Table of Contents1 . Cascade reactions of indi go with oxiranes and aziridines: efficient access to dihydropyrazinodiindoles and spiro -oxazocinodiindoles. Nicholas M. Butler, Rudi

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Cascade reactions of indigo with oxiranes and aziridines: efficient access to dihydropyrazinodiindoles and spiro-oxazocinodiindoles

Nicholas M. Butler, Rudi Hendra, John B. Bremner, Anthony C. Willis, Leonardo Lucantoni, Vicky M.

Avery, and Paul A. Keller

Supplementary Information Table of Contents Experimental Section ..................................................................................................................................... 3

General Experimental Information ............................................................................................................ 3

Products from the reaction of indigo with (S)-epichlorohydrin (7) ........................................................... 4

(S)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (8) ............................. 4

(7S, 9aR)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (9)................ 4

(7R,9aR)-14-((E)-3-hydroxyprop-1-en-1-yl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (10) ................................................................................................................... 5 14-((2-oxo-1,3-dioxolan-4-yl)methyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (±11)………………………………………………………………………… 5

(7S,9aR)-14-(((S)-2-oxo-1,3-dioxolan-4-yl)methyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (11) …………………………………………………………………… 6

(R)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (R-8) ......................... 6

(±)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (±8) ........................... 6

Products from the reaction of indigo with 2,2ꞌ-bioxirane .......................................................................... 6 (7S,8R,9aS)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (12) .................................................................................................................................... 6

(7S,8R,9aS)-14-((S)-2-hydroxy-2-((S)-oxiran-2-yl)ethyl)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (13) ................................................................ 7

Synthesis of (R)-N-tosyl-2-chloromethylaziridine (14) ............................................................................. 7

(S)-N-(3-chloro-2-hydroxypropyl)-4-methylbenzenesulfonamide (23) ................................................. 7

(S)-1-chloro-3-((4-methylphenyl)sulfonamido)propan-2-yl methanesulfonate (24) .............................. 8 (R)-N-tosyl-2-chloromethylaziridine (14) .............................................................................................. 8

Products from the reaction of indigo with (R)-N-tosyl-2-chloromethylaziridine (14) .................................. 9

(R)-N-((13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (15) ............................................................................................................. 9

N-(((R)-13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methyl-N-(((S)-1-tosylaziridin-2-yl)methyl)benzenesulfonamide (16) ................................................................ 10

N-(3-chloro-2-((4-methylphenyl)sulfonamido)propyl)-N-(((R)-13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (17) ................. 10 (±)-N-((13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (±15) ........................................................................................................ 11

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2018

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Reaction of indigo with N-Tosyl-2-phenylaziridine (18) ........................................................................ 11 (±)-N-tosyl-2-phenylaziridine (18) ....................................................................................................... 11

(E)-N-(2-(3,3'-dioxo-[2,2'-biindolinylidene]-1-yl)-1-phenylethyl)-4-methylbenzenesulfonamide (19) .............................................................................................................................................................. 11

(E)-N-(2-(3,3'-dioxo-[2,2'-biindolinylidene]-1-yl)-2-phenylethyl)-4-methylbenzenesulfonamide (20) .............................................................................................................................................................. 12

Reaction of indigo with 2,2ꞌ-biaziridine ...................................................................................................... 12

4-methyl-N-(((6R,7R)-7-((4-methylphenyl)sulfonamido)-14,15-dioxo-7,8,14,15-tetrahydro-6H-[1,4]diazepino[1,2-a:4,3-a']diindol-6-yl)methyl)benzenesulfonamide (21) ........................................ 12

Biological testing ..................................................................................................................................... 13

NMR data for isolated compounds .............................................................................................................. 15

RP-HPLC Traces for Compounds 16 and 17 ............................................................................................ 100

Structural Summary of Compounds Tested for Antiplasmodial Activity ................................................. 104 Crystallographic data for Compounds 9-12, and 20 .................................................................................. 106

Crystallographic data for compound 9: ................................................................................................. 106

Crystallographic data for compound 10: ............................................................................................... 106

Crystallographic data for compound 11: ............................................................................................... 107

Crystallographic data for compound 12: ............................................................................................... 107 Crystallographic data for compound 20: ............................................................................................... 108

References .................................................................................................................................................. 108

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Experimental Section General Experimental Information

Reagents and solvents were purchased reagent grade and used without further purification unless otherwise

stated. All reactions were performed in standard oven-dried glassware under a nitrogen atmosphere unless

otherwise stated. Melting point temperatures are expressed in degrees Celsius (˚C) and are uncorrected. 1H

and 13C NMR spectra (CDCl3/DMSO) were recorded either at 500 MHz and 125 MHz, or 400 MHz and

101 MHz, respectively, with chemical shifts (δ) reported as parts per million relative to TMS (δ = 0.00

ppm), CDCl3 (δ = 7.26; 77.0 ppm) or DMSO-d6 (δ = 2.50 ppm; 39.51 ppm). Coupling constants (J) are

reported in Hertz (Hz). Multiplicities are reported as singlets (s), doublets (d), triplets (t), doublet of

doublets (dd) or multiplet (m). Electrospray (ESI single quadrupole) mass spectra have their ion mass to

charge values (m/z) stated with their relative abundances as a percentage in parentheses. Peaks assigned to

the molecular ion are denoted as [M+H]+ or [M+Na]+. Infrared (IR) spectra were recorded neat. UV-visible

spectra were recorded as solutions in CH2Cl2 at 25 ˚C. Optical rotations were recorded in CHCl3 or CH2Cl2

at room temperature. Thin layer chromatography (TLC) was performed using silica gel F254 aluminium

sheets. Column chromatography was performed using silica gel 60 (0.063-0.200 mm). Eluents are reported

in volume to volume (v:v) ratios. Solvent extracts and chromatographic fractions were concentrated by

rotary evaporation in vacuo. Indigo dye was purchased from Sigma Aldrich and used without further

purification. N,Nꞌ-Ditosyl-2,2ꞌ-biaziridine,1 2,2ꞌ-bioxirane,2 and (±)-N-tosyl-2-bromomethylaziridine3 were

prepared by reported procedures. Pet. spirit had a bp range of 40-60 ˚C.

General Procedure A

A suspension of indigo (262 mg, 1.00 mmol) in anhydrous DMF (40 mL) was sonicated at 60 ˚C for one

hour. The resulting hot suspension was transferred by cannula under positive pressure of nitrogen into a

100 mL round-bottomed flask containing a magnetic stir bar, pre-dried and ground Cs2CO3 (1.206 g, 3.700

mmol), and fitted with a rubber septum. The resulting dark-green to amber solution was stirred for one hour

in an oil bath, pre-heated to strictly 85-87 ˚C under dry nitrogen. The nitrogen flow was cut, and the

electrophile (5.0 mmol) injected through the septum either neat, or as a solution in DMF (2 mL), and the

mixture stirred at 85-87 ˚C for the required time. The resulting intensely-coloured solution was quenched

over crushed ice (ca. 100 g), and the flask rinsed with EtOAc (ca. 10 mL) to remove adhered products.

Upon warming to RT, the emulsion was saturated with solid NaCl, and extracted with EtOAc (4×60 mL)

or CHCl3 (4×60 mL) until the aqueous phase became clear. The combined organic fractions were condensed

to ca. 150 mL, then extracted with brine (4×40 mL), dried (MgSO4), and filtered through a 2 cm plug of

celite, and the solvent removed in vacuo.

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Products from the reaction of indigo with (S)-epichlorohydrin (7)

(S)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (8)

Prepared following General Procedure A, using (S)-epichlorohydrin (462 mg, 5.0 mmol), and a 30 min

reaction time. The resulting crude residue was dissolved in a minimum of hot CHCl3, and gradual dilution

with a tenfold volume of hot pet. spirit precipitated an intense blue powder, which was collected on a Hirsch

funnel and combined with Fraction 5 (see below). The combined solids were dried in vacuo to furnish the

dihydropyrazinodiindole 8 (141 mg, 44%) as a dark blue powder, mp >350 ˚C. RF (9:1 CH2Cl2:EtOAc)

0.14. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 241 (12773), 303 (6339), 579 (2819). 1H-NMR (DMSO-d6,

500 MHz) 7.56-7.64 (m, 4H, ArH), 7.35 (d, J = 8.0 Hz, 2H, H4, H9), 6.99-7.06 (m, 2H, ArH), 5.28 (s, 1H,

CH2OH), 4.66 (s, 1H, H6), 4.44 (d, J = 12.0 Hz, 1H, H7a), 3.71 (dd, J = 3.5, 12.0 Hz, 1H, H7b), 3.47-3.57

(m, 2H, CH2OH). 13C-NMR (125 MHz) δ 179.9, 179.7, 150.2, 149.4, 135.3, 135.0, 123.9, 123.5, 123.1,

122.0, 121.5, 120.8, 120.6, 120.4, 110.9, 110.5, 59.9, 51.1, 39.5. IR (neat) 3432 (br, m), 2937 (w), 2876

(w), 1689 (s), 1607 (s), 1561 (s), 1470 (m), 1465 (m), 1369 (m), 1340 (w), 1298 (s), 1180 (s), 1158 (m),

1144 (s), 743 (s). HR-ESI-MS calcd. for C19H14N2O3Na+ = 341.0902 found 341.0888. [α]D29 = -6.4 ˚ (c

0.10, CHCl3).

(7S,9aR)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (9)

The mother liquor from compound 8 was concentrated, and subjected to flash chromatography (gradient

elution, 5-50% EtOAc:CH2Cl2) to afford five major fractions. Fraction 1 was subjected to PTLC (10%

EtOAc:CH2Cl2), and the selected band scraped off and soaked in EtOAc. The silica was removed by

filtration and the luminescent, yellow solution was combined with Fraction 2 and the solvent removed to

furnish the title compound 9 as small, bright orange crystals (101 mg, 32%), mp 189-191 ˚C (dec). X-ray

quality crystals were grown by slow evaporation of a saturated solution in EtOAc at -20 ˚C. RF (9:1

CH2Cl2:EtOAc) 0.57. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 236 (8964), 287 (5169), 364 (2651), 483

(3573). 1H NMR (500 MHz, CDCl3) δ 7.81 (d, J = 7.7 Hz, 1H, H1), 7.49 (m, 2H, ArH), 7.35 (t, J = 7.7 Hz,

1H, ArH), 7.08 (d, J = 8.3 Hz, 1H, ArH), 7.02 (t, J = 7.5 Hz, 2H, ArH), 6.92 (d, J = 7.9 Hz, 1H, ArH), 5.08

- 5.16 (m, 1H, H7), 4.37 (t, J = 7.2 Hz, 1H, H8b), 4.26 (dd, J = 7.0 Hz, 3 Hz, H8a) 4.20 (dd, J = 14.0, 3.0

Hz, 1H, H6b), 3.74 (d, J = 14 Hz, 1H, H6a).13C NMR (125 MHz, CDCl3) δ 188.1, 152.3, 146.8, 145.3,

144.7, 133.7, 132.2, 125.2, 124.3, 123.9, 122.8, 122.0, 119.9, 110.9, 110.6, 73.9, 65.4, 51.6. IR (neat) 3310

(br, w) 2966 (w), 2953 (w), 2893 (w), 1689 (s), 1614 (s), 1576 (s), 1098 (m), 995 (s), 747 (s). HR-ESI-MS

calcd. for C19H14N2O3Na+ = 341.0902 found 341.0900. [α]D27 = -6.5 ˚ (c 0.17, CHCl3).

