supporting information - wiley-vch · s-4 hexa-tert-butyl...

Supporting Information

© Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2008

S-1

Synthetic Receptors for the Differentiation of Phosphorylated Peptides with

Nanomolar Affinities

Andreas Grauer, Alexander Riechers, Stefan Ritter, and Burkhard König

Institute for Organic Chemistry, University of Regensburg,

D-93040 Regensburg, Germany

S-2

Supporting Information

1. Synthesis of Receptors 3 - 8 .............................................3

2. Solid-phase peptide synthesis ........................................34

3. Binding studies ...............................................................35

S-3

1. Synthesis of Receptors 3 – 8

General

All reactions were performed under an inert atmosphere of N2 using standard

Schlenk techniques if not otherwise stated. A Varian Cary BIO 50 UV/VIS/NIR

Spectrometer was used. A 1 cm quartz cell was purchased from Hellma and Uvasol

solvents from Merck or Baker. IR spectra were recorded on a Bio-Rad FT-IR FTS

155 and a Bio-Rad FTS 2000 MX FT-IR using a Specac Golden Gate Mk II ATR

accessory where stated. NMR spectrometers used were: Bruker Avance 600 (1H:

600.1 MHz, 13C: 150.1 MHz, T = 300 K), Bruker Avance 400 (1H: 400.1 MHz, 13C:

100.6 MHz, T = 300 K) and Bruker Avance 300 (1H: 300.1 MHz, 13C: 75.5 MHz, T =

300 K). The chemical shifts are reported in d [ppm] relative to external standards

(solvent residual peak). The spectra were analyzed by first order, the coupling

constants are given in Hertz [Hz]. Characterization of the signals: s = singlet, d =

doublet, t = triplet, q = quartet, m = multiplet, bs = broad singlet, psq = pseudo

quintet, dd = double doublet, dt = double triplet, ddd = double double doublet.

Integration is determined as the relative number of atoms. Assignment of signals in 13C-spectra was determined with DEPT-technique (pulse angle: 135 °) and given as

(+) for CH3 or CH, (-) for CH2 and (Cquat) for quaternary C. Error of reported values:

chemical shift: 0.01 ppm for 1H-NMR, 0.1 ppm for 13C-NMR and 0.1 Hz for coupling

constants. The solvent used is reported for each spectrum. Mass spectra were

recorded on Varian CH-5 (EI), Finnigan MAT 95 (CI; FAB and FD) and Finnigan MAT

TSQ 7000 (ESI). Xenon served as the ionization gas for FAB. Melting Points were

determined on a Büchi SMP-20 melting point apparatus and are uncorrected. TLC

analyses were performed on silica gel 60 F-254 with a 0.2 mm layer thickness.

Detection was via UV light at 254 nm / 366 nm or by staining with ninhydrin in EtOH.

For preparative column-chromatography, Merck Geduran SI 60 silica gel was used.

Commercially available solvents of standard quality were used. If otherwise stated,

purification and drying was done according to accepted general procedures.[1]

The following compounds were synthesized according to literature known

procedures: complex 10,[2] compound 14,[3] benzyl 2-aminoethylcarbamate 35,[4]

bis(Boc)-2-methyl-2-thiopseudourea 38,[5] tert-butyl 2-aminoethylcarbamate 45.[6]

S-4

Hexa-tert-butyl 10,10'-(6-(3-azidopropylamino)-1,3,5-triazine-2,4-diyl)bis(1,4,7,10-

tetraazacyclododecane-1,4,7-tricarboxylate (17):

The chloride 14 (1.0 g, 0.95 mmol) was dissolved in 40 ml of dioxane. Following,

potassium carbonate (2.0 g, 14 mmol, 15 eq) and 3-azido-propylamine hydrochloride

(19) (0.65 g, 4.7 mmol, 5 eq) were added. The suspension was stirred at 90 °C under

an atmosphere of N2 for 3 days. The suspension was filtered and the residue was

washed thoroughly with EtOAc. After evaporation of the solvent in vacuum the

obtained brown oil was purified by column chromatography twice. First with

PE:EtOAc = 7:3 and then with DCM:MeOH = 98:2. This gave the azide 17 as a

colorless solid in a yield of 1.0 g (0.90 mmol, 95 %). Rf (EtOAc:PE = 1:1) = 0.74.

MP: 140 °C. -1H-NMR (300 MHz, CDCl3): δ = 1.37–1.39 (m, 54 H, Boc-CH3), 1.76 (tt, 3JH,H = 6.6 Hz, 2 H, CH2), 3.24–3.58 (m, 36 H, 2 CH2 + cyclen-CH2), 4.78 (bs, 1 H,

NH). -13C-NMR (75 MHz, CDCl3): δ = 28.4 (+, Boc-CH3), 28.5 (+, Boc-CH3), 29.2 (–, 1

C), 37.9 (–, 1 C), 49.0 (–, 1 C), 50.3 (–, 16 C, cyclen), 79.7 (Cquat, 4 C, Boc), 79.8

(Cquat, 2 C, Boc), 156.3 (Cquat, 6 C, urethane), 165.8 (Cquat, 1 C, triazine), 166.5 (Cquat, 2

C, triazine). -MS (ES, CH2Cl2/MeOH): m/z (%) = 1121.0 (100) [MH+]. -

EA (C52H93N15O12) calc.: C 55.75, H 8.37, N 18.75, found: C 56.02, H 8.60, N 18.48. -

UV/Vis (CH3CN): λ (lg ε) = 231 nm (4.559). -IR (KBr) [cm-1]: ν~ = 3402, 3255, 2976,

2932, 2138, 1680, 1539, 1479, 1411, 1367, 1249, 1178, 777. -MF: C52H93N15O12. -

MW = 1120.40 g/mol.

N N

NN

NN

N

BocBoc

BocN N

NN

Boc

BocBoc

NH

N3

S-5

Hexa-tert-butyl 10,10'-(6-(prop-2-ynylamino)-1,3,5-triazine-2,4-diyl)bis(1,4,7,10-

tetraazacyclododecane-1,4,7-tricarboxylate) (20):

The chloride 14 (0.82 g, 0.77 mmol) was dissolved in 25 ml of dioxane. Potassium

carbonate (0.53 g, 3.9 mmol, 5 eq) and propargylamine (21) (0.25 ml, 0.21 g,

3.9 mmol, 5 eq) were added. The suspension was stirred at 90 °C in the dark under

an atmosphere of N2 for 3 days. The suspension was filtered and the residue was

washed thoroughly with EtOAc. After evaporation of the solvent in vacuum the

obtained brown oil was purified by column chromatography twice. (PE:EtOAc = 7:3,

then DCM:MeOH = 98:2). This gave the alkyne 20 as a colorless solid in a yield of

0.78 g (0.73 mmol, 94 %). Rf (EtOAc:PE = 1:1) = 0.74.

MP: 149 °C. -1H-NMR (300 MHz, CDCl3): δ = 1.37–1.39 (m, 54 H, Boc-CH3), 2.09 (t, 4JH,H = 2.3 Hz, 1 H, CH), 3.22–3.58 (m, 32 H, cyclen-CH2), 4.07–4.10 (m, 2 H, CH2),

4.86 (bs, 1 H, NH). -13C-NMR (75 MHz, CDCl3): δ = 28.4 (+, Boc-CH3), 28.5 (+, Boc-

CH3), 30.4 (–, 1 C), 50.3 (–, 16 C, cyclen), 70.5 (+, 1 C, CH), 79.7 (Cquat, 4 C, Boc),

79.8 (Cquat, 2 C, Boc), 81.1 (Cquat, 1 C, alkyne), 156.3 (Cquat, 6 C, urethane), 165.4

(Cquat, 1 C, triazine), 166.1 (Cquat, 2 C, triazine). -MS (ES, CH2Cl2/MeOH): m/z (%) =

1075.8 (100) [MH+]. -EA (C52H90N12O12) calc.: C 58.08, H 8.44, N 15.63, found: C

57.76, H 8.80, N 15.45. -UV/Vis (CH3CN): λ (lg ε) = 231 nm (4.564). -IR (KBr) [cm-1]:

ν~ = 3402, 3314, 3255, 2976, 2932, 2102, 1680, 1539, 1479, 1411, 1367, 1249,

1178, 777. -MF: C52H90N12O12. -MW = 1075.36 g/mol.

N N

NN

NN

N

BocBoc

BocN N

NN

Boc

BocBoc

NH

S-6

(S)-Benzyl 2-(tert-butoxycarbonylamino)-5-oxo-5-(prop-2-ynylamino)pentanoate (22):

Boc-Glu-OBn (23) (1.0 g, 3.0 mmol) was dissolved in 2 ml of dry DMF and cooled in

an ice bath. EDC (0.63 ml, 0.55 g, 3.6 mmol, 1.2 eq), HOBt (0.48 g, 3.6 mmol, 1.2

eq), DIPEA (0.33 ml, 0.46 g, 3.6 mmol, 1.2 eq) and propargylamine (21) (0.21 ml,

179 mg, 3.26 mmol, 1.1 eq) were added. The ice bath was removed and the solution

was stirred in the dark at room temperature for 24 h. The solution was cooled to 0 °C

and water at 0 °C was added in order to precipitate the product. After filtration, the

white solid was dissolved in 50 ml DCM. The solution was washed with 30 ml water,

30 ml 1 M aqueous citric acid, 30 ml water and 30 ml brine. The organic layer was

dried with Na2SO4 and the solvent was removed in vacuum. This gave 1.1 g of 22

(2.9 mmol, 98 %) as a white crystalline solid in sufficient purity for the following

reaction steps. For analytical data the crude product was purified by column

chromatography with EtOAc:PE in a ratio of 2:3. This gave pure 22 in a yield of 1.1 g

(2.9 mmol, 96 %). Rf (EtOAc:PE = 1:1) = 0.37.

MP: 122 °C. -1H-NMR (300 MHz, CDCl3): δ = 1.42 (s, 9 H, Boc-CH3), 1.91–2.03 (m, 1

H, CH2), 2.11–2.19 (m, 1 H, CH2), 2.22 (t, 4JH,H = 2.5 Hz, 1 H, CH), 2.25–2.30 (m, 2

H, CH2), 3.98 (dd, 4JH,H = 2.5 Hz, 3J = 5.2 Hz, 2 H, propargyl-CH2), 4.27–4.34 (m, 1

H, C*H), 5.12 (d, 2J = 12.4 Hz, 1 H, Bn-CH2), 5.18 (d, 2JH,H = 12.4 Hz, 1 H, Bn-CH2),

5.63 (d, 3JH,H = 8.2 Hz, 1 H, NH), 6.85 (t, 3JH,H = 5.2 Hz, 1 H, NH), 7.31–7.36 (m, 5 H,

arom. CH). -13C-NMR (75 MHz, CDCl3): δ = 28.3 (+, 3 C, Boc-CH3), 29.0 (–, 1 C),

29.2 (–, 1 C), 32.3 (–, 1 C), 53.0 (+, C*H), 67.3 (–, 1 C, Bn), 71.5 (+, alkyne-CH), 79.5

(Cquat, alkyne), 80.3 (Cquat, Boc), 128.4 (+, 2 C, arom.), 128.6 (+, 1 C, arom.), 128.7

(+, 2 C, arom.), 135.2 (Cquat, 1 C, arom.), 155.8 (Cquat, urethane), 171.5 (Cquat), 172.1

(Cquat). -MS (CI, NH3): m/z (%) = 275.2 (100) [MH+ - C4H8 + CO2], 319.2 (65) [(MH+ -

C4H8], 336.1 (20) [M + NH4+ - C4H8], 375.2 (40) [MH+]. -EA (C20H26N2O5) calc.: C

64.16, H 7.00, N 7.48, found: C 63.85, H 7.03, N 7.40. -IR (KBr) [cm-1]: ν~ = 3344,

3313, 3266, 2970, 1736, 1690, 1645, 1535, 1451, 1283, 1165, 727. -MF:

C20H26N2O5. -FW = 374.44 g/mol.

