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Preparation of 3-[4-(1-formylpiperazin-4-yl)benzylidenyl]-2-indolinone (SU4984) was as follows: A reaction mixture of 133.15 mg (1.0 mmol) of oxindole, 228.3 mg (1.2 mmol) of 4-(1-formylpiperazin-4-yl)benzaldehyde, and three drops of piperidine in 2 ml of ethanol was stirred at 90°C for 5 hours. After cooling, the precipitate was filtered, washed with cold ethanol, and dried to yield 199.5 mg (65%) of SU4984 as a yellow solid. Nuclear magnetic resonance (NMR) spectroscopy showed that SU4984 exists predominantly in the frans configuration (C-2, C-1 ′), although in the crystal structure, SU4984 is observed in the cis configuration. Preparation of 3-[(3-(2-carboxyethyl)-4-methylpyrrol-2-yl)methylene]-2-indolinone (SU5402) was as follows: A reaction mixture of 134.0 mg (1.0 mmol) of oxindole, 217.43 mg (1.2 mmol) of 3-(2-carboxyethyl)-4-methylpyrrol-2-carboxaldehyde, and three drops of piperidine in 3 ml of ethanol was stirred at 90°C for 3 hours. After cooling, the precipitate was filtered, washed with cold ethanol, and dried to yield 172.4 mg (58%) of SU5402 as a yellow solid. NMR spectroscopy showed that SU5402 exists predominantly in the cis configuration.
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33
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15444351117
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note
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32P incorporation was quantitated by Cerenkov counting with a β counter.
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34
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NIH 3T3 cells expressing endogenous FGF receptors were used. Cell culture, cell lysis, immunoprecipitation, and immunoblotting were carried out according to standard procedures. Anti-FGFR1 was described previously [M. Mohammadi ef al., Mol. Cell. Biol. 16, 977 (1996)].
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3H]Thymidine incorporation was done as described by Mohammadi et al. (19)
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3H]Thymidine incorporation was done as described by Mohammadi et al. (19).
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Equipment in the structural biology program at the Skirball Institute is partially supported by a grant from the Kresge Foundation. Coordinates have been deposited in the Brookhaven Protein Data Bank, entries 1FGI and 1AGW.
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