isolated in good purity and

yields as a single diastereomer. Purity (%) was determined by reversed phase HPLC, using UV

detection (215 nM).

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3. Characterization of compounds (8a-n).

(6R,12aS)-2-benzyl-6-phenyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8a):

8aNH

N

NO

O

Chemical Formula: C27H23N3O2Exact Mass: 421.18

Yield (171 mg, 81%); pale yellow solid; mp= 263-265 °C; [α]D = -30.0 ° (c 0.6, CHCl3); 1H NMR (250 MHz, CDCl3) δ 7.92 (br s, 1H, NH), 7.56 (d, J = 8.0 Hz, 1H), 7.40-7.14 (m, 13H), 7.04 (s, 1H), 4.87 (d, J = 14.75 Hz, 1H), 4.43 (dd, J = 12.0, 4.0 Hz, 1H), 4.34 (d, J = 14.5 Hz, 1H), 4.00 (d, J = 18.25 Hz, 1H), 3.95 (d, J = 18.25 Hz, 1H), 3.62 (dd, J = 16.0, 4.0 Hz, 1H), 2.99 (dd, J = 16.0, 12.0 Hz, 1H); 13C NMR (63 MHz, CDCl3) δ 165.46, 162.07, 137.99, 136.34, 134.74, 129.56, 129.07, 128.96, 128.83, 128.54, 128.33, 126.23, 122.78, 120.13, 118.44, 111.19, 108.95, 52.67, 52.19, 49.45, 48.78, 27.83; TLC: Rf = 0.55 (EtOAc /Hexane 9/1, v/v); rt(HPLC) = 17.20 min; HRMS-ESI (m/z): [M+Na]+ calcd for C27H23N3O2Na444.1682; found 444.1660.

(6R,12aS)-2-methyl-6-phenyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8b):

8bNH

N

NO

O


Yield (121 mg, 70%); brown solid; mp= 288-290 °C; [α]D = -73.3 ° (c 0.5, CHCl3); 1H NMR (250 MHz, CDCl3) δ 8.14 (br s, 1H, NH), 7.53 (d, J = 7.75 Hz, 1H), 7.32-7.12 (m, 8H), 7.06 (s, 1H), 4.32 (dd, J = 11.75, 4.5 Hz, 1H), 4.12 (d, J = 17.5 Hz, 1H), 3.95 (d, J = 17.75 Hz, 1H), 3.55 (dd, J = 15.25, 4.5 Hz, 1H), 3.01-2.89 (m, 1H), 2.96 (s, 3H); 13C NMR (63 MHz, CDCl3) δ 165.46, 161.64, 138.24, 136.36, 129.63, 128.89, 128.78, 126.26, 122.70, 120.06, 118.41, 111.18, 109.07, 52.55, 52.08, 51.42, 33.40, 27.65; TLC: Rf = 0.42 (EtOAc /Hexane 9/1, v/v); rt(HPLC) = 14.22 min; HRMS-ESI (m/z): [M+H]+ calcd for C21H20N3O2 346.1550; found 346.1558.

(6R,12aS)-2-methyl-6-(m-tolyl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8c):

8cNH

N

NO

O


Yield (136 mg, 76%); yellow oil; [α]D = -176.8 ° (c 1, CHCl3); 1H NMR (250 MHz, CDCl3) δ 7.88 (br s, 1H, NH), 7.55 (d, J = 8.0 Hz, 1H), 7.33-7.10 (m, 7H), 7.03 (s, 1H), 4.39 (dd, J = 12.5, 4.5 Hz, 1H), 4.15 (d, J = 17.5 Hz, 1H), 3.98 (d, J = 17.5 Hz, 1H), 3.57 (dd, J = 15.0, 4.5 Hz, 1H), 3.04-2.91 (m, 1H), 3.00 (s, 3H), 2.29 (s, 3H); 13C NMR (63 MHz, CDCl3) δ 165.53, 161.56, 138.83, 138.08, 136.31, 129.84, 129.71, 129.31, 128.81, 125.96, 122.70, 120.08, 118.43, 111.15, 109.12, 52.60, 52.06, 51.48, 33.40, 27.67, 21.38; TLC: Rf = 0.51 (EtOAc /Hexane 9/1, v/v); rt(HPLC) = 14.85 min; HRMS-ESI (m/z): [M+H]+ calcd for C22H22N3O2 360.1707; found 360.1695.

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(6R,12aS)-2-methyl-6-(naphthalen-2-yl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8d):

8dNH

N

NO

O


Yield (141 mg, 71%); brown solid; mp= 165-167 °C; [α]D = -87.5 ° (c 0.7, CHCl3); 1H NMR (250 MHz, CDCl3) δ 8.28 (br s, 1H, NH), 7.78-7.70 (m, 1H), 7.62-7.55 (m, 3H), 7.50-7.39 (m, 3H), 7.33-7.15 (m, 5H), 4.34 (dd, J = 12.0, 4.5 Hz, 1H), 4.05 (d, J = 17.5 Hz, 1H), 3.86 (d, J = 17.5 Hz, 1H), 3.57 (dd, J = 15.5, 4.5 Hz, 1H), 3.00-2.90 (m, 1H), 2.86 (s, 3H); 13C NMR (63 MHz, CDCl3) δ 165.40, 161.76, 136.43, 135.73, 133.32, 132.97, 129.73, 128.91, 128.14, 127.95, 127.61, 126.76, 126.63, 126.33, 126.22, 122.68, 120.04, 118.45, 111.31, 109.14, 52.54, 52.24, 51.35, 33.28, 27.69; TLC: Rf = 0.47 (EtOAc /Hexane 8/2, v/v); rt(HPLC) = 15.76 min; HRMS-ESI (m/z): [M+H]+ calcd for C25H22N3O2 396.1707; found 396.1706.