O

O

NHBoc

O

NH

S-7

(S)-Benzyl 5-(3-azidopropylamino)-2-(tert-butoxycarbonylamino)-5-oxopentanoate

(24):

Boc-Glu-OBn (23) (0.90 g, 2.7 mmol) was dissolved in 2 ml of dry DMF and cooled in

an ice bath. EDC (0.57 ml, 0.50 g, 2.9 mmol, 1.2 eq), HOBt (0.44 g, 3.2 mmol, 1.2

eq), DIPEA (0.54 ml, 0.76 g, 5.9 mmol, 2.2 eq) and 3-azido-propylamine

hydrochloride (19) (0.40 g, 2.9 mmol, 1.1 eq) were added. The ice bath was removed

and the solution was stirred at room temperature for 24 h. Then the solution was

cooled to 0 °C and water at 0 °C was added. A pale yellow oil separated. The

aqueous solution was discarded. The oil was dissolved in 50 ml of DCM. The solution

was washed with 30 ml water, 30 ml 1 M aqueous citric acid, 30 ml water and 30 ml

brine. The organic layer was dried with Na2SO4 and the solvent was removed in

vacuum. This gave 1.1 g of 21 (2.6 mmol, 98 %) as a pale yellow crystalline solid in

sufficient purity for the following reaction steps. For analytical data the crude product

was purified by column chromatography with EtOAc:PE in a ratio of 2:3. This gave

pure 24 as white crystalline solid in a yield of 1.1 g (2.6 mmol, 96 %). Rf (EtOAc:PE =

1:1) = 0.37.

MP: 116 °C. -1H-NMR (300 MHz, CDCl3): δ = 1.42 (s, 9 H, Boc-CH3), 1.72–1.81 (m, 2

H, CH2), 1.85–1.98 (m, 1 H, CH2), 2.11–2.24 (m, 3 H, CH2), 3.27–3.36 (m, 4 H, CH2),

4.26–4.33 (m, 1 H, C*H), 5.11 (d, 2JH,H = 12.1 Hz, 1 H, Bn-CH2), 5.20 (d, 2JH,H = 12.1

Hz, 1 H, Bn-CH2), 5.35 (d, 3JH,H = 7.7 Hz, 1 H, NH), 6.28–6.33 (m, 1 H, NH), 7.32–

7.37 (m, 5 H, arom. CH). -13C-NMR (75.5 MHz, CDCl3): δ = 28.3 (+, 3 C, Boc-CH3),

28.8 (–, 1 C), 29.1 (–, 1 C), 32.6 (–, 1 C), 37.1 (–, 1 C), 49.3 (–, 1 C), 53.1 (+, C*H),

67.3 (–, 1 C, Bn), 80.3 (Cquat, Boc), 128.4 (+, 2 C, arom.), 128.6 (+, 1 C, arom.), 128.7

(+, 2 C, arom.), 135.2 (Cquat, 1 C, arom.), 155.9 (Cquat, 1 C, urethane), 172.1 (Cquat, 2

C, amide + ester). -MS (pos. ES, DCM/MeOH): m/z (%) = 320.1 (30) [M - C5H9O2-],

364.1 (40) [(MH+ - C4H8], 420.2 (100) [MH+]. -MS (neg. ES, DCM/MeOH): m/z (%) =

236.0 (35) [M - H+ - BnOH - tBuOH], 310.1 (75) [M - H+ - BnOH], 454.3 (20) [M + Cl-],

478.3 (100) [M + Ac-]. -EA (C20H29N5O5) calc.: C 57.27, H 6.97, N 16.70, found: C

57.29, H 7.08, N 16.65. -IR (KBr) [cm-1]: ν~ = 3344, 3028, 2970, 2148, 1734, 1688,

1647, 1535, 1455, 1283. -MF: C20H29N5O5. -MW = 419.48 g/mol.

O

O

NHBoc

O

NH

N3

S-8

(S)-1-(Benzyloxy)-1,5-dioxo-5-(prop-2-ynylamino)pentan-2-aminium chloride (25):

The amide 22 (1.0 g, 2.7 mmol) was dissolved in little amount of dry EtOAc and 18 ml

of a saturated solution of HCl in dry diethyl ether were added (1 ml per 0.15 mmol

Boc). The mixture is stirred at room temperature under an atmosphere of N2 for 15 h.

After 30 min the product began to separate as colorless oil. The solvent was removed

in vacuum. This gave 25 as a colorless hygroscopic solid in quantitative yield (0.83 g,

2.7 mmol).

1H-NMR (300 MHz, CD3CN): δ = 2.27–2.34 (m, 2 H, CH2), 2.47 (t, 4JH,H = 2.5 Hz, 1 H,

alkyne), 2.47–2.56 (m, 2 H, CH2), 3.86–3.88 (m, 4JH,H = 2.5 Hz, 2 H, propargyl-CH2),

4.26–4.31 (m, 1 H, C*H), 5.19 (d, 2JH,H = 12.5 Hz, 1 H, Bn-CH2), 5.23 (d, 2JH,H = 12.5

Hz, 1 H, Bn-CH2), 7.31–7.42 (m, 5 H, arom. CH), 7.87 (t, 3JH,H = 5.5 Hz, 1 H, NH),

8.82 (bs, 3 H, NH3+). -13C-NMR (75.5 MHz, CD3CN): δ = 26.8 (–, 1 C), 29.5 (–, 1 C),

32.2 (–, 1 C), 53.7 (+, C*H), 69.1 (–, 1 C, Bn), 72.5 (+, alkyne), 81.3 (Cquat, alkyne),

129.5 (+, 2 C, arom.), 129.6 (+, 1 C, arom.), 129.7 (+, 2 C, arom.), 136.2 (Cquat, 1 C,

arom.), 170.3 (Cquat), 173.2 (Cquat). -MS (ES, H2O): m/z (%) = 275.0 (100) [MH+]. -IR

(KBr) [cm-1]: ν~ = 3324, 3069, 3028, 2970, 2121, 1731, 1681, 1643, 1533, 1449. -MF:

C15H19N2O3Cl. -MW = 310.78 g/mol.

5-(3-Azidopropylamino)-1-(benzyloxy)-1,5-dioxopentan-2-aminium chloride (26):

The amide 24 (1.4 g, 3.3 mmol) was dissolved in a little amount of dry EtOAc and 22

ml of a saturated solution of HCl in dry diethyl ether were added (1 ml per 0.15 mmol

Boc). The mixture was stirred at room temperature under an atmosphere of N2 for 15

h. After 30 min the product began to separate as colorless oil. The solvent was

removed in vacuum. This gave 26 as a pale yellow hygroscopic solid in quantitative

yield (1.2 g, 3.3 mmol).

O

O

N+

O

NH

HHH Cl-

O

O

N+

O

NH

HHH

N3

Cl-

S-9

1H-NMR (300 MHz, CD3CN): δ = 1.69 (tt, 3JH,H = 6.7 Hz, 2 H, CH2), 2.33–2.42 (m, 2

H, CH2), 2.61–2.67 (m, 2 H, CH2), 3.16–3.21 (m, 2 H, CH2), 3.30 (t, 3JH,H = 6.7 Hz, 2

H, CH2), 4.31–4.37 (m, 1 H, C*H), 5.24 (s, 2 H, Bn-CH2), 7.29–7.44 (m, 5 H, arom.

CH), 8.21–8.27 (m, 1 H, NH), 8.89 (bs, 3 H, NH3+). -13C-NMR (75.5 MHz, CD3CN): δ

= 27.2 (–, 1 C), 29.2 (–, 1 C), 32.2 (–, 1 C), 38.1 (–, 1 C), 49.9 (–, 1 C), 53.8 (+, C*H),

69.1 (–, 1 C, Bn), 129.5 (+, 2 C, arom.), 129.6 (+, 1 C, arom.), 129.7 (+, 2 C, arom.),

136.2 (Cquat, 1 C, arom.), 170.4 (Cquat), 174.3 (Cquat). -MS (ES, MeCN): m/z (%) =

320.1 (100) [MH+], 639.4 (7) [(2 M + H+]. -IR (KBr) [cm-1]: ν~ = 3334, 3069, 3028,

2967, 2141, 2118, 1732, 1681, 1642, 1533, 1450. -MF: C15H22N5O3Cl. -

MW = 355.82 g/mol.

General Procedure 1 (GP 1) Alkylation of glutamines under acidic conditions:

The ammonium chloride was dissolved in acetonitrile (4 ml per mmol). Bromo-ethyl-

acetate (60 eq), NaF (9 eq), SiO2 (5 eq), phase transfer-catalysts TBABr und

18-crown-6 (each approx. 0.10 g) were added. The mixture was refluxed under an

atmosphere of N2 for 20 h. After cooling to room temperature potassium carbonate (1

eq) was added. After stirring the suspension at room temperature for 1 h and

filtration, the residue was washed thoroughly with EtOAc. The solvents were

evaporated at reduced pressure and the excess bromo-ethylacetate was removed in

vacuum. This gave the crude product as dark brown oil, which was purified by

column chromatography twice. First EtOAc:PE in a ration of 2:3 and then

DCM:MeOH 98:2.

(S)-Diethyl 2,2'-(1-(benzyloxy)-1,5-dioxo-5-(prop-2-ynylamino)pentan-2-ylazanediyl)

diacetate (18):

Synthesis followed GP 1 using 25 (1.6 g, 5.2 mmol), bromo-ethylacetate (34 ml, 52 g,

0.31 mol), NaF (2.0 g, 46 mmol), SiO2 (1.6 g, 26 mmol) and potassium carbonate

(0.71 g, 5.2 mmol). This gave 2.1 g of 18 (4.7 mmol, 92 %) as a colorless oil.

1H-NMR (600 MHz, CDCl3): δ = 1.17 (t, 3JH,H = 7.2 Hz, 6 H, CH3), 1.89 (dddd, 2JH,H =

14.9 Hz , 3JH,H = 5.5 Hz, 3JH,H = 5.8 Hz, 3JH,H = 11.6 Hz, 1 H, CH2), 2.06 (dddd, 2JH,H =

O

O

N

O

NH

OOO

O

S-10

14.9 Hz, 3JH,H = 4.5 Hz, 3JH,H = 5.7 Hz, 3JH,H = 10.1 Hz, 1 H, CH2), 2.09 (dd, 4JH,H =

2.6 Hz, 4JH,H = 2.6 Hz, 1 H, CH), 2.30 (ddd, 3JH,H = 5.5 Hz, 3JH,H = 5.8 Hz, 2JH,H =

13.9 Hz, 1 H, CH2), 2.41 (ddd, 3JH,H = 5.8 Hz, 3JH,H = 10.1 Hz, 2JH,H = 13.9 Hz, 1 H,

CH2), 3.40 (dd, 3JH,H = 11.6 Hz, 3JH,H = 4.5 Hz, 1 H, C*H), 3.52 (s, 4 H, CH2), 3.83

(ddd, 2JH,H = 17.5 Hz, 3JH,H = 5.0 Hz, 4JH,H = 2.6 Hz, 1 H, CH2), 4.00 (ddd, 2JH,H = 17.5

Hz, 3JH,H = 5.9 Hz, 4JH,H = 2.6 Hz, 1 H, CH2), 4.05 (q, 3JH,H = 7.2 Hz, 4 H, CH2), 5.01

(d, 2JH,H = 12.3 Hz, 1 H, CH2), 5.05 (d, 2JH,H = 12.3 Hz, 1 H, CH2), 6.88 (dd, 3JH,H =

5.0 Hz, 3JH,H = 6.0 Hz, 1 H, NH), 7.22–7.30 (m, 5 H, arom. CH). -13C-NMR

(150.9 MHz, CDCl3): δ = 14.1 (+, 2 C, CH3), 25.8 (–, 1 C, Glu), 28.8 (–, 1 C), 32.3 (–,

1 C, Glu), 53.0 (–, 2 C, N-CH2), 60.8 (–, 2 C, Et-ester-CH2), 64.1 (+, 1 C, C*H), 66.5

(–, 1 C, Bn-CH2), 70.8 (+, 1 C, alkyne), 80.2 (Cquat, 1 C, alkyne), 128.2 (+, 2 C,

arom.), 128.3 (+, 1 C, arom.), 128.6 (+, 2 C, arom.), 135.4 (Cquat, 1 C, arom.), 171.5

(Cquat, 2 C, Et-ester), 171.9 (Cquat, 1 C, Bn-ester), 172.5 (Cquat, 1 C, amide). -MS (ES,

DCM/MeOH): m/z (%) = 447.1 (100) [MH+]. -MS-HR (EI-MS): [M+.] (calc.) =

446.2053, [M+.] (found) = 446.2053 ± 0.43 ppm. -IR (KBr) [cm-1]: ν~ = 3344, 3313,

3266, 3030, 2970, 2119, 1742, 1736, 1643, 1533, 1450. -MF: C23H30N2O7. -MW =

446.50 g/mol.

