. Mhz, 09 (s, 1H), 7.49 (br s, 1H), 7.39 (d, 2H, J = 8.8 Hz), 7.12 (br s, 1H), 4.88 (br s, 1H, vol.8, 2307.

, Biotinylated-Isoniconitamide 18. A suspension of NH 2 EGBiotine, TFA, vol.17

, Et 2 O (10 mL) was then added, and the resulting mixture was stirred for 15 min. The white solid was filtered off, washed several times with Et 2 O, and dried, CH 2 Cl 2 (5 mL) was vigorously stirred overnight at rt

H. Nmr,

. Hz, 1.78?1.61 (m, 4H), 1.47 (m, 2H); 13 C NMR, p.100

. Mhz, Biotinylated-Pyridinium Bromide 19. Biotinylated-isoniconitamide 18 (400 mg, 0.83 mmol) was dissolved in MeOH (15 mL). Methyl bromoacetate (0.41 mL, 4 mmol) was added, and the solution was stirred under microwave irradiation for 2 days. After evaporation of the solvent, the oily residue was stirred in acetone to eliminate the excess of reagent. The oil was separated and dried (472 mg, 0.74 mmol). The crude biotinylated

2. and J. =. , 87 (s, 3H), 3.71?3.64 (m, 8H), 3.54 (t, 2H, J = 5.6 Hz), 3.34 (t, 2H, J = 5.6 Hz), 3.20 (m, 1H), 2.92 (dd, 1H, 5.70 (s, 1H), 4.48 (m, 1H), 4.31 (m, 1H), vol.3, p.13

. Biotinylated-indolizine, Biotinylated-pyridinium bromide 19 (140 mg, 0.25 mmol) and para-nitrophenyl propiolate 5 (140 mg, 0.75 mmol) were dissolved in DMF (1.5 mL), vol.20

, 25 mmol) was then added and the solution was stirred at rt for 6 h. A large volume of Et 2 O was added to the solution, and the resulting suspension was stirred for 15 min and then filtered

2. Hz-;-d and J. =. , 90 (s, 3H), 3.69? 3.51 (m, 8H), 3.49?3.45 (m, 2H), 3.39?3.25 (m, 2H), 3.10? 2.99 (m, 1H), 2.78 (dd, 1H, J = 13.2, 5.2 Hz), 2.58 (d, 1H, J = 12.8 Hz), 2.07 (t, 1H, J = 7.6 Hz), 1.62?1.45 (m, 4H), 1.30 (m, 2H); 13 C NMR (100 MHz, Hz), 6.44 (br s, 1H), 6.18 (s, 1H), 5.32 (s, 1H), 4.37 (m, 1H), 4.18 (m, 1H), vol.3

, Biotinylated-COB223. Biotinylated-indolizine 20 was dissolved in CH 2 Cl 2 (2 mL), and N-dansyl-bis(aminopropyl)11 N HCl (1 drop), acetone was added to separate the resulting salt that was obtained as an oil in 44% yield. 1 H NMR (400 MHz, CD 3 OD) ? 9, vol.52

. Hz, 96 (br s, 1H), 8.66 (br s, 1H), 8.46 (d, 1H, J =, vol.8, issue.2

1. Hz-;-d, J. =-;-d, 1. , and J. =. , Hz), 7.96 (m, 2H, J = 9.2 Hz), vol.8, p.47

. Hz, 2.20 (m, 4H), 2.04 (m, 2H), 1.77?1.67 (m, 1H)

, HRMS (ESI) m/z: calcd for C 50 H 70 N 10 NaO 10 S 2

1. and J. , Biotinylated-COB236. This compound was prepared as described for biotinylated-COB223 from biotinylated-indolizine 20 and N-tosyl-bis(aminopropyl)piperazine 21b. 1 H NMR (400 MHz, CD 3 OD) ? 9.51 (dd, vol.8

2. Hz-;-t, J. =-;-t, 2. , and J. =. , , vol.2

, HRMS (ESI) m/ z: calcd for C 45 H 65 N 9 NaO 10 S 2 [M + Na] + 978, vol.4188

A. S. Goldmann, M. Glassner, A. J. Inglis, C. Barner-kowollik, C. S. Mckay et al., A universal and ready-to-use heterotrifunctional crosslinking reagent for facile synthetic access to sophisticated bioconjugates, Triton X-100, and the cytosolic extracts were prepared by centrifugation at 20 000 g. Fractions of 10 mL of placental cytosolic extracts (containing 35 mg of proteins) were diluted ? REFERENCES (1), vol.34, pp.4329-4345, 1075.

J. Jira?c?jira?c?-ek, M. Engelen, A. Lombardi, R. Vitale, L. Lista et al., Bioreductive fluorescent markers for hypoxic cells: a study of 2-nitroimidazoles with 1-substituents containing fluorescent, bridgehead-nitrogen, bicyclic systems, Biotechnol. Appl. Biochem, vol.62, issue.8, p.332, 1095.

. Indolizine-based-novel-fluorophore, . Seoul-fluor, N. Marangoci, L. Popovici, E. Ursu et al., Inhibition of heregulin mediated MCF-7 breast cancer cell growth by the ErbB3 binding protein EBP1. Cancer Lett. 2008, 265, 298?306. (29) Troll, T.; Beckel, H.; Lentner-Bo? hm, C. Electrochemical synthesis of substituted indolizines; UV and fluorescence spectra, Pyridyl-indolizine derivatives as DNA binders and pHsensitive fluorescent dyes, vol.48, pp.362-363, 1706.