Non-antigenic branched polymer conjugates
AssigneeEnzon, Inc.
InventorsAnthony Martinez, Richard B. Greenwald
Life of this patent
Estimating the term…
Abstract
Branched, substantially non-antigenic polymers are disclosed. Conjugates prepared with the polymers and biologically active molecules such as proteins and peptides demonstrate extended circulating life in vivo. Substantially fewer sites on the biologically active material are used as attachment sites. Methods of forming the polymer, conjugating the polymers with biologically active moieties and methods of using the conjugates are also disclosed.
Claims40
A branched substantially non-antigenic polymer comprising the formula:
- (R).sub.n L-A
- wherein (R) is a water soluble, substantially non-antigenic polymer containing a C.sub.1 -C.sub.4 terminal group;
- (n)=2 or 3;
- (A) is a functional group capable of bonding with a biologically active nucleophile selected from the group consisting of proteins, peptides, polypeptides, enzymes and chemotherapeutic molecules or a moiety capable of being functionalized to react with said nucleophile;
- and
- (L) is selected from the group consisting of ##STR16## wherein (.alpha.) is 1 to 5.
A method of forming a biologically active conjugate, comprising:
- contacting a biologically active nucleophile selected from the group consisting of proteins, peptides, polypeptides, enzymes and chemotherapeutic molecules with an activated branched non-antigenic polymer having a structure represented by:
- (R).sub.n L-A
- wherein:
- (R) is a water-soluble substantially non-antigenic polymer containing a C.sub.1 -C.sub.4 terminal group;
- (n)=2 or 3;
- (L) is selected from the group consisting of:
- ##STR17## wherein (.alpha.) is 1 to 5;
- and (A) is a functional group capable of forming a covalent bond with said biologically active nucleophile.
A polymer conjugate comprising the formula:
- [(R).sub.n L-A.sup.1 ].sub.z -(nucleophile)
- wherein:
- (R) is a water-soluble substantially non-antigenic polymer containing a C.sub.1 -C.sub.4 terminal group;
- (n)=2 or 3;
- (L) is selected from the group consisting of:
- ##STR18## wherein (.alpha.) is 1 to 5;
- (nucleophile) is a biologically active nucleophile selected from the group consisting of proteins, peptides, polypeptides, enzymes and chemotherapeutic molecules;
- (A.sup.1) represents a covalent linkage between (L) and the biologically active nucleophile;
- and
- (z) represents the number of polymers attached to the nucleophile.
A method of preparing a succinimidyl carbonate active ester of a branched non-antigenic polymer comprising the steps of:
- i) contacting a branched non-antigenic polymer having a structure represented by:
- (R).sub.n L-A
- wherein (R) independently comprises a water-soluble substantially non-antigenic polymer containing a C.sub.1 -C.sub.4 terminal group;
- (n)=2 or 3;
- (L) is selected from the group consisting of:
- ##STR19## wherein (.alpha.) is 1 to 5;
- and (A) is a functional group capable of forming a covalent bond with a nucleophile;
- with p-nitrophenyl chloroformate;
- and
- ii) reacting the p-nitrophenyl carbonate active ester of step i) with N-hydroxysuccinimide.
A method of preparing a succinimidyl carbonate active ester of a branched non-antigenic polymer comprising
- contacting a branched non-antigenic polymer having a structure represented by:
- (R).sub.n L-A
- wherein (R) independently comprises a water-soluble substantially non-antigenic polymer containing a C.sub.1 -C.sub.4 terminal group;
- (n)=2 or 3;
- (L) is selected from the group consisting of:
- ##STR20## wherein (.alpha.) is 1 to 5;
- and (A) is a functional group capable of forming a covalent bond with a nucleophile with bis-succinimidyl carbonate.
Dependencies are inferred from the claim text.