Diethyl 2,2'-(5-(3-azidopropylamino)-1-(benzyloxy)-1,5-dioxopentan-2-

ylazanediyl)diacetate (27):

Synthesis followed GP 1 using 26 (0.63 g, 1.8 mmol), bromo-ethylacetate (12 ml,

17.8 g, 0.1 mol), NaF (0.67 g, 16 mmol), SiO2 (0.53 g, 8.9 mmol) and potassium

carbonate (0.25 g, 1.8 mmol, 1 eq). This gave 0.80 g of 27 (1.6 mmol, 92 %) as a

colorless oil.

Compound 27 is not stable at room temperature and decomposes slowly within a few

days turning yellow.

1H-NMR (300 MHz, CDCl3): δ = 1.16 (t, 3JH,H = 7.2 Hz, 6 H, CH3), 1.68 (tt, 3JH,H =

6.9 Hz, 2 H, CH2), 1.79–1.93 (m, 1 H, Glu-CH2), 1.99–2.10 (m, 1 H, Glu-CH2), 2.24–

4.42 (m, 2 H, Glu-CH2), 3.13–3.27 (m, 4 H, 2 CH2), 3.40 (dd, 3JH,H = 11.1 Hz, 3JH,H =

4.5 Hz, 1 H, C*H), 3.47 (d, 2JH,H = 18.1 Hz, 2 H, N-CH2), 3.53 (d, 2JH,H = 18.1 Hz, 2 H,

O

O

N

O

NH

OOO

O

N3

S-11

CH2), 4.05 (q, 3JH,H = 7.2 Hz, 4 H, CH2), 5.00 (d, 2JH,H = 12.4 Hz, 1 H, Bn-CH2), 5.05

(d, 2JH,H = 12.4 Hz, 1 H, Bn-CH2), 6.75 (t, 3JH,H = 5.9 Hz, 1 H, NH), 7.22–7.28 (m, 5 H,

arom. CH). -13C-NMR (75.5 MHz, CDCl3): δ = 14.1 (+, 2 C, CH3), 25.9 (–, 1 C, Glu),

28.8 (–, 1 C), 32.4 (–, 1 C, Glu), 36.8 (–, 1 C), 49.1 (–, 1 C, N3-CH2), 52.9 (–, 2 C, N-

CH2), 60.7 (–, 2 C, Et-ester-CH2), 64.1 (+, 1 C, C*H), 66.4 (–, 1 C, Bn-CH2), 128.2 (+,

2 C, arom.), 128.3 (+, 1 C, arom.), 128.5 (+, 2 C, arom.), 135.6 (Cquat, 1 C, arom.),

171.5 (Cquat, 2 C, Et-ester), 172.0 (Cquat, 1 C, Bn-ester), 172.9 (Cquat, 1 C, amide). -

MS (ES, DCM/MeOH): m/z (%) = 492.4 (100) [MH+], 983.8 (5) [2 M + H+] (5). -

EA (C23H33N5O7) calc.: C 56.20, H 6.77, N 14.25, found: C 55.86, H 6.61, N 14.07. -

IR (KBr) [cm-1]: ν~ = 3344, 3310, 3268, 3025, 2968, 2139, 1742, 1737, 1642, 1532,

1449. -MF: C23H33N5O7. - MW = 491.54 g/mol.

General procedure 2 (GP 2) Cu(I)-catalyzed Huisgen reaction:

Equimolar amounts of the alkyne and the azide compound were dissolved in a 1:2

mixture of tBuOH:EtOH (10 ml per mmol alkyne). A solution of sodium ascorbate

(2 eq) in aqueous acetate buffer (pH 5, c = 0.5 mol/L) was added (4 ml buffer per

mmol alkyne). Finally an aqueous solution of CuSO4 x 5 H2O (c = 1 mol/L, 1 eq) was

added. The color of the mixture turned brown immediately and then slowly yellow.

The mixture was stirred at room temperature for 5 h and diluted with 50 ml of water

(an orange precipitate occurs). The suspension was extracted with EtOAc (3 x 50

ml). The organic layers were combined, together with the orange precipitate and

washed thoroughly with a saturated aqueous solution of EDTA di-sodium salt until

the orange solid dissolved completely. The blue aqueous layer was discarded and

the organic layer was dried with Na2SO4. Filtration and removal of the solvent in

vacuum yielded the crude product as a white solid, which was purified by column

chromatography. (first run: DCM:MeOH; second run: EtOAc:EtOH, gradient 98:2 to

96:4 for both mixtures.

S-12

(S)-Hexa-tert-butyl 10,10'-(6-(3-(4-((5-(benzyloxy)-4-(bis(2-ethoxy-2-oxoethyl)amino)-

5-oxopentanamido)methyl)-1H-1,2,3-triazol-1-yl)propylamino)-1,3,5-triazine-2,4-

diyl)bis(1,4,7,10-tetraazacyclododecane-1,4,7-tricarboxylate) (28):

Synthesis followed GP 2 using 17 (1.1 g, 0.97 mmol), 18 (0.43 g, 0.97 mmol), sodium

ascorbate (0.39 mg, 2.0 mmol) and CuSO4 x 5 H2O (0.24 g, 0.97 mmol). This gave

1.4 g of 28 (0.91 mmol, 94 %) as a colorless solid. Rf (DCM:MeOH = 95:5) = 0.30.

MP: 110 °C. - 1H-NMR (300 MHz, CDCl3): δ = 1.16 (t, 3JH,H = 7.2 Hz, 6 H, ester-CH3),

1.37 (bs, 36 H, Boc-CH3), 1.39 (bs, 18 H, Boc-CH3), 1.83–1.95 (m, 1 H, C2H2), 2.00–

2.10 (m, 3 H, C6H2 + C2H2), 2.28–2.47 (m, 2 H, C3H2), 3.22–3.62 (m, 39 H, C*H +

C7H2 + C1H2 + cyclen-CH2), 4.04 (q, 3JH,H = 7.2 Hz, 4 H, Et-ester-CH2), 4.31–4.41 (m,

4 H, C4H2 + C5H2), 4.91–5.00 (m, 1 H, NH), 5.01 (d, 2JH,H = 12.4 Hz, 1 H, Bn-CH2),

5.06 (d, 2JH,H = 12.4 Hz, 1 H, Bn-CH2), 7.10 (t, 3JH,H = 5.5 Hz, 1 H, amide-NH), 7.25–

7.30 (m, 5 H, arom. CH), 7.53 (bs, 1 H, triazole-CH). - 13C-NMR (75.5 MHz, CDCl3): δ

= 14.1 (+, 2 C, Et-ester), 25.9 (–, 1 C, C2), 28.4 (+, Boc-CH3), 28.5 (+, Boc-CH3), 30.7

(–, 1 C, C6), 32.3 (–, 1 C, C3), 35.0 (–, 1 C, C4), 37.5 (–, 1 C, C7), 47.6 (–, 1 C, C5),

50.3 (–, broad signal, 16 C, cyclen), 52.9 (–, 2 C, C1), 60.7 (–, 2 C, Et-ester), 64.1 (+,

1 C, C*H), 66.4 (–, 1 C, Bn-CH2), 79.7 (Cquat, 4 C, Boc), 79.9 (Cquat, 2 C, Boc), 122.3

(+, 1 C, triazole), 128.2 (+, 2 C, arom.), 128.3 (+, 1 C, arom.), 128.6 (+, 2 C, arom.),

135.6 (Cquat, 1 C, arom.), 145.1 (Cquat, 1 C, triazole), 156.3 (Cquat, 6 C, urethane),

165.3 (Cquat, 1 C, triazine), 166.1 (very broad signal, 2 C, triazine), 171.5 (Cquat, 2 C,

Et-ester), 172.0 (Cquat, 1 C, Bn-ester), 172.9 (Cquat, 1 C, amide). - MS (ES,

DCM/MeOH): m/z (%) = 1589.6 (7) [M + Na+], 1567.6 (37) [MH+], 784.2 (100) [(M +

2 H+]. - EA (C75H123N17O19) calc.: C 57.49, H 7.91, N 15.20, found: C 57.11, H 8.08,

N 15.05. - IR (KBr) [cm-1]: ν~ = 3387, 2974, 2933, 2361, 2340, 1740, 1691, 1539,

1410, 1366, 1165, 1028, 777. - MF: C75H123N17O19. - MW = 1566.90 g/mol.

N N

NN

NN

N

BocBoc

BocN N

NN

Boc

BocBoc

NH

N

NN NH

O

N

COOBnCOOEt

COOEt7

54

3

2 1

6

S-13

(S)-Hexa-tert-butyl 10,10'-(6-((1-(4-(5-(bis(2-ethoxy-2-oxoethyl)amino)-6-methoxy-6-

oxohexylamino)-4-oxobutyl)-1H-1,2,3-triazol-4-yl)methylamino)-1,3,5-triazine-2,4-


Synthesis followed GP 2 using 20 (0.75 g, 0.70 mmol), 27 (0.31 g, 0.70 mmol),

sodium ascorbate (0.28 g, 1.4 mmol) and CuSO4 x 5 H2O (0.17 g, 0.70 mmol). This

gave 0.95 g of 29 (0.63 mmol, 90 %) as a colourless solid. Rf (DCM:MeOH = 95:5) =

0.31.

MP: 108 °C. - 1H-NMR (400 MHz, CDCl3): δ = 1.19 (t, 3JH,H = 7.1 Hz, 6 H, Et-ester-

CH3), 1.37–1.52 (m, 58 H, C2H2, C3H2, Boc-CH3), 1.61–1.67 (m, 2 H, C1H2), 2.12–

2.18 (m, 4 H, C5H2, C6H2), 3.11–3.73 (m, 35 H, C*H, C4H2, cyclen-CH2), 3.54 (s, 2 H,

N-CH2), 3.55 (s, 2 H, N-CH2), 3.62 (s, 3 H, Me-ester-CH3), 4.07 (q, 3JH,H = 7.1 Hz,

4 H, Et-ester-CH2), 4.34 (t, 3JH,H = 6.1 Hz, 2 H, C7H2), 4.56–4.58 (m, 2 H, C8H2),

5.20–5.26 (m, 1 H, NH), 6.35–6.38 (m, 1 H, amide-NH), 7.63 (bs, 1 H, triazole). - 13C-

NMR (100.6 MHz, CDCl3): δ = 14.2 (+, 2 C, Et-ester-CH3), 22.9 (–, 1 C, C2), 26.3 (–,

1 C, C6), 28.4 – 28.5 (19 C, C3 and Boc-CH3), 29.7 (–, 1 C, C1), 32.6 (–, 1 C, C5),

36.3 (–, 1 C, C8), 39.2 (–, 1 C, C4), 49.3 (–, 1 C, C7), 50.2 (–, 16 C, cyclen), 51.4 (+,

1 C, Me-ester), 52.7 (–, 2 C, N-CH2), 60.6 (–, 2 C, Et-ester), 64.3 (+, 1 C, C*H), 79.8

(Cquat, 4 C, Boc), 79.8 (Cquat, 2 C, Boc), 122.0 (+, 1 C, triazole), 146.1 (Cquat, 1 C,

triazole), 156.3 (Cquat, 6 C, urethane), 165.6 (Cquat, 1 C, triazine), 166.4 (broad signal,

Cquat, 2 C, triazine), 171.4 (Cquat, 2 C, Et-ester), 171.5 (Cquat, 1 C, amide), 173.1

(Cquat, 1 C, Me-ester). - MS (ES, DCM/MeOH): m/z (%) = 1519.4 (22) [MH+], 760.1

(100) [M + 2 H+], 710.2 (20) [M + 3 H+ - CO2 - C4H8+] (20). - EA (C71H123N17O19) calc.:

C 56.15, H 8.16, N 15.68, found: C 56.01, H 8.23, N 15.57. - IR (KBr) [cm-1]: ν~ =

3390, 2978, 2934, 2359, 2340, 1739, 1689, 1538, 1411, 1366, 1166, 1028. -

MF: C71H123N17O19. - MW = 1518.86 g/mol.

N N

NN

NN

N

BocBoc

BocN N

NN

Boc

BocBoc

NH

NN N

O

NH

N

COOMe

COOEtCOOEt

3

2

1

8

5 4

67

S-14

General procedure 3 (GP 3) Cleavage of Boc-groups

The Boc-protected compound was dissolved in diethyl ether (20 ml per mmol). A

saturated solution of HCl in diethyl ether was added (1 ml per 0.15 mmol Boc). The

mixture was stirred at room temperature under an atmosphere of N2 for 48 h. After a

few minutes a white precipitate appeared. The solvent was removed at reduced

pressure and the remaining colorless solid was dried in vacuum.