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(7R,9aR)-14-((E)-3-hydroxyprop-1-en-1-yl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-

b']diindol-15(14H)-one (10)

Fraction 3 was subjected to PTLC (developed twice; 20% EtOAc:CH2Cl2, then EtOAc) and the selected

band scraped off and soaked in EtOAc. The silica was removed, and the solution combined with Fraction

4 to give the title compound 10 as red-orange crystals (75.2 mg, 21%), mp 152-154 ˚C (dec.). X-ray quality

crystals were grown by slow evaporation of a saturated solution in 9:1 CH2Cl2 : hexane at RT. RF 0.21 (20%

EtOAc:CH2Cl2). UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 243 (18482), 274 (20838), 355 (1472), 500 (1174). 1H NMR (500 MHz, CDCl3) δ 7.78 (d, J = 7.6 Hz, 1H, ArH), 7.48 – 7.55 (m, 2H, 2×ArH), 7.45 (t, J = 7.7

Hz, 1H, ArH), 7.30 – 7.36 (m, 2H, 2×ArH), 7.17 (t, J = 7.4 Hz, 1H, ArH), 7.06 (d, J = 8.3 Hz, 1H, ArH),

7.02 (t, J = 7.4 Hz, 1H ArH), 6.87 (d, J = 14.2 Hz, 1H, H1ꞌ), 6.07 (dt, J = 6.5, 14.0 Hz, 1H, H2ꞌ), 5.01 –

5.05 (m, 1H, H7), 4.21 – 4.44 (m, 6H, H6b, H8a, H8b, H3ꞌa, H3ꞌb, OH), 3.74 (d, J = 14.3 Hz, 1H, H6a). 13C NMR (125 MHz, CDCl3) δ 185.1, 152.7, 151.4, 149.7, 134.2, 131.9, 130.8, 125.5, 124.9, 124.5, 124.3,

123.6, 121.6, 120.0, 119.1, 111.7, 109.9, 73.4, 67.6, 61.9, 52.2. IR (neat) 3349 (br, m), 1726 (m), 1466 (s),

1265 (s), 702 (s). HR-ESI-MS calcd. for C22H19N2O4+ = 375.1345 found 375.1357. [α]D29 = -6.7 ˚ (c 0.11,

CHCl3).

14-((2-oxo-1,3-dioxolan-4-yl)methyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-

15(14H)-one (±11)

Prepared following General Procedure A, using (±)-epichlorohydrin (5.0 mmol, 462 mg), and a 2 h

reaction time. Following the above separation protocol, the racemic products (±)-3 (134.4 mg, 43%), (±)-4

(19.8 mg, 6%), and (±)-5 (27.9 mg, 7%) were isolated. Fraction 4 was condensed, and recrystallisation

from a minimum volume of EtOAc afforded cyclic carbonate 11 (27 mg, 6%) as luminescent orange

crystals. X-ray quality crystals were grown from slow evaporation of a saturated solution in 9:1 EtOAc:pet

spirit at 4 ˚C. mp 146-149 ˚C (dec.) UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 238 (17364), 344 (5999), 499

(2531). 1H NMR (400 MHz, CDCl3) δ 7.68 (d, J = 7.1 Hz, 1H), 7.52 – 7.41 (m, 3H), 7.33 (d, J = 8.1 Hz,

1H), 7.13 (td, J = 7.4, 0.8 Hz, 1H), 7.04 – 6.94 (m, J = 16.6, 7.8 Hz, 2H), 5.48 (ddd, J = 14.9, 8.2, 1.1 Hz,

1H), 5.20 (d, J = 9.2 Hz, 1H), 5.00 – 4.94 (m, 1H), 4.33 – 4.22 (m, 4H), 4.22 – 4.08 (m, 2H), 3.68 (d, J =

14.3 Hz, 1H). 13C NMR (101 MHz, CDCl3) δ 182.2, 178.3, 155.0, 152.9, 148.0, 146.6, 134.3, 132.7, 128.3,

124.8, 124.6, 124.2, 123.4, 119.9, 112.0, 111.1, 110.2, 81.3, 73.4, 69.0, 67.9, 55.5, 52.4. IR (neat) 3431

(w), 1799 (m), 1734 (s), 1611 (s), 1466 (s), 1065 (s), 774 (m). HRESI-MS calcd. for C23H18N2O6Na+ =

441.1063 found 441.1082 (+4.3 ppm); calcd. for C23H19N2O6+ = 419.1243 found 419.1257.

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(7R,9aR)-(((S)-2-oxo-1,3-dioxolan-4-yl)methyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-

b']diindol-15(14H)-one (11)

Prepared following General Procedure A, using (S)-epichlorohydrin 7 (462 mg, 5.0 mmol) and a 30 min

reaction time. Following the above separation protocol, products 8 (133 mg, 41%), and 9 (97 mg, 30%)

were isolated. Fraction 4 was subjected to PTLC (20% EtOAc:CH2Cl2) and the selected band scraped off

and soaked in EtOAc. Recrystallisation from a minimum volume of EtOAc afforded cyclic carbonate 11

(87 mg, 20%) as luminescent orange crystals. [α]D29 = -6.1 ˚ (c 0.09, CHCl3).

(R)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (R-8)

Prepared following General Procedure A, using of (R)-glycidyl tosylate (1.141 g, 5.0 mmol), and a 10 min

reaction time. The crude residue was dissolved in a minimum of hot CHCl3, and gradual dilution with a

tenfold volume of hot pet. spirit precipitated an intense blue powder, which was collected on a Hirsch

funnel. The mother liquor was condensed, and a second recrystallization of the residue afforded a further

crop of product. The combined solids were dried in vacuo to furnish the title compound (R)-8 as a dark blue

powder (242.3 mg, 76%). Spectral characteristics were identical to those obtained from its enantiomer.

[α]D29 = +3.7˚ (c 0.33, CH2Cl2).

(±)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (±8)

Prepared following General Procedure A, using (±)-epibromohydrin (685 mg, 5.0 mmol), and a 10 min

reaction time. The crude residue was dissolved in a minimum of hot CHCl3, and gradual dilution with a

tenfold volume of hot pet. spirit precipitated an intense blue powder, which was collected on a Hirsch

funnel. The mother liquor was condensed, and a second recrystallization of the residue afforded a second

crop of product. The combined solids were dried in vacuo to furnish the title compound as a dark blue

powder (266.9 mg, 84%). Spectral characteristics were identical to those noted for (S)-8.

Products from the reaction of indigo with 2,2ꞌ-bioxirane

(7S,8R,9aS)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-

one (12)

Prepared following General Procedure A, using (2S,2'S)-2,2ꞌ-bioxirane (281 mg, 3.3 mmol), and a 30 min

reaction time. The crude residue was separated into five fractions on a plug of silica (70 g), eluting

sequentially with 1) CH2Cl2 (250 mL each) 2) 20% EtOAc:CH2Cl2 3) 40% EtOAc:CH2Cl2 4) 60%

EtOAc:CH2Cl2 5) 80% EtOAc:CH2Cl2. Fractions 2 and 3 were combined, and subjected to flash

chromatography on 25 g silica, and elution with 20% EtOAc:CH2Cl2 afforded the mono-substituted

spirocycle 12 as a luminescent orange powder (216 mg, 62%), mp 155-156 ˚C (dec.). X-ray quality crystals

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were grown from slow evaporation of a hexane/ethyl acetate solution. RF (20% EtOAc:CH2Cl2) 0.33. UV-

Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 244 (12573), 365 (2296), 485 (2882). 1H NMR (500 MHz, CDCl3) δ

7.79 (d, J = 7.6 Hz, 1H, H1), 7.50 (dd, J = 10.4, 4.4 Hz, 2H, H3, H10), 7.34 (dd, J = 11.1, 4.4 Hz, 1H, H12),

7.10 (d, J = 8.4 Hz, 1H, H4), 7.01 (dd, J = 16.6, 7.7 Hz, 2H, H2, H11), 6.90 (d, J = 7.9 Hz, 1H, H13), 5.02

– 4.97 (m, 1H, H7), 4.74 (dd, J = 13.0, 6.1 Hz, 1H, H8), 4.37 (dd, J = 14.1, 2.7 Hz, 1H, H6b), 4.19 – 4.12

(m, 1H, H1ꞌa), 3.95 (dd, J = 11.3, 5.7 Hz, 1H, H1ꞌb), 3.69 (dd, J = 14.0, 2.3 Hz, 1H, H6a). 13C NMR (101

MHz, CDCl3) δ 184.1, 151.8, 145.2, 144.5, 138.7, 134.0, 132.4, 125.3, 124.2, 124.0, 123.5, 122.3, 121.0,

120.1, 117.8, 111.2, 110.7, 109.6, 77.8, 75.6, 60.4, 48.3. IR (neat) 3420 (w), 2922 (w), 1715 (m), 1612 (s),

1317 (m), 735 (s). HRESI-MS calcd. for C20H17N2O4+ 349.1188, found 349.1192. [α]D25 = -7.2˚ (c 0.21,

CH2Cl2).

(7S,8R,9aS)-14-((S)-2-hydroxy-2-((S)-oxiran-2-yl)ethyl)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-

epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (13)

Further elution of Fractions 2 and 3 with 40% EtOAc:CH2Cl2 gave a residue which was combined with

Fractions 4 and 5 and concentrated, and flash chromatography of this residue (20% EtOAc:CH2Cl2; 20 g

silica) gave the di-alkylated spirocycle 13 (82 mg, 19%) as a pale lime-green, amorphous solid. RF (40%

EtOAc:CH2Cl2) 0.63. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 323 (3820), 350 (4772), 425 (1723). 1H NMR

(400 MHz, DMSO) δ 7.65 – 7.46 (m, 1H), 7.26 – 7.19 (m, 1H), 7.12 – 7.00 (m, 1H), 6.82 (t, J = 7.3 Hz,

1H), 5.83 (d, J = 4.8 Hz, 1H), 5.22 (d, J = 5.7 Hz, 1H), 4.61 (t, J = 6.0 Hz, 1H), 4.54 (d, J = 12.6 Hz, 1H),

4.21 (dt, J = 11.6, 5.8 Hz, 1H), 4.11 – 3.96 (m, 1H), 3.83 (dd, J = 9.2, 6.4 Hz, 1H), 3.73 (dd, J = 9.2, 5.0

Hz, 1H), 3.24 (dt, J = 11.6, 5.9 Hz, 1H), 2.78 (dd, J = 12.3, 10.3 Hz, 1H), 2.73 – 2.68 (m, 1H), 2.61 – 2.54

(m, 1H), 2.33 (dd, J = 5.2, 2.7 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 193.5, 158.4, 139.2, 133.9, 131.9,

130.6, 125.5, 122.6, 120.2, 120.1, 118.8, 118.6, 118.2, 110.5, 109.4, 81.9, 75.9, 70.8, 70.6, 62.8, 53.7, 44.1,

43.2, 37.4. IR (neat) 3566 (w), 2922 (w), 1737 (m), 1608 (m), 1155 (s), 748 (s). HRESI-MS (+) calcd. for

C24H23N2O6+ 435.1556, found 435.1575. [α]D25 = -8.5˚ (c 0.32, CH2Cl2).