(6R,12aS)-6-isobutyl-2-methyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8e):

8eNH

N

NO

O


Yield (119 mg, 73%); yellow oil; [α]D = -29.2 ° (c 0.5, CHCl3); 1H NMR (250 MHz, MeOD) δ 7.42 (d, J = 8.75 Hz, 1H), 7.29 (d, J = 8.75 Hz, 1H), 7.10-6.94 (m, 2H), 6.43 (d, J = 16.5 Hz, 1H), 6.21 (dd, J = 16.5, 6.75 Hz, 1H), 4.23 (br s, 1H), 3.49-3.11 (m, 5H), 2.58-2.25 (m, 2H), 2.50 (s, 3H), 1.16 (d, J = 6.5 Hz, 3H), 1.15 (d, J = 6.5 Hz, 3H); 13C NMR (126 MHz, MeOD) δ 167.40, 165.77, 137.00, 136.39, 135.25, 128.41, 121.69, 118.65, 118.17, 115.59, 110.00, 105.29, 56.42, 50.01, 32.53, 31.58, 28.69, 21.43; TLC: Rf = 0.32 (EtOAc /Hexane 9/1, v/v); rt(HPLC) = 13.04 min; HRMS-ESI (m/z): [M+H]+ calcd for C19H24N3O2 326.1863; found 326.1871.

(6R,12aS)-6-(benzo[d][1,3]dioxol-5-yl)-2-methyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione {12a-epi-Cialis }(8f):

8fNH

N

NO

O

O

O


Yield (133 mg, 68%); pale yellow solid; mp= 291-293 °C (lit.1 mp = 287-289 °C); [α]D = -297.8 ° (c 1, CHCl3) {lit.2[α]D = -293.4 ° (c 1.2, CHCl3)}; 1H NMR (250 MHz, CDCl3) δ 8.00 (br s, 1H, NH), 7.53 (d, J = 7.5 Hz, 1H), 7.33-7.12 (m, 3H), 6.97 (s, 1H), 6.81 (s, 1H), 6.70 (s, 2H), 5.92 (s, 2H), 4.34 (dd, J = 12.0, 4.5 Hz, 1H), 4.14 (d, J = 17.5 Hz, 1H), 3.98 (d, J = 17.5 Hz, 1H), 3.55 (dd, J = 15.5, 4.5 Hz, 1H), 2.99 (s, 3H), 2.96-2.89 (m, 1H); 1H NMR (500 MHz, DMSO-d6) δ 11.02 (br s, 1H, NH), 7.47 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 7.08 (t, J = 7.5 Hz, 1H), ), 6.99 (t, J = 7.5 Hz, 1H), 6.85-6.80 (m, 2H), 6.74 (s, 1H), 6.58 (d, J = 8.0 Hz, 1H), 5.97 (d, J = 7.0 Hz, 2H), 4.22 (d, J = 18.0 Hz, 1H), 4.05 (dd, J = 11.5, 4.0 Hz, 1H), 4.01 (d, J = 18.0 Hz, 1H), 3.24 (dd, J = 15.5, 4.0 Hz, 1H), 2.93 (dd, J = 15.5, 11.5 Hz, 1H), 2.82 (s, 3H); 13C NMR (63 MHz, CDCl3) δ 165.46, 161.53, 148.15, 148.05, 136.32, 132.05, 129.71, 126.23, 122.77, 122.48, 120.10, 118.45, 111.15, 109.14, 108.31, 101.36, 52.42, 51.81, 51.47, 33.40, 27.60; TLC: Rf = 0.45 (EtOAc /Hexane 8/2, v/v); rt(HPLC) = 14.21 min; HRMS-ESI (m/z): [M+Na]+ calcd for C22H19N3O4Na 412.1268; found 412.1284.

(2) Daugan, A. C. –M. U.S. Patent 5, 859, 006.

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(6S,12aR)-6-(benzo[d][1,3]dioxol-5-yl)-2-methyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione{6-epi-Cialis } (8g):

8gNH

N

NO

O

O

O


Yield (142 mg, 73%); off-white solid; mp= 289-291 °C (lit.1mp = 287-288 °C); [α]D = +288.4 ° (c 1, CHCl3) {lit.20d [α]D