(S)-Diethyl 2,2'-(1-(benzyloxy)-5-((1-(3-(4,6-di(1,4,7,10-tetraazacyclododecan-1-yl)-

1,3,5-triazin-2-ylamino)propyl)-1H-1,2,3-triazol-4-yl)methylamino)-1,5-dioxopentan-2-


Synthesis according to GP 3 using 28 (0.63 g, 0.40 mmol) and 16 ml of HCl/Et2O

gave 0.47 g of 30 (0.40 mmol, quant.) as a colorless hygroscopic solid.

MP: > 250 °C. - 1H-NMR (300 MHz, D2O): δ = 1.01 (t, 3JH,H = 7.1 Hz, 6 H, Et-ester-

CH3), 1.79–1.92 (m, 2 H, C2H2), 2.04 (tt, 3JH,H = 6.7 Hz, 2 H, C6H2), 2.26 (t, 3JH,H = 6.9

Hz, 2 H, C3H2), 3.05–3.35 (m, 26 H, C7H2 + cyclen-CH2), 3.51 (t, 3JH,H = 7.6 Hz, 1 H,

C*H), 3.57 (s, 4 H, C1H2), 3.67–3.79 (m, 8 H, cyclen-CH2), 3.93 (q, 3JH,H = 7.1 Hz,

4 H, Et-ester-CH2), 4.25 (s, 2 H, C4H2), 4.32 (t, 3JH,H = 6.7 Hz, 2 H, C5H2), 4.90–4.98

(m, 2 H, Bn-CH2), 7.16–7.21 (m, 5 H, arom. CH), 7.83 (s, 1 H, triazole). - 13C-NMR

(75.5 MHz, D2O): δ = 13.3 (+, 2 C, Et-ester), 24.7 (–, 1 C, C2), 28.7 (–, 1 C, C6), 31.6

(–, 1 C, C3), 34.0 (–, 1 C, C4), 37.8 (–, 1 C, C7), 44.3 (–, broad signal, 8 C, cyclen),

46.4 (–, 4 C, cyclen), 48.1 (–, broad signal, 4 C, cyclen), 48.4 (–, 1 C, C5), 53.2 (–,

2 C, C1), 62.3 (–, 2 C, Et-ester), 64.6 (+, 1 C, C*H), 67.4 (–, 1 C, Bn-CH2), 124.5 (+, 1

C, triazole), 128.3 (+, 2 C, arom.), 128.4 (+, 1 C, arom.), 128.8 (+, 2 C, arom.), 135.0

(Cquat, 1 C, arom.), 144.0 (Cquat, 1 C, triazole), 155.2 (Cquat, 2 C, triazine), 163.4

(broad signal, 1 C, triazine), 171.9 (Cquat, 2 C, Et-ester), 172.1 (Cquat, 1 C, Bn-ester),

174.9 (Cquat, 1 C, amide). - MS (ES, MeCN/H2O): m/z (%) = 966.7 (4) [MH+], 483.9

N N

NN

NN

N

HH

HN N

NN

H

HH

NH

N

NN NH

O

NCOOBn COOEt

COOEt7

54

3

2 1

6

X 6 HCl

S-15

(100) [M + 2 H+], 323.0 (16) [M + 3 H+] (16). - IR (KBr) [cm-1]: ν~

= 3431, 2961, 2938,

2810, 2361, 2342, 1736, 1651, 1610, 1531, 1426, 1340, 751. - MF: C45H81N17O7Cl6. -

MW = 1184.96 g/mol.

(S)-Diethyl 2,2'-(6-(4-(4-((4,6-di(1,4,7,10-tetraazacyclododecan-1-yl)-1,3,5-triazin-2-

ylamino)methyl)-1H-1,2,3-triazol-1-yl)butanamido)-1-methoxy-1-oxohexan-2-


Synthesis according to GP 3 using 29 (0.79 g, 0.52 mmol) and 21 ml of HCl/Et2O

yielded 0.59 g of 31 (0.52 mmol, quant.) as a colorless hygroscopic solid.

MP: > 250 °C. - 1H-NMR (300 MHz, D2O): δ = 1.12 (t, 3JH,H = 7.1 Hz, 6 H, Et-ester-

CH3), 1.23–1.42 (m, 4 H, C2H2 + C3H2), 1.66–1.86 (m, 2 H, C1H2), 2.00–2.15 (m, 4 H,

C5H2 + C6H2), 2.98 (t, 3JH,H = 6.7 Hz, 2 H, C4H2), 3.08–3.33 (m, 24 H, cyclen-CH2),

3.65 (s, 3 H, Me-ester-CH3), 3.78–3.84 (m, 8 H, cyclen-CH2), 3.93–3.98 (m, 1 H,

C*H), 4.03–4.04 (m, 4 H, N-CH2), 4.11 (q, 3JH,H = 7.1 Hz, 4 H, Et-ester-CH2), 4.32 (t, 3JH,H = 6.5 Hz, 2 H, C7H2), 4.67 (s, 2 H, C8H2), 7.95 (bs, 1 H, triazole). - 13C-NMR

(75.5 MHz, D2O): δ = 13.3 (+, 2 C, Et-ester-CH3), 22.6 (–, 1 C, C2), 25.8 (–, 1 C, C6),

27.5 (–, 1 C, C3), 27.9 (–, 1 C, C1), 32.5 (–, 1 C, C5), 35.8 (–, 1 C, C8), 38.8 (–, 1 C,

C4), 44.4 (–, broad signal, 8 C, cyclen), 46.2 (–, 4 C, cyclen), 47.9 (–, broad signal, 4

C, cyclen), 50.2 (–, 1 C, C7), 53.2 (+, 1 C, Me-ester), 53.9 (–, 2 C, N-CH2), 63.3 (–, 2

C, Et-ester), 66.3 (+, 1 C, C*H), 124.4 (+, 1 C, triazole), 144.1 (Cquat, 1 C, triazole),

155.6 (Cquat, 2 C, triazine), 163.8 (very broad signal, Cquat, 1 C, triazine), 169.3 (Cquat,

2 C, Et-ester), 171.0 (Cquat, 1 C), 174.7 (Cquat, 1 C). - MS (ES, MeCN/H2O): m/z (%) =

918.7 (5) [MH+], 460.0 (100) [M + 2 H+], 307.0 (40) [M + 3 H+]. - IR (KBr) [cm-1]: ν~

=

3428, 2964, 2940, 2811, 2363, 2341, 1734, 1651, 1609, 1530, 1426, 1342. -

MF: C41H81N17O7Cl6. - MW = 1136.92 g/mol.

N N

NN

NN

N

HH

HN N

NN

H

HH

NH

NN N

O

NH

N

COOMe

COOEtCOOEt

32

1

8

5 4

67

x 6 HCl

S-16

General procedure 4 (GP 4): deprotonation and cleavage of ester-groups

The ammonium salt was dissolved in water (10 ml per mmol) and eluted over a

strongly basic anion exchanger (OH- -form, loading 0.9 mmol/ml, 16 eq). The resin

was washed with a small amount of MeOH. Both solutions were combined and LiOH

was added (3 eq). The solution was stirred at room temperature for 24 h.

Lyophilisation yielded the completely deprotected compounds. 1H-NMR as well as 13C-NMR spectra of compounds 32 and 33 could not be analyzed in detail because

of too slow movement of cyclen rings in NMR timescale. However, spectra showed

no signals that could be assigned to ester functions. MS spectroscopy also confirmed

the complete cleavage of all ester groups.

Lithium (S)-2,2'-(1-carboxylato-4-((1-(3-(4,6-di(1,4,7,10-tetraazacyclododecan-1-yl)-

1,3,5-triazin-2-ylamino)propyl)-1H-1,2,3-triazol-4-yl)methylamino)-4-

oxobutylazanediyl)diacetate (32):

Synthesis according to GP 4 using 30 (0.73 g, 0.40 mmol), 7.1 ml of ion-exchanger

resin (6.4 mmol) and LiOH (29 mg, 1.2 mmol) yielded 0.34 g of 32 (0.40 mmol,

quant.) as a colorless frothy solid.

MS (pos. ES, H2O/MeCN/TFA): m/z (%) = 410.8 (60) [A3- + 5 H+], 413.8 (100) [A3- +

Li+ + 4 H+], 416.8 (80) [A3- + 2 Li+ + 3 H+], 419.8 (40) [A3- + 3 Li+ + 2 H+], 820.7 (55)

[A3- + 4 H+], 826.7 (40) [A3- + Li+ + 3 H+], 832.7 (35) [A3- + 2 Li+ + 2 H+], 826.7 (25)

[A3- + 3 Li+ + H+]. - MS (neg. ES, H2O/MeCN/TFA): m/z (%) = 932.7 (100) [A3- +

CF3COO- + 3 H+], 938.7 (25) [A3- + CF3COO- + Li+ + 2 H+], 1046.7 (30) [A3- + 2

CF3COO- + Li+ + 3 H+], 1052.6 (20) [A3- + 2 CF3COO- + 2 Li+ + 2 H+], 1058.6 (10) [A3-

+ 2 CF3COO- + 3 Li+ + H+]. - MF: C34H58N17O7Li3. - MW = 837.76 g/mol.

N N

NN

NN

N

HH

HN N

NN

H

HH

NH

N

NN NH

O

NCOOLi COOLi

COOLi

S-17

Lithium (S)-2,2'-(1-carboxylato-5-(4-(4-((4,6-di(1,4,7,10-tetraazacyclododecan-1-yl)-

1,3,5-triazin-2-ylamino)methyl)-1H-1,2,3-triazol-1-yl)butanamido)pentylazanediyl)-

diacetate (33):

The synthesis followed GP 4 using 31 (0.59 g, 0.52 mmol), 9.2 ml of ion-exchanger

resin (8.3 mmol) and LiOH (38 mg, 1.6 mmol). This gave 0.45 g of 33 (0.52 mmol,

quant.) as a colorless frothy solid.

MS (pos. ESI, MeCN/H2O): m/z (%) = 424.8 (100) [A3- + 5 H+] (100), 427.8 (75) [A3- +

Li+ + 4 H+], 430.9 (70) [A3- + 2 Li+ + 3 H+], 433.9 (20) [A3- + 3 Li+ + 2 H+], 848.6 (15)

[A3- + 4 H+], 854.6 (10) [A3- + Li+ + 3 H+], 860.7 (5) [A3- + 2 Li+ + 2 H+]. - MS (neg. ES,

H2O/MeCN): m/z (%) = 422.9 (20) [A3- + H+], 846.8 (100) [A3- + 2 H+], 852.8 (30) [A3-

+ Li+ + H+] , 858.8 (20) [A3- + 2 Li+]. - MF: C36H62N17O7Li3. - MW = 865.82 g/mol.

General procdure 5 (GP 5): Synthesis of Zn(II) complexes

The ligand was dissolved in MeOH (10 ml per mmol). A slight excess of Zn(ClO4)2 x 6

H2O (3.3 eq) was dissolved in water (10 ml per mmol) and both solutions were added

simultaneously drop wise to water (5 ml per mmol) at 80 °C. In the case that a

precipitate appeared, water was added until the precipitate dissolved. After complete

addition of both solutions the pH value was checked. If it was lower that 8, an

aqueous solution of LiOH (0.1 mol/l) was added. The mixture was heated to reflux for

6 h. EtOH was subsequently added to the boiling solution until the product began to

precipitate. After cooling to room temperature the suspension was stored at -20 °C

over night in order to complete the precipitation. The product was separated by

centrifugation, washed with MeOH and EtOH, the solution was concentrated at

reduced pressure and dried in vacuum. The remaining solid was recrystallized from

water/MeOH/EtOH.

N N

NN

NN

N

HH

HN N

NN

H

HH

NH

NN N

O

NH

N

COOLi

COOLiCOOLi

S-18

Complex 7:

The synthesis according to GP 5 using 32 (0.34 g, 0.40 mmol) and Zn(ClO4)2 x 6 H2O

(0.49 g, 1.3 mmol) gave 1 (0.36 g, 0.29 mmol, 72 %) as a colorless solid.

