Method of producing fully carbamylated erythropoietin

Details Method of producing fully carbamylated erythropoietin Method of producing fully carbamylated erythropoietin

A - Human Necessities

61

K

A61K 38/00 (2006.01)

Patent

CA 2579813

The present invention relates to a method of carbamylating an erythropoietin such that the resulting carbamylated erythropoietin has less that about 10% free primary amines on the lysines and the N-terminal amino acids, is not digested when exposed to Lys-C proteolysis, exhibits no erythropoietic activity in a TF-1 or UT-7/EPOR cell viability assay at a concentration of 1 ~g/ml, and demonstrates a static sciatic index of less than about .65 within a Sciatic Nerve Assay. Additionally, the invention is related to pharmaceutical compositions containing carbamylated erythropoietins of the invention and the use of the pharmaceutical compositions for the treatment of conditions and diseases of excitable tissues.

L'invention concerne un procédé de carbamylation d'une érythropoïétine permettant d'obtenir une érythropoïétine carbamylée qui renferme moins de 10 % environ d'amines primaires libres sur les lysines et les acides aminés N-terminaux; n'est pas digérée lorsqu'elle est exposée à une protéolyse par Lys-C; ne possède aucune activité érythropoïétique dans une analyse de viabilité de cellules TF-1 ou UT-7/EPOR à une concentration de 1 ?g/ml; et présente à l'analyse du nerf sciatique un indice sciatique statique inférieur à environ 0,65. De plus, l'invention concerne des compositions pharmaceutiques contenant les érythropoïétines carbamylées de l'invention, et l'utilisation de ces compositions pour traiter des états pathologiques et des maladies des tissus excitables.

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