18F - MDPI

Supporting Information

A practical route to clinically relevant PET probes from nucleophilic [18F]fluoride and commercially available arylstannyl precursors

Fadi Zarrad1,2, Boris D. Zlatopolskiy1,2,3,*, Philipp Krapf1,2, Johannes Zischler1,2 and Bernd Neumaier1,2,3,* 1

Institute of Neuroscience and Medicine, Nuclear chemistry (INM-5), Forschungszentrum Jülich, Jülich, Germany. [email protected] (F.Z.), [email protected] (P.K.), [email protected] (J.Z.), b.neumaier@fz-juelich (B.N.) 2 Institute of Radiochemistry and Experimental Molecular Imaging, University Clinic Cologne, Cologne, Germany. [email protected] (B.D.Z.) 3 Max Planck Institute for Metabolism Research, Cologne, Germany * Correspondence: [email protected] ; Tel.: +49 221 478 82842; [email protected]; Tel.: +49 2461 614141

Contents NMR ................................................................................................................................. 3 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-bromophenyl)-3-hydroxyprop-2-en-1-one ............ 3 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one: ........... 5 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole................................... 7 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole: .................................. 8 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-(trimethylstannyl)phenyl)-1H-pyrazole .............. 11 Methyl 4-fluorobenzoate............................................................................................. 13 Methyl 4-(trimethylstannyl)benzoate ......................................................................... 14 3-(Trimethylstannyl)benzaldehyde ............................................................................. 15 (2-Methoxyphenyl)trimethylstannane ........................................................................ 16 (3-Methoxyphenyl)trimethylstannane ........................................................................ 17 (4-Methoxyphenyl)trimethylstannane ........................................................................ 17 tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5methoxy-2-(trimethylstannyl)phenyl}propanoate

[Boc2-4-Boc-3-Me-6-

(SnMe3)DOPA-OtBu] ................................................................................................. 18 Ethyl

(S)-3-{4,5-bis[(tert-butoxycarbonyl)oxy]-2-(trimethylstannyl)phenyl}-2-

[bis(tert-butoxycarbonyl)amino]propanoate [Boc2-6-(SnMe3)DOPA(Boc)2-OEt]: ... 20 Ethyl

(S)-2-[bis(tert-butoxycarbonyl)amino]-3-{5-[(tert-butoxycarbonyl)oxy]-2-

(trimethylstannyl)phenyl}propanoate [Boc-6-(SnMe3)mTyr(Boc)-OEt] ................... 22 Ethyl

(S)-2-[bis(tert-butoxycarbonyl)amino]-3-[4-(tert-butoxycarbonyl)oxy]-2-

(trimethylstannyl)phenylpropanoate [Boc2-2-(SnMe3)Tyr(Boc)-OEt]....................... 24 Radiochemistry ............................................................................................................... 26 Optimization of 18F-fluorodestannylation ................................................................... 26 Optimization of

18

F-fluorodestannylation of amino acid derivatives precursors using

N-mono and N,N-diBoc protected precursors of [18F]OMFD .................................... 31 Chromatograms ............................................................................................................... 37 Chromatogramms of the automated synthesis ............................................................ 41

NMR 5.89 2.00

0.92

1.01

1.03 1.04 2.05 1.11 1.09

7.91 7.91 7.90 7.74 7.73 7.72 7.70 7.69 7.68 7.50 7.49 7.49 7.48 7.46 7.45 7.42 7.41 7.30 7.29 7.23 7.19 7.15 6.74 6.70 6.57

3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-bromophenyl)-3-hydroxyprop-2-en-1-one

92.48

101.61

107.89 106.85

125.19 122.91 122.42

129.91 129.50

134.57

147.91

151.34

Figure S 1:1H-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-bromophenyl)-3-hydroxyprop-2-en-1-one

Figure S 2: 13C-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-bromophenyl)-3-hydroxyprop-2-en-1-one

OH O O

Br

O

Figure S 3: MS of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-bromophenyl)-3-hydroxyprop-2-en-1-one

92.88

102.08

108.41 107.41

114.28 113.82

123.28 122.76 122.70 119.40 118.98

130.45 130.29 130.02

148.44

151.79

160.59

165.36

182.20

186.52

2.26

0.98 2.15 2.25 1.26 1.07 1.00

0.87

7.76 7.75 7.72 7.71 7.69 7.67 7.67 7.64 7.63 7.62 7.59 7.58 7.51 7.47 7.46 7.44 7.43 7.40 7.28 7.27 7.24 7.24 7.23 7.22 7.20 7.20 7.19 6.92 6.88 6.71 6.07

