Claims for Patent: 5,824,784

Summary for Patent: 5,824,784

Title:	N-terminally chemically modified protein compositions and methods
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.
Inventor(s):	Kinstler; Olaf B. (Thousand Oaks, CA), Gabriel; Nancy E. (Newbury Park, CA), Farrar; Christine E. (Newbury Park, CA), DePrince; Randolph B. (Raleigh, NC)
Assignee:	Amgen Inc. (Thousand Oaks, CA)
Application Number:	08/321,510
Patent Claims:	1. A substantially homogenous preparation of N-terminally PEGylated G-CSF or analog thereof, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant, said preparation being essentially free of G-CSF or analog thereof PEGylated at sites other than the N-terminus. 2. A preparation of claim 1 wherein the polyethylene glycol used for conjugation has a molecular weight of between about 2 kDa and 100 kDa. 3. A preparation of claim 2 wherein said polyethylene glycol has a molecular weight of between about 6 kDa and 25 kDa. 4. A preparation of claim 1 wherein said preparation is comprised of at least 90% N-terminally monoPEGylated G-CSF or analog thereof and at most 10% unPEGylated G-CSF or analog thereof. 5. A preparation of claim 4 wherein said preparation is comprised of at least 95% N-terminally monoPEGylated G-CSF or analog thereof and at most 5% unPEGylated G-CSF or analog thereof. 6. A preparation of claim 1 whererein said G-CSF has the sequence identified in SEQ. ID No. 1. 7. 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 between about 6 kDa and about 25 kDa. 8. 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 6 kDa and about 25 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 pharmaceutially acceptable diluent, adjuvant or carrier. 9. A method of treating a hematopoietic disorder comprising administering a therapeutically effective dose of a preparation of any one of claims 1-8. 10. 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 alpha-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 from non-PEGylated G-CSF. 11. A method of claim 10 wherein said polyethylene glycol molecule has a molecular weight of about 6 kDa to about 25 kDa. 12. The PEGylated G-CSF product produced by the process of claim 10.

Details for Patent 5,824,784

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Amgen Inc.	NEULASTA	pegfilgrastim	Injection	125031	January 31, 2002		2039-02-26
Amgen Inc.	NEULASTA ONPRO	pegfilgrastim	Injection	125031	December 23, 2014		2039-02-26

International Patent Family for US Patent 5,824,784

Country	Patent Number	Estimated Expiration
Australia	1841995
Australia	706700
Austria	E179991
Austria	E277078
Canada	2178752
Canada	2307142