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C. Wängler et al. / Bioorg. Med. Chem. 19 (2011) 3864–3874
0–40% MeCN + 0.1% TFA in 8 min as the gradient (Rt = 3.2 min). The
145
lmol) in DMF (200
l
L) and reacted for 60 min. After acidificat-
product was obtained as white, highly hygroscopic solid after
ion with 1 M HCl (60
lL), the product was purified by semi-pre-
lyophilization (46 mg, 70
lmol, 37% yield).
parative HPLC with 10–65% MeCN + 0.1% TFA in 8 min as the
1H NMR (500 MHz, DMSO-d6, 25 °C, TMS): d = 10.09 (s, 1H);
9.35 (t, 1H, 3J(H,H) = 6.0 Hz); 8.97 (br s, 1H); 8.09–8.07 (m, 2H);
8.02–8.00 (m, 2H); 7.32–7.31 (m, 2H); 7.28–7.26 (m, 2H); 4.48
(br d, 2H, 3J(H,H) = 5.9 Hz); 4.32 (br d, 2H, 3J(H,H) = 5.5 Hz); 4.01
(br s, 2H); 3.94 (br s, 2H); 3.63 (br s, 4H); 3.33 (br s, 8H); 3.12
(br s, 8H). 13C NMR (125 MHz, DMSO-d6, 25 °C, TMS): d = 192.84;
165.29; 158.13; 157.87; 139.24; 138.20; 137.73; 136.91; 129.36;
127.89; 127.49; 127.31; 54.60; 54.58; 53.89; 53.86; 52.79; 52.77;
52.76; 50.50; 50.48; 50.46; 50.33; 50.31; 48.53; 48.51; 48.49;
48.42; 48.39; 42.46; 42.05. ESI-MS (m/z) for [M+H]+ (calculated):
655.31 (655.30).
gradient (Rt = 4.2 min). The product was obtained as brownish,
highly hygroscopic solid after lyophilization (13 mg, 17
11% yield).
lmol,
1H NMR (500 MHz, acetone-d6, 25 °C, TMS): d = 8.94 (br s, 1H);
7.62 (br s, 1H); 7.30–7.28 (m, 2H); 7.26–7.24 (m, 2H); 4.44 (br d,
2H, 3J(H,H) = 5.5 Hz); 4.36 (br d, 2H, 3J(H,H) = 5.6 Hz); 4.33–4.04
(br s, 8H); 3.77–3.62 (br s, 4H); 3.59–3.40 (br s, 4H); 3.35–3.10
(br s, 8H); 2.36 (t, 1H, 4J(H,H) = 2.7 Hz); 2.35 (t, 2H,
3J(H,H) = 7.4 Hz); 2.23 (dt, 2H, 3J(H,H) = 7.1 Hz, 4J(H,H) = 2.7 Hz);
1.81 (p, 2H, 3J(H,H) = 7.2 Hz); 1.51 (s, 9H); 1.47 (s, 18H). 13C NMR
(125 MHz, acetone-d6, 25 °C, TMS): d = 172.33; 160.33; 160.06;
139.72; 128.55; 128.38; 84.46; 82.20; 70.29; 58.03; 57.93; 56.17;
56.07; 56.00; 55.88; 52.53; 52.45; 52.31; 52.28; 50.37; 50.08;
49.97; 49.85; 43.13; 43.00; 35.19; 28.34; 25.44; 18.40. ESI-MS
(m/z) for [M+H]+ (calculated): 785.52 (785.51).
