Reaction #48472

Reaction equation

Cystamine hydrochloride

triethylamine

HPMA

→

HPMA

cystamine

Solvents

methanol

methanol

Conditions

Workup

1. 1
   workup.STIRRINGThe mixture was stirred overnight at room temperature
2. 2
   ConcentrationThe solution was concentrated under vacuum
3. 3
   Otherto separate the polymer conjugate
4. 4
   Washeluted with methanol
5. 5
   OtherThe polymer fraction was collected
6. 6
   Othersolvent was removed under vacuum

Procedure

Cystamine hydrochloride (400 mg excess) and triethylamine (1.0 ml, excess) were dissolved in 20 ml methanol, and HPMA copolymer with p-nitrophenyl active ester (Mn=18,300 Da and Mw=22,500 Da, ONp %=9.6 mol-%) (420 mg) in 5 ml methanol was added dropwise to above solution with stirring. The mixture was stirred overnight at room temperature. The solution was concentrated under vacuum and applied to Sephadex LH-20 column to separate the polymer conjugate, eluted with methanol. The polymer fraction was collected and solvent was removed under vacuum giving HPMA copolymer-cystamine conjugates. The product was dissolved in water, and DTPA dianhydride (300 mg, excess) was added to the solution. The mixture was stirred for 30 minutes, and pH was adjusted to 7.0 with NaHCO3. The mixture was stirred at room temperature overnight. The polymer was separated from excess DTPA by passing through a Sephadex G-25 column, eluted with water. GdCl3 was added with xylene orange as an indicator. The solution was neutralized with NaOH to pH 5.0. The resulting solution was passed through a Sephadex G-25 column to remove excess GdCl3 and salt. Finally, the product was lyophilized giving a colorless product, HPMA copolymer-[DTPA-Gd] conjugate, indicated as (C) below. The yield was 340 mg, or 85%.

Source