MP: > 250 °C. - MS (pos. ES, H2O/MeCN): m/z (%) = 503.8 (100) [K3+ - H+], 512.8

(70) [K3+ + OH-], 553.8 (20) [K3+ + ClO4-] (20). - EA (C34H63N17O18Zn3Cl2) calc.:

C 32.28, H 5.02, N 18.82, found: C 31.93, H 5.13, N 18.56. - UV/Vis (H2O): λ (lg ε) =

222 nm (4.663). - IR (KBr): ν [cm-1] = 3422, 3272, 2934, 2889, 2362, 2341, 1596,

1561, 1425, 1347, 1286, 1090, 978, 805. - MF: [C34H58N17O7Zn3]3+(ClO4)2(OH) x

2 H2O. - MW = 1265.02 g/mol.

Complex 8:

The synthesis following GP 5 using 33 (0.45 g, 0.52 mmol) and Zn(ClO4)2 x 6 H2O

(0.64 g, 1.7 mmol) yielded 2 (0.50 g, 0.38 mmol, 74 %) as a colorless solid.

MP: > 250 °C. - MS (pos. ES, H2O/MeCN): m/z (%) = 517.9 (100) [K3+ - H+], 526.9

(30) [K3+ + OH-], 547.9 (20) [K3+ + CH3COO-], 567.9 (20) [K3+ + ClO4-]. -

EA (C36H67N17O18Zn3Cl2) calc.: C 33.44, H 5.22, N 18.41, found: C 33.15, H 5.33,

N 18.23. - UV/Vis (H2O): λ (lg ε) = 222 nm (4.658). - IR (KBr) [cm-1]: ν~

= 3423, 3272,

2934, 2889, 2361, 2341, 1596, 1561, 1426, 1347, 1286, 1090, 978, 805. -

MF: [C36H62N17O7Zn3]3+(ClO4)2(OH) x 2 H2O. - MW = 1293.08 g/mol.

Zn2+

Zn2+

OHZn2+

H2OOH2

O-

O

NO-

O

OO-

NHO

NN N

N

N

N

N

NN

N

NN

NN

NH

H

HH

H

H

H

2+

(ClO4)2

Zn2+

Zn2+

OH

Zn2+H2O

OH2

O-

O

N O

NH

O-

O

OO-N

NN

N

N

N

N

N

NN

N

NN

NN

H

H

HH

H

H

H

2+

(ClO4)2

S-19

Hexa-tert-butyl 10,10'-(6-(2-(benzyloxycarbonylamino)ethylamino)-1,3,5-triazine-2,4-


Benzyl 2-aminoethylcarbamate 35 (1.47 g, 7.6 mmol) was suspended in dioxane and

potassium carbonate (42 g, 0.30 mol) was added. The mixture was stirred for half an

h, then 14 (5.0 g, 4.7 mmol) was added and the solution was stirred for three days at

145 °C. The potassium carbonate was filtered off and the solution was evaporated to

dryness. The crude product was purified by chromatography on silica gel with

PE:EtOAc 80:20 (Rf (PE:EtOAc 1:1) = 0.47) as eluent to give the product as a

colorless solid (4.8 g, 4.0 mmol, 84 %).

MP: 134-135 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.22-1.56 (m, 54 H, Boc-CH3),

2.81-3.88 (m, 36 H, Cyclen-CH2 + NH-CH2-CH2), 4.88 (s, 1 H, NH), 5.05 (s, 2 H, CH2-

Cbz), 6.67 (s, 1 H, NH), 7.27-7.37 (m, 5 H, CH-Cbz). - 13C-NMR (75 MHz, CDCl3): d

= 28.5 (+, 18 C, Boc-CH3), 40.2 (-, 1 C, NH-CH2), 41.0 (-, 1 C, NH-CH2), 49.5-51.1 (-,

16 C, Cyclen-CH2), 66.4 (-, 1 C, CH2-Cbz), 79.8 (Cquat, 6 C, OC(CH3)3), 127.9 (+, 1 C,

CH-Cbz), 128.2 (+, 1 C, CH-Cbz), 128.3 (+, 1 C, CH-Cbz), 136.9 (Cquat, 1 C, C-Ar-

Cbz), 155.9-157.4 (Cquat, 7 C, Boc-CO + Cbz-CO), 166.0 (Cquat, 3 C, Ar-C). - MS (ES,

DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 1214.9 (100) [MH+]. - IR (neat) [cm-1]: ν~ =

3350, 2974, 2932, 1685, 1538, 1409, 1364, 1246, 1158, 1105, 972, 776. -

MF: C49H99N13O14. - MW = 1214.50 g/mol.

CbzNH

HN

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

S-20

Hexa-tert-butyl 10,10'-(6-(2-aminoethylamino)-1,3,5-triazine-2,4-diyl)bis(1,4,7,10-

tetraazacyclododecane-1,4,7-tricarboxylate) (11):

Compound 34 (4.8 g, 4.0 mmol) was dissolved in methanol. To this mixture a spatula

of palladium on activated charcoal was added. The mixture was stirred at 30 bars of

H2 pressure for three days. The Pd was filtered off; the mixture was concentrated

under reduced pressure and dried under high vacuum to obtain the product as a

colorless solid in 79 % yield (3.4 g, 3.1 mmol).

MP: 126-127 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.29-1.58 (m, 54 H, Boc-CH3),

2.85-2.96 (m, 2 H, CH2-NH2), 3.13-3.86 (m, 34 H, Cyclen-CH2 + CH2-CH2-NH2). -13C-

NMR (75 MHz, CDCl3): d = 28.5 (+, 18 C, Boc-CH3), 41.7 (-, 1 C, CH2), 43.0 (-, 1 C,

CH2), 50.3 (-, 16, Cyclen-CH2), 79.9 (Cquat, 6 C, Boc- Cquat), 156.5 (Cquat, 6 C, Boc-

CO), 166.1 (Cquat, 3 C, C-Ar). - MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%)

= 1081 (100) [MH+]. - IR (neat) [cm-1]: ν~ = 2973, 2930, 1686, 1539, 14,66, 1408,

1364, 1247, 1159, 1105, 971, 776. - MF: C51H93N13O12. - MW = 1080.36 g/mol.

Compound 36:

H-Gly-OMe · HCl (37) (1.5 g, 12.1 mmol) was dissolved in dioxane and triethylamine

(3.3 ml, 24 mmol) was added. The mixture was stirred for 10 minutes. Then

1,3-bis(Boc)-2-methyl-2-thiopseudourea (38) (2.5 g, 8.6 mmol) was added and the

mixture was stirred for 30 h at room temperature. The reaction progress was

monitored by TLC. The reaction mixture was dissolved in water and extracted three

times with DCM. Afterwards the combined organic layers were dried over MgSO4 and

H2N

HN

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

NH

O

OHN

N

O

O

OO1 2

34

56

78

910

11

1213

1415

16

S-21

concentrated under reduced pressure. The crude product was purified by column

chromatography on silica gel using PE:EtOAc 2:1 (Rf(PE: EtOAc 1:1) = 0,78) as

eluent. The pure product was obtained as a colorless solid in 67 % yield. (1.9 g, 5.8

mmol)

MP: 132-133 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.49 (s, 9 H, 1/15), 1.50 (s, 9 H,

1/15), 3.77 (s, 3 H, 11), 4.23 (s, 3JH,H = 4.9, 2 H, 8), 8.86 (s, 1 H, NH), 11.43 (s, 1 H,

NH). - 13C-NMR (75 MHz, CDCl3): d = 28.2 (+, 6 C, 1 + 16), 42.7 (-, 1 C, 8), 52.5 (+, 1

C, 11), 79.6 (Cquat, 1 C, 2), 83.4 (Cquat, 1 C, 15), 152.9 (Cquat, 1 C, 4), 156.0 (Cquat, 1

C, 13), 163.2 (Cquat, 1 C, 6), 169.9 (Cquat, 1 C, 9). - MS (ES, DCM/MeOH + 10 mmol/l

NH4OAc): m/z (%) = 332.2 (100) [MH+], 663.5 (42) [2 M + H+]. - IR (neat) [cm-1]: ν~ =

3318, 2982, 2952, 2362, 1757, 1725, 1614, 1292, 1138, 1094, 1060, 975, 811, 761. -

MF: C14H25N3O6. - MW = 331.36 g/mol.

Compound 12:

Compound 36 (1.6 g, 4.7 mmol) was dissolved in a mixture of water and acetone 1:4.

LiOH (0.17 g, 7.1 mmol) was added and the mixture was stirred at room temperature

over night. The solution was acidified with saturated ammonium chloride solution and

was extracted with DCM. The combined organic layers were dried over MgSO4 and

concentrated under reduced pressure to give the product as a colorless salt in

quantitative yield. (1.5 g, 4.7 mmol)

MP: 104-105 °C. - 1H-NMR (300 MHz, DMSO): d = 1.39 (s, 9 H, Boc-CH3), 1.49 (s, 9

H, Boc-CH3), 3.98 (d, 3JH,H = 5.4, 2 H, CH2), 8.62 (s, 1 H, NH), 11.47 (s, 1 H, NH. - 13C-NMR (75 MHz, DMSO): d = 27.5 (+, 3 C, Boc-CH3), 27.8 (+, 3 C, Boc-CH3), 42.2

(-, 1 C, CH2), 78.3 (Cquat, 1 C, C(CH3)3), 83.0 (Cquat, 1 C, C(CH3)3), 151.7 (Cquat, 1 C,

Boc-CO), 155.2 (Cquat, 1 C, Boc-CO), 162.7 (Cquat, 1 C, C-NH), 170.4 (Cquat, 1 C,

COO). - MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 206 (60) [MH+ - 2

C4H8], 262 (32) [MH+ - C4H8], 318 (100) [MH+], 635 (40) [2 M + H+]. - IR (neat) [cm-1]:

ν~ = 3329, 2978, 2932, 2362, 1795, 1726, 1633, 1398, 1319, 1229, 1144, 1102,

1060, 987, 883, 763. - MF: C13H23N3O6. - MW = 317.34 g/mol.

NH

OH

ONH

NBoc

Boc

S-22

Compound 39:

The acid 12 (0.46 g, 1.4 mmol), HOBt (0.28 g, 2.1 mmol) and DIPEA (0.71 ml, 4.2

mmol) were dissolved under nitrogen atmosphere in dry DMF (10 ml). The mixture

was cooled to 0 °C in an ice bath and EDC (0.37 ml, 2.1 mmol) was added. Then the

amine 11 (1.0 g, 0.93 mmol) was added slowly in portions. The reaction was

monitored by TLC (EtOAc). After 24 h water was added and extracted with DCM. The

combined organic layers were then washed three times with an aqueous citric acid

solution (10 %) and twice with brine. Afterwards the solution was dried over MgSO4

and concentrated under reduced pressure. The crude product was purified by flash

chromatography on silica gel (PE:EtOAc 1:1, Rf = 0.43) to give the product as a


MP: 158-159 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.25-1.61 (m, 72 H, 1 + 22 +

cyclen Boc-CH3), 2.74-3.91 (m, 38 H, 8 + 11 + 12 + cyclen-CH2), 4.92 (s, 1 H, NH),

7.53 (bs, 1 H, NH), 8.92 (s, 1 H, NH), 11.21 (s, 1 H, NH). - 13C-NMR (75 MHz,

CDCl3): d = 28.1-29.4 (+, 34 C, 1 + 22 + cyclen Boc-CH3), 44.4 (-, 2 C, 8 + 11), 50.3

(-, 16 C, cyclen-CH2), 54.9.2 (-, 2 C, 12), 79.9 (Cquat, 7 C, 2 + cyclen Boc-Cquat), 83.1

(Cquat, 1 C, 21), 152.7 (Cquat, 1 C, 4), 154.6-145.7 (Cquat, 7 C, 19 + cyclen Boc-CO),

163.2 (Cquat, 1 C, 6), 165.9 (Cquat, 3 C, 14 + 16), 168.4 (Cquat, 1 C, 9). - MS (ES,

DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 690 (76) [M + 2 H+), 1380 (100) [MH+].

- Elemental analysis calcd. (%) for C64H114N16O17 (1379.69) + 2 H2O: C 54.30, H

8.40, N 15.83. - found: C 54.27, H 8.47, N 15.01. - IR (neat) [cm-1]: ν~ = 3318, 2981,

2942, 2362, 1757, 1726, 1638, 1615, 1367, 1292, 1249, 1141, 1095, 1059, 975, 812,

774. - MF: C64H114N16O17. - MW = 1379.69 g/mol.

