Novel g-csf conjugates

Details Novel g-csf conjugates Novel g-csf conjugates

A - Human Necessities

61

K

A61K 47/48 (2006.01) A61P 35/00 (2006.01) A61P 43/00 (2006.01)

Patent

CA 2562890

The present application relates to novel PEG-G-CSF conjugates in which a PEG molecule is linked to the cyteine residue in position 17 of native G-CSF primary sequence or to the cysteine residue of the corresponding position of a G-CSF analogue. The present application also describes a process for the manufacture of such conjugates, such process comprising the following steps: (i) subjecting the G-CSF protein to conditions inducing reversible denaturation of the protein, (ii) conjugation of the denatured protein obtained is step i with a thiol-reactive PEG under denaturing conditions, (iii) subjecting the conjugates obtained in step ii to conditions promoting renaturation of the conjugate yielding biologically active G-CSF-PEG conjugate.

L'invention concerne des nouveaux conjugués de PEG-G-CSF dans lesquels une molécule de PEG est liée au résidu cystéine en position 17 de la séquence primaire de G-CSF native ou au résidu cystéine de la position correspondante d'un analogue de G-CSGF. L'invention concerne également un procédé de fabrication desdits conjugués, comprenant les étapes qui consistent à : i) soumettre la protéine G-CSF à des conditions induisant la dénaturation réversible de la protéine ; ii) conjuguer la protéine dénaturée obtenue dans l'étape I avec un PEG réagissant au thiol dans des conditions de dénaturation ; iii) soumettre les conjugués obtenus dans l'étape ii à des conditions favorisant la renaturation du conjugué produisant un conjugué de G-CSF-PEG actif au plan biologique.

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