28 = +278 ° (c 1.03, CHCl3)};1H NMR (500 MHz, DMSO-d6) δ 11.02 (br s, 1H, NH), 7.47 (d, J = 8.0 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 7.08 (t, J = 7.5 Hz, 1H), ), 6.99 (t, J = 7.5 Hz, 1H), 6.85-6.81 (m, 2H), 6.75 (s, 1H), 6.59 (d, J = 8.0 Hz, 1H), 5.97 (d, J = 6.0 Hz, 2H), 4.22 (d, J = 18.0 Hz, 1H), 4.06 (dd, J = 12.0, 4.0 Hz, 1H), 4.01 (d, J = 18.0 Hz, 1H), 3.24 (dd, J = 15.5, 4.0 Hz, 1H), 2.93 (dd, J = 15.5, 12.0 Hz, 1H), 2.82 (s, 3H); 13C NMR (126 MHz, DMSO-d6) δ 165.13, 162.74, 148.11, 147.73, 136.75, 133.38, 130.83, 126.39, 122.19, 122.15, 119.36, 118.59, 111.80, 108.83, 108.62, 107.99, 101.73, 52.49, 51.33, 51.15, 33.09, 27.17; TLC: Rf = 0.45 (EtOAc /Hexane 8/2, v/v); rt(HPLC) = 14.20 min; HRMS-ESI (m/z): [M+Na]+ calcd for C22H19N3O4Na 412.1268; found 412.1288.

(6R,12aS)-2-benzyl-6-(4-bromophenyl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8h):

8hNH

N

NO

O

Br

Chemical Formula: C27H22BrN3O2Exact Mass: 499.09

Yield (185 mg, 74%); pale yellow oil; [α]D = -80.0 ° (c 0.5, CHCl3); 1H NMR (250 MHz, CDCl3) δ 8.23 (br s, 1H, NH), 7.54 (d, J = 10.0 Hz, 1H), 7.40-7.21 (m, 8H), 7.13-7.03 (m, 4H), 6.95 (s, 1H), 4.82 (d, J = 14.25 Hz, 1H), 4.32 (d, J = 14.25 Hz, 1H), 4.31 (dd, J = 12.0, 4.5 Hz, 1H), 3.96 (d, J = 18.0 Hz, 1H), 3.86 (d, J = 18.0 Hz, 1H), 3.59 (dd, J = 16.0, 4.5 Hz, 1H), 2.95 (dd, J = 16.0, 12.0 Hz, 1H); 13C NMR (63 MHz, CDCl3) δ 165.17, 161.94, 137.22, 136.39, 134.81, 132.04, 130.40, 129.07, 128.52, 128.32, 126.19, 123.05, 122.88, 120.18, 118.49, 111.25, 109.15, 52.65, 51.45, 49.39, 48.78, 27.75; TLC: Rf = 0.56(EtOAc /Hexane 9/1, v/v); rt(HPLC) = 18.33 min; HRMS-ESI (m/z): [M+H]+ calcd for C27H23BrN3O2 500.0968; found 500.0976.

(6R,12aS)-6-phenyl-2-((S)-1-phenylethyl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8i):

8iNH

N

NO

O


Yield (158 mg, 73%); yellow oil; [α]D = -184.4 ° (c 0.6, CHCl3); 1H NMR (250 MHz, CDCl3) δ 7.97 (br s, 1H, NH), 7.55 (d, J = 9.0 Hz, 1H), 7.38-7.12 (m, 13H), 7.00 (s, 1H), 6.01 (q, J = 7.0 Hz, 1H), 4.44 (dd, J = 12.25, 4.5 Hz, 1H), 4.01 (d, J = 18.0 Hz, 1H), 3.65 (d, J = 18.0 Hz, 1H), 3.62 (dd, J = 16.0, 4.5 Hz, 1H), 3.00 (dd, J = 16.0, 12.25 Hz, 1H), 1.58 (d, J = 6.5 Hz, 3H); 13C NMR (63 MHz, CDCl3) δ 165.49, 162.92, 137.82, 137.38, 136.65, 129.46, 128.99, 128.94, 128.41, 127.82, 127.40, 126.83, 126.21, 122.96, 120.31, 118.47, 111.26, 52.83, 52.39, 50.92, 43.64, 27.59, 15.08; TLC: Rf = 0.35 (EtOAc /Hexane 1/1, v/v); rt(HPLC) = 17.66 min; HRMS-ESI (m/z): [M+H]+ calcd for C28H26N3O2 436.2020; found 436.2018.

(20d)Vedantham, R.; Shanmugam, S.; Vetukuri, P.V.N.K.V.; Khagga, M.; Bandichhor, R. Arkivoc2013, 22.

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(6S,12aR)-6-phenyl-2-((S)-1-phenylethyl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8j):

8jNH

N

NO

O


Yield (163 mg, 75%); pale yellow solid; mp= 195-197 °C; [α]D = +75. 7 ° (c 0.6, CHCl3); 1H NMR (250 MHz, CDCl3) δ 7.80 (br s, 1H, NH), 7.56 (d, J = 7.75 Hz, 1H), 7.39-7.14 (m, 13H), 7.03 (s, 1H), 6.09 (q, J = 7.5 Hz, 1H), 4.45 (dd, J = 12.0, 3.75 Hz, 1H), 3.90 (d, J = 18.0 Hz, 1H), 3.63 (dd, J = 15.5, 3.75 Hz, 1H), 3.53 (d, J = 18.0 Hz, 1H), 2.94 (dd, J = 15.5, 12.0 Hz, 1H), 1.55 (d, J = 6.75 Hz, 3H); 13C NMR (63 MHz, CDCl3) δ 165.26, 162.58, 138.08, 136.41, 129.59, 128.93, 128.88, 128.75, 128.28, 127.44, 126.21, 122.78, 120.10, 118.42, 111.17, 108.93, 52.79, 52.18, 50.26, 43.82, 27.72, 14.54; TLC: Rf = 0.47 (EtOAc /Hexane 6/4, v/v); rt(HPLC) = 17.65 min; HRMS-ESI (m/z): [M+H]+ calcd for C28H26N3O2 436.2020; found 436.2010.