1

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

NH

HN

ONH

HN

N

O

O O

O

23

4

56

7 89

10 11

1213

14

1516

17

181921

22

20

S-23

Complex 3:

Compound 39 (0.25 g, 0.18 mmol) was dissolved in methanol and cooled to 0 °C in

an ice bath and HCl-saturated ether (0.7 ml/mmol Boc-group) was added. The

mixture was allowed to warm to room temperature and was stirred over night. The

solvent was evaporated, the crude product was dissolved in water and purified over

an ion exchanger column using a strongly basic ion exchanger to give a colorless

solid after freeze drying. Two aqueous solutions, one containing the obtained

colorless solid from the column and a second one containing Zn(ClO4)2 · 6 H2O (0.15

g, 0.40 mmol) were prepared. These two solutions were simultaneously added to

water at 80 °C. The resulting mixture was stirred at 90 °C over night. The solution

was lyophilized and the product was recrystallized from a water methanol mixture to

give the complex as a colorless solid in 83 % yield. (0.16 g, 0.15 mmol)

MP: > 250 °C. - MS (ES, H2O/MeOH + 10 mmol/l NH4OAc) (K3+ = C24H51N16O2ClZn2)

: m/z (%) = 353 (62) [K3+ - H+ - H2O], 363 (100) [K3+ - H+], 385 (71) [K3+ + OH- +

MeOH], 822 (19) [K3+ - H+ + ClO4-]. - IR (neat) [cm-1]: ν~ = 3568, 3527, 3370, 2942,

1625, 1548, 1480, 1421, 1342, 1285, 1063, 963, 814, 744. - MF:

[C24H50N16OZn2]4+(ClO4)3(OH). - MW = 1024.89 g/mol.

NN

N

N N

NN

N N

NN

NH

NH

HN

ONH2

NH

H H

H

H

HH

Zn2+

Zn2+

-HO

3+

3ClO4-

S-24

Compound 40:

Peptide H-Gly-Gly-OMe · HCl (41) (1.1 g, 6.2 mmol) was dissolved in CHCl3 :

methanol 5:1 and triethylamine (1.7 ml, 12 mmol) was added. The mixture was stirred

for 10 minutes. Then 1,3-bis(Boc)-2-methyl-2-thiopseudourea (38) (2.0 g, 6.9 mmol)

was added and the mixture was stirred for 6 h at room temperature. The reaction

process was monitored by TLC. The reaction mixture was added to water and

extracted three times with DCM. Afterwards the combined organic layers were dried

over MgSO4 and concentrated under reduced pressure. The crude product was

purified by column chromatography on silica gel using PE:EtOAc 1:1 (Rf = 0,35) as

eluent. The pure product was obtained as a colorless solid in 46 % yield. (1.1 g, 2.8

mmol)


1/19), 3.69 (s, 3 H, 14), 4.00 (d, 3JH,H = 5.2, 2 H, 9), 4.10 (d, 3JH,H = 4.9, 2 H, 11), 6.98

(d, 3JH,H = 4.9, 1 H, NH), 8.88 (t, 3JH,H = 4.8, 1 H, NH), 11.32 (s, 1 H, NH). - 13C-NMR

(75 MHz, CDCl3): d = 28.2 (+, 6 C, 1 + 19), 41.3 (-, 1 C, 11), 44.4 (-, 1 C, 8), 52.4 (+,

1 C, 14), 79.6 (Cquat, 1 C, 2), 83.5 (Cquat, 1 C, 18), 152.8 (Cquat, 1 C, 4), 156.2 (Cquat, 1

C, 16), 163.0 (Cquat, 1 C, 6), 168.6 (Cquat, 1 C, 12), 170.0 (Cquat, 1 C, 9). - MS (CI,

NH3): m/z (%) = 389 (100) [MH+]. - IR (neat) [cm-1]: ν~ = 3347, 3305, 2977, 2936,

2363, 1723, 1678, 1641, 1525, 1430, 1395, 1349, 1303, 1220, 1148, 1123, 1091,

1055, 971, 811. - MF: C16H28N4O7. - MW = 388.42 g/mol.

Compound 13:

Compound 40 (1.0 g, 2.6 mmol) was dissolved in a mixture of water and THF 1:4.

LiOH (74 mg, 3.1 mmol) was added and the mixture was stirred at room temperature

over night. The solution was acidified with 5 % aqueous KHSO4 solution and was

extracted with DCM. The combined organic layers were dried over MgSO4 and

NH

HN

ONH

NBoc

BocOH

O

NH

HN

OHN

N

O

O

OO1 2

34

56

78

910

1112

13

14

1516

O

O17

18

19

S-25

concentrated under reduced pressure to give the product as a colorless salt in 68 %

yield. (0.64 g, 1.7 mmol)

MP: 111-112 °C. - 1H-NMR (300 MHz, MeOD): d = 1.46 (s, 9 H, Boc-CH3), 1.53 (s, 9

H, Boc-CH3), 3.78 (s, 2 H, CH2), 4.09 (s, 2 H, CH2). - 13C-NMR (75 MHz, MeOD): d =

28.6 (+, 3 C, Boc-CH3), 29.0 (+, 3 C, Boc-CH3), 44.6 (-, 1 C, CH2), 45.0 (-, 1 C, CH2),

78.9 (Cquat, 1 C, C(CH3)3), 82.8 (Cquat, 1 C, C(CH3)3), 162.7 (Cquat, 1 C, Boc-CO),

164.2 (Cquat, 1 C, Boc-CO), 171.8 (Cquat, 1 C, C-NH), 176.1 (Cquat, 1 C, COO). - MS

(ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 206 (60) [MH+ - 2 C4H8], 262 (32)

[MH+ - C4H8], 318 (100) [MH+], 635 (40) [2 M + H+]. - IR (neat) [cm-1]: ν~ = 2975,

2935, 1559, 1541, 1499, 1408, 1366, 1303, 1249, 1148, 1056, 973, 864. -

MF: C15H26N4O7. - MW = 374.39 g/mol.

Compound 42:

The acid 13 (0.26 g, 0.69 mmol), HOBt (0.14 g, 1.0 mmol) and DIPEA (0.36 ml, 2.0

mmol) were dissolved under nitrogen atmosphere in dry DMF (10 ml). The mixture

was cooled to 0 °C in an ice bath and EDC (0.18 ml, 1.0 mmol) was added. Then the

amine 11 (0.50 g, 0.46 mmol) was added slowly in portions. The reaction was

monitored by TLC (EtOAc). After 24 h the solution was added to water and extracted

with DCM. The combined organic layers were then washed three times with an

aqueous citric acid solution (10 %) and twice with brine. Afterwards the solution was

dried over MgSO4 and concentrated under reduced pressure. The crude product was

then purified by flash chromatography on silica gel (EtOAc, Rf = 0.26) to give the

product as a colorless solid (0.15 g, 0.11 mmol, 23 %).

MP: 162-163 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.24-1.57 (m, 72 H, 1 + 25 +

Cyclen Boc-CH3), 3.02-3.73 (m, 36 H, 14 + 15 + Cyclen-CH2), 3.89 (d, 3JH,H = 4.7, 2

H, 8), 4.10 (d, 3JH,H = 4.9, 2 H, 11), 5.00 (s, 1 H, NH), 6.91 (s, 1 H, NH), 7.49 (s, 1 H,

1

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

HN

HN

NH

O

ONH

HN

N

O

O O

O

23

4

56

7 89

1011

12

1314

1516 17

18 19

20

21222324

25

S-26

NH), 8.87 (s, 1 H, NH), 11.34 (s, 1 H, NH). - 13C-NMR (75 MHz, CDCl3): d = 28.0-

28.5 (+, 34 C, 1 + 25 + Cyclen Boc-CH3), 40.5 (-, 1 C, 14), 42.8 (-, 1 C, 15), 44.2 (-, 2

C, 8 + 11), 50.3 (-, 16 C, Cyclen-CH2), 79.4 (Cquat, 1 C, 2), 79.9 (Cquat, 6 C, Cyclen

Boc-Cquat), 83.2 (Cquat, 1 C, 24), 152.8 (Cquat, 1 C, 4), 155.9 (Cquat, 6 C, Cyclen Boc-

CO), 156.4 (Cquat, 1 C, 22), 163.2 (Cquat, 1 C, 6), 168.1-168.5 (Cquat, 3 C, 17 + 18),

171.1 (Cquat, 2 C, 9 + 12). - MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) =

619 (67) [M + 2 H+ - 2 Boc], 719 (26) [M + 2H+], 1437 (100) [MH+]. - Elemental

analysis calcd. (%) for C66H117N17O18 (1436.74) + 2 H2O: C 53.82, H 8.28, N 16.17. -

found: C 53.69, H 8.31, N 15.61. - IR (neat) [cm-1]: ν~ = 3318, 3109, 2982, 2952,

1757, 1725, 1637, 1614, 1292, 1138, 1094, 1060, 975, 874, 811. -

MF: C66H117N17O18 - MW = 1436.74 g/mol.

Complex 4:


an ice bath. HCl saturated ether (0.7 ml/mmol Boc-group) was added to the solution.

The mixture was allowed to warm to room temperature and was stirred over night.

The solvent was evaporated and the crude product was dissolved in water and

purified over an ion exchanger column using a strongly basic ion exchanger (OH- -

form, loading 0.9 mmol/ml, 4 eq. per protonated nitrogen) to give a colorless solid

after lyophylization. Then two aqueous solutions, one containing the obtained

colorless solid from the column and a second one containing Zn(ClO4)2 · 6 H2O (90

mg, 0.24 mmol, 2.2 eq. per deprotected receptor) were prepared. These two

solutions were then simultaneously added drop wise to water at 80 °C under vigorous

stirring. The resulting mixture was stirred at 90 °C over night. The solution was freeze

dried and the product was recrystallized from a water methanol mixture to give the

complex as colorless solid in 63 % yield. (75 mg, 0.07 mmol).

NN

N

N N

NN

N N

NN

NH

NH

HN

O

O HN NH2

NH

HH

H

H

HH

Zn2+

Zn2+

-HO 3 ClO4-

3+

S-27

MP > 250 °C. - MS (ES, H2O/MeOH + 10 mmol/l NH4OAc) (K4+ = C26H53N17O2Zn2) :

m/z (%) = 353 (29) [K4+ + ClO4- + 2 HClO4], 392 (100) [K4+ + OH- - H+], 413 (41) [K4+

+ CH3COO- - H+], 909 (8) [K4+ + OH- + 2 Cl- + CH3COOH] . - IR (neat) [cm-1]: ν~ =

3282, 2950, 2896, 2364, 2343, 1652, 1558, 1429, 1347, 1285, 1084, 970, 815, 777. -

MF: [C26H53N17O2Zn2]4+(ClO4)3(OH). - MW = 1081.94 g/mol.

2-(Boc-amino)-N-(2-(Boc-amino)ethyl)acetamide (43):

Boc-Gly-OH (44) (1.0 g, 5.7 mmol), HOBt (0.85 g, 6.3 mmol) and DIPEA (1.9 ml, 11

mmol) were dissolved in DCM (15 ml). To the solution tert-butyl

2-aminoethylcarbamate (45) (1.1 g, 6.9 mmol) was added and the mixture was

cooled to 0 °C in an ice bath. Then EDC (1.1 ml, 6.3 mmol) was added. The reaction

was monitored by TLC (EtOAc). After 24 h the solution was added to water and

extracted with DCM. The combined organic layers were washed twice with an

aqueous citric acid solution (10 %) and twice with brine, the solution was dried over

MgSO4 and concentrated under reduced pressure. The crude product was purified by

column chromatography on silica gel (EtOAc, Rf = 0.28) to give the product as a


MP: 110-111 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.44 (s, 18 H, 1 + 15), 3.24 (s,2 H,

10), 3.32-3.41 (m, 2 H, 9), 3.78 (s, 2 H, 6), 5.15 (s, 1 H, 11), 5.34 (s, 1 H, 5), 6.91 (s,

1 H, 8). - 13C-NMR (75 MHz, CDCl3): d = 28.3 (+, 3 C, 1/15), 28.4 (+, 3 C, 1/15), 40.4

(-, 2 C, 9 + 10), 44.3 (-, 1 C, 6), 79.7 (Cquat, 1 C, 2/14), 80.3 (Cquat, 1 C, 2/14), 156.1

(Cquat, 1 C, 4/12) , 156.7 (Cquat, 1 C, 4/12), 170.3 (Cquat, 1 C, 7). - MS (ES,

DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 317 (100) [MH+], 635 (16) [2 M + H+]. -

IR (neat) [cm-1]: ν~ = 3334, 2980, 2936, 2360, 1685, 1650, 1521, 1365, 1281, 1244,

1167, 1066, 864. - MF: C51H93N13O12. - MF: C14H27N3O5. - MW = 317.38 g/mol.

