Molecules with extended half-lives, compositions and uses thereof
View Patent ↗The present invention provides molecules, including IgGs, non-IgG immunoglobulin, proteins and non-protein agents, that have increased in vivo half-lives due to the presence of an IgG constant domain, or a portion thereof that binds the FcRn, having one or more amino acid modifications that increase the affinity of the constant domain or fragment for FcRn. Such proteins and molecules with increased half-lives have the advantage that smaller amounts and or less frequent dosing is required in the therapeutic, prophylactic or diagnostic use of such molecules.
1. A modified IgG comprising a human IgG constant domain comprising one or more amino acid substitutions relative to a wild-type human IgG constant domain at one or more of amino acid residues 251–256, 285–290, 308–314, 385–389 and 428–436, numbered according to the EU index as in Kabat, wherein the modified IgG has an increased half-life compared to the half-life of an IgG having the wild-type human IgG constant domain, and wherein at least one of the amino acid substitutions is at amino acid residue 252, 254, 256, 309, 311, 433 or 434, and an amino acid substitution at amino acid residue 252 is a substitution with tyrosine, phenylalanine, tryptophan or threonine, an amino acid substitution at amino acid residue 254 is a substitution with threonine, an amino acid substitution at amino acid residue 256 is a substitution with serine, arginine, glutamine, glutamic acid, aspartic acid, or threonine, an amino acid substitution at amino acid residue 309 is a substitution with proline, an amino acid substitution at amino acid residue 311 is a substitution with serine, an amino acid substitution at amino acid residue 433 is a substitution with arginine, serine, isoleucine, proline, or glutamine, and an amino acid substitution at amino acid residue 434 is a substitution with histidine, phenylalanine, or tyrosine.
2. A modified IgG comprising a human IgG constant domain comprising amino acid substitutions relative to a wild-type human IgG constant domain at amino acid residues 252, 254 and 256, numbered according to the EU index as in Kabat, wherein the modified IgG has an increased half-life compared to the half-life of an IgG having the wild-type human IgG constant domain, and wherein the amino acid substitution at amino acid residue 252 is a substitution with tyrosine, the amino acid substitution at amino acid residue 254 is a substitution with threonine, and the amino acid substitution at amino acid residue 256 is a substitution with glutamic acid.
3. A modified IgG comprising a human IgG constant domain comprising amino acid substitutions relative to a wild-type human IgG constant domain at amino acid residues 252, 254, 256, 433, 434 and 436, numbered according to the EU index as in Kabat, wherein the modified IgG has an increased half-life compared to the half-life of an IgG having the wild-type human IgG constant domain, and wherein the amino acid substitution at amino acid residue 252 is a substitution with tyrosine, the amino acid substitution at amino acid residue 254 is a substitution with threonine, the amino acid substitution at amino acid residue 256 is glutamic acid, the amino acid substitution at amino acid residue 433 is a substitution with lysine, the amino acid substitution at amino acid residue 434 is a substitution with phenyalanine, and the amino acid substitution at amino acid residue 436 is a substitution with histidine.
4. A modified IgG comprising a non-human LgG constant domain comprising one or more amino acid substitutions relative to a wild-type non-human IgG constant domain at one or more amino acid residues 251–256, 285–290, 308–314, 385–389 and 428–436, numbered according to the EU index as in Kabat, wherein the modified IgG has an increased half-life compared to the half-life of an IgG having the wild-type non-human IgG constant domain, and wherein at least one of the amino acid substitutions is at amino acid residue 252, 254, 256, 309, 311, 433 or 434, and an amino acid substitution at amino acid residue 252 is a substitution with tyrosine, phenylalanine, tryptophan or threonine, an amino acid substitution at amino acid residue 254 is a substitution with threonine, an amino acid substitution at amino acid residue 256 is a substitution with serine, arginine, glutamine, glutamic acid, aspartic acid, alanine, asparagine or threonine, an amino acid substitution at amino acid residue 309 is a substitution with proline, an amino acid substitution at amino acid residue 311 is a substitution with serine, an amino acid substitution at amino acid residue 433 is a substitution with arginine, serine, isoleucine, proline, or glutamine, and an amino acid substitution at amino acid residue 434 is a substitution with histidine, phenylalanine, or tyrosine.
5. The modified IgG of claim 1 , 2 , 3 , or 4 which has a higher affinity for FcRn than the IgG having the wild-type constant domain.
6. The modified IgG of claim 1 , wherein an amino acid substitution at amino acid residue 385 is a substitution with arginine, aspartic acid, serine, threonine, histidine, lysine, alanine or glycine, an amino acid substitution at amino acid residue 386 is a substitution with threonine, proline, aspartic acid, serine, lysine, arginine, isoleucine, or methionine, an amino acid substitution at amino acid residue 387 is a substitution with arginine, proline, histidine, serine, threonine, or alanine, and an amino acid substitution at amino acid residue 389 is a substitution with proline, serine or asparagine.