(6R,12aS)-6-phenyl-2-((R)-1-phenylethyl)-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8k):

8kNH

N

NO

O


Yield (171mg, 79%); pale yellow solid; mp= 200-203 °C; [α]D = -82.4 ° (c 0.55, CHCl3); 1H NMR (250 MHz, CDCl3) δ 8.28 (br s, 1H, NH), 7.52 (d, J = 7.75 Hz, 1H), 7.40-7.09 (m, 13H), 6.96 (s, 1H), 6.02 (q, J = 7.0 Hz, 1H), 4.33 (dd, J = 12.5, 4.5 Hz, 1H), 3.81 (d, J = 18.0 Hz, 1H), 3.55 (dd, J = 15.5, 4.5 Hz, 1H), 3.46 (d, J = 18.0 Hz, 1H), 2.86 (dd, J = 15.5, 12.5 Hz, 1H), 1.49 (d, J = 7.5 Hz, 3H); 13C NMR (126 MHz, CDCl3) δ 165.20, 162.30, 138.27, 136.40, 129.75, 128.98, 128.89, 128.83, 128.21, 127.35, 126.31, 122.64, 120.01, 118.41, 111.19, 108.90, 52.85, 52.07, 50.10, 43.91, 27.76, 14.61; TLC: Rf = 0.48 (EtOAc /Hexane 6/4, v/v); rt(HPLC) = 17.65 min; HRMS-ESI (m/z): [M+H]+ calcd for C28H26N3O2 436.2020; found 436.2027.

(3R,6R,12aS)-2-benzyl-3,6-diphenyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8l):

8lNH

N

NO

O

Ph


Yield (178 mg, 72%); white solid; mp= 267-269 °C; [α]D = -26.6 ° (c 0.3, CHCl3); 1H NMR (250 MHz, DMSO-d6) δ 11.04 (br s, 1H, NH), 7.58 (d, J = 7.25 Hz, 1H), 7.35-7.01 (m, 18H), 6.90 (s, 1H), 5.05 (d, J = 14.50 Hz, 1H), 5.02 (s, 1H), 4.53 (dd, J = 11.25, 4.75 Hz, 1H), 3.59 (dd, J = 16.0, 4.75 Hz, 1H), 3.56 (d, J = 14.50 Hz, 1H), 3.10 (dd, J = 16.0, 11.25 Hz, 1H); 13C NMR (126 MHz, DMSO-d6) δ 165.70, 163.56, 139.66, 138.02, 136.90, 136.23, 130.42, 129.63, 129.28, 129.05, 128.99, 128.73, 128.16, 128.13, 128.03, 127.86, 126.40, 122.17, 119.42, 118.72, 111.84, 107.38, 63.48, 52.42, 52.07, 47.16, 28.91; TLC: Rf = 0.53 (EtOAc /Hexane 3/7, v/v); rt(HPLC) = 19.25 min; HRMS-ESI (m/z): [M+Na]+ calcd for C23H27N3O2Na520.1995; found 520.2089.

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(S)-2'-benzyl-2',3',12',12a'-tetrahydro-4'H-spiro[cyclohexane-1,6'-pyrazino[1',2':1,6]pyrido[3,4-b]indole]-1',4'(7'H)-dione (8m):

8mNH

N

NO

O


Yield (166 mg, 80%); white solid; mp= 281-283 °C; 1H NMR (500 MHz, DMSO-d6) δ 10.83 (br s, 1H, NH), 8.04 (s, 1H); 7.46 (d, J = 8.0 Hz, 1H), 7.31-7.22 (m, 4H), 7.05-7.02 (m, 3H), 6.95 (t, J = 8.0 Hz, 1H), 6.00 (s, 1H), 4.60 (d, J = 15.0 Hz, 1H), 4.10-4.07 (m, 1H); 3.91 (d, J = 15. 0 Hz, 1H), 3.31-3.21 (m, 3H), 3.05 (d, J = 17.0 Hz, 1H), 2.40-2.35 (m, 2H), 2.20-2.14 (m, 2H), 1.75-1.68 (m, 2H), 1.66-1.59 (m, 2H); 13C NMR (126 MHz, DMSO-d6) δ 166.99, 165.07, 139.45, 136.55, 135.65, 130.43, 129.13, 129.04, 128.17, 127.96, 127.85, 121.40, 118.97, 111.17, 104.68, 56.72, 48.92, 48.72, 29.81, 27.96, 25.69, 22.88, 22.00; TLC: Rf = 0.31 (EtOAc /Hexane 8/2, v/v); rt(HPLC) = 15.72 min; HRMS-ESI (m/z): [M+Na]+ calcd for C26H27N3O2Na436.1996; found 436.1970.

