N-terminally chemically modified protein compositions and methods
View Patent ↗Provided herein are methods and compositions relating to the attachment of water soluble polymers to proteins. Provided are novel methods for N-terminally modifying proteins or analogs thereof, and resultant compositions, including novel N-terminally chemically modified G-CSF compositions and related methods of preparation. Also provided is chemically modified consensus interferon.
1. A preparation comprising a predominant amount of a monopolymer/protein conjugate prepared from a hematopoietic protein having one or more available epsilon amino reactive groups,
wherein the monopolymer/protein conjugate has a polyethylene glycol polymer moiety located at the N-terminus but not on an epsilon amino group, and the polyethylene glycol polymer moiety is conjugated via an alpha amino group and,
wherein the monopolymer/protein conjugate has a prolonged in vivo biological activity relative to the non-conjugated hematopoietic protein.
2. The preparation of claim 1 wherein the preparation comprises greater than 80% monopolymer/protein conjugate.
3. The preparation of claim 1 wherein the preparation comprises at least 90% monopolymer/protein conjugate.
4. The preparation of claim 1 wherein the preparation comprises greater than 95% monopolymer/protein conjugate.
5. The preparation of claim 1 wherein the preparation comprises at least 99% monopolymer/protein conjugate.
6. A preparation of a monopegylated hematopoietic protein prepared from a hematopoietic protein having one or more available epsilon amino reactive groups,
wherein a polyethylene glycol polymer moiety is conjugated predominantly at the N-terminus of the hematopoietic protein and is conjugated via an alpha amino group and
wherein the monopegylated hematopoietic protein has a prolonged in vivo biological activity relative to the non-conjugated hematopoietic protein.
7. The hematopoietic protein preparation of claim 1 or 6 wherein the polyethylene glycol has a molecular weight of between about 2 kDa and about 100 kDa.
8. The hematopoietic protein preparation of claim 7 wherein the polyethylene glycol has a molecular weight of about 6 kDa.
9. The hematopoietic protein preparation of claim 7 wherein the polyethylene glycol has a molecular weight of about 12 kDa.
10. The hematopoietic protein preparation of claim 7 wherein the polyethylene glycol has a molecular weight of about 20 kDa.
11. The hematopoietic protein preparation of claim 7 wherein the polyethylene glycol has a molecular weight of about 25 kDa.
12. A pharmaceutical composition comprising (a) the preparation of claim 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 or 11 and (b) a pharmaceutically acceptable diluent, carrier or adjuvant.
13. A pharmaceutical composition comprising (a) the preparation of claim 7 and (b) a pharmaceutically acceptable diluent, carrier or adjuvant.