Synthesis of (R)-N-tosyl-2-chloromethylaziridine (14)

(S)-N-(3-chloro-2-hydroxypropyl)-4-methylbenzenesulfonamide (23)

To a well-stirred suspension of chloramine-T trihydrate (3.59 g, 12.8 mmol) in dry acetonitrile (20 mL) in

a 50 mL round-bottomed flask fitted with a rubber septum was injected (S)-epichlorohydrin (2.36 g, 25.5

mmol). The mixture was warmed to 50 ˚C in an oil bath, and stirred vigorously for 24 h. Sat. Na2S2O3

solution (5 mL) was added and the acetonitrile removed, then the wet residue was diluted with EtOAc (40

mL) and extracted with water (40 mL). The aqueous phase was extracted with further EtOAc (3×20 mL)

and the combined organic fractions washed sequentially with sat. Na2S2O3 solution (2×10 mL), water (2×10

mL) and brine (2×40 mL). The organic fraction was dried (MgSO4) and the solvent removed to give a clear

8

syrup, which solidified upon standing to give a pale ivory solid. The resulting chlorohydrin 23 (3.25 g,

97%) was found to be >90% pure by NMR, and was used directly for subsequent steps. RF (9:1

CH2Cl2:EtOAc) 0.25. 1H NMR (400 MHz, CDCl3) δ 7.75 (d, J = 8.3 Hz, 2H, 2×ArH), 7.33 (d, J = 8.0 Hz,

2H, 2×ArH), 4.79 (t, J = 6.2 Hz, 1H, NH), 3.94 (dq, J = 10.7, 5.2 Hz, 1H, H2), 3.63 – 3.47 (m, 2H, CH2Cl),

3.22 – 3.14 (m, 1H, H1a), 3.07 – 2.99 (m, 1H, H1b), 2.58 (d, J = 5.2 Hz, 1H, OH), 2.44 (s, 3H, ArCH3). 13C NMR (101 MHz, CDCl3) δ 143.9 (Cq-SO2), 136.4 (Cq-CH3), 130.1 (ArCH), 127.2 (ArCH), 70.0 (C2),

46.8 (C3), 45.9 (C1), 21.6 (TsCH3). ESI-MS (+) 286 (100%, [35Cl]M+Na+), 288 (33%, [37Cl]M+Na+].

[α]D21 = -19.7 ˚ (c 0.64, CH2Cl2).

(S)-1-chloro-3-((4-methylphenyl)sulfonamido)propan-2-yl methanesulfonate (24)

To a solution of 23 (1.331 g, 5.05 mmol) and NEt3 (1.1 mL, 7.9 mmol) in dry CH2Cl2 (30 mL) was added

slowly dropwise methanesulfonyl chloride (0.58 mL, 7.5 mmol) at 0 ˚C, and the resulting pale golden

solution allowed to warm to RT overnight with stirring. The mixture was partitioned with 1N HCl solution

(10 mL) and the resulting organic phase washed with 1N HCl (2×20 mL), then sat. NaHCO3 solution (1×20

mL), and the resulting solution dried (MgSO4) and the solvent removed to give chloromesylate 24 (1.566

g, 91%) as a clear yellow oil. NMR showed the mesylate to be >90% pure, and was used directly for the

next step. RF (9:1 CH2Cl2:EtOAc) 0.62. 1H NMR (400 MHz, CDCl3) δ 7.74 (d, J = 8.3 Hz, 2H, 2×ArH),

7.34 (d, J = 8.1 Hz, 2H 2×ArH), 4.93 (d, J = 6.5 Hz, 1H, NH), 4.89 – 4.80 (m, 1H, H2), 3.77 (d, J = 5.4 Hz,

2H, H1a, H1b), 3.68 (s, 3H, SO2CH3), 3.37 – 3.29 (m, 2H, H3a, H3b), 2.44 (s, 3H, ArCH3). 13C NMR (101

MHz, CDCl3) δ 144.2 (Cq-SO2), 135.7 (Cq-CH3), 130.2 (ArCH), 127.1 (ArCH), 78.9 (C2), 52.6 (MsCH3),

44.5 (C3), 43.1 (C1), 21.6 (TsCH3). ESI-MS (+) 364 (100%, [35Cl]M+Na+), 366 (33%, [37Cl]M+Na+].

[α]D21 = +14.1˚ (c 0.22, CH2Cl2).

(R)-N-tosyl-2-chloromethylaziridine (14)

To a 0 ˚C solution of 24 (621 mg, 1.82 mmol) in dry CH2Cl2 (160 mL) was added pre-dried and ground

cesium carbonate (976 mg, 3.00 mmol), and the mixture stirred vigorously for 2 h at 0 ˚C, then allowed to

warm to room temperature overnight. The mixture was partitioned with water (100 mL), and the resulting

organic layer separated and washed with brine (2×50 mL), dried (MgSO4), and the solvent removed to give

a clear, bluish oil. Flash chromatography (20 g silica, 20% EtOAc:pet spirit) gave the desired aziridine 14

(325 mg, 73%) as a colourless, viscous oil which slowly crystallised on standing, mp 42-44 ˚C. RF (20%

EtOAc:Pet Spirit) 0.40. 1H-NMR (CDCl3, 500 MHz) δ 7.82 (d, J = 8.0 Hz, 2H, H2ꞌ, H6ꞌ), 7.33 (d, J = 8.0

Hz, 2H, H3ꞌ, H5ꞌ), 3.39-3.50 (m, 2H, CH2Cl), 3.04 (quint, J = 5.0 Hz, 1H, H2), 2.74 (d, J = 7.0 Hz, 1H,

H1b) 2.43 (s, 3H, TsCH3), 2.24 (d, J = 4.5 Hz, 1H, H1a). 13C-NMR (125 MHz) δ 145.2 (Ar C1ꞌ), 134.6 (Ar

C4ꞌ), 130.0 (Ar C2ꞌ, C6ꞌ), 128.4 (Ar C3ꞌ, C5ꞌ), 43.7 (CH2Cl), 39.9 (C3), 33.1 (C2), 21.9 (TsCH3). ESI-MS

(+) 268 (100%, [35Cl]M+Na+), 270 (33%, [37Cl]M+Na+]. [α]D21 = +61.5˚ (c 0.71, CH2Cl2).

9

Products from the reaction of indigo with (R)-N-tosyl-2-chloromethylaziridine (14) (R)-N-((13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-

methylbenzenesulfonamide (15)

A suspension of indigo (131 mg, 0.500 mmol) in anhydrous DMF (20 mL) was sonicated at 60 ˚C for one

hour. The resulting hot suspension was transferred by cannula under positive pressure of nitrogen into a 50

mL round-bottomed flask containing a magnetic stir bar, pre-dried and ground Cs2CO3 (603 mg, 1.85

mmol), and fitted with a rubber septum. The resulting dark-green to amber solution was stirred for one hour

in an oil bath, pre-heated to strictly 85-87 ˚C under dry nitrogen. The nitrogen flow was cut, and a solution

of (R)-N-tosyl-2-chloromethylaziridine 14 (627 mg, 2.56 mmol) in DMF (2 mL) was injected through the

septum, and the mixture stirred at 85-87 ˚C for 30 min. The resulting intensely-coloured solution was

quenched over crushed ice (ca. 50 g), and the flask rinsed with EtOAc (ca. 10 mL) to remove adhered

products. Upon warming to RT, the emulsion extracted with EtOAc (5×40 mL) until the aqueous phase

became clear. The combined organic fractions were washed with brine (4×40mL), dried (MgSO4), and

filtered through a 2 cm plug of celite, and the solvent removed in vacuo. The crude residue was fractionated

on silica (60 g) using CHCl3 as eluent to afford two major fractions. Removal of the solvent from Fraction

2 afforded the N,N-cyclised mono-adduct 15 (108.2 mg, 46%) as a glassy, intense-blue solid, mp 165-167

˚C (dec.). RF (9:1 CHCl3:MeCN) 0.23. UV-VIS (CH2Cl2) λmax/nm (ε, M-1cm-1) 236 (18847), 302 (14297),

380 (2329), 576 (8879). 1H NMR (500 MHz, DMSO) δ 8.07 (d, J = 6.0 Hz, 1H, NH), 7.66 – 7.59 (m, 6H,

2×TsH, 4×ArH), 7.32 (d, J = 7.8 Hz, 3H, 2×TsH, ArH), 7.25 (d, J = 8.3 Hz, 1H, ArH), 7.05 (dd, J = 14.5,

7.2 Hz, 2H, 2×ArH), 4.67 (s, 1H, H6), 4.43 (d, J = 12.3 Hz, 1H, H7a), 3.71 (d, J = 9.4 Hz, 1H, H7b), 3.08

– 2.96 (m, 1H, CH2a), 2.93 – 2.83 (m, 1H, CH2b), 2.32 (s, 3H, TsCH3). 13C NMR (126 MHz, DMSO) δ

180.43, 180.40, 151.1, 149.6, 143.5, 138.0, 136.0, 135.9, 130.5, 130.3, 127.1, 126.9, 124.6, 124.5, 124.3,

124.2, 123.6, 122.9, 122.4, 121.6, 121.4, 121.3, 111.6, 110.9, 106.9, 88.1, 73.8, 50.5, 50.4, 21.5. IR (neat)

3566 (w), 2987 (w), 1737 (w), 1697 (m), 1609 (s), 1159 (s), 1132 (s) 754 (s). HR-ESI-MS calcd. for

C26H21N3O4SNa+ 472.1331, found 472.1319. [α]D25 = -9.3˚ (c 0.41, CHCl3).

Fraction 1 was concentrated to dryness then dissolved in acetonitrile (20.0 mL), of which 10.0 mL was

subjected to preparative RP-HPLC over 5×2.0 mL injections, using a Prominence Preparatory Liquid

Chromatograph (Shimadzu) with PDA detector, and a Shim-Pack GIS C18 5 μm column (Shimadzu) with

150×20 mm I.D. Gradient elution from 80% to 0% Solvent A (0.1% TFA in H2O) in 60 minutes with

Solvent B (0.1% TFA in MeCN) at a flow rate of 10.0 mL/min afforded three major fractions at TR = 35

min, 37 min, and 47.7 min, and their purity was confirmed by analytical RP-HPLC, using a Prominence

LC-2030C 3D system (Shimadzu) with PDA detector, and a Shim-Pack GIS C18 5 μm column (Shimadzu)

with 4.6×150 mm I.D. Gradient elution from 80% to 0% Solvent A (0.1% TFA in H2O) in 30 minutes with

10

Solvent B (0.1% TFA in MeCN) at a flow rate of 1.0 mL/min showed the three fractions (TR = 16.5, 17.2,

and 21.4 min, respectively) to be free of neighbouring impurities.

