GLP-1 fusion proteins

1. A heterologous fusion protein comprising a first polypeptide with a N-terminus and a C-terminus fused to a second polypeptide with a N-terminus and a C-terminus wherein the first polypeptide is a GLP-1 compound and the second polypeptide is selected from the group consisting of

a) human albumin;

b) human albumin analogs; and

c) fragments of human albumin,

and wherein the C-terminus of the first polypeptide is fused to the N-terminus of the second polypeptide.

2. A heterologous fusion protein comprising a first polypeptide with a N-terminus terminus and a C-terminus fused to a second polypeptide with a N-terminus and a C-terminus wherein the first polypeptide is a GLP-1 compound and the second polypeptide is selected from the group consisting of

a) human albumin;

b) human albumin analogs; and

c) fragments of human albumin,

and wherein the C-terminus of the first polypeptide is fused to the N-terminus of the second polypeptide via a peptide linker.

3. The heterologous fusion protein of the claim 2 wherein the peptide linker is selected from the group consisting of:

a) a glycine rich peptide;

b) a peptide having the sequence [Gly-Gly-Gly-Gly-Ser] n where n is 1, 2, 3, 4, 5 or 6; and

c) a peptide having the sequence [Gly-Gly-Gly-Gly-Ser] 3 .

4. The heterologous fusion protein of claims 1 , 2 , or 3 wherein the GLP-1 compound comprises the sequence of formula 1 [SEQ ID NO: 2]

(SEQ ID NO: 2)

7   8   9   10  11  12  13  14  15  16  17

His-Xaa-Xaa-Gly-Xaa-Phe-Thr-Xaa-Asp-Xaa-Xaa-

18  19  20  21  22  23  24  25  26  27  28

Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Phe-

29  30  31  32  33  34  35  36  37  38  39

Ile-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-

40  41  42  43  44  45

Xaa-Xaa-Xaa-Xaa-Xaa-Xaa

Formula I

wherein:

Xaa at position 8 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 9 is Glu, Asp, or Lys;

Xaa at position 11 is Thr, Ala, Gly, Ser, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 14 is Ser, Ala, Gly, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 16 is Val, Ala, Gly, Ser, Thr, Leu, Ile, Tyr, Glu, Asp, Trp, or Lys;

Xaa at position 17 is Ser, Ala, Gly, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 18 is Ser, Ala, Gly, Thr, Leu, Ile, Val, Glu, Asp, Trp, Tyr, or Lys;

Xaa at position 19 is Tyr, Phe, Trp, Glu, Asp, Gln, or Lys;

Xaa at position 20 is Leu, Ala, Gly, Ser, Thr, Ile, Val, Glu, Asp, Met, Trp, Tyr, or Lys;

Xaa at position 21 is Glu, Asp, or Lys;

Xaa at position 22 is Gly, Ala, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 23 is Gln, Asn, Arg, Glu, Asp, or Lys;

Xaa at position 24 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Arg, Glu, Asp, or Lys;

Xaa at position 25 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 26 is Lys, Arg, Gln, Glu, Asp, or His;

Xaa at position 27 is Leu, Glu, Asp, or Lys;

Xaa at position 30 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 31 is Trp, Phe, Tyr, Glu, Asp, or Lys;

Xaa at position 32 is Leu, Gly, Ala, Ser, Thr, Ile, Val, Glu, Asp, or Lys;

Xaa at position 33 is Val, Gly, Ala, Ser, Thr, Leu, Ile, Glu, Asp, or Lys;

Xaa at position 34 is Asn, Lys, Arg, Glu, Asp, or His;

Xaa at position 35 is Gly, Ala, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys;

Xaa at position 36 is Gly, Arg, Lys, Glu, Asp, or His;

Xaa at position 37 is Pro, Gly, Ala, Ser, Thr, Leu, Ile, Val, Glu, Asp, or Lys, or is deleted;

Xaa at position 38 is Ser, Arg, Lys, Glu, Asp, or His, or is deleted;

Xaa at position 39 is Ser, Arg, Lys, Glu, Asp, or His, or is deleted;

Xaa at position 40 is Gly, Asp, Glu, or Lys, or is deleted;

Xaa at position 41 is Ala, Phe, Trp, Tyr, Glu, Asp, or Lys, or is deleted;

Xaa at position 42 is Ser, Pro, Lys, Glu, or Asp, or is deleted;

Xaa at position 43 is Ser, Pro, Glu, Asp, or Lys, or is deleted;

Xaa at position 44 is Gly, Pro, Glu, Asp, or Lys, or is deleted; and

Xaa at position 45 is Ala, Ser, Val, Glu, Asp, or Lys, or is deleted;

provided that when the amino acid at position 37, 38, 39, 40, 41, 42, 43, or 44 is deleted, then each amino acid downstream of that amino acid is also deleted.