(S)-2-benzyl-2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (8n):

8nNH

N

NO

O


Yield (130 mg, 75%); off-white solid; mp= 185-188 °C;1H NMR (500 MHz, CDCl3) δ 7.93 (br s, 1H, NH), 7.50 (d, J = 7.0 Hz, 1H), 7.41-7.22 (m, 6H), 7.21-7.13 (m, 2H), 5.62 (d, J = 17.50 Hz, 1H), 4.74 (d, J = 14.0 Hz, 1H), 4.66 (d, J = 14. 0 Hz, 1H), 4.45 (d, J = 10.50 Hz, 1H), 4.21 (d, J = 16.50 Hz, 1H), 4.02 (s, 2H), 3.59 (d, J = 15.0 Hz, 1H), 2.96 (t, J = 13.5 Hz, 1H); 13C NMR (126 MHz, CDCl3) δ 165.44, 163.06, 136.22, 134.84, 129.09, 128.53, 128.83, 128.33, 128.02, 126.65, 122.58, 120.20, 118.20, 111.00, 107.44, 56.88, 49.49, 48.68, 40.05, 27.41; TLC: Rf = 0.45 (EtOAc /Hexane 19/1, v/v); rt(HPLC) = 14.95 min; HRMS-ESI (m/z): [M+H]+ calcd for C21H20N3O2346.1550; found 346.1553.

7

S

4. NOESY Studies of 8a and 8l

The relative configuration of the THBC-based DKP 8a was confirmed by NOESY studies. As

shown in Figure 3, the configuration of the trans isomer 8a was determined by the presence of a

nOe between the proton at position C-12a and the aryl ortho protons at position C-6, thus

confirming a trans-stereochemistry for 8a. Since the C-12a configuration was known (L-

tryptophan was used as starting material) the absolute configuration of the major isomers

corresponds to (6R,12aS).

NH

N

NO

O

THBC-based DKP (8a)

H

HH

NO NOENOE

6

12a

d = 4.0 A°

d = 2.8 A°

Figure 1. Identification of the relative configuration of 8a by 2D NOESY experiments

The relative configuration of the THBC-based DKP 8l was confirmed by NOESY studies. As

shown in Figure 4, the configuration of the trans isomer 8l was determined by the presence of a

nOe between the proton at position C-12a and the aryl ortho protons at position C-3, thus

confirming a trans-stereochemistry for 8l. Since the C-12a configuration was known (L-

tryptophan was used as starting material) the absolute configuration of the major isomers

corresponds to (3R, 6R,12aS).

NH

N

NO

O

THBC-based DKP (8l)

HNO NOE

6

12a

d = 4.0 A°H

NOEd = 2.7 A°

3H

Figure 2. Identification of the relative configuration of 8l by 2D NOESY experiments

8

S

5. NMR Spectra (8a): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.932

2.982

2.997

3.043

3.584

3.599

3.646

3.663

3.876

3.948

3.965

4.039

4.304

4.362

4.402

4.420

4.451

4.467

4.840

4.899

7.039

7.136

7.403

7.543

7.575

7.923

1.0

1.1

2.1

1.0

1.0

1.0

1.1

13.31.1

1.0

(8a): 13C NMR (63 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

27.83

48.78

49.45

52.19

52.67

76.53

77.04

77.55

108.95

111.19

118.44

120.13

122.78

126.23

128.33

128.54

128.83

128.96

129.07

129.56

134.74

136.34

137.99

162.07

165.46

9

C

DC

l 3

8aNH

N

NO

O

Chemical Formula: C27H23N3O2

S

(8b): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.895

2.962

3.015

3.512

3.531

3.573

3.592

3.914

3.983

4.081

4.152

4.289

4.307

4.336

4.354

7.059

7.124

7.320

7.522

7.553

8.137

4.1

1.1

1.0

1.0

1.0

1.1

8.3

1.0

1.0

(8b): 13C NMR (63 MHz, CDCl3)

220 200 180 160 140 120 100 80 60 40 20 0 ppm

27.65

33.40

51.42

52.08

52.55

76.54

77.05

77.55

109.07

111.18

118.41

120.06

122.70

126.26

128.78

128.89

129.63

136.36

138.24

161.64

165.46

(8c): 1H NMR (250 MHz, CDCl3)

10

C

DC

l 3

8bNH

N

NO

O


S

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.292

2.910

2.996

3.038

3.533

3.551

3.592

3.612

3.958

4.029

4.116

4.184

4.350

4.367

4.399

4.417

7.033

7.104

7.334

7.535

7.564

7.881

3.0

4.2

1.2

1.2

1.2

1.1

8.5

1.0

1.0

(8c): 13C NMR (63 MHz, CDCl3)

102030405060708090100110120130140150160170180190 ppm

21.38

27.67

33.40

51.48

52.06

52.60

76.51

77.02

77.53

109.12

111.15

118.43

120.08

122.70

125.96

128.81

129.31

129.71

129.84

136.31

138.08

138.83

161.56

165.53

11

C

DC

l 3

8cNH

N

NO

O


S

(8d): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.859

2.896

3.008

3.533

3.550

3.595

3.611

3.821

3.893

4.010

4.080

4.311

4.328

4.358

4.375

7.152

7.325

7.391

7.502

7.547

7.623

7.705

7.785

8.284

2.9

1.0

0.9

1.0

0.9

1.0

5.0

3.1

3.1

1.3

0.9

(8d): 13C NMR (63 MHz, CDCl3)