N-(((R)-13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methyl-N-(((S)-1-

tosylaziridin-2-yl)methyl)benzenesulfonamide (16)

Removal of the solvent from the fraction collected at TR = 35 min afforded the di-substituted N,Nꞌ-cyclised

indigo derivative 16 (27.4 mg, 8%) as an intense blue-purple solid, mp 121-123 ˚C (dec.). RF (9:1

CHCl3:MeCN) 0.492. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 281 (37018), 360 (2022), 578 (7659). 1H

NMR (400 MHz, CDCl3) δ 7.81 – 7.71 (m, 2H, 2×ArH), 7.60 (d, J = 8.1 Hz, 2H, 2×TsH), 7.57 – 7.48 (m,

J = 8.0 Hz, 4H, 2×TsH, 2×ArH), 7.29 (d, J = 8.1 Hz, 2H, 2×TsH), 7.21 (d, J = 8.0 Hz, 2H, 2×TsH), 7.16

(s, 2H, 2×ArH), 7.02 (s, 2H, 2×ArH), 4.63 (s, 1H, H6), 4.47 (d, J = 10.8 Hz, 1H, H7a), 3.64 (d, J = 15.8

Hz, 1H, H4ꞌa), 3.51 – 3.37 (m, 1H, H7b), 3.31 – 3.19 (m, 1H, H6aa), 3.05 – 2.95 (m, 1H, H4ꞌb), 2.95 – 2.86

(m, 2H, H6ab, H3ꞌ), 2.57 (d, J = 6.9 Hz, 1H, H2ꞌa), 2.38 (d, J = 3.1 Hz, 6H, 2×TsCH3), 2.21 (d, J = 4.1 Hz,

1H, H2ꞌb). 13C NMR (126 MHz, DMSO) δ 180.4, 180.1, 150.9, 149.0, 145.4, 144.5, 143.1, 135.7, 133.9,

130.6, 130.3, 130.1, 128.0, 127.7, 127.0, 124.1, 121.4, 111.6, 110.9, 60.6, 54.4, 49.5, 48.9, 47.3, 40.8, 21.5,

21.4. IR (neat) 2987 (w), 1738 (w), 1697 (m), 1609 (s), 1132 (s), 754 (s). HRESI-MS calcd. for

C36H32N4O6S2Na+ 703.1655 found 703.1661. [α]D25 = -12.6˚ (c 0.11, CHCl3).

N-(3-chloro-2-((4-methylphenyl)sulfonamido)propyl)-N-(((R)-13,14-dioxo-6,7,13,14-

tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (17)

Removal of the solvent from the fraction collected at TR = 37 min afforded the di-substituted N,Nꞌ-cyclised

indigo derivative 17 (33.3 mg, 9%) as an intense blue-purple solid, mp 132-134 ˚C (dec.). RF (9:1

CHCl3:MeCN) 0.493. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 574 (8850). 1H NMR (500 MHz, CDCl3) δ

7.82 (d, J = 7.9 Hz, 2H), 7.75 (d, J = 7.3 Hz, 2H), 7.54 (d, J = 8.2 Hz, 3H), 7.49 (t, J = 7.7 Hz, 1H), 7.36

(d, J = 7.9 Hz, 2H), 7.27 (s, 1H), 7.12 (d, J = 8.1 Hz, 1H), 7.01 (dt, J = 22.7, 7.6 Hz, 3H), 5.69 (s, 1H), 4.60

(d, J = 10.3 Hz, 1H), 4.39 (d, J = 12.1 Hz, 1H), 4.00 (s, 1H), 3.86 (d, J = 11.6 Hz, 1H), 3.57 – 3.37 (m, 4H),

3.19 (dd, J = 15.1, 6.8 Hz, 1H), 2.65 (d, J = 14.3 Hz, 1H), 2.42 (s, 3H), 2.38 (s, 3H). 13C NMR (126 MHz,

CDCl3) δ 181.2, 180.5, 150.7, 148.4, 145.2, 144.3, 137.3, 135.7, 133.2, 130.4, 130.2, 127.7, 127.4, 125.7,

125.2, 125.1, 123.0, 121.8, 121.6, 110.7, 108.9, 77.2, 53.9, 53.1, 51.0, 49.5, 45.4, 40.2, 21.8, 21.7. IR (neat)

3013 (w), 1743 (w), 1608 (m), 1304 (m), 756 (s). HRESI-MS calcd. for C36H33N4O6S235ClNa+ 739.1428

found 739.1448. [α]D25 = -11.4˚ (c 0.12, CHCl3)

11

(±)-N-((13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-

methylbenzenesulfonamide (±15)

Prepared following General Procedure A, using (±)-N-tosyl-2-bromomethylaziridine (1.00 g, 3.45 mmol),

and a 5 min reaction time.* The crude residue was fractionated on silica (80 g) using 5% MeCN:CHCl3 as

eluent. Removal of the solvent from Fraction B afforded the N,N-cyclised mono-adduct ±15 (247.2 mg,

52%) as a glassy, intense-blue solid. Spectral and physical characteristics (excepting optical rotation) were

identical to those reported above for the R-isomer.

Fraction A was condensed and assessed by analytical RP-HPLC (80%-0% H2O:MeCN; 1.0 mL/min),

which showed the presence of small amounts of 11, though this was not quantified.

Reaction of indigo with N-Tosyl-2-phenylaziridine (18)

(±)-N-tosyl-2-phenylaziridine (18)†

To a solution of chloramine-T trihydrate (8.46 g, 30 mmol) and iodine (381 mg, 1.50 mmol, 5 mol%) in

dry acetonitrile (90 mL) was added styrene (3.6 mL, 31.6 mmol) with stirring under nitrogen at RT. The

flask was wrapped in aluminium foil, and stirred vigorously under static nitrogen for 2 days. The resulting

orange mixture was filtered through a thin pad of celite, eluted with acetonitrile (3×20 mL), and the filtrate

extracted with pet. spirit (3×50 mL) to remove unreacted styrene. The solvent was removed in vacuo, then

the residue dissolved in ethyl acetate (100 mL) and washed with sat. Na2S2O3 solution (2×40 mL) and brine

(50 mL), dried (MgSO4) and the solvent removed. The resulting crystalline solid was triturated in hexane

(50 mL) and collected by vacuum filtration to give phenylaziridine 18 (7.66 g, 93%) as off-white crystals.

Spectral and physical characteristics were identical to those reported previously.4

(E)-N-(2-(3,3'-dioxo-[2,2'-biindolinylidene]-1-yl)-1-phenylethyl)-4-methylbenzenesulfonamide (19)

Prepared following General Procedure A, using phenylaziridine 18 (1.363 g, 4.99 mmol) and a 5 min

reaction time. The crude residue was fractionated on silica (60 g) by sequential elution with 1: 10% pet.

spirit:CH2Cl2, 2: CH2Cl2, and 3: 10% EtOAc:CH2Cl2 to afford two major fractions. The solvent was

removed from Fraction 1 to give a dark blue residue, which was re-dissolved in a minimum of hot CHCl3,

and precipitated with a tenfold volume of pet. spirit. The resulting solid was collected on a Hirsch funnel,

and dried in vacuo to give the terminally ring-opened aziridine adduct 19 (333.4 mg, 62%) as a dark teal

powder, mp 211-213 ˚C RF (10% Pet Spirit:CH2Cl2) 0.43. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 294

(24346), 341 (9142), 645 (13774). 1H NMR (500 MHz, CDCl3) δ 10.65 (s, 1H), 7.72 (d, J = 7.6 Hz, 1H),

7.64 (d, J = 7.5 Hz, 1H), 7.57 (t, J = 5.9 Hz, 3H), 7.50 (t, J = 7.4 Hz, 1H), 7.40 (t, J = 7.5 Hz, 2H), 7.32 (t,

* Longer reaction times typically led to extensive polymerisation † Adapted from a literature procedure, with modifications made for easier scalability. 1H and 13C NMR spectra are reported in the SI.

12

J = 7.3 Hz, 1H), 7.28 – 7.19 (m, 2H), 7.06 (t, J = 7.4 Hz, 1H), 7.04 – 6.94 (m, 3H), 6.72 (d, J = 7.9 Hz, 2H),

5.23 – 5.11 (m, 1H), 4.96 – 4.85 (m, 1H), 3.91 (s, 1H), 2.22 (s, 3H). 13C NMR (125 MHz, CDCl3) δ 189.0,

188.8, 151.8, 151.3, 142.7, 139.7, 138.1, 137.0, 136.2, 129.2, 128.3, 126.8, 126.1, 125.5, 125.0, 124.1,

121.4, 120.9, 120.8, 120.0, 112.0, 110.2, 77.2, 55.6, 51.9, 21.6. IR (neat) 3288 (w), 1630 (m), 1607 (s),

1026 (s). HRESI-MS calcd. for C31H26N3O4S+ = 536.1644, found 536.1664.


Fraction 2 was condensed to give a dark blue-green residue, which was dissolved in a minimum of hot

CHCl3, then diluted with a tenfold volume of hot pet. spirit. The solution was chilled, and the resulting

precipitate collected by vacuum filtration and dried in vacuo to give the title compound 20 (191.9 mg, 36%)

as a dark blue powder, mp 194-196 ˚C. X-ray quality crystals were grown by gradual diffusion of pet spirit

into a saturated solution in 1:1 CHCl3:MeOH in a sealed vapour chamber at room temperature. RF (10%

pet spirit:CH2Cl2) 0.31. UV-Vis (CH2Cl2) λmax/nm (ε, M-1cm-1) 292 (23907), 340 (7457), 637 (9167). 1H

NMR (500 MHz, CDCl3) δ 10.74 (s, 1H, N1ꞌH), 7.64 (dd, J = 15.8, 7.6 Hz, 2H, H4, H4ꞌ), 7.50 (t, J = 8.2

Hz, 3H, 2×TsH, 1×ArH), 7.29 – 7.16 (m, 7H, 7×ArH), 7.02 (d, J = 8.0 Hz, 1H, ArH), 6.94 (dt, J = 14.6,

8.0 Hz, 5H, 3×ArH, 2×TsH), 6.57 (dd, J = 17.5, 8.1 Hz, 2H, ArH, H2ꞌꞌ), 5.89 (s, 1H, C1ꞌꞌNH), 4.11 (dd, J

= 17.1, 11.2 Hz, 2H, C1ꞌꞌH2), 2.26 (s, 3H, TsCH3). 13C NMR (126 MHz, CDCl3) δ 189.3, 188.2, 151.8,

150.0, 143.1, 137.2, 136.9, 136.7, 135.4, 129.5, 129.0, 128.0, 126.7, 126.7, 125.9, 125.3, 124.2, 122.4,

121.3, 121.1, 120.1, 114.1, 112.1, 77.2, 60.0, 43.4, 21.6. IR (neat) 3254 (w), 1628 (m), 1605 (s), 1069 (s).

HRESI-MS calcd. for C31H25N3O4SNa+ = 558.1463, found 558.1481.