16.80

3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one:

Figure S 4: 1H-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one

Figure S 5: 13C-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one

-111.97

Figure S 6: 19F-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one

OH O O

F

O

Figure S 7 MS of 3-(Benzo[d][1,3]dioxol-5-yl)-1-(3-fluorophenyl)-3-hydroxyprop-2-en-1-one

5.96 2.00

6.91 6.87 1.06

4.10

1.09

1.08

1.06

7.97 7.81 7.77 7.51 7.47 7.38 7.34 7.29

3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole

Figure S 9: 13C-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole

12.33

109.56 106.84 102.31 101.64

132.67 131.93 131.75 129.01 125.66 123.08 122.73 121.20

148.94 148.55 147.18 146.64

Figure S 8: 1H-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole

N NH

Br

O O

Figure S 10: MS of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole

5.94 2.00

2.15 1.07 3.10 1.24 1.14

7.60 7.42 7.39 7.31 7.25 7.16 7.11 6.87 6.83

3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole:

Figure S 11: 1H-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole

102.28 101.65

113.53 113.07 109.48 106.79

122.78 122.61 121.23

131.98 131.80 131.59 131.42

148.95 148.50 147.24 146.84 146.78

160.65

165.50

-112.07

Figure S 12: 13C-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole

Figure S 13: 13F-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole

Figure S 14: MS of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-fluorophenyl)-1H-pyrazole

-9.29

108.59 105.60 101.12 99.30

118.82

132.25 128.28 125.03

147.74 146.86

3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-(trimethylstannyl)phenyl)-1H-pyrazole

13.20

7.93 7.74 7.42 7.39 7.35 7.12 7.01 6.97

6.06

0.31

0.98

0.90 1.13 4.08 1.03 1.04

2.00

9.02

Figure S 15: 1H-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-(trimethylstannyl)phenyl)-1H-pyrazole

Figure S 16: 13C-NMR of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-(trimethylstannyl)phenyl)-1H-pyrazole

Figure S 17: MS of 3-(Benzo[d][1,3]dioxol-5-yl)-5-(3-(trimethylstannyl)phenyl)-1H-pyrazole

Figure S 19: 13C-NMR of Methyl 4-fluorobenzoate 52.31

115.85 115.42

132.34 132.16 126.58 126.52

168.41 166.27 163.36

3.00

2.00

2.01

3.92

8.11 8.10 8.09 8.07 8.06 8.06 8.05 8.03 8.02 8.01 7.17 7.15 7.14 7.12 7.11 7.11 7.10 7.08 7.07 7.05

Methyl 4-fluorobenzoate

Figure S 18: 1H-NMR of Methyl 4-fluorobenzoate

-105.79

Figure S 20: 19F-NMR of Methyl 4-fluorobenzoate

3.92

0.33

3.06

9.05

2.10

2.00

8.02 8.00 8.00 7.99 7.98 7.98 7.97 7.96 7.96 7.95 7.94 7.71 7.71 7.67 7.61 7.60 7.60 7.57 7.57 7.57 7.50 7.46

Methyl 4-(trimethylstannyl)benzoate

Figure S 21: 1H-NMR of Methyl 4-(trimethylstannyl)benzoate

Figure S 23: 1H-NMR of 3-(Trimethylstannyl)benzaldehyde

9.06

1.13

2.05

1.05

1.00

0.37

8.13 8.02 7.91 7.86 7.85 7.84 7.82 7.81 7.80 7.80 7.79 7.77 7.76 7.76 7.76 7.66 7.56 7.52 7.48

10.05

Figure S 22: 13C-NMR of Methyl 4-(trimethylstannyl)benzoate

3-(Trimethylstannyl)benzaldehyde

-9.50

52.08

135.82 135.82 129.86 128.55 128.55

149.69

167.48

-9.33

143.88 142.05 137.09 135.75 129.87 128.61

193.18

Figure S 24: 13C-NMR of 3-(Trimethylstannyl)benzaldehyde

3.80

0.27

3.00

8.95

1.05

1.06

1.98

7.41 7.40 7.37 7.36 7.33 7.33 7.29 7.29 7.02 7.01 6.98 6.95 6.94 6.90 6.86 6.82 6.77

