Modified human acid sphingomyelinase having increased...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/55 (2006.01) A61K 38/46 (2006.01) C12N 9/16 (2006.01) C12N 9/20 (2006.01)

Patent

CA 2528427

Deficiencies in the enzymatic activity of acid sphingomyelinase (ASM) result in Niemann-Pick disease. A variety of modifications which eliminate the activity of the free thiol on the C-terminal cysteine residue of ASM all result in substantially increased specific activity of the enzyme. Methods used to alter the activity of this residue include site-directed mutagenesis to delete or alter the residue, enzymatic degradation of the ASM to remove the residue, copper-promoted dimerization of ASM (via the terminal cysteine residues) and chemical modification of the free thiol group on this residue.

L'invention concerne des déficiences au niveau de l'activité enzymatique de la sphingomyélinase acide (ASM) à l'origine de la maladie de Niemann-Pick. Diverses modifications permettant d'éliminer l'activité du thiol libre sur le résidu de cystéine à terminaison C de l'ASM résultent toutes en une activité sensiblement accrue de l'enzyme. Les procédés utilisés pour modifier l'activité de ce résidu comportent notamment une mutagenèse dirigée pour supprimer ou modifier le résidu, une dégradation enzymatique de l'ASM pour évacuer le résidu, une dimérisation activée au cuivre de l'ASM (par le biais des résidus de cystéine à terminaison) et une modification chimique du groupe thiol libre sur ce résidu.

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