Reaction of indigo with 2,2ꞌ-biaziridine 4-methyl-N-(((6R,7R)-7-((4-methylphenyl)sulfonamido)-14,15-dioxo-7,8,14,15-tetrahydro-6H-

[1,4]diazepino[1,2-a:4,3-a']diindol-6-yl)methyl)benzenesulfonamide (21)

Prepared following General Procedure A using (2R,2ꞌR)-N,Nꞌ-ditosyl-2,2ꞌ-biaziridine (512 mg, 1.30 mmol)

in DMF (3 mL) over a 20 min reaction time. The crude residue was dissolved in a minimum of hot CHCl3,

and gradual dilution with a tenfold volume of hot pet. spirit precipitated an intense dark blue powder. The

suspension was cooled to 0 ̊ C overnight, and the fine dark blue precipitate was collected on a Hirsch funnel

and dried in vacuo to give the 1,3-di-ring-opened biaziridine adduct 21 (610 mg, 93%) as an intense dark

blue-black powder, mp 135-136˚ (dec.). RF (10% EtOAc:CH2Cl2) 0.29. UV-Vis (CH2Cl2) λmax/nm (ε, M-

1cm-1) 573 (6647). 1H NMR (400 MHz, CDCl3) δ 7.61 (d, J = 8.1 Hz, 2H, 2×TsH), 7.56 (t, J = 7.6 Hz, 1H,

H10), 7.48 (t, J = 8.4 Hz, 3H, 2×TsH, 1×ArH), 7.34 (d, J = 8.2 Hz, 3H, 2×TsH, 1×ArH), 7.18 (t, J = 8.6

Hz, 3H, 2×TsH, 1×ArH), 7.01 (dd, J = 14.8, 7.3 Hz, 1H, ArH), 6.94 (t, J = 9.1 Hz, 3H, 2×TsH, 1×ArH),

6.69 (d, J = 5.9 Hz, 1H, C7NH), 5.71 (s, 1H, C6aNH), 4.95 (t, J = 4.7 Hz, 1H, H6), 4.63 (d, J = 12.9 Hz,

1H, H8a), 3.62 (d, J = 8.1 Hz, 1H, H8b), 3.14 – 2.98 (m, 1H, C6aHa), 2.83 (d, J = 14.8 Hz, 1H, C6aHb),

13

2.35 (s, 3H, TsCH3), 2.23 (s, 3H, TsCH3). 13C NMR (101 MHz, CDCl3) δ 180.9, 180.8, 149.9, 149.3, 144.1,

143.8, 136.0, 135.4, 130.1, 130.0, 129.7, 128.1, 127.2, 127.1, 126.8, 125.0, 124.5, 124.2, 123.0, 122.2,

121.8, 121.5, 110.3, 110.1, 54.3, 52.5, 42.4, 39.4, 21.7, 21.6. IR (neat) 3253 (w), 1741 (w), 1697 (w), 1609

(m), 1304 (m), 745 (s). HRESI-MS calcd. for C34H30N4O6S2Na+ = 677.1499, found 677.1513. [α]D29 = -

15.0 ˚ (c 0.04, CHCl3)

Biological testing

The antiplasmodial activity of the experimental compounds was assessed against Plasmodium falciparum

3D7 (chloroquine sensitive) and Dd2 (drug resistant) parasite strains. Preliminary cytotoxicity assessment

was carried out using Human Embryonic Kidney (HEK293) cells, sourced from the American Type Culture

Collection (ATCC, Manassas, VA, USA). The compounds were tested in a 22-point concentration-response

range of 80 µM – 0.01 nM (160 µM – 0.02 nM for Compound 10). Artesunate, dihydroartemisinin (DHA),

chloroquine, puromycin and pyrimethamine were used as reference compounds / drugs, and were tested in

both experiments using 22-point concentration-response range of 40 µM – 0.01 nM (10 µM – 0.003 nM for

DHA and artesunate). 0.4% DMSO and 5 µM puromycin were used as negative and positive in-plate

controls, respectively.

Antimalarial assay. Plasmodium falciparum 3D7 (chloroquine sensitive) and Dd2 (drug resistant) parasite

strains were maintained in RPMI 1640 supplemented with 25 mM HEPES, 5% AB human male serum, 2.5

mg/ml Albumax II, and 0.37 mM hypoxanthine.

Parasites were subjected to two rounds of sorbitol synchronization before undergoing compound treatment.

Ring stage parasites at 2% parasitemia and 0.3% hematocrit were exposed to the experimental compounds

in 384-wells imaging CellCarrier microplates (PerkinElmer), as previously described.5 Plates were

incubated for 72 h at 37 °C, 90% N2, 5% CO2, 5% O2, parasites then stained with 2-(4-amidinophenyl)-1H-

indole-6-carboxamidine (DAPI) in permeabilization buffer (PBS, 5mM EDTA, 0.5ug/ml DAPI, 0.01%

Triton X-100, and 0.001% saponin), and imaged with an Opera QEHS micro-plate confocal imaging system

(PerkinElmer) using 20x water-immersion objective and 405 nm excitation, 450/50 nm emission filters.

Images were analysed using a custom Acapella spot detection script, to quantify DAPI-stained parasites in

each well, as previously described.5

Cytotoxicity Assay. Human Embryonic Kidney cells (HEK293) were maintained in DMEM medium

supplemented with 10% FBS. Cells were seeded at 2,000 cells/well in TC-treated 384-wells clear-bottom

plates (Greiner), 24h before the addition of compounds. Compounds were added to the plates as described

above, then the plates were incubated for 72h at 37 °C, 5% CO2. At the end of the incubation, the media

was removed from the wells and replaced with an equal volume of 44 µM resazurin. After an additional 5-

14

6 hours incubation at standard conditions, the total fluorescence (excitation/emission: 530 nm / 595 nm)

was measured using an Envision plate reader (PerkinElmer).

Raw data from each assay was normalized using the in-plate positive and negative controls to obtain

normalized % inhibition data, which was then used to calculate IC50 values, through a 4 parameter logistic

curve fitting in Prism v 6.0 (GraphPad).

The experiments were carried out in two biological replicates, each consisting of two technical repeats.

15

NMR data for isolated compounds

(S)-6-(hydroxymethyl)-6,7-dihydropyrazino[1,2-a:4,3-a']diindole-13,14-dione (8)

Figure 1: 1H spectrum of compound 8

-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

A (m)7.59

B (d)7.35

C (m)7.02

D (t)5.25

E (dd)4.66

G (dd)3.70

H (m)3.53

F (d)4.44

2.40

1.27

1.16

1.14

1.00

2.44

2.52

4.35

2.50

DM

SO-d

63.

463.

483.

493.

513.

523.

533.

543.

663.

673.

693.

704.

404.

434.

624.

634.

644.

655.

225.

235.

24

6.97

6.99

7.00

7.01

7.03

7.32

7.33

7.51

7.54

7.56

7.57

7.58

7.59

7.60

NN

O O

OH

16

Figure 2: 13C spectrum of compound 8

0102030405060708090100110120130140150160170180190200f1 (ppm)

39.5

2 D

imet

hyl S

ulfo

xide

-d6

51.0

851

.23

59.8

8

110.

5111

0.87

120.

4112

0.65

120.

8212

1.51

122.

0312

3.07

123.

5312

3.88

135.

0013

5.29

149.

4415

0.23

179.

7317

9.89

17

Figure 3: gCOSY spectrum of compound 8

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

f1 (

ppm

)

{3.52,4.68}

{3.69,4.32}

{4.44,3.68}{5.25,3.52} {4.66,3.50}

18

Figure 4: NOESY spectrum of compound 8

1.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.01,7.58}{4.44,7.35}

{4.65,7.34}

{7.61,7.04}

{3.70,4.45}{7.34,4.44}

{4.44,3.68}{5.24,3.53}

19

Figure 5: gHMBC spectrum of compound 8

3.54.04.55.05.56.06.57.07.58.0f2 (ppm)

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

f1 (

ppm

)

20

(7S,9aR)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (9)


-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.010.5f1 (ppm)

A (d)7.82

B (m)7.49

C (t)7.35

D (d)7.08

E (t)7.02

F (d)6.92

G (m)5.13

H (t)4.38

I (d)3.74

J (dd)4.25

K (dd)4.20

1.22

1.28

1.22

1.25

1.16

1.04

2.06

1.15

1.08

1.98

1.00

0.00

TM

S

3.72

3.75

4.19

4.19

4.21

4.22

4.24

4.25

4.26

4.26

4.36

4.38

4.39

5.13

5.13

5.14

6.91

6.93

7.00

7.02

7.03

7.07

7.09

7.34

7.35

7.37

7.48

7.49

7.51

7.81

7.83

N

HN

OO

O

21


-100102030405060708090100110120130140150160170180190200210220f1 (ppm)

51.8

0

65.5

7

74.0

877

.16

Chlo

rofo

rm-d

110.

7911

1.09

120.

0112

2.15

122.

9512

4.01

124.

4312

5.31

132.

3713

3.82

144.

9014

5.46

146.

8215

2.47

188.

21

22


3.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.4f2 (ppm)

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.84,7.83} {7.09,7.50}

{7.51,7.49}{6.93,7.37}{7.35,7.34}

{7.02,7.02}{7.82,7.01}{6.92,6.88}

{4.19,5.17}{5.13,5.14}

{4.37,5.14}

{4.38,4.38}{5.14,4.35}{4.27,4.26} {3.72,4.21}{4.19,4.20}

{3.75,3.74}{4.19,3.74}

23

Figure 9: gHSQC spectrum of compound 9

3.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.45,133.91}{7.35,132.44}

{7.48,125.34}

{7.82,123.92}

{7.02,122.18}{7.02,119.98}

{6.92,111.12}

{7.08,110.74}

{5.13,74.15}

{4.25,65.53}

{4.37,65.53}

{4.18,51.87} {3.72,51.83}

24


3.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

f1 (

ppm

)

{7.82,183.71}

{7.50,152.38}

{7.82,151.54}

{7.02,145.84}

{7.48,145.02}

{7.82,133.47} {7.01,132.86}

{7.48,131.76}

{7.35,125.08} {6.93,124.32}

{7.02,123.18}

{6.92,120.15}{4.18,118.05}

{4.37,111.15}{5.13,111.08}{7.02,110.83}

{7.49,110.50}

{4.37,74.52} {3.73,73.26}

{3.73,65.70} {3.73,65.45}{4.22,65.12}

{4.37,51.32} {4.25,50.98}

25


3.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.2f2 (ppm)

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{5.14,7.38}

{4.22,7.04}{5.14,7.02}

{7.02,5.15} {3.74,5.15}

{7.37,5.15}

{4.16,5.14}

{3.75,4.39}

{4.22,4.38}

{5.14,4.37}{7.03,4.27} {5.14,4.23} {3.72,4.23}{4.37,4.22}

{7.08,4.18}

{4.38,3.76}{5.14,3.75} {4.20,3.71}

26

(7R,9aR)-14-((E)-3-hydroxyprop-1-en-1-yl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (10)

Figure 12: 1H spectrum of compound 10. The presence of the grease contaminant in the 1.0 – 2.0 ppm region of the spectrum could not be removed, despite repeated attempts.