(2-Methoxyphenyl)trimethylstannane

Figure S 25: 1H-NMR of (2-Methoxyphenyl)trimethylstannane

3.82

0.28

3.00

8.60

2.01

2.00

7.55 7.51 7.45 7.44 7.44 7.43 7.41 7.40 7.40 7.39 7.33 7.29 6.97 6.96 6.95 6.95 6.94 6.93 6.92 6.91

Figure S 27: 1H-NMR of (4-Methoxyphenyl)trimethylstannane 9.00

3.01

1.13

2.03

1.11

0.33

3.85

7.36 7.36 7.33 7.33 7.32 7.29 7.13 7.12 7.12 7.09 7.08 7.08 6.92 6.92 6.91 6.90 6.88 6.87 6.87 6.86

(3-Methoxyphenyl)trimethylstannane

Figure S 26: 1H-NMR of (3-Methoxyphenyl)trimethylstannane

(4-Methoxyphenyl)trimethylstannane

tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5methoxy-2-(trimethylstannyl)phenyl}propanoate

[Boc2-4-Boc-3-Me-6-

0.30

1.53 1.48 1.38

9.00

9.13 9.31 18.05

3.52 3.49 3.47 3.45 3.42

4.92 4.89 4.87 4.84 1.07

2.42

5.29

6.79 6.75 6.71 1.13

3.10

7.09 0.86

(SnMe3)DOPA-OtBu]

-8.22

28.13 28.01 27.77

55.74

60.52

83.26 82.84 81.86

114.13

129.41

143.78

152.27

169.01

Figure S 28: 1H-NMR of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5-methoxy2-(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Figure S 29: 13C-NMR of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5methoxy-2-(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Figure S 30: MS of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5-methoxy-2(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Ethyl

(S)-3-{4,5-bis[(tert-butoxycarbonyl)oxy]-2-(trimethylstannyl)phenyl}-2-

0.33

2.00

8.88

3.43 3.38 3.36 3.32 3.24

2.01

1.53 1.51 1.38 1.30 1.27 1.24

4.27 4.25 4.23 4.21 4.20 4.18 4.16 4.14

18.62 18.19 3.70

5.06 5.04 5.01 4.99 1.00

1.00

0.98

7.23 7.07 7.03 6.99

[bis(tert-butoxycarbonyl)amino]propanoate [Boc2-6-(SnMe3)DOPA(Boc)2-OEt]:

-8.03

14.26

29.80 27.96 27.72

37.93

61.57 59.48

83.54 83.45 83.19

123.94

130.08

143.22 142.58 141.37 140.61

152.04 150.87 150.72

170.17

Figure S 31: 1H-NMR of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5-methoxy2-(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Figure S 32: 13C-NMR of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5methoxy-2-(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Figure S 33: MS of tert-Butyl (S)-2-(bis(tert-butoxycarbonyl)amino)-3-{4-[(tert-butoxycarbonyl)oxy]-5-methoxy-2(trimethylstannyl)phenyl}propanoate [Boc2-4-Boc-3-Me-6-(SnMe3)DOPA-OtBu]

Ethyl

(S)-2-[bis(tert-butoxycarbonyl)amino]-3-{5-[(tert-butoxycarbonyl)oxy]-2-

0.32

3.49 3.47 3.42 3.39 3.38 3.33 3.26 2.18

1.53 1.37 1.28

4.28 4.26 4.24 4.23 4.20 4.19 4.17 4.16 2.21

9.71

10.02 18.47 3.17

5.05 5.03 5.00 4.98 1.00

1.14 1.10

1.02

7.41 7.37 7.33 7.02 7.01 6.98 6.97 6.92 6.91

(trimethylstannyl)phenyl}propanoate [Boc-6-(SnMe3)mTyr(Boc)-OEt]

-8.10

14.28

29.82 27.96

38.42

61.58 59.69

83.34 83.09

118.86

122.32

140.06 137.19

151.98 151.90 151.72 146.34

170.21

Figure S 34: 1H-NMR of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-{5-[(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenyl}propanoate [Boc-6-(SnMe3)mTyr(Boc)-OEt]

Figure S 35: 13C-NMR of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-{5-[(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenyl}propanoate [Boc-6-(SnMe3)mTyr(Boc)-OEt]

