Modified IL-2 polypeptides and uses thereof
The present disclosure relates to modified IL-2 polypeptides, compositions comprising modified IL-2 polypeptides, methods of making the same, and methods of using the modified IL-2 polypeptides for treatment of diseases. In one aspect, the disclosure relates to the treatment of cancer using the modified IL-2 polypeptides. In some embodiments, the disclosed IL-2 polypeptides exhibit preferential binding characteristics to IL-2 receptor βγ complex (IL-2Rβ) over IL-2 receptor αβγ complex (IL-2Rα). In some embodiments, the molecular weight distribution of the modified IL-2 polypeptides is monodispersed.
1. A modified interleukin-2 (IL-2) polypeptide, comprising:
a first polymer covalently attached at residue F42Y and a second polymer covalently attached at residue Y45 of the modified IL-2 polypeptide, and wherein residue position numbering of the modified IL-2 polypeptide is based on SEQ ID NO:1 as a reference sequence.
2. The modified IL-2 polypeptide of claim 1 , wherein the first polymer comprises a water-soluble polymer.
3. The modified IL-2 polypeptide of claim 1 , wherein the first polymer has a weight average molecular weight of at most 50,000 Daltons, at most 25,000 Daltons, at most 10,000 Daltons, or at most 6,000 Daltons.
4. The modified IL-2 polypeptide of claim 1 , wherein the first polymer has a weight average molecular weight of from 250 Daltons to 50,000 Daltons.
5. The modified IL-2 polypeptide of claim 4 , wherein the second polymer has a weight average molecular weight of from 250 Daltons to 50,000 Daltons.
6. The modified IL-2 polypeptide of claim 1 , wherein the first polymer has a weight average molecular weight of from 5,000 Daltons to 40,000 Daltons and the second polymer has a weight average molecular weight of from 250 Daltons to 1,000 Daltons.
7. The modified IL-2 polypeptide of claim 1 , wherein each of the first polymer and the second polymer independently comprises a water-soluble polymer.
8. The modified IL-2 polypeptide of claim 7 , wherein each water-soluble polymer comprises poly(alkylene oxide), polysaccharide, poly(vinyl pyrrolidone), poly(vinyl alcohol), polyoxazoline, poly(acryloylmorpholine), or a combination thereof.
9. The modified IL-2 polypeptide of claim 1 , wherein the modified IL-2 polypeptide comprises one or more PEGylated tyrosine having a structure of formula (I),
wherein n is an integer selected from 4 to 30.
10. The modified IL-2 polypeptide of claim 9 , wherein the one or more PEGylated tyrosine is located at residue 42, residue 45, or both.
11. The modified IL-2 polypeptide of claim 1 , wherein the modified IL-2 polypeptide comprises at least two amino acid substitutions, wherein the at least two amino acid substitutions are selected from:
a) a homoserine (Hse) residue located in any one of residues 35-45;
b) a homoserine residue located in any one of residues 61-81; and
c) a homoserine residue located in any one of residues 94-114.
12. The modified IL-2 polypeptide of claim 11 , wherein the modified IL-2 polypeptide comprises Hse41, Hse71, Hse104, or a combination thereof.
13. The modified IL-2 polypeptide of claim 11 , wherein the modified IL-2 polypeptide further comprises a norleucine (Nle) substitution, wherein the Nle substitution is located at residue 23, 39, 46, or any combination thereof.
14. The modified IL-2 polypeptide of claim 13 , wherein the modified IL-2 polypeptide comprises a polypeptide sequence identical to that of SEQ ID NO: 3.
15. The modified IL-2 polypeptide of claim 1 , wherein binding between the modified IL-2 polypeptide and IL-2 Receptor β (IL-2Rβ) is increased by at least 0.1%, at least 1%, at least 2%, at least 5%, at least 10%, at least 15%, or at least 20% compared to the binding between a wild-type IL-2 polypeptide and IL-2Rβ.
16. The modified IL-2 polypeptide of claim 1 , wherein binding between the modified IL-2 polypeptide and IL-2 Receptor α (IL-2Rα) is reduced by at least 99% compared to the binding between a wild-type IL-2 polypeptide and IL-2Rα.
17. The modified IL-2 polypeptide of claim 14 , wherein the first polymer has a weight average molecular weight of from 250 Daltons to 50,000 Daltons and the second polymer has a weight average molecular weight of from 250 Daltons to 50,000 Daltons, and wherein each polymer independently comprises a water-soluble polymer.
18. The modified IL-2 polypeptide of claim 14 , wherein the first polymer has a weight average molecular weight of from 120 Daltons to 1000 Daltons and the second polymer has a weight average molecular weight of from 120 Daltons to 1000 Daltons, and wherein each polymer comprises polyethylene glycol.
19. The modified IL-2 polypeptide of claim 14 , wherein residue F42Y with the first polymer attached and residue Y45 with the second polymer attached each comprise a structure of Formula D,
20. The modified IL-2 polypeptide of claim 14 , wherein the first polymer has a weight average molecular weight of from 5,000 Daltons to 40,000 Daltons and the second polymer has a weight average molecular weight of from 250 Daltons to 1,000 Daltons, and wherein each polymer comprises polyethylene glycol.