-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

A (d)7.74

B (dd)7.49

C (t)7.42

D (m)7.29

E (t)7.13

F (m)7.00

G (d)6.83

H (m)6.04

I (s)4.99

J (m)4.28

K (d)3.71

1.62

7.20

1.23

1.14

1.18

2.75

1.44

1.37

1.45

2.36

1.00

-0.0

0 TM

S

3.69

3.72

4.22

4.23

4.24

4.26

4.27

4.29

4.32

4.33

4.99

6.01

6.02

6.04

6.05

6.82

6.84

6.97

6.98

7.00

7.02

7.03

7.12

7.13

7.15

7.27

7.29

7.30

7.40

7.42

7.43

7.46

7.48

7.49

7.51

7.73

7.75

N

N

OO

O

OH

27


-100102030405060708090100110120130140150160170180190200210220230f1 (ppm)

52.1

1

61.9

367

.62

73.3

5

109.

9211

1.68

111.

7011

9.09

119.

1011

9.95

121.

5512

3.63

123.

6412

4.32

124.

3312

4.35

124.

4712

4.85

124.

8612

5.53

130.

7813

0.79

131.

9313

1.95

134.

1413

4.16

149.

6615

2.68

28


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

0

1

2

3

4

5

6

7

8

f1 (

ppm

)

{7.77,7.00} {6.07,6.86}

{6.86,6.06} {4.36,6.01}

{6.09,4.35} {3.73,4.25}{5.02,4.24}

29


3.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.4f2 (ppm)

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

f1 (

ppm

)

{7.75,181.76}

{7.50,153.02}

{7.75,152.86}

{7.48,145.53} {7.42,145.49} {6.85,145.42}

{7.75,134.46}

{7.48,132.13}

{4.30,131.75}{6.04,130.95}{7.14,125.67}

{7.42,125.46}

{7.03,125.17}{7.03,120.04}

{7.14,111.88} {4.24,111.06}{6.99,110.11}

{3.71,73.48}

{3.71,67.71}

{6.05,61.81}{6.84,61.72}

{4.23,52.08} {4.28,52.06}

30


2.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f2 (ppm)

40

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.54,134.24}{7.46,131.85}

{6.86,130.35}

{7.50,124.71}

{7.79,124.38}

{6.99,119.95} {6.02,119.22}

{7.30,111.00} {7.07,109.84}

{5.04,73.18}

{4.27,67.51} {4.30,67.45}

{4.36,61.70}

{4.29,52.16} {3.74,51.82}

31

Figure 17: ROESY spectrum of compound 10

0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

0

1

2

3

4

5

6

7

8

f1 (

ppm

)

{4.25,7.04}

{4.32,6.09}

{3.74,5.03}

{4.31,5.01}

{6.07,4.31} {5.02,4.27} {3.73,4.25}{7.05,4.25}

{4.27,3.71}

32

Figure 18: zTOCSY spectrum of compound 10

3.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.8f2 (ppm)

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.05,7.79}

{7.32,7.57} {7.01,7.51}

{7.78,7.07}

{7.52,7.02}

{4.37,6.87}{6.07,6.86}

{4.36,6.07}{6.85,6.06}

{3.73,5.03}{4.27,5.03}

{6.85,4.37} {6.06,4.37}{5.02,4.28}

{5.02,3.74} {4.25,3.74} {3.65,3.70}

33

(7S,9aR)-14-(((S)-2-oxo-1,3-dioxolan-4-yl)methyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (11)


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

A (d)7.68

B (m)7.48

C (d)7.33

D (td)7.13

E (m)6.99

F (ddd)5.48

H (m)4.97

G (d)5.20

I (d)3.68

J (m)4.16

K (m)4.28

1.08

2.27

3.91

1.02

1.00

1.10

1.99

1.02

1.07

2.97

1.00

0.00

TM

S

3.67

3.70

4.10

4.12

4.14

4.16

4.17

4.18

4.20

4.21

4.23

4.24

4.25

4.25

4.28

4.30

4.31

4.32

4.32

4.96

4.97

4.98

5.18

5.20

5.22

5.45

5.46

5.47

5.48

5.49

5.49

5.51

5.51

6.96

6.98

7.00

7.02

7.11

7.11

7.13

7.13

7.15

7.15

7.32

7.34

7.42

7.44

7.44

7.46

7.47

7.48

7.48

7.49

7.50

7.50

7.51

7.52

7.67

7.69

N

N

OO

O

O

OO

34

Figure 20: 13C spectrum of compound 11, with expansion of the region 185-105 ppm.

0102030405060708090100110120130140150160170180190f1 (ppm)

52.5

155

.63

68.0

469

.17

73.5

377

.16

Chlo

rofo

rm-d

81.4

5

110.

3811

1.24

112.

1112

0.09

123.

5512

4.28

124.

6312

4.89

128.

3913

2.86

134.

44

146.

7614

8.10

153.

0115

5.15

178.

41

182.

30

110120130140150160170180f1 (ppm)

110.

3811

1.24

112.

11

120.

0912

3.55

124.

2812

4.63

124.

8912

8.39

132.

8613

4.44

146.

7614

8.10

153.

0115

5.15

178.

41

182.

30

35


3.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.0f2 (ppm)

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

f1 (

ppm

)

{6.97,7.69}

{7.00,7.69}

{7.02,7.51}

{7.35,7.49}

{7.12,7.45}{7.49,7.35}

{7.46,7.16}{7.52,7.01}

{5.19,5.52} {4.10,5.52}

{4.25,5.47}

{5.48,5.24} {4.25,5.23}{4.15,5.05}

{4.29,4.98}

{4.11,4.33}{4.99,4.32}

{5.19,4.31}

{5.48,4.26} {3.68,4.21}{4.97,4.18}{5.49,4.14} {4.29,4.12}

{4.20,3.71}

36


3.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.0f2 (ppm)

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.50,134.45}{7.46,132.64}

{7.43,124.44}

{7.69,124.11} {7.12,123.43}

{6.98,119.99}

{7.33,111.05} {7.02,110.24}

{5.49,81.32}

{7.26,77.16}Chloroform-d

{4.97,73.38}

{5.20,68.98} {4.27,68.94} {4.28,67.87}

{4.25,67.86}

{4.28,55.46}{4.16,52.46} {3.70,52.23}

37


3.54.04.55.05.56.06.57.07.5f2 (ppm)

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{6.97,7.69}{7.33,7.51} {7.13,7.50}

{7.00,7.50}

{7.12,7.37}

{7.45,7.12}

{7.50,7.06}

{4.17,7.01}{7.68,6.98}

{5.20,5.49} {4.26,5.47}

{5.51,5.20} {4.26,5.20}

{3.66,4.99}{4.31,4.95}

{5.21,4.27}

{5.48,4.27}

{3.69,4.19}{4.97,4.18}

{4.97,3.68} {4.20,3.68}

38

(7S,8R,9aS)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (12)


-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

A (dd)7.81

B (ddd)7.51

C (td)7.35

D (d)7.11

E (qd)7.02

F (d)6.92

G (dt)5.01

H (dt)4.75

I (dd)4.38

J (dd)4.17

K (dd)3.96

L (dd)3.70

1.10

1.16

1.16

1.07

1.46

1.04

1.09

3.90

1.08

1.08

2.09

1.00

0.00

TM

S3.

683.

693.

713.

723.

923.

943.

953.

963.

984.

154.

164.

174.

194.

194.

364.

374.

394.

394.

734.

734.

744.

744.

744.

754.

754.

754.

764.

775.

005.

005.

015.

015.

015.

026.

916.

937.

007.

007.

027.

027.

037.

037.

047.

057.

057.

107.

127.

347.

347.

357.

357.

377.

377.

497.

497.

517.

517.

517.

527.

537.

807.

817.

827.

82

N

HN

OO

O

OH

39

Figure 25: 13C spectrum of compound 12, and expansion of the region 81-73 ppm

-100102030405060708090100110120130140150160170180190200f1 (ppm)

48.2

9

60.3

8

75.5

977

.16

Chlo

rofo

rm-d

77.7

5

109.

5811

0.70

111.

1611

7.81

120.

0912

1.00

122.

2912

3.52

124.

0312

4.19

125.

2913

2.40

134.

0313

8.84

144.

5214

5.22

151.

76

184.

11

737475767778798081f1 (ppm)

75.5

9

77.1

6 Ch

loro

form

-d

77.7

5

40


1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

f1 (

ppm

)

{7.03,7.79}

{7.09,7.52}

{7.02,7.50}

{7.01,7.36}{7.49,7.11} {7.02,7.10}

{7.81,7.00}

{4.74,5.00}

{4.36,4.99} {3.69,4.99}{3.95,4.74}{5.01,4.74}

{3.71,4.39}{5.01,4.39}{3.97,4.17}{4.75,4.16}

{4.17,3.97}

{4.74,3.95}

{4.99,3.69} {4.36,3.68}

41


4.04.55.05.56.06.57.07.58.0f2 (ppm)

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

42


3.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.0f2 (ppm)

40

50

60

70

80

90

100

110

120

130

f1 (

ppm

)

{7.50,133.99}{7.34,132.31}

{7.49,125.18}

{7.80,123.90}

{7.03,122.20}{7.00,119.97}

{6.91,111.00}

{7.10,110.54}

{4.74,77.53}{5.00,75.38}

{4.14,60.15} {3.96,60.11}

{3.68,48.54}{4.35,47.97}

43


1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

f1 (

ppm

)

{7.79,184.11}

{7.50,151.74}

{7.47,145.33}

{7.33,145.12}

{7.78,133.85}

{7.11,124.10} {6.97,123.99}

{7.01,123.13}

{4.35,117.80}

{5.00,111.48}{7.48,111.32}

{7.52,77.28}

{7.00,77.23}

{4.16,75.66} {3.67,75.59}

{4.75,60.24}

{4.75,47.86}

44

(7S,8R,9aS)-14-((S)-2-hydroxy-2-((S)-oxiran-2-yl)ethyl)-8-(hydroxymethyl)-7,8-dihydro-6H-7,9a-epoxy[1,5]oxazocino[5,4-a:3,2-b']diindol-15(14H)-one (13)

Figure 30: 1H NMR spectrum of compound 13

0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f1 (ppm)

A (d)5.82

B (d)5.21

C (t)4.60

D (d)4.53

E (dt)4.20

F (m)4.03

G (m)3.76

H (dt)3.23

I (dt)2.76

J (m)2.69

K (dd)2.56

L (dd)2.32

M (t)6.81

N (d)7.03

O (dd)7.08

P (m)7.21

Q (dd)7.60

R (m)7.56

S (d)7.52

T (d)7.49

0.78

0.74

0.84

0.97

1.02

1.64

2.12

1.09

1.04

1.01

0.75

1.00

0.83

0.85

0.87

0.86

0.42

0.81

1.25

1.23

2.31

2.31

2.32

2.33

2.56

2.57

2.58

2.68

2.68

2.69

2.69

2.70

2.70

2.77

2.77

3.21

3.23

3.24

3.33

DM

SO-d

63.

