Journal of Synthetic rganic Chemistry SUPPORTING INFORMATIONSYNTHESIS

Thieme

Supporting Information for DOI: 10.1055/s-0033-1340556

© Georg Thieme Verlag KG Stuttgart · New York 2013

Strategies for Large-Scale Synthesis of

Coelenterazine for in Vivo Applications

Tej B. Shrestha1,2*, Deryl L. Troyer2, Stefan H. Bossmann1*

TS-6-17-34gscale

9.5 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 0.5 0Chemical Shif t (ppm)

2.001.02

CHLOROFORM-d

5.05

7.27

8.05

N

NH2N

BrBr

TS-4-63-fr1-2-13C

192 184 176 168 160 152 144 136 128 120 112 104 96 88 80 72 64 56 48 40 32 24 16 8 0Chemical Shif t (ppm)

CHLOROFORM-d

76.6

977

.00

77.3

2

123.

6212

3.90

143.

12

151.

85

N

NH2N

BrBr

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

Chloroform-d

152.

33

142.

6414

1.92 13

5.90

129.

2912

8.60

127.

5012

6.43

77.5

577

.23

76.9

2

40.9

2

TS-4-80-fr14-28

9 8 7 6 5 4 3 2 1 0Chemical Shift (ppm)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Nor

mal

ized

Inte

nsity

2.202.025.940.99

CHLOROFORM-d8.00

7.31

7.29 7.

217.

20

4.49

4.06

)

BrN

NH2N

BrN

NH2N

TS-4-80-fr14-28

8.5 8.0 7.5 7.0Chemical Shift (ppm)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Nor

mal

ized

Inte

nsity

5.940.99

CHLOROFORM-d8.00

7.31

7.29

7.27

7.25

7.22

7.21

7.20

TS-4-85

9 8 7 6 5 4 3 2 1 0Chemical Shift (ppm)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Nor

mal

ized

Inte

nsity

6.009.211.941.931.906.042.010.92

CHLOROFORM-d

8.34 7.84

7.82

7.33 7.

32 7.28

7.27

6.95

6.93

4.37

4.19

1.02

0.24

TS-4-85-13C

200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10Chemical Shif t (ppm)

CHLOROFORM-d-4

.38

18.2

6

25.7

0

41.2

7

76.6

977.0

077

.31

120.

48

126.

9412

8.55

128.

9513

0.55

136.

8113

6.94

140.

4114

2.69

151.

22

155.

97

N

NH2N

OTBDMS

TS-4-85

8.5 8.0 7.5 7.0 6.5Chemical Shift (ppm)

0

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

No

rma

lize

d I

nte

nsi

ty

1.906.042.010.92

CHLOROFORM-d

8.3

4

7.8

47

.82

7.3

3 7.3

27

.28

7.2

7

6.9

56

.93

N

NH2N

OTBDMS

TS-6-18-2highvaccum

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 0.5 0 -0.5Chemical Shift (ppm)

5.959.342.022.01

CHLOROFORM-d

0.19

0.97

0.98

6.716.

73

7.27

7.31

7.34

Br

OTBDMS

TS-4-82-13C

220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Nor

mal

ized

Inte

nsity

155.

06

132.

51

122.

12

113.

83

77.5

577

.23

76.9

1

25.8

6

18.4

2

-4.2

6

Br

OTBDMS

TS-6-8-3

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 0.5 0Chemical Shif t (ppm)

2.052.092.002.274.30

CHLOROFORM-d

4.58

5.08

6.966.

987.

317.

34

7.43

TS-5-15new13C

200 192 184 176 168 160 152 144 136 128 120 112 104 96 88 80 72 64 56 48 40 32 24 16 8 0Chemical Shif t (ppm)

CHLOROFORM-d

46.2

5

70.0

5

76.6

877

.00

77.3

2

115.

05

127.

4312

8.6113

0.06

136.

74

158.

87

OBn

Cl

OBn

Cl

TS-5-15new13C

143 142 141 140 139 138 137 136 135 134 133 132 131 130 129 128 127 126 125 124 123Chemical Shif t (ppm)

12

7.4

3

12

8.0

3

12

8.6

1

12

9.9

61

30

.06

13

6.7

4

TS-6-8-3

8.3 8.2 8.1 8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1Chemical Shif t (ppm)

2.002.274.30

CHLOROFORM-d

6.966.

98

7.31

7.34

7.407.

43

TS-6-31

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 0.5Chemical Shift (ppm)

6.002.032.002.010.922.032.012.014.98

CHLOROFORM-d

OOEt

OEt

BnO

TS-6-31-13C

220 210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 -20Chemical Shift (ppm)

CHLOROFORM-d

15.1

6

42.8

0

63.3

4

70.0

0

76.6

977

.00

77.3

1

102.

26

114.

88

126.

0012

7.45

130.

76

137.

03

157.

75

203.

50

OOEt

OEt

BnO

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

Chloroform-d

204.

04

154.

81

131.

1212

5.85

115.

64

102.

40

77.5

477

.23

76.9

1

63.6

0

43.0

7

15.3

6

TS-5-23-columndry

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 0Chemical Shift (ppm)

6.461.971.992.081.001.002.012.00

OOEt

OEt

HO

OOEt

OEt

HO

NH

NN

OH

HOO

13 12 11 10 9 8 7 6 5 4 3 2 1 0 -1 -2Chemical Shift (ppm)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Nor

mal

ized

Inte

nsity

2.001.681.99

METHANOL-d4

METHANOL-d4

7.13

27.

029

6.92

96.

806

6.79

16.

533

6.52

76.

511

6.34

66.

331

4.50

74.

044

3.71

83.

303

2.96

32.

959

8 11 1

N

NH2N

OTBDMS8

HO O

OEt

OEt

11

+HCl / ethanol, reflux

N

NN

OTBDMS

HO

OEtOEt

N

NN

OTBDMS

HO

OEtH

H

- H2O

OEt

ring closure

1) hydrolysis2) H-shift

NH

N

OH

N

OHO

1

coelenterazine