5. The heterologous fusion protein of claims 1 , 2 , or 3 wherein the GLP-1 compound comprises the sequence of formula II (SEQ ID NO: 3):

(SEQ ID NO: 3)

7   8   9   10  11  12  13  14  15  16  17

Xaa-Xaa-Xaa-Gly-Xaa-Xaa-Thr-Ser-Asp-Xaa-Ser-

18  19  20  21  22  23  24  25  26  27  28

Xaa-Tyr-Leu-Glu-Xaa-Xaa-Xaa-Ala-Xaa-Xaa-Phe-

29  30  31  32  33  34  35  36  37

Ile-Xaa-Xaa-Leu-Xaa-Xaa-Xaa-Xaa-Xaa

Formula II

wherein:

Xaa at position 7 is: L-histidine, D-histidine, desamino-histidine, 2-amino-histidine,

β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine;

Xaa at position 8 is: Gly, Ala, Val, Leu, Ile, Ser, or Thr;

Xaa at position 9 is: Thr, Ser, Arg, Lys, Trp, Phe, Tyr, Glu, or His;

Xaa at position 11 is: Asp, Glu, Arg, Thr, Ala, Lys, or His;

Xaa at position 12 is: His, Trp, Phe, or Tyr;

Xaa at position 16 is: Leu, Ser, Thr, Trp, His, Phe, Asp, Val, Tyr, Glu, or Ala;

Xaa at position 18 is: His, Pro, Asp, Glu, Arg, Ser, Ala, or Lys;

Xaa at position 22 is: Gly, Asp, Glu, Gln, Asn, Lys, Arg, or Cys;

Xaa at position 23 is: His, Asp, Lys, Glu, Gln, or Arg;

Xaa at position 24 is: Glu, Arg, Ala, or Lys;

Xaa at position 26 is: Trp, Tyr, Phe, Asp, Lys, Glu, or His;

Xaa at position 27 is: Ala, Glu, His, Phe, Tyr, Trp, Arg, or Lys;

Xaa at position 30 is: Ala, Glu, Asp, Ser, or His;

Xaa at position 31 is: Asp, Glu, Ser, Thr, Arg, Trp, or Lys;

Xaa at position 33 is: Asp, Arg, Val, Lys, Ala, Gly, or Glu;

Xaa at position 34 is: Glu, Lys, or Asp;

Xaa at position 35 is: Thr, Ser, Lys, Arg, Trp, Tyr, Phe, Asp, Gly, Pro, His, or Glu;

Xaa at position 36 is: Thr, Ser, Asp, Trp, Tyr, Phe, Arg, Glu, or His;

Xaa at position 37 is: Lys, Arg, Thr, Ser, Glu, Asp, Trp, Tyr, Phe, His, Gly, Gly-Pro, or is deleted.

6. The heterologous fusion protein of claims 1 , 2 , or 3 wherein the GLP-1 compound comprises the sequence of formula III (SEQ ID NO: 4):

(SEQ ID NO: 4)

7   8   9   10  11  12  13  14  15  16  17

Xaa-Xaa-Glu-Gly-Xaa-Xaa-Thr-Ser-Asp-Xaa-Ser-

18  19  20  21  22  23  24  25  26  27  28

Ser-Tyr-Leu-Glu-Xaa-Xaa-Xaa-Xaa-Lys-Xaa-Phe-

29  30  31  32  33  34  35  36  37

Ile-Xaa-Trp-Leu-Xaa-Xaa-Xaa-Xaa-Xaa

formula III

wherein:

Xaa at position 7 is: L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine;

Xaa at position 8 is: Gly, Ala, Val, Leu, Ile, Ser, or Thr;

Xaa at position 11 is: Asp, Glu, Arg, Thr, Ala, Lys, or His;

Xaa at position 12 is: His, Trp, Phe, or Tyr;

Xaa at position 16 is: Leu, Ser, Thr, Trp, His, Phe, Asp, Val, Glu, or Ala;

Xaa at position 22: Gly, Asp, Glu, Gln, Asn, Lys, Arg, or Cys;

Xaa at position 23 is: His, Asp, Lys, Glu, or Gln;

Xaa at position 24 is: Glu, His, Ala, or Lys;

Xaa at position 25 is: Asp, Lys, Glu, or His;

Xaa at position 27 is: Ala, Glu, His, Phe, Tyr, Trp, Arg, or Lys;

Xaa at position 30 is: Ala, Glu, Asp, Ser, or His;

Xaa at position 33 is: Asp, Arg, Val, Lys, Ala, Gly, or Glu;

Xaa at position 34 is: Glu, Lys, or Asp;

Xaa at position 35 is: Thr, Ser, Lys, Arg, Trp, Tyr, Phe, Asp, Gly, Pro, His, or Glu;

Xaa at position 36 is: Arg, Glu, or His;

Xaa at position 37 is: Lys, Arg, Thr, Ser, Glu, Asp, Trp,

Tyr, Phe, His, Gly, Gly-Pro, or is deleted.

7. The heterologous fusion protein of claim 1 wherein the GLP-1 compound has no more than 6 amino acids that are different from the corresponding amino acid in GLP-1(7-37)OH, GLP-1(7-36)OH, or Exendin-4.

8. The heterologous fusion protein of claim 7 wherein the GLP-1 compound has no more than 5 amino acids that differ from the corresponding amino acid in GLP-1(7-37)OH, GLP-1(7-36)OH, or Exendin-4.

9. The heterologous fusion protein of claim 8 wherein the GLP-1 compound has no more than 4 amino acids that differ from the corresponding amino acid in GLP-1(7-37)OH, GLP-1(7-36)OH, or Exendin-4.

10. The heterologous fusion protein of claim 9 wherein the GLP-1 compound has no more than 3 amino acids that differ from the corresponding amino acid in GLP-1(7-37)OH, GLP-1(7-36)OH, or Exendin-4.

11. The heterologous fusion protein of claim 10 wherein the GLP-1 compound has no more than 2 amino acids that differ from the corresponding amino acid in GLP-1(7-37)OH, GLP-1(7-36)OH, or Exendin-4.

12. A method of treating a patient with non-insulin dependent diabetes mellitus comprising the administration of a therapeutically effective amount of the heterologous fusion protein of claim 1 .

13. A method of treating obesity comprising the administration of a therapeutically effective amount of the heterologous fusion protein of claim 1 .