713.

723.

733.

743.

803.

823.

833.

844.

004.

014.

034.

044.

074.

194.

204.

224.

244.

514.

554.

594.

604.

625.

205.

215.

825.

836.

796.

816.

837.

027.

057.

077.

097.

107.

197.

207.

217.

237.

237.

487.

507.

517.

537.

557.

577.

577.

587.

597.

607.

62

N

N

OO

O

OH

OHO

45


-100102030405060708090100110120130140150160170180190200f1 (ppm)

37.0

139

.52

DM

SO-d

642

.75

43.6

9

53.2

6

62.3

4

70.2

070

.37

75.4

781

.44

109.

0111

0.07

117.

7711

8.16

118.

3411

9.71

119.

7712

2.18

125.

1113

0.12

131.

4813

3.46

138.

82

158.

00

193.

02

46

Figure 32: COSY spectrum of compound 13

1.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.10,7.64}{7.04,7.58}

{6.82,7.57}{7.23,7.53}

{7.10,7.23}{7.62,7.10}

{7.56,6.81}

{4.07,5.84} {4.00,5.83}{5.87,5.83}

{3.22,5.22}

{4.02,4.62} {4.06,4.61}

{4.23,4.53}

{4.61,4.23} {4.53,4.21}{2.74,4.09}{4.62,4.06}{5.84,4.06} {4.60,4.01}{5.83,4.00}

{3.73,3.84}{3.25,3.83}

{3.25,3.74}

{3.82,3.72}

{5.22,3.25} {2.70,3.25}{3.83,3.25}

{3.75,3.24}

{4.09,2.79} {4.05,2.79}

{2.59,2.70}{3.23,2.70}

{2.32,2.69}{2.32,2.59}

47

Figure 33: TOCSY spectrum of compound 13

2.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.53,7.62}

{7.22,7.62}{7.10,7.62}

{6.82,7.60}

{6.82,7.05}{7.50,6.88} {6.82,6.82}

{2.78,5.83}{5.83,5.83} {4.62,5.79}

{2.58,5.25}{5.23,5.22} {3.22,5.22}{3.81,5.21} {2.69,5.21}

{3.74,5.21}

{4.23,4.61}

{4.61,4.61}

{4.02,4.61} {2.78,4.60}{5.83,4.60}

{4.61,4.52}

{4.61,4.22}{4.07,4.07}{5.83,4.05}

{2.78,4.04}{4.02,4.03}

{3.83,3.88} {3.72,3.83}{3.24,3.83}{5.22,3.83} {2.70,3.82}

{3.81,3.74}

{5.22,3.74}

{2.70,3.74}{3.23,3.73}

{3.73,3.69}

{2.59,3.29}{3.83,3.24} {3.72,3.24}{5.21,3.24} {2.70,3.24}

{2.32,3.23}{3.23,3.19}

{5.83,2.78} {4.05,2.77} {2.76,2.77} {2.32,2.70}{5.21,2.70}

{2.56,2.70}

{5.19,2.58}

{2.32,2.57}

{3.22,2.32} {2.58,2.32} {2.69,2.32}

48

Figure 34: HMBC spectrum of compound 13

2.53.03.54.04.55.05.56.06.57.07.5f2 (ppm)

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

f1 (

ppm

)

{7.56,193.54}

{7.57,158.33}

{6.80,139.30}

{7.57,139.22}

{7.22,133.81}

{7.62,133.55}

{7.11,120.24}{7.23,118.72}

{7.05,118.54}

{4.52,118.01} {4.55,117.87}

{7.09,110.22} {6.81,109.57}

{4.61,81.71}{3.23,76.12}

{3.23,75.90}

{5.22,75.78}{5.83,70.93} {4.22,70.66} {3.81,70.60}{3.72,70.46}{5.21,70.30} {2.58,69.98}

{2.75,62.56}{4.05,62.51}{4.60,62.45} {2.81,62.44}{4.06,62.37}{5.83,62.32}

{3.23,53.70}{3.82,53.68} {2.57,53.58}{5.22,53.50}

{4.60,43.18} {4.62,43.11}{5.83,43.08}{2.50,39.82}Dimethyl Sulfoxide-d6

49

Figure 35: HSQC spectrum of compound 13

2.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

30

40

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.61,139.18}

{7.58,125.47}

{7.11,120.10} {6.83,118.67}

{7.51,110.48}

{7.06,109.32}

{3.84,75.82} {3.70,75.81}

{4.58,70.70} {3.24,70.39}

{3.98,62.52}

{2.71,53.49}

{4.10,43.12} {2.80,43.07}

{4.24,37.40}

50


2.02.53.03.54.04.55.05.56.06.57.07.5

f2 (ppm)

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (p

pm)

{2.50,2.50}Dimethyl Sulfoxide-d6

{7.61,3.83}

{7.61,3.73}{7.52,4.22}

{7.49,4.53}

{7.03,4.05}

{7.58,6.81}

{7.58,7.07}

{7.22,7.07}{7.21,7.54} {7.03,7.61}

{6.75,7.51}

{5.84,4.54}

{5.83,4.04}

{5.83,3.34}

{5.81,2.80}

{5.21,3.25}

{5.16,3.73}

{4.52,7.53}{3.81,7.62}

{4.05,7.04}

{4.01,5.83}{4.52,5.83}

{4.61,4.21}{4.61,4.02}

{4.52,4.21}

{4.22,4.61} {4.05,4.61}

{4.02,2.75}

{2.75,4.04}

{2.32,3.22}{3.73,3.21} {3.81,3.20}

{2.32,2.58}

{3.21,5.18}

51

(S)-N-(3-chloro-2-hydroxypropyl)-4-methylbenzenesulfonamide (23)


-1-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)

A (d)7.75

B (d)7.33

C (s)4.79

D (dq)3.94

E (m)3.57

F (m)3.12

G (d)2.58

H (s)2.44

3.48

1.04

2.25

2.21

1.00

0.98

2.46

2.18

2.44

2.57

2.59

3.00

3.01

3.02

3.03

3.03

3.05

3.05

3.07

3.16

3.16

3.17

3.18

3.19

3.20

3.21

3.22

3.53

3.55

3.56

3.57

3.58

3.59

3.60

3.61

3.91

3.92

3.93

3.95

3.96

3.97

4.79

7.26

CD

Cl3

7.32

7.34

7.74

7.75

7.76

OHTsHN Cl

52


2.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.33,7.76}

{7.75,7.33} {2.44,7.33}

{3.02,4.82}{4.79,4.80}

{3.19,4.79}

{3.56,3.96} {3.01,3.94}

{3.18,3.94}

{3.96,3.57}

{4.81,3.20} {3.94,3.20} {3.02,3.19}

{4.80,3.04}

{3.18,3.03}{3.93,3.00}

{7.33,2.45}

53


1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.29,129.79}{7.73,127.01}

{7.26,77.16}CDCl3

{3.93,69.94}

{3.52,46.25} {3.15,45.73}

{3.00,45.72}

{2.41,21.42}

54

Figure 40: 13C-DEPTQ spectrum of Compound 23, with CH/CH3 phased up, and CH2/Cq phased down

-100102030405060708090100110120130140150160170180190200f1 (ppm)

21.6

4

45.9

846

.48

70.1

5

77.1

6 CD

Cl3

127.

2113

0.00

136.

41

143.

93

55

(S)-1-chloro-3-((4-methylphenyl)sulfonamido)propan-2-yl methanesulfonate (24)

Figure 41: 1H NMR spectrum of compound 24

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f1 (ppm)

A (m)7.74

B (d)7.34

C (t)4.94

E (d)3.77

F (s)3.68

G (m)3.33

I (s)2.44

K (p)4.85

3.61

1.88

2.08

2.00

1.00

1.07

2.40

2.30

2.44

3.31

3.32

3.33

3.34

3.35

3.68

3.76

3.78

4.82

4.84

4.85

4.86

4.88

4.92

4.94

4.96

7.33

7.35

7.72

7.73

7.74

7.75

7.76

D-NEt3+

D-NEt3+

OMsTsHN Cl

56


010203040506070809010011012013014015060f1 (ppm)

-0.0

0 TM

S

21.5

7

43.0

744

.48

52.5

6

78.8

4

127.

1013

0.02

57


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

f1 (

ppm

)

{2.45,144.24}{7.75,144.20}

{7.34,136.13}

{2.44,130.07}{7.35,129.97}

{3.77,78.85} {3.32,78.75}

{3.77,44.44} {3.33,43.08}

{7.35,21.65}

{0.00,-0.00}TMS

58


2.42.62.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.0f2 (ppm)

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.33,7.76}

{2.43,7.34}{7.75,7.34}{7.26,7.26}CDCl3

{3.33,4.93}{3.77,4.88}{3.32,4.84}

{4.83,3.78}

{4.83,3.33} {4.92,3.31}

59

(R)-N-tosyl-2-chloromethylaziridine (14)


-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)

A (d)7.83

B (d)7.35

C (ddd)3.46

D (tdd)3.04

E (d)2.74

F (t)2.26

G (s)2.44

1.03

3.21

1.03

1.00

2.08

2.01

1.98

0.00

TM

S

2.25

2.26

2.28

2.44

2.74

2.75

3.01

3.03

3.03

3.03

3.04

3.04

3.04

3.05

3.06

3.06

3.06

3.07

3.40

3.41

3.43

3.44

3.48

3.49

3.51

3.52

7.34

7.36

7.82

7.84

TsN Cl

60

Figure 46: 13C-DEPTQ spectrum of compound 14, with CH/CH3 phased up, and CH2/Cq phased down.

0102030405060708090100110120130140150160170f1 (ppm)

21.6

5

32.8

3

39.7

0

43.5

3

77.1

6 Ch

loro

form

-d

128.

1412

9.75

134.

36

144.

95

61

Figure 47: COSY spectrum of compound 14.

1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{7.35,7.83}

{7.82,7.36} {2.43,7.34}

{3.03,3.51}

{3.41,3.50}

{3.02,3.45}

{3.50,3.42}

{3.47,3.05} {2.74,3.04}

{2.25,3.04}

{3.04,2.74}

{7.35,2.43}{3.04,2.25}

62

Figure 48: HSQC spectrum of compound 14.

2.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

f1 (

ppm

)

{7.32,130.11}

{7.80,128.48}

{7.26,77.16}CDCl3

{3.40,43.62}

{3.46,43.59}

{3.02,39.76}

{2.23,33.08}

{2.72,33.07}

{2.41,21.89}

63

(R)-N-((13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (15)


-1-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5.0f1 (ppm)

A (t)8.06

B (dd)7.61

C (d)7.30

D (d)7.24

E (q)7.03

F (m)4.65

G (d)4.42

H (dd)3.70

I (m)3.00

J (dd)2.87

K (s)2.32

3.31

1.10

1.02

1.09

1.08

1.09

2.16

1.15

3.15

6.00

1.14

2.32

2.50

DM

SO-d

62.

852.

862.

882.

892.

982.

993.

013.

023.

683.

693.

703.

