Synthesis

O

O

O

Br

Br

N

O

O

Br

BrO

N

O

O

I

IO

4 5

a b

Scheme S1. Synthesis of compound 4 and 5. (a) furfurylamine, acetic acid, 22 h; (b)

KI, acetic acid, 5 h.

3,4-dibromo-1-(furan-2-ylmethyl)-1H-pyrrole-2,5-dione (Compound 4). 3,4-

dibromomaleic anhydride (6.14 g, 24.3 mmol) was dissolved in acetic acid (60 mL)

with slowly addition of furfurylamine (2.748 g, 28.3 mmol), the solution was stirred

at room temperature until the white smoke disappeared, then the system was heated at

120 ºC for 22 h. After removal of solvent, the crude residue was separated by column

chromatography on silica gel (hexane/ethyl acetate = 15/1) to give compound 4 as a

light yellow solid (5.645 g, 70.1 %). 1H NMR (DMSO-d6, 400 MHz, ppm): 7.33-7.32

(d, -O-CH=CH, 1H), 6.35-6.34 (m, -CH=CH-, 1H), 6.30-6.29 (d, -C=CH-, 1H), 4.75

(s, -N-CH2-, 2H). 13C NMR (DMSO-d6, 100MHz, ppm): 163.1, 148.0, 142.7, 129.5,

110.5, 109.4, 35.6.

3,4-diiodo-1-(furan-2-ylmethyl)-1H-pyrrole-2,5-dione (Compound 5). A

solution of compound 4 (3.723 g, 11.18 mmol) and potassium iodide (7.427 g, 44.74

mmol) in 50 mL acetic acid was heated at 120 ºC and stirred for 5 h. After removal of

acetic acid, the crude residue was separated by column chromatography on silica gel

(hexane/ethyl acetate = 10/1) to give compound 5 as a light yellow solid (3.03 g, 63.2

%). 1H NMR (DMSO-d6, 400MHz, ppm): 7.27-7.20 (d, -O-CH=, 1H), 6.28 (m, -

CH=CH-, 1H), 6.24-6.23 (d, -C=CH-, 1H), 4.72 (s, -N-CH2-, 2H). 13C NMR (DMSO-

d6, 100MHz, ppm): 166.6, 148.9, 142.7, 119.2, 110.6, 108.3, 35.7. HRMS (ESI): m/z

cald. for C9H5I2NO3Na (M+Na)+: 451.8256; found: 451.8254.

Electronic Supplementary Material (ESI) for Journal of Materials Chemistry B.This journal is © The Royal Society of Chemistry 2015

Sc

N

O

O

I

I

N

O

OO

+O

7NO O

N

O

ON NO

NNN

O

O

8

(PTAD)

N

O

O

I

I

N

O

O

9

5

OEtOEt

OEt

N

O

O

EtO

OEtOEt

EtO

OEtOEt

10

heme S2. Synthesis of fluorescent enediyne compounds.

ppm (t1)4.505.005.506.006.507.007.508.00

0

5000

7.3317.3297.327

6.3476.3396.3046.2996.2966.291

4.756

1.00

0.97

0.94

2.00

N

O

O

Br

BrO

Figure S1. 1H NMR spectrum of compound 4

ppm (t1)50100150

0

5000

10000

163.111

148.041

142.749

129.495

110.533

109.406

35.639

N

O

O

Br

BrO

Figure S2. 13C NMR spectrum of compound 4

ppm (t1)4.505.005.506.006.507.007.508.00

0

1000

2000

3000

4000

7.567

6.3926.3846.3806.3606.352

4.650

2.00

2.05

0.95

N

I

I

O

O

O

Figure S3. 1H NMR spectrum of compound 5

ppm (t1)50100150

0

1000

2000

3000

4000

5000

6000

166.623

148.873

142.708

119.156

110.543108.298

35.696N

I

I

O

O

O

Figure S4. 13C NMR spectrum of compound 5

ppm (t1)1.02.03.04.05.06.07.0

0

2500

5000

7500

10000

7.265

6.2676.2596.2406.2366.228

4.646

3.6913.6743.6563.638

1.2031.1851.167

12.9

3

2.00

1.01

0.98

1.06

23.7

7

N

O

O

O

OO

O

OO

O

Figure S5. 1H NMR spectrum of enediyne 1

ppm (t1)255075100125150

0

2500

5000

7500

10000

165.158

148.166

142.738

128.826

110.495

109.205109.061103.714

72.806

59.419

35.149

14.829

N

O

OO

O

OO

O

OO

Figure S6. 13C NMR spectrum of enediyne 1

ppm (t1)4.505.005.506.006.507.007.508.00

0

5000

100007.6757.6587.6047.5847.5667.5597.5497.5377.5187.501

6.418

4.688

2.02

4.08

1.03

5.88

2.00

N

O

OO

Figure S7. 1H NMR spectrum of enediyne 7

ppm (t1)5075100125150175

0

1000

2000

3000

4000

5000

6000

7000

8000165.763

148.736

142.737

132.085

130.911129.132127.304120.252

110.579108.700108.286

80.295

34.970

N

O

OO

Figure S8. 13C NMR spectrum of enediyne 7

3470 3480 3490 3500 3510 3520 3530 3540 3550 3560

Magnetic Field (G)

Figure S9. EPR spectrum of DA-product (enediyne 2) in CHCl3. Enediyne 2 was

dissolved in CHCl3 with the addition of one drop of TFA.

0.6 1.3 2.5 5 10 20 400

20

40

60

80

100

Cytotoxicity of DA-product 2 on A549 cell

Concentration(µM)

Cell

viab

ility

(% o

f con

trol

)

24h 48h 72h

Figure S10.Effect of DA-product (enediyne 2) on cell viability of tumor model cell

lines A549 cells

1.5 3.1 6.25 12.5 25 50 100 2000

20

40

60

80

100

Concentration(µM)

Cell

viab

ility

(% o

f con

trol

)

48 h L-02 A549

Figure S11. Effect of enediyne 1 on cell viability of tumor model cell lines A549 cells

and normal model cell lines L-02 cells. Cells were incubated with enediyne 1 for 48 h

and cell viability was determined and analyzed by MTT assay.

1.5 3.1 6.25 12.5 25 50 100 2000

20

40

60

80

100

Concentration(µM)

Cell

viab

ility

(% o

f con

trol

)

72 h L-02 A549

Figure S12. Effect of enediyne 1 on cell viability of tumor model cell lines A549 cells

and normal model cell lines L-02 cells. Cells were incubated with enediyne 1 for 72 h

and cell viability was determined and analyzed by MTT assay.

24 48 720

20

40

60

80

100

Effect of acetone on cells

Time (h)

Cell

viab

ility

(% o

f con

trol

)

A549cell L-02cell

Figure S13. Effects of 0.1% acetone on cell viability of tumor and normal cells. The

cells were incubated with acetone for 24 h, 48 h, and 72 h respectively. Cell viability

was determined and analyzed by MTT assay.

Figure. S14. Cellular distribution of enediyne 7 entering in A549 cell at 12 h observed

by CLSM. Blue-nucleus; green-enediyne; red-lysosome.

Figure. S15. Cellular distribution of enediyne 8 in A549 cell at 12 h observed by

CLSM. Blue-nucleus; green-enediyne; red-lysosome.

Figure. S16. Cellular distribution of enediyne 10 entering in A549 cell at 12 h

observed by CLSM. Blue-nucleus; green-enediyne; red-lysosome.