7. The modified IgG of claim 4 , wherein an amino acid substitution at amino acid residue 385 is a substitution with arginine, aspartic acid, serine, threonine, histidine, lysine, alanine or glycine, an amino acid substitution at amino acid residue 386 is a substitution with threonine, proline, aspartic acid, serine, lysine, arginine, isoleucine, or methionine, an amino acid substitution at amino acid residue 387 is a substitution with arginine, proline, histidine, serine, threonine, or alanine, and an amino acid substitution at amino acid residue 389 is a substitution with proline, serine or asparagine.
8. The modified IgG of claim 1 , 2 , 3 or 6 which is a human or humanized IgG.
9. The modified IgG of claim 8 which is IgG 1 , IgG 2 , IgG 3 or IgG 4 .
10. The modified IgG of claim 4 or 7 which is a non-human IgG.
11. The modified IgG of claim 10 which is IgG 1 , IgG 2a , IgG 2b , IgG 2c or IgG 3 .
12. The modified IgG of claim 1 , wherein the IgG constant domain is an IgG 1 constant domain.
13. The modified IgG of claim 2 , 3 or 6 , wherein the IgG constant domain is an IgG 1 constant domain.
14. The modified IgG of claim 1 , 2 , 3 or 6 , wherein the IgG constant domain is an IgG 1 , IgG 2 , IgG 3 or IgG 4 constant domain.
15. The modified IgG of claim 12 , wherein an amino acid substitution at amino acid residue 252 is a substitution with tyrosine, phenylalanine, tryptophan or threonine, an amino acid substitution at amino acid residue 254 is a substitution with threonine, an amino acid substitution at amino acid residue 256 is a substitution with serine, arginine, glutamine, glutamic acid, or aspartic acid, an amino acid substitution at amino acid residue 309 is a substitution with proline, an amino acid substitution at amino acid residue 311 is a substitution with serine, an amino acid substitution at amino acid residue 433 is a substitution with arginine, serine, isoleucine, proline or glutamine, and an amino acid substitution at amino acid residue 434 is a substitution with histidine, phenylalanine, or tyrosine.
16. The modified IgG of claim 15 , wherein an amino acid substitution at amino acid residue 385 is a substitution with arginine, aspartic acid, serine, threonine, histidine, lysine or alanine, an amino acid substitution at amino acid residue 386 is a substitution with threonine, proline, aspartic acid, serine, lysine, arginine, isoleucine, or methionine, an amino acid substitution at amino acid residue 387 is a substitution with arginine, histidine, serine, threonine, or alanine, and an amino acid substitution at amino acid residue 389 is a substitution with proline or serine.
17. The modified IgG of claim 10 which is a rodent, donkey, sheep, rabbit, goat, guinea pig, camel, horse or chicken IgG.
18. The modified IgG of claim 4 , wherein the non-human IgG constant domain is a rodent, donkey, sheep, rabbit, goat, guinea pig, camel, horse or chicken IgG constant domain.
19. The modified IgG of claim 1 , 2 , 3 , 4 , 6 , 7 , 15 or 16 which inimunospecifically binds to an RSV antigen.
20. The modified IgG of claim 1 , 2 , 3 , 6 , 15 or 16 which comprises the heavy chain variable domain and light chain variable domain of palivizumab (SEQ ID NOS.: 7 and 8).
21. The modified IgG of claim 1 , 2 , 3 , 6 , 15 or 16 which comprises the heavy chain variable domain and light chain variable domain of A4B4L1FR-S28R (SEQ ID NOS.:48 and 11).
22. A pharmaceutical composition comprising the modified IgG of claim 1 , 2 , 3 , 4 , 6 , 7 , 15 or 16 and a pharmaceutically acceptable carrier.
23. A pharmaceutical composition comprising the modified IgG of claim 19 and a pharmaceutically acceptable carrier.
24. A pharmaceutical composition comprising the modified IgG of claim 20 and a pharmaceutically acceptable carrier.
25. A pharmaceutical composition comprising the modified IgG of claim 21 and a pharmaceutically acceptable carrier.
26. A kit comprising the modified IgG of claim 1 , 2 , 2 , 4 , 6 , 7 , 15 or 16 , in a container, and instructions for use.
27. A kit comprising the modified IgG of claim 19 , in a container, and instructions for use.
28. A kit comprising the modified IgG of claim 20 , in a container, and instructions for use.
29. A kit comprising the modified IgG of claim 21 , in a container, and instructions for use.
30. The modified IgG of claim 1 , 2 , 3 , 6 , 15 or 16 which comprises the variable heavy (VH) complementarily determining region (CDR)1, VH CDR2, VH CDR3, variable light (VL) CDR1, VL CDR2 and VL CDR3 of palivizumab (SEQ ID NOS.: 1–6, respectively).
31. The modified IgG of claim 1 , 2 , 3 , 6 , 15 or 16 which comprises the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2 and VL CDR3 of A4B4L1FR-S28R (SEQ ID NOS.: 10, 19, 20, 39, 5, and 6, respectively).