2.2.5. [N-(4-(Aminomethyl)benzyl)-2-(aminooxy)acetamido]-
DOTA (aminooxy-DOTA) (7)
A solution of HBTU (87.2 mg, 230
added to Boc-aminooxy-acetic acid (45.8 mg, 239
by DIPEA (41 L, 239 mol) and reacted for 2 min. Subsequently,
the mixture was added to a solution of amino-DOTA (5, 100 mg,
192 mol) in DMF (250 L) and reacted for 30 min. After acidificat-
ion with 1 M HCl (ꢂ70 L), the product was purified by semi-pre-
l
mol) in DMF (500
lL) was
l
mol) followed
l
l
2.2.8. tert-Butyl 2,20,200-(10-(2-(4-((5-azidopentanamido)methyl)-
benzylamino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-
1,4,7-triyl)triacetate (P-DOTA-azide) (10)
l
l
l
A solution of HBTU (65.9 mg, 174
added to 5-azidopentanoic acid (25.9 mg, 181
DIPEA (30.9 L, 181 mol) and reacted for 2 min. Subsequently, the
mixture was added to a solution of P-amino-DOTA (8, 100 mg,
145 mol) in DMF (200 L) and reacted for 60 min. After acidification
L), the product was purified by semi-preparative
lmol) in DMF (500
lL) was
parative HPLC with 0–20% MeCN + 0.1% TFA in 8 min as the
gradient (Rt = 2.5 min). After solvent removal, the obtained color-
less solid was dissolved in a mixture of neat TFA (3 mL) and TIS
(100 lL) and reacted for 15 min. The product was purified by
semi-preparative HPLC with 0–20% MeCN + 0.1% TFA in 8 min as
lmol) followed by
l
l
l
l
with 1 M HCl (ꢂ50
l
the gradient (Rt = 2.0 min). The product was obtained as white,
HPLC with 0–60% MeOH + 0.1% TFA in 8 min as the gradient
highly hygroscopic solid after lyophilization (32 mg, 53.8
28% overall yield).
lmol,
(Rt = 6.6 min). The product was obtained as colorless, highly hygro-
scopic solid after lyophilization (46 mg, 56 lmol, 39% yield).
1H NMR (500 MHz, water-d2, 25 °C, TMS): d = 7.23 (s, 4H); 4.58
(s, 2H); 4.35 (s, 2H); 4.30 (s, 2H); 3.66 (br s, 8H); 3.21 (br s, 16H).
13C NMR (125 MHz, water-d2, 25 °C, TMS): d = 168.81; 163.26;
162.90; 136.73; 136.70; 127.91; 127.87; 71.97; 55.16; 54.07;
53.85; 53.24; 52.72; 49.90; 49.81; 49.50; 49.26; 49.23; 49.11;
48.64; 45.25; 45.15; 42.96; 42.57. ESI-MS (m/z) for [M+K+H]+
(calculated): 635.34 (635.26).
1H NMR (500 MHz, acetone-d6, 25 °C, TMS): d = 8.58 (br s, 1H);
7.72 (br s, 1H); 7.30–7.24 (m, 4H); 4.47 (br d, 2H, 3J(H,H) = 5.2 Hz);
4.37 (br d, 2H, 3J(H,H) = 5.8 Hz); 4.30 (br s, 2H); 3.93–3.16 (br m,
22H); 3.35 (t, 2H, 3J(H,H) = 6.6 Hz); 2.28 (t, 2H, 3J(H,H) = 7.1 Hz);
1.74–1.58 (m, 4H); 1.54 (s, 9H); 1.47 (s, 18H). 13C NMR (125 MHz,
acetone-d6, 25 °C, TMS): d = 172.76; 160.33; 159.96; 139.84;
137.99; 128.61; 128.42; 82.93; 56.00; 55.96; 55.48; 55.42; 54.96;
54.93; 54.92; 52.21; 52.19; 52.14; 52.09; 51.77; 49.97; 49.95;
49.91; 49.86; 49.77; 43.50; 42.99; 35.91; 29.11; 28.35; 23.63. ESI-
MS (m/z) for [M+H]+ (calculated): 816.53 (816.53).