102030405060708090100110120130140150160170 ppm

27.69

33.28

51.35

52.24

52.54

76.56

77.06

77.57

109.14

111.31

118.45

120.04

122.68

126.22

126.33

126.63

126.76

127.61

127.95

128.14

128.91

129.73

132.97

133.32

135.73

136.43

161.76

165.40

12

C

DC

l 3

8dNH

N

NO

O


S

(8e): 1H NMR (250 MHz, MeOD)

8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm

1.146

1.151

1.172

1.178

2.245

2.499

2.577

3.111

3.494

4.193

4.261

6.172

6.198

6.237

6.264

6.397

6.463

6.944

7.100

7.270

7.305

7.405

7.440

6.0

5.1

5.0

1.2

0.9

0.8

2.1

1.0

1.1

(8e): 13C NMR (63 MHz, MeOD)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

21.431

28.695

31.582

32.530

46.471

46.812

47.151

47.491

47.831

48.170

48.510

50.009

56.425

105.295

110.004

115.592

118.174

118.646

121.693

128.409

135.251

136.394

137.002

165.777

167.402

13

8eNH

N

NO

O


S

(8f): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.886

2.957

2.994

3.507

3.525

3.569

3.587

3.946

4.017

4.102

4.169

4.316

4.332

4.362

4.380

5.923

6.703

6.806

6.972

7.124

7.328

7.515

7.545

8.002

1.1

3.0

1.0

1.2

1.2

1.0

2.1

2.1

1.0

1.1

3.5

1.1

0.8

(8f): 13C NMR (63 MHz, CDCl3)

102030405060708090100110120130140150160170180190 ppm

27.60

33.40

51.47

51.81

52.42

76.52

77.03

77.53

101.36

108.31

109.14

111.15

118.45

120.10

122.48

122.77

126.23

129.71

132.05

136.32

148.05

148.15

161.53

165.46

14

C

DC

l 3

8fNH

N

NO

O

O

O


S

(8f): 1H NMR (500 MHz, (CD3)2SO)

11 10 9 8 7 6 5 4 3 2 1 ppm

2.822

2.906

2.926

2.936

2.957

3.218

3.226

3.248

3.256

3.992

4.028

4.031

4.041

4.056

4.064

4.199

4.234

5.967

5.981

6.577

6.593

6.735

6.801

6.835

6.850

6.976

6.991

7.006

7.062

7.078

7.092

7.279

7.295

7.464

7.482

11.024

3.0

1.2

1.0

2.0

1.0

2.0

1.0

1.0

1.1

1.0

1.2

1.1

1.1

1.0

1.0

15

8fNH

N

NO

O

O

O


Water

DMSO-d6

S

(8g): 1H NMR (500 MHz, (CD3)2SO)

11 10 9 8 7 6 5 4 3 2 1 ppm

2.823

2.906

2.930

2.935

2.960

3.226

3.234

3.257

3.265

3.994

4.029

4.044

4.052

4.068

4.075

4.201

4.237

5.968

5.980

6.585

6.601

6.746

6.813

6.833

6.849

6.978

6.993

7.008

7.064

7.079

7.094

7.285

7.301

7.468

7.484

11.024

3.0

1.1

1.0

2.1

1.0

2.1

1.0

1.0

2.0

1.0

1.0

1.0

1.0

0.7

(8g): 13C NMR (126 MHz, (CD3)2SO)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

27.17

33.09

39.51

39.68

39.85

40.01

40.18

40.35

40.51

51.15

51.33

52.49

101.73

107.99

108.62

108.83

111.80

118.59

119.36

122.15

122.19

126.39

130.83

133.38

136.75

147.73

148.11

162.74

165.13

16

8gNH

N

NO

O

O

O


S

(8h): 1H NMR (250 MHz, CDCl3)

8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm

2.899

2.946

2.961

3.010

3.552

3.567

3.614

3.631

3.824

3.896

3.920

3.987

4.267

4.288

4.313

4.333

4.345

4.788

4.847

6.950

7.028

7.112

7.128

7.215

7.405

7.521

7.561

8.235

0.9

0.9

1.9

1.9

0.9

1.0

2.0

1.9

8.3

1.0

0.8

(8h): 13C NMR (63 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

27.75

48.78

49.39

51.45

52.65

76.56

77.06

77.57

109.15

111.25

118.49

120.18

122.88

123.05

126.19

128.32

128.52

129.07

130.40

132.04

134.81

136.39

137.22

161.94

165.17

17

C

DC

l 3

8hNH

N

NO

O

Br

Chemical Formula: C27H22BrN3O2

S

(8i): 1H NMR (250 MHz, CDCl3)

8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm

1.566

1.592

2.955

2.996

3.011

3.063

3.566

3.582

3.624

3.644

3.696

3.970

4.042

4.410

4.430

4.459

4.479

5.970

5.999

6.028

6.054

7.007

7.124

7.385

7.532

7.568

7.976

3.0

1.0

2.1

1.0

1.0

1.1

1.1

13.31.1

0.9

(8i): 13C NMR (63 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

15.08

27.59

43.64

50.92

52.39

52.83

76.53

77.19

77.63

111.26

118.47

120.31

122.96

126.21

126.83

127.40

127.82

128.41

128.94

128.99

129.46

136.65

137.38

137.82

162.92

165.49

18

C

DC

l 3

8iNH

N

NO

O


S

(8j): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

1.537

1.564

2.892

2.941

2.956

3.004

3.487

3.559

3.595

3.609

3.655

3.671

3.860

3.932

4.415

4.430

4.463

4.478

6.045

6.071

6.100

6.130

7.029

7.140

7.229

7.277

7.387

7.549

7.580

7.803

2.9

1.0

1.0

1.2

1.0

1.0

1.0

1.2

2.1

11.41.1

1.0

(8j): 13C NMR (63 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

14.54

27.72

43.82

50.26

52.18

52.79

76.56

77.07

77.59

108.93

111.17

118.42

120.10

122.78

126.21

127.44

128.28

128.75

128.88

128.93

129.59

136.41

137.99

138.08

162.58

165.26

19

C

DC

l 3

8jNH

N

NO

O


S

(8k): 1H NMR (250 MHz, CDCl3)

