Ultra-long acting insulin-Fc fusion protein
The present disclosure relates to compositions of insulin-Fc fusion proteins and their use to treat diabetes.
1. A fusion protein comprising an insulin polypeptide and an Fc fragment connected by a peptide linker, wherein the Fc fragment comprises the following sequence:
(SEQ ID NO: 19)
DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP
EVKFNWYVDGVEVHNAKTKPREEQYKSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPG.
2. The fusion protein of claim 1 , said fusion protein comprising domains in the following orientation from N- to C-terminus:
(N-terminus)-insulin polypeptide-peptide linker-Fc fragment-(C-terminus).
3. The fusion protein of claim 1 , wherein the insulin polypeptide is a human insulin polypeptide.
4. The fusion protein of claim 1 , wherein said insulin polypeptide is a single chain insulin polypeptide.
5. The fusion protein of claim 4 , wherein said single chain insulin polypeptide comprises an A-chain, B-chain, and C-chain, wherein the A-chain and the B-chain are linked by the C-chain.
6. The fusion protein of claim 5 , wherein said C-chain comprises the following sequence:
(SEQ ID NO: 7)
GGGPRR.
7. The fusion protein of claim 5 , wherein the insulin polypeptide is an insulin analog having an aspartic acid mutation at B10 and a histidine mutation at A8.
8. The fusion protein of claim 4 , wherein the insulin polypeptide comprises the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6:
(SEQ ID NO: 4)
FVNQHLCGSDLVEALALVCGERGFFYTDPTGGGPRRGIVEQCCHSICSLYQ
LENYCN
(SEQ ID NO: 5)
FVNQHLCGSHLVEALYLVCGERGFFYTPKAAAAAAAKGIVEQCCTSICSLY
QLENYCN
(SEQ ID NO: 6)
FVNQHLCGSHLVEALYLVCGERGFFYTPKAGGGPRRGIVEQCCTSICSLYQ
LENYCN
or an analog thereof.
9. The fusion protein of claim 1 , wherein the peptide linker comprises the following sequence:
(SEQ ID NO: 8)
GGGGAGGGG.
10. The fusion protein of claim 1 , wherein the fusion protein is a homodimer.
11. The fusion protein of claim 10 , wherein the percentage homodimer of the fusion protein is greater than 90%.
12. The fusion protein of claim 10 , wherein the fusion protein is recombinantly expressed in HEK293 or CHO cells, and the resulting homodimer titer after purification using Protein A beads or a Protein A column is greater than 150 mg/L.
13. The fusion protein of claim 1 , wherein the insulin receptor IC50 for the fusion protein is less than or equal to 5000 nM.
14. A pharmaceutical composition comprising a fusion protein according to claim 1 .
15. The pharmaceutical composition of claim 14 , wherein the fusion protein is present in the pharmaceutical composition at a concentration of about 3 mg/mL or greater.
16. A method for lowering the blood glucose of a target patient, the method comprising administering a physiologically effective amount of the fusion protein of claim 1 or a pharmaceutical composition thereof to the target patient.
17. The method of claim 16 , in which the target patient is diagnosed with diabetes.
18. The method of claim 16 , wherein the fusion protein is administered daily, twice weekly, or once weekly to the target patient.
19. The method of claim 16 , wherein the fusion protein is administered once weekly to the target patient at a dose between 0.025 and 0.5 mg/kg/week.
20. The method of claim 16 , wherein the serum half-life of the fusion protein in the blood or serum of the target patient upon administration is longer than about 3 days.
21. The method of claim 16 , wherein the time during which there is a statistically significant decrease in blood glucose level in the target patient relative to a pre-dose level is longer than one of 2 hours, 6 hours, 9 hours, 12 hours, 18 hours, 1 day, 1.5 days, 2 days, 2.5 days, 3 days, 4 days, 5 days, 6 days, 7 days, or longer.
22. The method of claim 16 , wherein the fusion protein is administered subcutaneously.
23. The method of claim 22 , wherein the NAOC after a first subcutaneous injection in a target patient is greater than 150% FBGL days·kg/mg.
24. The method of claim 22 , wherein the ratio of the NAOC after a third weekly subcutaneous injection of the fusion protein in the target patient to the NAOC after a first subcutaneous injection of the fusion protein in the target patient is greater than 0.50.