Figure S 36: MS of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-{5-[(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenyl}propanoate [Boc-6-(SnMe3)mTyr(Boc)-OEt]

Ethyl

(S)-2-[bis(tert-butoxycarbonyl)amino]-3-[4-(tert-butoxycarbonyl)oxy]-2-

9.22

0.01

0.33

1.54 1.36 1.27 9.06 18.05 3.03

3.37 3.36 3.34 3.31 2.12

2.09

4.27 4.26 4.23 4.22 4.19 4.19 4.16 4.15

5.01 4.99 4.96 4.94 1.00

0.96 2.12

7.18 7.16 7.10 7.06 7.04 7.03 7.00 6.99

(trimethylstannyl)phenylpropanoate [Boc2-2-(SnMe3)Tyr(Boc)-OEt]

-8.10

14.26

29.79 27.97 27.80

37.87

61.54 59.89

83.39 83.11

121.39

130.20 128.49

151.95 151.81 149.48 144.66 142.01

170.19

Figure S 37: 1H-NMR of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-[4-(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenylpropanoate [Boc2-2-(SnMe3)Tyr(Boc)-OEt]

Figure S 38: 13C-NMR of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-[4-(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenylpropanoate [Boc2-2-(SnMe3)Tyr(Boc)-OEt]

Figure S 39: MS of ethyl (S)-2-[bis(tert-butoxycarbonyl)amino]-3-[4-(tert-butoxycarbonyl)oxy]-2(trimethylstannyl)phenylpropanoate [Boc2-2-(SnMe3)Tyr(Boc)-OEt]

Radiochemistry Optimization of 18F-fluorodestannylation

18

[µmol]

F-Recovery [%]

Et4NHCO3 Et4NOTf Et4NBF4 Et4NI

60

98

98

30

94

94

93

99

15

96

95

97

97

5

94

96

97

92

2.5

95

96

97

96

0.5

81

76

80

89

Table S 1: Recovery of 18F– from anion exchange resin with MeOH solutions of different tetramethylammonium salts

Et4NOTf Et4NBC KOTf/K222 Bu4POMs Rest on the cartridge [%] 18

F recovery [%] RCC [%]

Table S 2:

18F

recovery and RCCs of

[18F]FPh

19±1

18±2

9±2

14±4

72±8

71±1

81±5

76±2

69±4

63±9

9±1

60±11

using different salts in nBuOH

QMA-

Strata X-

Strata X-

Chromafix PS-

CO3

CO3

HCO3

HCO3

19±1

14±1

18±2

35±2

73±8

81±1

68±9

57±3

69±4

24±15

37±10

42±5

Rest on the cartridge [%] 18

F recovery [%] RCC [%]

Table S 3: Dependence of [18F]fluoride recovery and cartridge

18

18F-incorporation

yields on the type of an anion exchange

ROH

RCC [%]

MeOH

11±4

85±1

1±0

EtOH

45±8

83±1

5±1

TFE

0

84±2

4±2

nPrOH

62±6

77±4

11±2

iPrOH

70±3

72±5

16±3

nBuOH

72±8

73±8

19±1

sBuOH

66±7

47±6

37±7

tBuOH

80±4

10±1

78±1

nAmOH

75±10

68±4

18±5

nHexOH

55±2

64±4

21±4

F Recovery [%] Rest on cartridge [%]

Table S 4: Effect of alcohol on 18F-recovery and 18F-fluorodestannylation

Water content [µL/1 mL] RCC [%] 0

73±8

5

35±2

10

16±1

15

9±1

Table S 5: Effect of water on [18F]fluorodestannylation

nBuOH (%) RCC [%] 0

84±4

10

85±2

20

78±4

30

72±8

40

58±9

50

42±2

Table S 6: Dependency of RCC on alcohol content

Table S 7: Optimization of aprotic solvent

Solvent

RCC [%]

DMA

72±8

NMP

73±3

TMU

28±2

DMF

9±3

DMSO

7±2

Pyridine

0

NMF

0

tBuOH

0

Reaction temperature [°C] RCC [%] 80

34±2

90

45±11

100

72±8

110

65±5

120

60±5

Reaction time [min]

RCC [%]

5

76±4

10

72±8

15

72±9

20

75±2

25

60±1

Table S 8: Dependence of RCCs on temperature and on time

Precursor amount [µmol]

RCC [%]