2.2.6. Tris-tBu-(phenylene-1,4-dimethylamino-)DOTA (P-amino-
DOTA) (8)
A solution of HBTU (314.5 mg, 830
added to tris-tBu-DOTA (500 mg, 874
(149 L, 874 mol) and reacted for 2 min. Subsequently, the mix-
ture was added to a solution of 1,4-bis(aminomethyl)benzene
l
mol) in DMF (1.5 mL) was
lmol) followed by DIPEA
2.2.9. 2,20,200-(10-(2-(4-(Hex-5-ynamidomethyl)benzylamino)-2-
oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic
acid (DOTA-alkyne) (11)
l
l
(297 mg, 2.18 mmol) in DMF (250
lL) and reacted for 30 min. After
A solution of HBTU (43.6 mg, 115
added to 5-hexynoic acid (13.4 mg, 120
(22 L, 120 mol) and reacted for 2 min. Subsequently, the mix-
ture was added to a solution of amino-DOTA (5, 50 mg, 96 mol)
in DMF (250 L) and reacted for 30 min. At this time, water
(250 L) was added and the reaction was continued for further
15 min. After acidification with HCl (1 M, 35 L), the product was
purified by semi-preparative HPLC with 0–20% MeCN + 0.1% TFA
in 8 min as the gradient (Rt = 3.5 min). The product was obtained
as colorless, highly hygroscopic solid after lyophilization
l
mol) in DMF (500
lL) was
acidification with 1 M HCl (ꢂ200
lL), the product was purified by
lmol) followed by DIPEA
semi-preparative HPLC with 0–50% MeCN + 0.1% TFA in 8 min as
the gradient (Rt = 4.3 min). The product was obtained as slightly
green, highly hygroscopic solid after lyophilization (390 mg,
l
l
l
l
565 lmol, 68% yield).
1H NMR (500 MHz, acetone-d6, 25 °C, TMS): d = 7.47 (d, 2H,
3J(H,H) = 8.1 Hz); 7.39 (d, 2H, 3J(H,H) = 7.9 Hz); 5.01 (br s, 2H);
4.61–3.13 (br m, 24H); 4.49 (br s, 2H); 1.53 (s, 9H); 1.46 (s, 18H).
13C NMR (125 MHz, acetone-d6, 25 °C, TMS): d = 160.60; 160.25;
140.10; 130.29; 129.53; 128.85; 55.70; 55.68; 55.22; 55.19;
55.16; 52.27; 52.24; 50.80; 50.66; 50.09; 49.97; 28.34; 28.32.
ESI-MS (m/z) for [M+H]+ (calculated): 691.48 (691.47).
l
l
(17.2 mg, 28 lmol, 29% yield).
1H NMR (500 MHz, water-d2, 25 °C, TMS): d = 7.27–7.22 (m,
4H); 4.79–4.78 (m, 4H); 4.29 (br s, 2H); 4.01–3.53 (br m, 6H);
3.50–2.94 (br s, 16H); 2.34 (t, 2H, 3J(H,H) = 7.4 Hz); 2.26 (t, 1H,
4J(H,H) = 2.6); 2.15 (dt, 2H, 3J(H,H) = 6.9, 4J(H,H) = 2.6); 1.75 (p,
2H, 3J(H,H) = 7.1). 13C NMR (125 MHz, water-d2, 25 °C, TMS):
d = 176.19; 172.32; 137.37; 127.89; 127.70; 84.74; 70.04; 57.33;
57.25; 55.12; 54.04; 51.82; 49.56; 42.97; 42.80; 34.72; 24.24;
17.19. ESI-MS (m/z) for [M+H]+ (calculated): 617.33 (617.32); (m/
z) for [M+Na+K+H]+ (calculated): 679.24 (679.27).
2.2.7. tert-Butyl 2,20,200-(10-(2-(4-(hex-5-ynamidomethyl)
benzylamino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-
1,4,7-triyl)triacetate (P-DOTA-alkyne) (9)
A solution of HBTU (65.9 mg, 174
added to 5-hexynoic acid (20.3 mg, 181
(30.9 L, 181 mol) and reacted for 2 min. Subsequently, the mix-
ture was added to a solution of P-amino-DOTA (8, 100 mg,
lmol) in DMF (500
lL) was
lmol) followed by DIPEA
l
l