Method for producing polyol-IFN-β conjugate
View Patent ↗PEG-IFN-β conjugates, where a PEG moiety is covalently bound to Cys 17 of human IFN-β, are produced by a process of site specific PEGylation with a thiol reactive PEGylating agent. A pharmaceutical composition and a method for treating infections, tumors and autoimmune and inflammatory diseases are also provided. The invention further relates to a method for the stepwise attachment of PEG moieties in series to a polypeptide, and more particularly to IFN-β.
1. A method for producing a polyol-interferon-β conjugate having a polyol moiety covalently bound to Cys 17 of human interferon-β(fibroblast interferon), comprising:
reacting interferon-β with a thiol-reactive polyol agent to site specifically and covalently attach a polyol moiety to Cys 17 of human interferon-β [fibroblast interferon] to produce a polyol-interferon-β [human fibroblast interferon] conjugate; and recovering the produced polyol-interferon-β conjugate.
2. The method according to claim 1 , wherein the thiol-reactive polyol agent is a thiol-reactive PEGylating agent.
3. The method according to either claim 1 or claim 2 , wherein the thiol-reactive polyol agent is mono-methoxylated.
4. The method according to either claim 1 or claim 2 , wherein the thiol-reactive polyol agent is bifunctional.
5. The method according to either claim 1 or claim 2 , wherein the thiol-reactive polyol agent is a polyol derivative having a functional group selected from the group consisting of orthopyridyl disulfide, vinyl sulfone, maleimide, and iodoacetimide.
6. The method according to either claim 1 or claim 2 , wherein the thiol-reactive polyol agent is an orthopyridyl disulfide derivative of a mono-methoxylated polyol.
7. The process according to claim 1 , wherein the reacting step is carried out at an acidic pH where interferon-ss is stable.