Method for the production of conjugates of insulin-like...

Details Method for the production of conjugates of insulin-like... Method for the production of conjugates of insulin-like...

A - Human Necessities

61

K

A61K 47/48 (2006.01) C07K 14/65 (2006.01) C12N 15/12 (2006.01) C12N 15/62 (2006.01) C12N 9/52 (2006.01)

Patent

CA 2662062

Method for the production of a lysine-PEGylated IGF-I or IGF-I variant, said variant comprising one or two amino acid(s) selected from the group consisting of lysine 27, 65 and/or 68 substituted independently by another polar amino acid, characterized in cultivating a prokaryotic host cell comprising an expression vector containing a nucleic acid encoding a fusion protein comprising said IGF-I or IGF-I variant N-terminally linked to the C-terminus of a propeptide, that said propeptide ends C-terminally with amino acids -Y-Pro, wherein Y is selected from the group consisting of Pro, Pro-Ala, Pro-Gly, Pro-Thr, Ala-Pro, Gly-Pro, Thr-Pro, Arg-Pro, or Pro-Arg-Pro, recovering and PEGylating said fusion protein, cleaving said PEGylated fusion protein with IgA protease, and recovering said PEGylated IGF-I or IGF-I variant. The PEGylated IGF-I or IGF-I variant is useful for the treatment of neurodegenerative disorders like Alzheimer's Disease.

L'invention concerne un procédé de production de facteurs de croissance de type insuline PEG-lysine ou de variant de IGF-I. Ce variant comprend un ou plusieurs acides aminés sélectionnés dans le groupe constitué de lysine 27, 65 et/ou 68 indépendamment substitué par un autre acide aminé polaire, caractérisé par la culture d'une cellule hôte procaryote comprenant un vecteur d'expression contenant un acide nucléique codant pour une protéine de fusion comprenant ce IGF-I ou ce variant d'IGF-I, lié en N-terminus au C-terminus d'un propeptide. Ce propeptide se termine en C- terminus avec des acides aminés Y-PRO, Y étant sélectionné dans le groupe constitué de Pro, Pro-Ala, Pro-Gly, Pro-Thr, Ala-Pro, Gly-Pro, Thr-Pro, Arg-Pro, or Pro-Arg-Pro,, recouvrant et PEGylant cette protéine de fusion, clivant cette protéine de fusion PEGylée avec une protéase IgA, et recouvrant ce IGF-I PEGylé ou ce variant d'IGF-I. Ce IGF-I PEGylé ou ce variant d'IGF-I convient pour le traitement de troubles neurodégénératifs tels que la maladie d'Alzheimer.

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