HN

NH

O HN O

O

O

O

1 34

56

7

8 910

1112

132

14

15

S-28

2-Amino-N-(2-aminoethyl)acetamide·2 HCl (15):

Compound 43 (1.0 g, 3.2 mmol) was dissolved in methanol and cooled to 0 °C in an

ice bath. To this mixture six pipettes of HCl saturated ether were added. The mixture

was allowed to warm to room temperature and stirred over night. The reaction

progress was controlled by 1H-NMR. The mixture was concentrated under reduced

pressure and dried at high vacuum to obtain the product as a colorless solid in

quantitative yield (0.60 g, 3.2 mmol).

1H-NMR (300 MHz, D2O): d = 3.10 (t, 3JH,H = 5.9, 2 H, 6), 3.51 (t, 3JH,H = 6.0, 2 H, 5),

3.78 (s, 2 H, 2). - IR (neat) [cm-1]: ν~ = 3166, 2913, 2757, 2360, 2061, 1673, 1567,

1494, 1391, 1274, 1176, 1070, 901, 757. - MF: C4H11N3O x 2 HCl. - MW = 254.07

g/mol.

Hexa-tert-butyl 10,10'-(6-(2-(2-(4,6-bis(4,7,10-tris(tert-butoxycarbonyl)-1,4,7,10-

tetraazacyclodo-decan-1-yl)-1,3,5-triazin-2-ylamino)acetamido)ethylamino)-1,3,5-

triazine-2,4-diyl)bis(1,4,7,10-tetraazacyclododecane-1,4,7-tricarboxylate) (46):

Compound 12 (0.20 g, 1.1 mmol) was suspended in dioxane and potassium

carbonate (5.8 g, 42 mmol) was added. The mixture was stirred for 30 min then

substance 14 (2.2 g, 2.1 mmol) was added and the solution was stirred for three days

at 145 °C. The potassium carbonate was filtered off, the solution was evaporated to

dryness and the crude product was purified by chromatography on silica gel with

EtOAc (Rf = 0.76) as eluent to give the product as colorless solid (1.5 g, 0.71 mmol,

68 %).

MP: 179-180 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.43 (s, 108 H, Boc-CH3), 2.85-

3.93 (m, 70 H, Cyclen-CH2 + NH-CH2), 5.27 (bs, 2 H, NH), 7.34 (s, 1 H, NH). - 13C-

H2NNH

ONH2

1 34

56 72

* 2 HCl

NN

N

N N

NN

BocBoc

Boc

N N

NNBoc

Boc

Boc

HN

NH

O HN

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

S-29

NMR (75 MHz, CDCl3): d = 28.5 (+, 36 C, Boc-CH3), 40.6 (-, 1 C, NH-CH2), 42.3 (-, 1

C, NH-CH2-CH2), 46.4 (-, 1 C, CH2-CO), 50.3 (-, 32 C, Cyclen-CH2), 79.9 (Cquat, 12 C,

OC(CH3)3), 156.3 (Cquat, 12 C, Boc-CO), 165.8 (Cquat, 6 C, Ar-C), 169.5 (Cquat, 1 C,

CH2-CO). - MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 1079 (100) [M + 2

H+], 2158 (13) [MH+]. - IR (neat) [cm-1]: ν~ = 2974, 2932, 2359, 1686, 1539, 1466,

1408, 1364, 1246, 1158, 1105, 971, 776. - MF: C102H181N25O25. - MW = 2157.68

g/mol.

Complex 5:


an ice bath. To the solution HCl saturated ether (0.7 ml/mmol Boc-group) was added.

The mixture was allowed to warm to room temperature, stirred over night, the solvent

was evaporated, the crude product was dissolved in water and purified over an ion

exchanger column using a strongly basic ion exchanger to deliver a colorless solid

after lyophylization. Two aqueous solutions were prepared: one containing the

obtained colorless solid from the column and a second containing Zn(ClO4)2 · 6 H2O

(0.66 g, 1.8 mmol). The two solutions were simultaneously added to hot water

(80 °C). The resulting mixture was stirred at 90 °C over night. The solution was

lyophilized and the product was recrystallized from a water methanol mixture to give

the complex as colorless solid in 84% yield. (0.69 g, 0.37 mmol)

MP: > 250 °C. - MS (ES, H2O/MeOH + 10 mmol/l NH4OAc) (K6+ = C48H93N19O3ClZn4)

: m/z (%) = 252 (50) [K6+ + OH-], 369 (41) [K6+ + Cl- + ClO4- + HClO4], 484 (93) [K6+ +

OH- + 2 ClO4-], 518 (100) [K6+ + OH- + 2 ClO4

- + HClO4], 826 (29) [K6+ + OH- + 3

ClO4- + HClO4] . - IR (neat) [cm-1]: ν~ = 3567, 3527, 3284, 2942, 2890, 2357, 1549,

1480, 1423, 1343, 1067, 812. - MF: [C42H85N25OZn4]8+(ClO4)6(OH-)2. - MW = 1848.54

g/mol.

NN

NN

NH

NN

NNH

H

N N

NH

H H

H

NN

NN

NH

N N

N NH

H

NN

NH

HH

H

Zn2+

Zn2+ Zn2+

Zn2+

OH--HO

6+

6 ClO4-

HN

O

S-30

Methyl 2-(2-(tert-butoxycarbonylamino)acetamido)acetate (47):

Boc-Gly-OH (44) (1.2 g, 6.7 mmol), HOBt (1.1 g, 8.0 mmol) and DIPEA (1.9 ml, 11

mmol) were dissolved in DCM (20 ml). The solution was cooled to 0 °C in an ice bath

and then EDC (1.4 ml, 8.0 mmol) was added. After several minutes H-Gly-OMe·HCl

(37) was added in portions. The solution was allowed to warm to room temperature

and was stirred over night. The reaction was monitored by TLC (EtOAc). After 24 h

the solution was added to water, extracted with DCM, the combined organic layers

were washed twice with an aqueous citric acid solution (10 %) and twice with brine.

The organic layer was dried over MgSO4, concentrated at reduced pressure and the

crude product was purified by column chromatography on silica gel (EtOAc, Rf =

0.34) to give the product as a colorless solid (0.93 g, 3.8 mmol, 57 %).

1H-NMR (300 MHz, CDCl3): d = 1.45 (s, 9 H, 1), 3.75 (s, 3 H, 12), 3.85 (s, 2 H, 6),

4.06 (d, 3JH,H = 5.5, 2 H, 9), 5.23 (s, 1 H, 5), 6.73 (s, 1 H, 8). - 13C-NMR (75 MHz,

CDCl3): d = 28.3 (+, 3 C, 1), 41.1 (-, 1 C, 9), 44.2 (-, 1 C, ), 52.5 (+, 1 C, 12), 80.5

(Cquat, 1 C, 2), 156.1 (Cquat, 1 C, 4), 169.8 (Cquat, 1 C, 4/7), 171.8 (Cquat, 1 C, 4/7) . - IR

(neat) [cm-1]: ν~ = 3281, 2979, 2359, 2338, 1660, 1525, 1366, 1250, 1161, 1030, 953.

- MF: C10H18N2O5. - MW = 246.26 g/mol.

2-(2-(Tert-butoxycarbonylamino)acetamido)acetic acid (48):

Boc-Gly-Gly-OMe (47) (0.50 g, 2.0 mmol) was dissolved in 10 ml of methanol and 4

ml of an 1 M aqueous sodium hydroxide solution was added. The solution was stirred

over night and the methanol was evaporated under reduced pressure. The residue

was acidified with aqueous KHSO4 solution (5 %), the mixture was extracted three

times with EtOAc, the combined organic layers were dried over MgSO4 and

concentrated under reduced pressure to give the product as a colorless solid. (0.42

g, 1.8 mmol, 88 %).

HN

NH

OOO

O

1 34

56

7

8 910

11122

O

HN

NH

OOHO

O

1 34

56

7

8 910

112

O

S-31

MP: 115-116 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.28 (s, 9 H, 1), 3.63 (s, 2 H, 6),

3.81 (s, 2 H, 9). - 13C-NMR (75 MHz, CDCl3): d = 23.5 (+, 3 C, 1), 40.7 (+, 1 C, 6/9),

43.4 (+, 1 C, 6/9), 80.1 (Cquat, 1 C, 2), 156.5 (Cquat, 1 C, 4), 170.7 (Cquat, 1 C, 7/10),

171.4 (Cquat, 1 C, 7/10). - MS (CI, NH3): m/z (%) = 231 (100) [M - H+], 267 (20) [MCl-],

463 (22) [2 M – H+]. - IR (neat) [cm-1]: ν~ = 3357, 2986, 2933, 2359, 1735, 1686,

1620, 1523, 1439, 1221, 1156, 1056, 946, 857. - MF: C9H16N2O5. - MW = 232.23

g/mol.

Tert-butyl 2-(2-(2-(Boc-amino)acetamido)acetamido)ethylcarbamat (49):

Boc-Gly-Gly-OH (48) (1.2 g, 5.2 mmol), HOBt (0.91 g, 6.7 mmol) and DIPEA (2.7 ml,

15 mmol) were dissolved in DCM (15 ml). To the solution

tert-butyl 2-aminoethylcarbamate (45) (1.2 g, 7.8 mmol) was added and the mixture

was cooled to 0 °C in an ice bath. Then EDC (1.1 ml, 6.7 mmol) was added and the

reaction was monitored by TLC (EtOAc). After 24 h the solution was added to water

and extracted with DCM. The combined organic layers were washed twice with an

aqueous citric acid solution (10 %), twice with brine, the organic layer was dried over

MgSO4 and concentrated under reduced pressure. The crude product was purified by

column chromatography on silica gel (EtOAc:EtOH 80:20, Rf = 0.09) to give the

product as a colorless oil (0.81 g, 2.2 mmol, 42 %).


1/18), 3.19-3.30 (s, 2 H, 13), 3.32-3.43 (m, 2 H, 12), 3.85 (s, 2 H, 6), 3.97 (d, 3JH,H =

5.5, 2 H, 9), 5.31 (bs, 1 H, 14), 5.56 (bs, 1 H, 5), 7.19 (bs, 2 H, 8 + 11). - 13C-NMR

(75 MHz, CDCl3): d = 28.3 (+, 3 C, 1/18), 28.4 (+, 3 C, 1/18), 40.4 (-, 2 C, 9 + 10),

42.9 (-, 1 C, 9), 44.4 (-, 1 C, 6), 79.8 (Cquat, 1 C, 2/117), 80.5 (Cquat, 1 C, 2/17), 156.4

(Cquat, 1 C, 4/15) , 156.8 (Cquat, 1 C, 4/15), 169.4 (Cquat, 1 C, 10), 170.4 (Cquat, 1 C, 7).

- MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 319 (36) [MH+ - C4H8], 375

(100) [MH+], 392 (32) [MNH4+]. - IR (neat) [cm-1]: ν~ = 3280, 3184, 2983, 2360, 1677,

1660, 1569, 1464, 1262, 1178, 1108, 954, 878. - MF: C16H30N4O6. - MW = 374.43

g/mol.

HN

NH

O HNO

O

1 34

56

7

8 910

11122

ONH

O

O1716

15

1413

18

S-32

2-Amino-N-(2-(2-aminoethylamino)-2-oxoethyl)acetamide · 2 HCl (16):

Compound 49 (0.81 g, 2.2 mmol) was dissolved in methanol and cooled to 0 °C in an

ice bath. To this mixture six pipettes of HCl saturated ether were added. The mixture

was allowed to warm to room temperature and stirred over night. The reaction

progress was controlled by 1H-NMR. The mixture was concentrated under reduced

pressure and dried under high vacuum to obtain the product as a colorless solid in

90% yield (0.48 g, 1.9 mmol).

MP: > 250 °C. - 1H-NMR (300 MHz, D2O): d = 3.07 (t, 3JH,H = 5.8, 2 H, 9), 3.45 (t, 3JH,H = 5.9, 2 H, 8), 3.82 (s, 2 H, 2), 3.92 (s, 2 H, 5). - 13C-NMR (75 MHz, D2O): d =

36.9 (-, 1 C, 9), 39.2 (-, 1 C, 8), 40.5 (-, 1 C, 2), 42.5 (-, 1 C, 5), 168.0 (Cquat, 1 C,

CO), 172.2 (Cquat, 1 C, CO). - IR (neat) [cm-1]: ν~ = 3280, 2990, 2879, 2359, 2016,

1677, 1660, 1570, 1465, 1264, 1178, 1108, 1009, 906, 788. - MF: C6H14N4O2 x 2

HCl. - MW = 247.12 g/mol.

