Activated polyethylene glycol compounds
View Patent ↗Novel arylisothiocyanate compounds are described that are useful for activating alcohol-containing macromolecules, for example polyethyleneglycols and cellulose, for covalent linkage to amino-groups of biomolecules, for example polypeptides such as antibodies, enzymes, and proteins.
1. A method for making a macromolecule M that is linked to a biomolecule B comprising the following steps:
(a) providing a linking reagent represented by the formula
where n=1 or 2;
R is —NHC(O)—O-M;
M is a reacted alcohol-containing macromolecule; and
R is in a para-, meta- or di-meta position relative to —NCS;
(b) providing an amine-containing biomolecule B in an admixture with the linking reagent provided in step (a) to form a linking mixture; and
(c) maintaining said linking mixture for a time period sufficient to form a urethane compound represented by the chemical formula
where n=1 or 2;
R is —NHC(O)—O-M;
M is a reacted alcohol-containing macromolecule; and
R is in a para-, meta- or di-meta position relative to —N(H)C(S)N(H)—B
thereby making a macromolecule M that is linked to a biomolecule B.
2. The method according to claim 1 wherein said macromolecule M is a polyethylene glycol.
3. The method according to claim 1 wherein said biomolecule B is a polypeptide.
4. The method according to claim 2 wherein said polyethylene glycol M is wherein M is methoxy polyethylene glycol —CH 2 CH 2 —(OCH 2 CH 2 ) x —O—CH 3 ; and x is an average value that is about 5 to about 500.
5. The method according to claim 1 wherein
the linking reagent of step (a) is represented by the formula shown above; and
the urethane compound formed in step (c) is represented by the chemical formula shown above.
6. The method according to claim 1 wherein
the linking reagent of step (a) is represented by the formula shown above; and
the urethane compound formed in step (c) is represented by the chemical formula shown above.