Glycopolysialylation of non-blood coagulation proteins

Patent metadata

Field	Value
Publication number	US-2018200380-A1
Application number	US-201715707844-A
Country	US
Kind code	A1
Filing date	Sep 18, 2017
Priority date	Jul 27, 2009
Publication date	Jul 19, 2018
Grant date	—

Abstract

A water soluble polymer, in particular polysialic acid (PSA) or a modified PSA (mPSA), is conjugated to an oxidized carbohydrate moiety of a glycoprotein other than a blood coagulation protein or to a ganglioside or drug delivery system by contacting the oxidized carbohydrate moiety with the water soluble polymer, wherein said water soluble polymer contains an aminooxy group and an oxime linkage is formed between the oxidized carbohydrate moiety and the aminooxy group on the water soluble polymer or wherein said water soluble polymer contains a hydrazide group and a hydrazone linkage is formed between the oxidized carbohydrate moiety and the hydrazide group on the water soluble polymer. Conjugates of aminooxy- or hydrazide-water soluble polymer, such as PSA and mPSA, are thus obtained in which the PSA or mPSA is attached via a carbohydrate moiety.

First claim

1 - 28 . (canceled)

29 . A method of conjugating a polysialic acid (PSA) or a modified PSA (mPSA) to an oxidized carbohydrate moiety of a glycoprotein other than a blood coagulation protein, comprising a carbohydrate group, comprising contacting the oxidized carbohydrate moiety with PSA or mPSA under conditions that allow conjugation, thereby forming a conjugated glycoprotein,
wherein said PSA or mPSA contains an aminooxy group, and an oxime linkage is formed between the oxidized carbohydrate moiety and the aminooxy group on the PSA or mPSA.

30 . The method of claim 29 , wherein said conjugated glycoprotein has a biological activity of at least 80% relative to a native glycoprotein.

31 . The method of claim 29 , wherein PSA or mPSA is mPSA, and said mPSA is PSA comprising a moiety derived from a terminal N-acetylneuraminic acid moiety by oxidation or reduction.

32 . The method of claim 29 , wherein PSA or mPSA is colominic acid or modified colominic acid.

33 . The method of claim 29 , wherein PSA or mPSA comprises 2-500 sialic acid units.

34 . The method of claim 29 , wherein the glycoprotein is selected from the group consisting of cytokines such as interleukins, alpha-, beta-, and gamma-interferons, colony stimulating factors including granulocyte colony stimulating factors, fibroblast growth factors, platelet-derived growth factors, phospholipase-activating protein (PUP), insulin, plant proteins such as lectins and ricins, tumor necrosis factors and related alleles, soluble forms of tumor necrosis factor receptors, interleukin receptors and soluble forms of interleukin receptors, growth factors, tissue growth factors, transforming growth factors such as TGFα or TGFβ and epidermal growth factors, hormones, somatomedins, pigmentary hormones, hypothalamic releasing factors, antidiuretic hormones, prolactin, chorionic gonadotropin, follicle-stimulating hormone, thyroid-stimulating hormone, tissue plasminogen activator, immunoglobulins such as IgG, IgE, IgM, IgA, and IgD, monoclonal antibodies, erythropoietin (EPO), blood factors other than blood coagulation proteins, galactosidases, α-galactosidases, β-galactosidases, DNAses, fetuin and fragments thereof, and fusion proteins comprising any of the above mentioned proteins or fragments thereof.

35 . The method of claim 29 , wherein the glycoprotein is selected from the group consisting of tumour necrosis factors and related alleles, soluble forms of tumor necrosis factor receptors, immunoglobulins such as IgG, IgE, IgM, IgA and IgD, monoclonal antibodies, erythropoietin (EPO), DNAses, fetuin, fragments thereof and fusion proteins comprising any of the above mentioned proteins or fragments thereof.

36 . The method of claim 29 , comprising oxidizing the carbohydrate moiety by incubating the glycoprotein with sodium periodate (NaIO 4 ).

37 . The method of claim 29 , comprising oxidizing the PSA or mPSA to form an aldehyde group on a terminal unit of the PSA or mPSA, and reacting the oxidized PSA or mPSA with an aminooxy linker.

38 . The method of claim 37 , comprising oxidizing the PSA or mPSA using NaIO 4 .

39 . The method of claim 29 , comprising contacting the oxidized carbohydrate moiety with the PSA or mPSA in a buffer comprising a nucleophilic catalyst selected from aniline or aniline derivatives.

40 . The method of claim 29 , wherein said PSA or mPSA contains an aminooxy group and an oxime linkage is formed between the oxidized carbohydrate moiety and the aminooxy group on the PSA or mPSA.

41 . The method of claim 40 , wherein the aminooxy group is formed by reacting oxidized PSA or mPSA with an aminooxy linker, and the aminooxy linker is 3-oxa-pentane-1,5-dioxyamine or 3,6,9-trioxa-undecane-1,11-dioxyamine.

42 . The method of claim 29 , further comprising reducing an oxime linkage in the conjugated glycoprotein by incubation in the presence of a reducing compound.

43 . The method of claim 42 , wherein the reducing compound is sodium cyanoborohydride (NaCNBH 3 ) or ascorbic acid (vitamin C).

44 . A conjugated glycoprotein, obtained by the method of claim 29 .

45 . The conjugated glycoprotein other than a blood coagulation protein, comprising:
(a) the glycoprotein and (b) at least one aminooxy-PSA or -mPSA bound to the glycoprotein of (a), wherein said aminooxy-PSA or -mPSA is attached to the glycoprotein via one or more carbohydrate moieties.

Assignees

• Lipoxen Tech Limited
• Baxalta Inc
• Baxalta GmbH

Inventors

• Jain Sanjay
• Gregoriadis Gregory
• Dwivedi Archana
• Nath Srijit
• Siekmann Juergen
• Haider Stefan
• Rottensteiner Hanspeter
• Turecek Peter

Classifications

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P5/14

  of the thyroid hormones, e.g. T3, T4 · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P37/02

  Immunomodulators · CPC title