Abstract: 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-reactivc 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.

Abstract: A method is described for the site-specific preparation of hGRF-PEG conjugates containing one or more PEG units (per mole of hGRF) covalently bound to Lys12 and/or Lys21 and/or N?, characterized in that the conjugation reaction between the hGRF peptide and activated PEG is carried out in solution and the desired hGRF-PEG conjugate can be purified by chromatography. The conjugates prepared by this method, as well as their use in the treatment, prevention of diagnosis of growth hormone deficiency, are also an object of the present invention.

Abstract: Multi-armed, monofunctional, and hydrolytically stable polymers are described having the structure wherein Z is a moiety that can be activated for attachment to biologically active molecules such as proteins and wherein P and Q represent linkage fragments that join polymer arms polya and polyb, respectively, to central carbon atom, C, by hydrolytically stable linkages in the absence of aromatic rings in the linkage fragments. R typically is hydrogen or methyl, but can be a linkage fragment that includes another polymer arm. A specific example is an mPEG disubstituted lysine having the structure where mPEGa and mPEGb have the structure CH3O—(CH2CH2O)nCH2CH2— wherein n may be the same or different for polya- and polyb- and can be from 1 to about 1,150 to provide molecular weights of from about 100 to 100,000.

Abstract: Biologically active drug polymer derivatives, namely peptides or protein derivatives, are useful medicaments and are represented by the generic formula: RO—(CH2—CH2O)n—(CO)—NH—X—(CO)—NH—Z wherein R represents a lower alkyl group, n is an integer between 25 and 250, X when combined with adjcacent NH and CO groups represents a dipeptide residue, and Z when combined with the adjacent group represents a biologically active peptide or protein.

Abstract: A method for preparing protease-polyethylene glycol adducts is presented wherein the immobilized reversible inhibitor, benzamidine, prevents reaction of activated polyethylene glycol with the active site of the protease. Improved activity against macromolecular substrates is obtained compared to when the benzamidine is in solution during the conjugation reaction.

Abstract: New biologically active drug polymer derivatives, namely peptides or protein derivatives, are useful medicaments and are represented by the generic formula:RO--(CH.sub.2 --CH.sub.2 O).sub.n --(CO)--NH--X--(CO)--NH--Z (I)whereinR represents a lower alkyl group,n is an integer comprised between 25 and 250,X when combined with adjacent NH and CO groups represents an amino acid or a dipeptide or tripeptide residue, andZ when combined with the adjacent NH group represents a biologically active peptide or protein or NH or NH.sub.2 containing drug residue.