71

4.40

4.43

4.63

4.64

4.65

4.66

7.01

7.03

7.04

7.05

7.23

7.24

7.30

7.31

7.59

7.60

7.61

7.62

8.05

8.06

8.07

NN

O O

NHTs

64

Figure 50: 13C spectrum of compound 15, with expansion of the region 155-100 ppm

-100102030405060708090100110120130140150160170180190200210220f1 (ppm)

21.5

1

41.1

643

.21

50.4

3

73.7

5

88.0

6

106.

8911

0.92

111.

5512

1.29

121.

4112

1.62

122.

3912

2.86

123.

5712

4.23

124.

2712

4.46

124.

6412

6.92

127.

1313

0.29

130.

5113

5.91

136.

0413

7.97

143.

4814

9.60

151.

1118

0.40

180.

43

105110115120125130135140145150155f1 (ppm)

106.

89

110.

9211

1.55

121.

4112

2.39

122.

8612

3.57

124.

2312

4.46

124.

6412

6.92

127.

13

130.

2913

0.51

135.

9113

6.04

137.

97

143.

48

149.

60

151.

11

65


1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f2 (ppm)

1

2

3

4

5

6

7

8

f1 (

ppm

)

66


1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

f1 (

ppm

)

67


1.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.60,135.93}

{7.30,130.42}{7.61,126.96}

{7.61,124.38}

{7.04,121.47}

{7.29,111.57} {7.24,110.84}

{4.64,50.42}

{3.02,43.15}

{2.91,43.12}

{4.42,41.17} {3.70,41.17}

{2.33,23.84}

68


2.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

f1 (

ppm

)

69

N-(((R)-13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methyl-N-(((S)-1-tosylaziridin-2-yl)methyl)benzenesulfonamide (16)

Figure 55: 1H spectrum of compound 16 (CDCl3). The impurities present at 1.8 – 2.0 ppm could not be removed despite repeated attempts. HPLC analysis indicated the presence of one compound only (Figure 87).

-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)

A (m)7.77

D (d)7.29

E (d)7.21

F (s)7.16

G (s)7.03

H (s)4.63

I (d)4.47

J (d)3.64

K (m)3.46

L (m)3.25

M (m)3.01

N (m)2.91

O (d)2.57

Q (d)2.38

R (d)2.21

B (d)7.60

C (m)7.54

1.27

6.29

1.20

2.12

1.20

1.20

1.31

1.18

1.01

1.00

2.07

1.97

2.46

2.71

4.24

2.74

2.07

0.00

TM

S

2.20

2.21

2.38

2.39

2.56

2.58

2.90

2.92

2.98

2.99

3.02

3.03

3.23

3.25

3.28

3.43

3.46

3.47

3.49

3.62

3.66

4.45

4.48

4.63

7.03

7.16

7.20

7.22

7.28

7.30

7.53

7.55

7.59

7.61

7.77

TsN

NTs

NN

OO

70

Figure 56: 13C spectrum of compound 16 (CDCl3)

-100102030405060708090100110120130140150160170180190200f1 (ppm)

21.6

9

31.1

3

37.7

340

.82

49.6

749

.88

51.7

9

77.1

6 Ch

loro

form

-d

109.

6211

0.72

121.

7112

5.04

127.

4912

8.22

134.

0113

4.88

135.

76

144.

9114

5.48

71

Figure 57: 13C spectrum of compound 16 (DMSO)

010203040506070809010011012013014015016017018090f1 (ppm)

21.3

921

.46

40.7

6

47.3

348

.85

49.4

954

.35

60.5

7

110.

9211

1.60

121.

4112

4.11

126.

9912

7.67

127.

9513

0.06

130.

3413

0.61

133.

8713

5.72

143.

0714

4.47

145.

3614

8.97

150.

91

180.

1118

0.37

72


2.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

)

{3.24,4.64}{3.43,4.45}

{2.99,3.62}{4.47,3.45}

{4.64,3.27} {2.88,3.25}

{3.62,2.99} {2.57,2.92} {2.21,2.90}

{2.91,2.56}

{2.92,2.21}

73


1.52.02.53.03.54.04.55.05.56.06.57.07.58.0f2 (ppm)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

f1 (

ppm

){3.42,4.63}

{3.25,4.63}{4.46,4.63}

{2.91,4.63}{4.63,4.46} {2.92,4.46}

{2.20,3.64}

{2.56,3.64}{3.00,3.64}{3.25,3.44}

{4.63,3.43} {2.89,3.43}{4.47,3.25}{4.63,3.25} {2.90,3.25}

{3.45,3.25}

{2.56,3.00} {2.20,3.00}{3.65,3.00}{4.45,2.91}

{2.20,2.90}

{3.65,2.57} {2.90,2.56} {3.02,2.56} {2.20,2.56}

{2.90,2.20} {3.02,2.20}{3.65,2.20}

74

Figure 60: HSQC spectrum of Compound 16 (CDCl3)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

f1 (

ppm

)

{7.55,135.60}

{7.29,130.24}

{7.21,129.95}

{7.59,127.35}

{7.03,121.50}

{7.16,110.55} {7.15,109.49}

{7.26,77.16}CDCl3

{3.65,51.64}{3.03,51.57}{4.63,49.66}{2.92,49.42}

{3.24,49.34}

{4.47,40.57} {3.43,40.53}{2.91,37.43}

{2.21,30.87}

{2.38,21.44}

75

N-((R)-3-chloro-2-((4-methylphenyl)sulfonamido)propyl)-N-(((R)-13,14-dioxo-6,7,13,14-tetrahydropyrazino[1,2-a:4,3-a']diindol-6-yl)methyl)-4-methylbenzenesulfonamide (17)


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)

A (d)7.82

B (d)7.75

C (d)7.54

D (t)7.49

E (d)7.36

F (dt)7.01

G (d)7.12

H (s)7.27

I (s)5.69

J (d)4.60

K (d)4.39

L (s)4.00

M (d)3.86

N (m)3.46

O (dd)3.19

P (d)2.65

Q (s)2.42

R (s)2.38

2.77

2.93

0.94

0.81

4.01

0.93

0.97

1.01

1.00

0.94

3.09

1.05

1.03

2.06

1.02

2.84

1.78

2.11

-0.0

0 TM

S

2.38

2.42

2.64

2.67

3.17

3.18

3.20

3.21

3.40

3.41

3.42

3.43

3.44

3.46

3.47

3.50

3.52

3.85

3.87

4.00

4.38

4.41

4.59

4.61

5.69

6.98

6.99

7.01

7.02

7.04

7.05

7.11

7.13

7.27

7.35

7.37

7.48

7.49

7.51

7.53

7.55

7.74

7.76

7.76

7.81

7.83

NTs

NN

OO

NHTs

Cl

76

Figure 62: 13C spectrum of compound 17.

-100102030405060708090100110120130140150160170180190200210f1 (ppm)

21.7

121

.80

40.2

345

.43

49.5

050

.95

53.1

353

.90

77.1

6 CD

Cl3

108.

8711

0.72

121.

6212

1.82

122.

9812

5.06

125.

2312

5.65

127.

3512

7.74

130.

1813

0.41

133.

1513

5.69

137.

2314

4.32

145.

1814

8.39

150.

67

180.

5018

1.19

77

Figure 63: COSY spectrum of compound 17.

-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

-1

0

1

2

3

4

5

6

7

8

f1 (

ppm

)

{4.60,4.60} {3.43,4.59}{3.51,4.38}

{3.44,4.07}

{4.41,3.51} {2.66,3.47}{4.62,3.46}

{3.22,3.44}

{3.43,3.17}

{3.49,2.67}

78


-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

f1 (

ppm

)

{7.75,150.43} {7.55,150.34}

{7.49,148.38}

{7.82,144.29}

{7.35,135.73}{7.73,135.53}

{7.56,127.68} {4.41,125.22}{7.56,125.21}

{7.06,110.39}

{3.44,54.12}

{3.18,50.95}

{4.38,49.68}

79


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f2 (ppm)

0

1

2

3

4

5

6

7

8

9

f1 (

ppm

)

{3.48,7.52}

{4.39,7.09}

{4.62,7.01}

{4.61,4.37}{7.14,4.37} {3.53,4.37}

{3.41,3.97}{4.59,3.97} {3.20,3.96}

{3.46,3.82}

{4.59,3.48} {4.42,3.48} {2.65,3.45}{7.53,3.40}{2.64,3.15}{3.43,3.15}

{3.20,2.63}{4.59,2.62}

{0.00,-0.03}TMS

80


0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

140

f1 (

ppm

)

{7.54,135.82}

{7.50,135.72}

{7.26,130.52}

{7.36,130.32}

{7.53,127.82}

{7.04,121.88} {7.00,121.74}

{7.13,110.84} {7.03,108.92}

{7.26,76.92}Chloroform-d

{3.99,53.86}

{3.41,53.10} {3.20,52.94}

{2.64,50.91}

{4.61,49.36}{3.41,45.13}

{4.40,40.28} {3.52,40.11}

{2.42,21.70} {2.38,21.59}

81


-1.0-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

-1

0

1

2

3

4

5

6

7

8

f1 (

ppm

)

{3.41,5.65}

{3.99,5.65}{3.83,5.65}

{3.17,5.65}{5.65,5.65}

{3.43,4.62}

{4.38,4.60}

{4.61,4.40}

{3.47,4.40}

{3.17,4.09}{5.64,4.00} {3.44,3.99}{5.65,3.86}

{3.17,3.85}

{3.83,3.50} {2.65,3.47}{4.59,3.47}{5.65,3.42}{3.80,3.22}{5.65,3.19} {3.99,3.19}

{4.36,2.68} {3.47,2.66}

82

N-tosyl-2-phenylaziridine (18)‡,4


-1.-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5.0f1 (ppm)

A (m)7.87

B (m)7.26

C (dd)3.77

D (d)2.97

E (s)2.42

F (d)2.38

1.50

3.97

1.47

1.03

7.20

2.00

-0.0

0 TM

S

2.38

2.38

2.42

2.97

2.98

3.76

3.77

3.77

3.78

7.20

7.20

7.21

7.21

7.21

7.22

7.26

7.27

7.27

7.28

7.28

7.29

7.31

7.33

7.86

7.86

7.87

7.87

‡ Prepared by the method described in the main text, as a modification of the cited procedure.

TsN

83

Figure 69: 13C spectrum of compound 18, with expansion of the region downfield from 125 ppm

10203040506070809010011012013014015060f1 (ppm)

21.7

5

36.0

4

41.1

4

126.

6512

8.03

128.

4012

8.66

129.

8613

5.07

135.

14

144.

76

126128130132134136138140142144146148150f1 (ppm)

126.

65

128.

0312

8.40

128.

6612

9.86

135.

0713

5.14

144.

76

84



-0.0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.010.51.0f1 (ppm)

A (s)10.65

B (d)7.72

C (d)7.64

D (t)7.57

E (t)

Documents

Table of Contents1 . Cascade reactions of indi go with oxiranes and aziridines: efficient access to dihydropyrazinodiindoles and spiro -oxazocinodiindoles. Nicholas M. Butler, Rudi