60

73±8

40

78±1

30

70±4

20

63±2

10

44±2

Table S 9: Dependence of

18F-incorporation

rate on the precursor amount

Cu(OTf)2(py)4 [µmol]

RCC [%]

40

61±9

30

70±4

20

66±5

10

54±2

Table S 10: Dependence of 18F-incorporation rate on the Cu(py)4(OTf)2 amount

Compound

RCC [%]

HPLC conditions tR=4.87 min, column: SpeedRod, eluent:

72±4(SnMe3) 88±1(SnBu3)

0–2 min: 5% MeCN, 2–2.5 min: 5–20% MeCN, 2.5–6 min; column: SpeedRod, 20% MeCN, 6–7 min: 20–70% MeCN, 7–9 min: 70% MeCN

17±1

16±1

84±2

tR=6.35 min, column: SpeedRod, eluent: 25% MeCN, flow rate: 3 mL/min tR=4.32 min, column: SpeedRod, eluent: 25% MeCN, flow rate: 1.5 mL/min tR=6.03 min, column: SpeedRod, eluent: 25% MeCN, flow rate: 1.5 mL/min

59±3(SnMe3)

tR=5.12 min, column: SpeedRod, eluent:

44±2(SnBu3)

25% MeCN, flow rate: 1.5 mL/min

34±3

tR=4.92 min, column: SpeedRod, eluent: 25% MeCN, flow rate: 1.5 mL/min

tR=7.3 min, column: Gemini 62±8

250×4.6 mm, eluent: 50% MeCN(0.1% TFA), flow rate: 2 mL/min

Table S 11: Substrate scope of the improved protocol for [18F]fluorodestannylation.

Optimization of 18F-fluorodestannylation of amino acid derivatives precursors using N-mono and N,N-diBoc protected precursors of [18F]OMFD

Cu(OTf)2(py)4:precursor [eq.] RCC [%] 1:1

22

1.5:1

24

2:1

27

3:1

25

Table S 12: Cu(OTf)2(py)4 to precursor ratio

Solvent

RCC [%] OMFD di-Boc OMFD tri-Boc

DMA

6

78

n-BuOH:DMA (1:9)

20

74

n-BuOH:DMA (3:7)

22

66

Table S 13: Solvent optimization

Compound

RCC

HPLC conditions

[%]

tR = 4.1 min, column: ProntoSil, eluent: 4% EtOH 37

in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 5.88 min, column: ProntoSil, eluent: 4%

55

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 5.66 min, column: ProntoSil, eluent: 4%

60

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 7.05 min, column: ProntoSil, eluent: 4%

78

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min

Table S 14: Manual synthesis of radiolabeled amino acids

Figure S 40: Flow scheme for the automated radiosynthesis of [18F]OMFD, 2-[18F]FTyr, 6-[18F]FMT and 6[18F]FDOPA. A: MeOH (2 mL); B: TEAOTf (5 mg, 18 µmol) in MeOH (700 µL); C: Cu(py)4(OTf)2 (40.7 mg, 60 µmol)

and radiolabeling precursor (30 µmol) in DMA (1 mL); D: synthetic air supply; E: H2O (1 mL); F: CH2Cl2 (2 mL); G: H2O (9 mL); H: 48% HBr (1 mL) (38% HCl in the case of [18F]OMFD); I: 45% NaOH (300 µL) and sodium phosphate buffer (3 mL, pH 4.5).

Compound

RCY [%]

HPLC conditions tR = 4.1 min, column: ProntoSil, eluent: 4% EtOH

54±5

in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 5.88 min, column: ProntoSil, eluent: 4%

42±2

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 5.66 min, column: ProntoSil, eluent: 4%

48±7

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min tR = 7.05 min, column: ProntoSil, eluent: 4%

32

EtOH in 25mM Na phosphate buffer (pH 2.5), flow rate: 1 mL/min

Table S 15: Automated radiosynthesis of [18F]OMFD, 2-[18F]FTyr, 6-[18F]FMT and 6-[18F]FDOPA

Molar activity of 6-[18F]FDOPA 300

UV 225 nm [mAU]

250 y = 47.191x 200 150 measured sample 100 50 0 0

1

2

3

4

5

6

[µg/mL]

Figure S 41: Calibration curve fort he determintation of the molar activity of 6-[18F]FDOPA

Molar activity of 6-[18F]FMT 60

UV 2230 nm [mAU]

