(2,5-dioxopyrrolidin-1-yl) 2-iodoacetate

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 117.4 mg of N-hydroxysuccinimidyl iodoacetate in solution in 3 ml of DCM are added to 100 mg of 3-(2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethoxy)propanoic acid. After 2 h at AT, 330 μl of MeOH are added and the mixture is cooled to 0° C. 360 μl of a 2M solution of trimethylsilyldiazomethane in hexane are added. After 1 h, the mixture is neutralized by addition of 50 μl of acetic acid and then a saturated aqueous NaHCO3 solution is added until pH=8 is obtained. The organic phase is dried over MgSO4 and concentrated under RP, and the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G) using a gradient from 0 to 10% of methanol in DCM. 132 mg of methyl 3-[2-(2-{2-[2-(2-iodoacetylamino)ethoxy]ethoxy}ethoxy)ethoxy]propanoate are thus obtained. is obtained The organic phase is dried over MgSO4 concentrated under RP the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G)

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 117.4 mg of N-hydroxysuccinimidyl iodoacetate in solution in 3 ml of DCM are added to 100 mg of 3-(2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethoxy)propanoic acid. After 2 h at AT, 330 μl of MeOH are added and the mixture is cooled to 0° C. 360 μl of a 2M solution of trimethylsilyldiazomethane in hexane are added. After 1 h, the mixture is neutralized by addition of 50 μl of acetic acid and then a saturated aqueous NaHCO3 solution is added until pH=8 is obtained. The organic phase is dried over MgSO4 and concentrated under RP, and the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G) using a gradient from 0 to 10% of methanol in DCM. 132 mg of methyl 3-[2-(2-{2-[2-(2-iodoacetylamino)ethoxy]ethoxy}ethoxy)ethoxy]propanoate are thus obtained. is obtained The organic phase is dried over MgSO4 concentrated under RP the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: DACB was synthesized by covalently conjugating iodoacetyl-deferoxamine and N-cysteinyl-biotin by the following Scheme II. ##STR20## Solid deferoxamine mesylate was added to methanol for a concentration of 20 mM, followed by addition of NaOH to reach a final concentration of 20 mM NaOH. The solution was heated and maintained at 65°. Solid iodoacetic acid N-hydroxysuccinimide ester (NHS-IA) was added to the deferoxamine solution to a final molarity of 50mM NHS-IA and incubated for at least one hour at 65°. The resulting iodoacetyl-deferoxamine (IA-DFO) was purified by HPLC with an EM LiChroCART reverse phase column using a linear gradient of 0-10 ml 100% Buffer A (0.025M phosphate, pH 6, 2 mM NTA) and 10-60 ml 0-100% Buffer B (0.025M phosphate, pH 6, 2 mM NTA, 75% methanol). IA-DFO, but not unconjugated DFO, has an absorbance at 260 nm and thus was purified by measuring the absorbance of the eluate and collecting the fraction with absorbance at 260 nm. The solution was heated maintained at 65° incubated for at least one hour at 65° The resulting iodoacetyl-deferoxamine (IA-DFO) was purified by HPLC with an EM LiChroCART reverse phase column thus was purified collecting the fraction with absorbance at 260 nm

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: FIG. 11 illustrates the synthesis of Mass Tag (2). 4-Amino-benzylamine (Aldrich, 1 mmol), N-succinimidyl iodoacetate (Pierce, 1 mmol), and N,N-diisopropyl ethylamine (DIEA, 100 μL) were mixed in dichloromethane (10 mL) and stirred at room temperature for 1 hr. Solvent was removed under reduced pressure. The product was purified by silica gel column chromatography, eluting with hexane, ethyl acetate (20%-60%), to give 4-amino-N-iodoacetylbenzylamine (62.2% yield). 1H NMR (MeOD): 7.1 (d, 2H), 6.75 (d, 2H), 4.21 (s, 2H), 3.65 (s, 2H). [M+H]+ in MS: 291.0, calculated; 291.0, found. Solvent was removed under reduced pressure The product was purified by silica gel column chromatography eluting with hexane, ethyl acetate (20%-60%)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 78 mg (0.1 mmole) of L-thyroxine was dissolved in 1.0 ml of methanol on addition of 34 mls (0.2 mmole) of 6N NaOH and sonication in a bath sonicator. 48 mg of iodoacetic acid N-hydroxysuccinimide ester was added and the mixture vortexed. 8.5 mls of 6N NaOH was added and the mixture vortexed. TLC (chloroform:methanol:acetic acid 50:6:2 on silica gel) indicated complete reaction. 50 mls of 6N HCl was added and then 4 mls of water and formed a precipitate. The precipitate was collected by centrifugation and washed with 1 ml of methanol and 4 mls of water. After centrifugation the pellet was dried under vacuum to 82 mg of an off-white solid. reaction The precipitate was collected by centrifugation washed with 1 ml of methanol and 4 mls of water After centrifugation the pellet was dried under vacuum to 82 mg of an off-white solid

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N-Hydroxysuccinimide (1 g, 8.7 mmole), iodoacetic acid (8.7 mmole, 1.6 g), and DCC (8.7 mmole, 1.7 g) were added to 250 mL of EtOAc and the mixture was stirred at room temperature for 5 hours. The mixture was filtered and the filtrate concentrated to dryness in vacuo. The residue was crystallized from ethanol to yield 1.74 g (71%) of white crystals, mp 148-150° C. The mixture was filtered the filtrate concentrated to dryness in vacuo The residue was crystallized from ethanol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 40 nm dextran-coated iron oxide nanoparticles (NP; 115,000 g/mole per iron core) were dissolved in 1× phosphate buffered saline (PBS; 137 mM NaCl, 10 mM Phosphate, 2.7 mM KCl, pH 7.4) at a concentration of 2 mg/mL. Vivotag-750 fluorophore was labeled on the NPs such that each NP has around 2 VT-750 fluorophores. The linker N-succinimidyl iodoacetate (SIA) was dissolved in DMSO at 20 mg/mL. The two solutions were mixed to obtain a 1-to-7 mass ratio between iron oxide NPs and SIA for 2 hr at room temperature with shaking. Size exclusion chromatography (column diameter×height=1 cm×30 cm; media: Sephadex G-50-coarse) was used to separate out the excess SIA and to exchange NPs into borate buffer (50 mM sodium borate, 5 mM EDTA, pH 8.3). The two solutions were mixed

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 117.4 mg of N-hydroxysuccinimidyl iodoacetate in solution in 3 ml of DCM are added to 100 mg of 3-(2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethoxy)propanoic acid. After 2 h at AT, 330 μl of MeOH are added and the mixture is cooled to 0° C. 360 μl of a 2M solution of trimethylsilyldiazomethane in hexane are added. After 1 h, the mixture is neutralized by addition of 50 μl of acetic acid and then a saturated aqueous NaHCO3 solution is added until pH=8 is obtained. The organic phase is dried over MgSO4 and concentrated under RP, and the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G) using a gradient from 0 to 10% of methanol in DCM. 132 mg of methyl 3-[2-(2-{2-[2-(2-iodoacetylamino)ethoxy]ethoxy}ethoxy)ethoxy]propanoate are thus obtained. is obtained The organic phase is dried over MgSO4 concentrated under RP the residue is purified by flash chromatography on silica (Interchrom Puriflash Silica 15/35U 10G)