USE OF PLP WITH PEG-rMETase IN VIVO FOR ENHANCED EFFICACY
This invention relates to methods of modifying pyridoxal 5′ phosphate (PLP) dependent enzymes to extend the serum half-life of the enzyme, extend the in vivo period of methionine depletion in a host, and decrease the immunogenicity of the enzyme. A preferred PLP-dependent enzyme to be modified is a methioninase, preferably a recombinant methioninase (rMETase). The invention further relates to compositions comprising a modified PLP-dependent enzyme and methods of using the same.
1 . A method of increasing the serum half-life of a methioninase by coupling the methioninase to a polyalkene glycol, wherein at least 33% of free lysine residues in the methioninase are coupled with polyalkene glycol units.
2 . The method of claim 1 , wherein the half-life of recombinant methioninase is adjusted by altering the amount of polyalkene glycol that is coupled to the methioninase.
3 . The method of claim 2 , wherein the molar ratio of polyalkene glycol to methioninase is approximately 30:1.
4 . The method of claim 2 , wherein the molar ratio of polyalkene glycol to methioninase is approximately 60:1.
5 . The method of claim 2 , wherein the molar ratio of polyalkene glycol to methioninase is approximately 120:1.
6 . The method of claim 1 , wherein the methioninase is recombinantly produced.
7 . The method of claim 1 , wherein the methioninase is L-methionine α-deamino-γ-mercaptomethane lyase.
8 . The method of claim 1 , wherein the methioninase coupled with polyalkene glycol is administered with pyridoxal 5′-phosphate (PLP) to a subject.