8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm

1.476

1.506

2.814

2.865

2.877

2.922

3.418

3.490

3.510

3.528

3.572

3.588

3.770

3.841

4.298

4.316

4.348

4.366

5.981

6.009

6.037

6.065

6.958

7.093

7.165

7.180

7.405

7.482

7.549

8.284

3.1

1.0

1.0

1.0

1.0

1.0

1.0

1.0

2.0

11.21.0

1.0

(8k): 13C NMR (126 MHz, CDCl3)

200 180 160 140 120 100 80 60 40 20 0 ppm

14.61

27.76

43.91

50.10

52.07

52.85

76.85

77.11

77.36

108.90

111.19

118.41

120.01

122.64

126.31

127.35

128.21

128.83

128.89

128.98

129.75

136.40

138.27

162.30

165.20

20

C

DC

l 3

8kNH

N

NO

O


S

(8l): 1H NMR (250 MHz, (CD3)2SO)

11 10 9 8 7 6 5 4 3 2 1 ppm

3.041

3.092

3.108

3.152

3.532

3.551

3.570

3.591

3.614

3.635

4.498

4.517

4.543

4.561

5.026

5.081

6.898

7.014

7.180

7.222

7.351

7.567

7.596

11.040

1.0

2.1

1.0

2.1

1.1

8.4

10.71.1

1.0

(8l): 13C NMR (126 MHz, (CD3)2SO)

200 180 160 140 120 100 80 60 40 20 0 ppm

28.91

39.52

39.69

39.86

40.02

40.19

40.36

40.52

47.16

52.07

52.42

63.48

107.38

111.84

118.72

119.42

122.17

126.40

127.86

128.03

128.13

128.16

128.73

128.99

129.05

129.28

129.63

130.42

136.23

136.90

138.02

139.66

163.56

165.70

21

8lNH

N

NO

O

Ph


S

(8m): 1H NMR (500 MHz, (CD3)2SO)

11 10 9 8 7 6 5 4 3 2 1 ppm

1.594

1.663

1.681

1.744

2.137

2.204

2.346

2.403

3.035

3.069

3.208

3.310

3.894

3.924

4.072

4.100

4.582

4.612

6.000

6.934

6.948

6.964

7.021

7.052

7.222

7.306

7.450

7.466

8.038

10.826

2.1

2.1

2.0

2.0

1.0

3.1

1.0

1.0

1.0

1.0

1.1

3.0

4.1

1.0

1.0

0.9

(8m): 13C NMR (126 MHz, (CD3)2SO)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

22.00

22.88

25.69

27.96

29.81

39.52

39.68

39.85

40.02

40.18

40.35

40.52

48.72

48.92

56.72

104.68

111.17

118.97

121.40

127.85

127.96

128.17

129.04

129.13

130.43

135.65

136.55

139.45

165.07

166.99

22

8mNH

N

NO

O


S

(8n): 1H NMR (500 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

2.935

2.961

2.988

3.575

3.605

4.021

4.198

4.231

4.440

4.461

4.642

4.670

4.730

4.758

5.601

5.636

7.135

7.149

7.165

7.196

7.212

7.225

7.323

7.411

7.495

7.509

7.930

1.3

1.0

1.9

0.9

1.0

1.0

1.0

1.0

1.1

1.2

6.5

1.0

0.9

(8n): 13C NMR (126 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

27.41

40.05

48.68

49.49

56.88

107.44

111.00

118.20

120.20

122.58

126.65

128.02

128.33

128.53

129.09

134.84

136.22

163.06

165.44

23

C

DC

l 3

8nNH

N

NO

O


S

(8i/8i’ : 70/30 dr): 1H NMR (250 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

1.473

1.501

1.526

1.555

2.788

2.843

2.858

2.896

2.940

2.955

3.003

3.395

3.463

3.499

3.520

3.566

3.579

3.636

3.809

3.882

3.897

3.969

4.340

4.359

4.388

4.405

5.887

6.055

6.919

6.953

7.113

7.385

7.453

7.590

8.260

8.319

1.0

2.4

0.2

0.9

0.2

0.7

1.4

0.3

0.9

1.1

1.2

0.3

0.8

16.11.9

0.3

0.8

(8i/8i’ : 70/30 dr): 13C NMR (63 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

14.57

15.10

23.24

27.61

43.64

44.83

50.41

50.81

52.29

52.83

56.30

56.54

76.65

77.16

77.67

106.17

108.60

109.69

111.30

118.43

120.07

122.38

122.73

126.24

126.77

127.39

127.74

128.37

128.77

128.94

129.64

132.84

136.42

137.51

138.39

141.15

162.74

165.40

166.61

167.86

24

C

DC

l 3

8i/8i' (70/30 dr)

