Claims for Patent: 5,637,749

Summary for Patent: 5,637,749

Title:	Aryl imidate activated polyalkylene oxides
Abstract:	Water-soluble aryl imidate activated polyalkylene oxides having improved hydrolytic stability and conjugates of the aryl imidate activated polyalkylene oxides with biologically active nucleophiles are disclosed. Methods of preparing the activated polyalkylene oxides and conjugates thereof are also disclosed.
Inventor(s):	Greenwald; Richard B. (Somerset, NJ)
Assignee:	Enzon, Inc. (Piscataway, NJ)
Application Number:	08/507,156
Patent Claims:	1. A polyalkylene oxide conjugate comprising: a nucleophile having biological activity; and at least one water-soluble polyalkylene oxide covalently bonded thereto by a hydrolyrically stable linkage formed by reacting said nucleophile with an aryl imidate activated polyalkylene oxide, having a structure represented by (I): ##STR5## wherein R is a water-soluble polyalkylene oxide; L is NHC(O) such that the carbonyl is attached to the phenyl ring; R.sub.1 is a moiety selected from the group consisting of alkyl, phenyl, phenyl alkyl, and cycloalkyl moieties; and X is a terminal moiety of said polyalkylene oxide selected from the group consisting of OH,C.sub.1-4 alkoxy moieties and aryl imidates corresponding to the structure: ##STR6## wherein L and R.sub.1 are defined as above. 2. The polyalkylene oxide conjugate of claim 1, wherein said polyalkylene oxide is selected from the group consisting of polyethylene glycol and block copolymers of polyethylene glycol and polypropylene glycol. 3. The polyalkylene oxide conjugate of claim 2, wherein said polyalkylene oxide comprises polyethylene glycol. 4. The polyalkylene oxide conjugate of claim 1, wherein said polyalkylene oxide has a number average molecular weight between about 600 and about 100,000 daltons. 5. The polyalkylene oxide conjugate of claim 4, wherein said polyalkylene oxide has a number average molecular weight between about 2,000 and about 20,000 daltons. 6. The polyalkylene oxide conjugate of claim 5, wherein said polyalkylene oxide has a 5,000 dalton number average molecular weight. 7. The polyalkylene oxide conjugate of claim 1, wherein said nucleophile is an enzyme selected from the group consisting of asparaginase, arginase, adenosine deaminase, superoxide dismutase, catalase, chymotrypsim, lipase, uricase, bilirubin oxidase, glucose oxidase, glucosidase, galactosidase, glucocerebrosidase and glucuronidase. 8. The polyalkylene oxide conjugate of claim 1, wherein said nucleophile is a serum protein selected from the group consisting of Factor VIII, Factor IX, interleukins, interferons, colony stimulating factors, immunoglobulins and lectins. 9. The polyalkylene oxide conjugate of claim 8, wherein said nucleophile is an immunoglobulin selected from the group consisting of IgG, IgE, IgM, IgA, IgD and fragments thereof. 10. The polyalkylene oxide conjugate of claim 1, wherein said nucleophile is a hormone selected from the group consisting of insulin, ACTH, glucagon, somatostatin, somatotropins, thymosin, parathyroid hormone, pigmentary hormones, somatomedins, erythropoietin, luteinizining hormone, hypothalmic releasing factors, antidiuretic hormones, prolactin, chorionic gonadotropin, follicle stimulating hormone, tissue plasminogen activator and thyroid-stimulating hormone. 11. The polyalkylene oxide conjugate of claim 1, wherein said nucleophile is a protein selected from the group consisting of hemoglobin, ovalbumin and bovines serum albumin. 12. The polyalkylene oxide conjugate of claim 1, wherein said nucleophile is a polypeptide comprising lysine residues, and at least one of said polyalkylene oxides is covalently bonded to the epsilon-NH.sub.2 moiety of said lysine residue via a linkage forming an amidate moiety with said epsilon-NH.sub.2 moiety. 13. The polyalkylene oxide conjugate of claim 1, comprising a plurality of polyalkylene oxides covalently bonded to said nucleophile. 14. The polyalkylene oxide conjugate of claim 1, wherein R.sub.1 is a moiety selected from the group consisting of methyl, ethyl, phenyl, benzyl and cyclohexyl moieties. 15. The polyalkylene oxide conjugate of claim 1, wherein X is a methoxy moiety. 16. A method of forming a biologically active conjugate of a biologically active nucleophile and one or more water-soluble polyalkylene oxides covalently bonded thereto, said method comprising the steps of: providing a biologically active nucleophile; contacting said nucleophile with one or more aryl imidate activated polyalkylene oxides, so that a biologically active conjugate of said biologically active nucleophile and said polyalkylene oxides is formed; and recovering said biologically active conjugate. 17. The method of claim 16, wherein said contacting is carried out in an aqueous reaction medium. 18. The method of claim 17, wherein the pH of said aqueous reaction medium is between about 6.5 and about 8.0. 19. The method of claim 18, wherein the pH of said reaction medium is about 7.4. 20. The method of claim 16, wherein said contacting is carried out at a temperature of from about 4 to about 37 degrees C.

Details for Patent 5,637,749

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	01/15/1974		2012-12-22
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	12/27/1984		2012-12-22
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	02/15/1985		2012-12-22
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	02/16/1990		2012-12-22