50

y = 5.3991x

40

30 measured sample 20

10

0 0

2

4

6

8

10

12

[µmol]

Figure S 42: Calibration curve fort he determintation of the molar activity of 6-[18F]FMT

Molar activity of 2-[18F]FTyr

350 300

y = 61.541x

UV 230 nm [mAU]

250 200 measured sample

150 100 50 0 0

1

2

3

4

5

6

[µmol] Figure S 43: Calibration curve fort he determintation of the molar activity of 2-[18F]FTyr

Molar activity of [18F]OMFD 700 600

UV 230 nm[mAU]

500 y = 70.64x 400 300

measured sample

200 100 0 0

2

4

6

8

10

[µmol]

Figure S 44: Calibration curve fort he determintation of the molar activity of [18F]OMFD

6-[18F]DOPA

6-[18F]FMT

2-[18F]FTyr

6-[18F]OMFD

Sn [µg/batch]

0.324±0.016

0.066±0.003 0.292±0.008

0.050±0.001

Cu [µg/batch]

2.56±0.12

Table S 16: Determination of tin and copper content

0.072±3

4.22±0.22

0.208±0.012

Chromatograms

250

200

SnBu3 SnMe3

150

100

50

0 0

2

4

6

8

10

Time [min]

Figure S 45: Radiochromatogramm of [18F]fluorobenzene

80 70 60 50

Bu3Sn

40

Me3Sn

30 20 10 0 0

2

4

6

8

Time [min]

Figure S 46: Radiochromatogramm of 4-[18F]fluoroanisole

10

12

14

80

60

40

20

0 0

2

4

6

8

10

12

14

16

Time [min]

Figure S 47: Radiochromatogramm of 3-[18F]fluoroanisole

100

80

60

40

20

0 0

2

4

6

8

Time [min]

Figure S 48: Radiochromatogramm of 2-[18F]fluoroanisole

10

12

14

350 300 250 200 150 100 50 0 0

5

10

15

Time [min]

Figure S 49: Radiochromatogramm of 3-[18F]fluorobenzaldehyde

140 120 100 80 60 40 20 0 0

2

4

6

8

10

Time [min]

Figure S 50: Radiochromatogramm of methyl 4-[18F]fluorobenzoate

12

14

1000

800

600

400

200

0 0

2

4

6

8

10

12

14

16

18

20

22

Time [min]

Figure S 51: Radiochromatogramm of 3-(benzo[d][1,3]dioxol-5-yl)-5-(3-[18F]fluorophenyl)-1H-pyrazole

Chromatogramms of the automated synthesis

Figure S 52: Purification of 2-[18F]FTyr

2000

1000

radio-HPLC UV-trace (230 nm)

0 0

2

4

6

8

10

12

14

Retention time [min]

-28,5 -29,0 -29,5 -30,0 -30,5 -31,0 -31,5

magnified UV-trace (230 nm)

-32,0 -32,5 0

2

4

6

8

Retention time [min]

Figure S 53: 2-[18F]FTyr: quality control.

10

12

14

Figure S 54: Purification of 2-[18F]FMT

2000

1000

radio-HPLC UV-trace (230 nm)

0 0

2

4

6

8

10

12

Retention time [min]

35 30 25 20 15 10

magnified UV-trace (230 nm)

5 0 0

2

4

6

8

Retention time [min]

Figure S 55: 2-[18F]FMT: quality control

10

12

Figure S 56: Purification of 6-[18F]FDOPA using Synergi™ 4 µm Hydro-RP 80 Å, 250×10 mm

Figure S 57: Purification of 6-[18F]FDOPA using Synergi™ 4 µm Hydro-RP 80 Å, 150×21.2 mm

2000

radio-HPLC UV-trace (230 nm)

1000

0 0

1

2

3

4

5

6

7

8

Retention time [min]

8

6

4

2

0

magnified UV-trace (230 nm)

-2

-4 0

1

2

3

4

5

Retention time [min]

Figure S 58: 6-[18F]FDOPA: quality control

6

7

8

Figure S 59: Purification of [18F]OMFD

800 700 600 500 400 300

radio-HPLC UV-trace (230 nm

200 100 0 0

2

4

6

8

10

12

14

16

18

Retention time [min]

-8

-10

-12

-14

magnified UV-trace (230 nm) -16

0

2

4

6

8

10

12

Retention time [min]

Figure S 60: [18F]OMFD: quality control

14

16

18