N-terminally chemically modified protein compositions and methods
AssigneeAmgen Inc.
InventorsOlaf Kinstler, Nancy Gabriel, Christine Farrar, Randolph DePrince
Life of this patent
Estimating the term…
Abstract
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.
Claims38
A substantially homogenous preparation of N-terminally chemically modified G-CSF or analog thereof, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant.
A substantially homogenous preparation of N-terminally monopegylated G-CSF, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant, wherein:
- (a) said G-CSF has the amino acid sequence identified in SEQ. ID No. 1;
- (b) said G-CSF is monopegylated with a polyethylene glycol moiety having a molecular weight of about 12 kDa.
A pharmaceutical composition comprising:
- (a) a substantially homogenous preparation of monopegylated G-CSF, said monopegylated G-CSF consisting of a polyethylene glycol moiety having a molecular weight of about 12 kDa connected to a G-CSF moiety solely at the N-terminus thereof via an amine linkage;
- (b) fewer than 5% non-pegylated G-CSF molecules;
- and (c) a pharmaceutically acceptable diluent, adjuvant or carrier.
A method for attaching a water soluble polymer to a protein or analog thereof, wherein said water soluble polymer has a single reactive aldehyde group, said method comprising:
- (a) reacting a protein moiety with a water soluble polymer moiety under reducing alkylation conditions, at a pH sufficiently acidic to selectively activate the α-amino group at the amino terminus of said protein moiety so that said water soluble polymer selectively attaches to said α-amino group;
- and
- (b) obtaining the reaction product and
- (c) optionally, separating the reaction products from unreacted moieties.
A method for attaching a polyethylene glycol molecule to a G-CSF molecule, wherein said polyethylene glycol molecule has a single reactive aldehyde group, said method comprising:
- (a) reacting said G-CSF with said polyethylene glycol molecule under reducing alkylation conditions, at a pH sufficiently acidic to selectively activate the α-amino group at the amino terminus of said G-CSF;
- and
- (b) obtaining the pegylated G-CSF and
- (c) optionally, separating the pegylated G-CSF from non-pegylated G-CSF.
Chemically modified consensus interferon comprised of a consensus interferon protein moiety connected to at least one water soluble polymer moiety.
A chemically modified consensus interferon comprised of IFN-con1 connected to at least one polyethylene glycol moiety.
Pegylated consensus interferon.
A method for attaching a water soluble polymer to consensus interferon, wherein said water soluble polymer has a single reactive aldehyde group, said method comprising:
- (a) reacting a consensus interferon moiety with a water soluble polymer moiety under reducing alkylation conditions, at a pH sufficiently acidic to selectively activate the α-amino group at the amino terminus of said consensus interferon moiety;
- and
- (b) obtaining the reaction product and
- (c) optionally, separating the reaction products from unreacted moieties.
A method for attaching a polyethylene glycol molecule to a consensus interferon molecule, wherein said polyethylene glycol molecule has a single reactive aldehyde group, said method comprising:
- (a) reacting said consensus interferon with said polyethylene glycol molecule under reducing alkylation conditions, at a pH sufficiently acidic to selectively activate the α-amino group at the amino terminus of said consensus interferon;
- and
- (b) obtaining the pegylated consensus interferon and
- (c) optionally, separating the pegylated consensus interferon from non-pegylated consensus interferon.
A substantially homogenous preparation of monopegylated consensus interferon.
A pharmaceutical composition comprising:
- (a) a substantially homogenous preparation of monopegylated consensus interferon, said monopegylated consensus interferon consisting of a polyethylene glycol moiety connected to a consensus interferon moiety solely at the N-terminus thereof via an amine linkage;
- (b) fewer than 5% non-pegylated consensus interferon molecules;
- and (c) a pharmaceutically acceptable diluent, adjuvant or carrier.
Dependencies are inferred from the claim text.