Hexa-tert-butyl 10,10'-(6-(2-(2-(2-(4,6-bis(4,7,10-tris(tert-butoxycarbonyl)-1,4,7,10-

tetraazacyclododecan-1-yl)-1,3,5-triazin-2-ylamino)acetamido)acetamido)-

ethylamino)-1,3,5-triazine-2,4-diyl)bis(1,4,7,10-tetraazacyclododecane-1,4,7-

tricarboxylate) (50)

Compound 16 (0.35 g, 1.4 mmol) was suspended in dioxane and potassium

carbonate (7.9 g, 57 mmol) was added. The mixture was stirred for 30 min, then

substance 14 (3.0 g, 2.8 mmol) was added and the solution was stirred for three days

at 145 °C. Potassium carbonate was filtered off, the solution was evaporated to

dryness and the crude product was purified by chromatography on silica gel with

H2NNH

O HN

ONH2

* 2HCl1 2

34 5

67 8

99

NN

N

N N

NN

BocBoc

Boc

N N

NNBoc

Boc

Boc

HN

O HN

NN

N

NN

N N

Boc Boc

Boc

NN

N NBoc

Boc

Boc

NH O

NH

S-33

EtOAc (Rf = 0.30) as eluent to give the product as a colorless solid (1.9 g, 0.62 mmol,

44 %).

MP: 162-163 °C. - 1H-NMR (300 MHz, CDCl3): d = 1.43 (s, 108 H, Boc-CH3), 2.85-

4.08 (m, 72 H, Cyclen-CH2 + NH-CH2), 5.05 (bs, 2 H, NH). - 13C-NMR (75 MHz,

CDCl3): d = 28.5 (+, 36 C, Boc-CH3), 39.9 (-, 1 C, NH-CH2), 42.8 (-, 1 C, NH-CH2-

CH2), 44.8 (-, 1 C, CH2-CO), 47.4 (-, 1 C, CH2-CO), 50.3 (-, 32 C, Cyclen-CH2), 79.8

(Cquat, 12 C, OC(CH3)3), 156.8 (Cquat, 12 C, Boc-CO), 165.8 (Cquat, 6 C, Ar-C), 169.5

(Cquat, 1 C, CH2-CO). - MS (ES, DCM/MeOH + 10 mmol/l NH4OAc): m/z (%) = 1108

(100) [M + 2 H+], 2115 (8) [MH+]. - IR (neat) [cm-1]: ν~ = 2974, 2933, 2360, 2344,

1686, 1539, 1466, 1409, 1364, 1246, 1158, 971, 858. - MF: C102H184N26O26. - MW =

2214.73 g/mol.

Complex 6:

The Boc protected compound 50 (1.4 g, 0.62 mmol) was dissolved in methanol and

cooled to 0 °C in an ice bath. To this solution HCl saturated ether (0.7 ml/mmol

Boc-group) was added. The mixture was allowed to warm to room temperature and

was stirred over night. The solvent was evaporated, the crude product was dissolved

in water and purified over an ion exchanger column using a strongly basic ion

exchanger (OH- -form, loading 0.9 mmol/ml, 4 eq. per protonated nitrogen) to give a

colorless solid after lyophylization. Then two aqueous solutions were prepared: one

containing the obtained colorless solid from the column and a second one containing

Zn(ClO4)2 · 6 H2O (0.90 g, 2.4 mmol, 4.4 eq. per deprotected receptor). These two

solutions were simultaneously added drop wise to water at 80 °C under vigorous

stirring. The resulting mixture was stirred at 90 °C over night, and then lyophilized,

and the product was recrystallized from a water methanol mixture to give the complex

as a violet red solid in 63 % yield. (0.75 g, 0.39 mmol).

NN

NN

NH

NN

NNH

H

N N

NH

H H

H

NN

NN

HN

N N

N NH

H

NN

NH

HH

H

Zn2+

Zn2+

Zn2+

Zn2+

OH--HO

6+

6 ClO4-

NHO

HN

O

S-34

MP > 250 °C. - MS (ES, H2O/AcN/MeOH + 10 mmol/l NH4OAc) (K8+ =

C44H88N26O2Zn4) : m/z (%) = 277 (82) [K8+ + 4 CH3COO-], 503 (52) [K8+ + OH- + Cl- +

3 CH3COO-], 536 (100) [K8+ + 4 CH3COO- + ClO4-], 854 (15) [K8+ + OH- + 2 CH3COO-

+ 3 ClO4-], 875 (14) [K8+ + OH- + CH3COO- + 4 ClO4

-], 896 (16) [K8+ + OH- + 5 ClO4-].

- IR (neat) [cm-1]: ν~ = 3585, 3387, 3296, 2943, 2894, 2360, 1652, 1548, 1480, 1424,

1343, 1286, 1232, 1067, 962, 808. - MF: [C44H90N26O2Zn4]8+(ClO4)6(OH)2. - MW =

1905.59 g/mol.

2. Solid-phase peptide synthesis

The peptides were synthesized on an Advanced Chemtech 496 MOS synthesizer.

Rink Amide MBHA resin and Fmoc protecting group strategy were used throughout

the syntheses. Coupling was achieved by TBTU / HOBt / DIPEA. HOBt was used as

a 0.45 M solution, TBTU as a 0.44 M solution and DIPEA as a 1.2 M solution, all in

DMF. The Fmoc protected amino acids were dissolved in NMP as 0.4 M solutions.

Carboxyfluorescein was coupled as a normal amino acid. The syntheses were

carried out in a 96 well reactor. All peptides were synthesized on 50 mg of resin. The

lot of the resin used had a loading of 0.72 mmole/g (manufacturer’s claims). Before

each synthesis the resin was allowed to preswell in DMF for 30 min. Each coupling

was done twice using a 5 fold excess of HOBt and slightly less than 5 fold excess of

TBTU. DIPEA was used in 10 fold excess. Fmoc deprotection was done by shaking

the resin with 40 % piperidine in DMF for 3 minutes, subsequent washing and

addition of 20 % piperidine in DMF followed by shaking for 10 minutes. After

completion, the resin was washed with MeOH, DCM and diethylether (5 x 2 ml each).

Phosphorylated serine was introduced using Fmoc-Ser(PO(OBzl)OH)-OH as the

phosphorylated building block. Cleavage of the peptides from the resin was afforded

by addition of 1.5 ml TFA / phenol / H2O / thioanisole / EDT (82.5 : 5 : 5 : 2.5) and

subsequent shaking for 3 h. After filtering off the resin, the TFA solution was reduced

in volume to about 0.5 ml. It was then transferred to a Falcon tube and precipitated

with cold diethylether. The precipitate was centrifuged at -4 °C for 10 minutes. The

solution was then carefully decanted off and the precipitate resuspended in cold

diethylether before being centrifuged again. This resuspending/centrifuging step was

repeated five times. Finally, the diethylether was decanted off again and the peptide

S-35

dried under vacuum. The peptides were analyzed by ES-MS, LC-MS and HPLC and

purified by preparative HPLC.

3. Binding studies

Fluorescence titrations

Instrument Parameters

Excitation wavelength: λex = 494 nm

Detection wavelength: λem?500 – 600 nm

Temperature: T = 298 K

PMT voltage: depending on concentration

Titration conditions

Solvent: 50 mM HEPES buffer, pH 7.5, 154 mM NaCl

Starting volume: 0.80 ml

Procedure

A Varian Cary Eclipse Fluorometer was used for the emission titrations. The cuvette

with 0.80 ml of peptide in HEPES buffer was titrated stepwise with small amounts

(beginning with 0.13 eq) of the receptor solution. After each addition the solution was

allowed to equilibrate for 2 min before the fluorescence intensity and the UV

spectrum (where permitted by the concentration range) were recorded. The

stoichiometries were determined by Job’s plots calculated from the titration data. To

determine the binding constants, the obtained fluorescence intensities at 520 nm

were volume corrected, plotted against the concentration of receptor and evaluated

by nonlinear fitting.

Fluorescence polarization titrations were conducted under identical conditions using

an ISS K2 Multifrequency Phase Fluorometer.

S-36

Fluorescence titration results of Flu-GpSAAHV-NH2 (2)

0,0 1,0x10-4 2,0x10-4-10

0

10

20

30

40

50

60

70

∆ in

tens

ity

c (3) [M]

0,0 1,0x10-4 2,0x10-4-10

0

10

20

30

40

50

60

70

80

∆ in

tens

ity

c (4) [M]

Figure S-1. Emission response of peptide 2 upon addition of receptor 3 (left) and 4

(right)

0,0 1,0x10-7 2,0x10-7

0

50

100

150

200

250

300

350

∆ in

tens

ity

c (5) [M]

0,0 5,0x10-7 1,0x10-6 1,5x10-6

0

20

40

60

80

100

120

∆ in

tens

ity

c (6) [M]


(right)

S-37

0,0 1,0x10-5 2,0x10-5 3,0x10-5 4,0x10-5

0

20

40

60

80

100∆

inte

nsity

c (7) [M]

0,0 5,0x10-6 1,0x10-5 1,5x10-5

0

10

20

30

40

50

60

∆ in

tens

ity

c (8) [M]


(right)

0,0 1,0x10-4 2,0x10-4

0

10

20

30

40

50

60

∆ in

tens

ity

c (9) [M]

0,0 5,0x10-5 1,0x10-4 1,5x10-4 2,0x10-4-20

020406080

100120140160180

∆ in

tens

ity

c (10) [M]

Figure S-4. Emission response of peptide 2 upon addition of Complex 9 (left) and 10

(right)

S-38

Fluorescence titration results of Flu-GpSAAEV-NH2 (1)

0,0 1,0x10-8 2,0x10-8

0

20

40

60

80

∆ in

tens

ity

c (3) [M]

0,0 1,0x10-8 2,0x10-8

0

20

40

60

80

100

∆ in

tens

ity

c (4) [M]


(right)

0,0 2,0x10-5 4,0x10-5 6,0x10-5

0

20

40

60

80

∆ in

tens

ity

c (5) [M]

0,0 2,0x10-5 4,0x10-5 6,0x10-5

0

20

40

60

80

100

120

∆ in

tens

ity

c (6) [M]


(right)

S-39

0,0 1,0x10-8 2,0x10-8

0

20

40

60

80

100∆

inte

nsity

c (7) [M]

0,0 1,0x10-8 2,0x10-8

0

10

20

30

40

50

60

∆ in

tens

ity

c (8) [M]


(right)

0,0 5,0x10-5 1,0x10-4

0

5

10

15

20

25

∆ in

tens

ity

c (10) [M]

Figure S-8. Emission response of peptide 1 upon addition of complex 10

S-40

Fluorescence polarization results with Flu-GpSAAEV-NH2 (1)

0,0 1,0x10-8 2,0x10-8

0,00

0,05

0,10

0,15

0,20

∆ po

lariz

atio

n

c (3) [M]

0,0 1,0x10-8 2,0x10-8

0,00

0,05

0,10

0,15

0,20

0,25

∆ po

lariz

atio

n

c (7) [M]

Figure S-9. Fluorescence polarization response of peptide 1 upon addition of

receptor 3 (left) and 7 (right)

References

[1] S. Hünig, G. Märkl, J. Sauer, Einführung in die apparativen Methoden in der

Organischen Chemie, 2nd Edition, Würzburg, Regensburg, 1994; Organikum, 17th

Edition, VEB Deutscher Verlag der Wissenschaften, Berlin, 1988. [2] M. Subat, K. Woinaroschy, S. Anthofer, B. Malterer, B. Koenig, Inorg. Chem.

2007, 46, 4336-4356. [3] D. S. Turygin, M. Subat, O. A. Raitman, V. V. Arslanov, B. Koenig, M. A. Kalinina,

Angew. Chem. Int. Ed. 2006, 45, 5340-5344. [4] L. Huang, J. C. Quada Jr., J. W. Lown, Bioconjugate Chem. 1995, 6, 21-33. [5] J. R. Bergeron, J. S. McManis, J. Org. Chem. 1987, 52, 1700-1703. [6] C. Dardonville, C. Fernandez-Fernandez, S.-L. Gibbons, G. J. Ryan, N. Jagerovic,

A. M. Gabilondo, J. J. Meana, L. F. Callado, Bioorg. Med. Chem. 2006, 14, 6570-

6580.

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