PEGYLATED OXM VARIANTS
A composition which includes oxyntomodulin and polyethylene glycol polymer (PEG polymer) linked via a reversible linker such as 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS) is disclosed. Pharmaceutical compositions comprising the reverse pegylated oxyntomodulin and methods of using same are also disclosed.
1 . A method of improving the area under the curve (AUC) of oxyntomodulin in a subject in need of treatment therewith, the method comprising administering to said subject an oxyntomodulin conjugate represented by the structure of:
2 . A method of reducing the dosing frequency of oxyntomodulin in a subject in need of treatment therewith, said method comprising administering to said subject an oxyntomodulin conjugate represented by the structure of:
3 . A method for extending the biological half-life of oxyntomodulin in a subject in need of treatment therewith, the method comprising administering to said subject an oxyntomodulin conjugate represented by the structure of:
4 . The method of claim 3 , wherein said oxyntomodulin is released into a biological fluid by chemically hydrolyzing the FMS or Fmoc linker from said oxyntomodulin conjugate.
5 . The method of claim 4 , wherein the released oxyntomodulin is intact and regains GLP-1 and glucagon receptor binding activity.
6 . The method of claim 4 , wherein said biological fluid is blood, sera, or cerebrospinal fluid.
7 . A method of inducing glucose tolerance, inducing glycemic control, increasing insulin sensitivity, or reducing insulin resistance in a subject in need thereof, the method comprising administering to the subject an oxyntomodulin conjugate represented by the structure of:
8 . A method of improving the cholesterol levels in a subject in need thereof, the method comprising administering to the subject an oxyntomodulin conjugate represented by the structure of:
9 . A homogeneous intermediate conjugate represented by the structure of:
wherein R 2 is H and said His 1 is conjugated to said Fmoc linker or R 2 is SO 3 H and said His 1 is conjugated to said FMS linker; and
wherein the amino side groups of Lys 12 and Lys 30 of said oxyntomodulin are protected.
10 . The conjugate of claim 9 , wherein said PEG is represented by (CH 2 CH 2 O) n CH 3 , wherein n is 30 (PEG30), 40 (PEG40) or 60 (PEG60).
11 . A homogeneous intermediate conjugate represented by the structure of:
wherein R 2 is H and Lys 30 is conjugated to the Fmoc linker or R 2 is SO 3 H and Lys 30 is conjugated to the FMS linker; and
wherein the terminal amino group of His 1 and the amino side group of Lys 12 of oxyntomodulin are protected.
12 . The conjugate of claim 11 , wherein said PEG is represented by (CH 2 CH 2 O) n CH 3 , wherein n is 30 (PEG30), 40 (PEG40) or 60 (PEG60).
13 . A homogeneous intermediate conjugate represented by the structure of:
wherein R 2 is H and Lys 12 is conjugated to the Fmoc linker or R 2 is SO 3 H and Lys 12 is conjugated to the FMS linker; and
wherein the terminal amino group of His' and the amino side group of Lys 30 of oxyntomodulin are protected.
14 . The conjugate of claim 13 , wherein said PEG is represented by (CH 2 CH 2 O) n CH 3 , wherein n is 30 (PEG30), 40 (PEG40) or 60 (PEG60).