NH

N

NO

ONH

N

NO

O

S

(8i/8j : 40/60 dr): 1H NMR (500 MHz, CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5 ppm

1.524

1.539

1.559

1.573

2.907

2.932

2.937

2.962

2.999

3.024

3.497

3.531

3.581

3.596

3.605

3.619

3.627

3.635

3.864

3.898

3.938

3.973

4.401

5.999

6.013

6.027

6.047

6.062

6.076

7.016

7.138

7.146

7.152

7.160

7.168

7.175

7.189

7.203

7.216

7.230

7.240

7.253

7.273

7.287

7.300

7.315

7.324

7.334

7.351

7.365

7.381

7.537

7.552

7.788

7.804

3.0

2.0

1.7

1.0

2.2

0.9

0.6

1.5

0.6

0.9

1.5

4.3

16.70.7

0.7

0.8

0.5

(8i/8j : 40/60 dr): 13C NMR (126 MHz, CDCl3)

190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm

14.58

15.06

27.67

27.76

29.69

43.72

43.90

50.20

50.56

52.13

52.17

52.89

76.75

77.01

77.26

109.10

109.17

111.14

118.45

120.19

122.82

126.29

127.34

127.72

128.24

128.89

128.97

129.62

129.71

136.34

137.72

137.95

138.02

138.15

162.53

162.62

165.20

165.26

25

C

DC

l 3

NH

N

NO

O

8i/8j (40/60 dr)

NH

N

NO

O

S

(7i : 53/47 dr): 1H NMR (250 MHz, CDCl3)

8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm

1.136

1.258

1.489

1.517

1.738

1.766

2.861

3.265

3.327

3.375

3.879

3.899

3.922

3.945

4.039

4.101

4.344

4.360

4.381

4.408

5.182

5.283

5.422

5.446

5.472

5.502

6.103

6.128

6.156

6.189

6.270

7.101

7.424

7.507

7.531

7.640

9.0

8.2

3.5

3.0

8.0

1.9

1.2

1.0

1.2

0.9

1.0

3.3

8.0

23.22.5

1.0

(7i : 53/47 dr):13C NMR (126 MHz, CDCl3)

220 200 180 160 140 120 100 80 60 40 20 0 ppm

16.11

18.27

25.18

26.15

28.36

28.60

47.87

48.10

50.89

51.37

51.74

54.52

55.08

55.34

57.99

58.51

109.33

111.06

111.17

118.11

118.22

119.58

119.66

122.08

122.18

127.05

127.17

127.25

127.82

128.03

128.13

128.49

128.62

128.70

128.84

129.02

129.15

134.17

134.40

136.24

139.24

139.99

140.34

140.53

168.35

168.78

173.35

173.78

26

NH

NH

NO

O

NH

Ph

7i (95%, 53/47 dr)Chemical Formula: C32H36N4O2

S

(7a : 75/25 dr): 1H NMR (500 MHz, CDCl3 at 293 K) (4rotamers : 34/32/20/14)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0 ppm

0.905

1.172

1.231

1.289

3.054

3.500

3.556

3.718

3.886

3.960

3.997

4.114

4.202

4.258

4.355

4.392

4.546

4.704

4.741

4.890

5.536

5.931

6.052

6.149

6.996

7.152

7.188

7.215

7.230

7.244

7.258

7.286

7.301

7.322

7.337

7.353

7.368

7.383

7.398

7.492

7.990

8.436

8.938

5.7

9.0

8.8

3.1

1.7

5.0

1.9

2.8

0.6

1.6

2.6

1.8

0.7

1.7

0.8

1.3

6.0

32.50.9

0.6

0.6

(7a : 75/25 dr): 1H NMR (500 MHz, CDCl3 at 323 K)

9 8 7 6 5 4 3 2 1 ppm

1.001

1.233

1.310

2.976

3.860

4.051

4.232

4.343

4.473

4.624

4.916

5.650

5.830

6.182

6.976

7.529

7.962

8.233

8.786

9.0

2.7

3.2

1.1

0.8

1.1

1.9

0.3

1.9

17.50.4

0.3

0.2

0.2

27

7a ( 75/25 dr)

NH

NH

N

O

O

HN


S

(8a): COSY

ppm

9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm9

8

7

6

5

4

3

2

1

ppm

2.83.03.23.43.63.84.04.24.44.64.85.05.25.4 ppm

2.6

2.8

3.0

3.2

3.4

3.6

3.8

4.0

4.2

4.4

4.6

4.8

5.0

5.2

5.4

28

S

(8a): NOESY

ppm

3.03.54.04.55.05.56.06.57.07.58.0 ppm

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

ppm

2.83.03.23.43.63.84.04.24.44.64.85.05.25.4 ppm

2.6

2.8

3.0

3.2

3.4

3.6

3.8

4.0

4.2

4.4

4.6

4.8

5.0

5.2

5.4

29

S

(8a): HSQC

ppm

9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm

180

160

140

120

100

80

60

40

20

ppm

3.03.54.04.55.05.56.06.57.07.58.08.59.0 ppm

25

30

35

40

45

50

55

30

S

(8a): HPLC

31

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