IL2 agonists
The invention relates to variants of interleukin-2 (IL2). In one embodiment, the IL2 variants activate effector T cells over regulatory T cells. In particular, the invention relates to a polypeptide comprising a mutein of human IL2 or of a functional variant of human IL2, wherein the human IL2 or functional variant thereof is substituted at at least a position having an acidic or basic amino acid residue in wild type human IL2 that contacts the alpha subunit of the αβγ IL2 receptor complex (I12Kαβγ). Alternatively, the mutein of human IL2 or of a functional variant of human IL2 comprises at least (i) one or more amino acid substitutions which reduce the affinity for the alpha subunit of II_2Kαβγ and (ii) one or more amino acid substitutions which enhance the affinity for II_2Kβγ. The invention also relates to polynucleotides coding for the polypeptides of the invention, host cells comprising the polynucleotides, pharmaceutical compositions comprising the polypeptides, polynucleotides or host cells, therapeutic or prophylactic methods of treatment using the polypeptides, polynucleotides, host cells or pharmaceutical compositions and medical preparations comprising the polypeptides, polynucleotides, host cells or pharmaceutical compositions.
1 . A polypeptide comprising a mutein of human interleukin-2 (IL2) or of a functional variant of human IL2, wherein the human IL2 or functional variant thereof is substituted at at least a position having an acidic or basic amino acid residue in wild type human IL2 that contacts the alpha subunit of the αβγ IL2 receptor complex (IL2Rαβγ), wherein if the amino acid residue is an acidic amino acid residue in wild type human IL2 the substitution is by a basic amino acid residue and if the amino acid residue is a basic amino acid residue in wild type human IL2 the substitution is by an acidic amino acid residue, wherein the human IL2 or functional variant thereof is substituted with at least a glutamic acid at lysine 43, and a lysine at glutamic acid 61, said amino acid substitutions numbered in accordance with wild type human IL2.
2 . The polypeptide of claim 1 , wherein wild type human IL2 has the amino acid sequence according to SEQ ID NO: 17.
3 . The polypeptide of claim 1 , wherein the acidic amino acid residue in wild type human IL2 contacts a basic amino acid residue in the alpha subunit of IL2Rαβγ.
4 . The polypeptide of claim 1 , wherein the basic amino acid residue in wild type human IL2 contacts an acidic amino acid residue in the alpha subunit of IL2Rαβγ.
5 . The polypeptide of claim 1 , wherein the substitution reduces the affinity for IL2Rα βγ or wherein the substitution reduces the affinity for IL2Rαβγ to a greater extent than for the βγ IL2 receptor complex (IL2Rβγ).
6 . The polypeptide of claim 1 , wherein the polypeptide preferentially activates effector T cells over regulatory T cells.
7 . The polypeptide of claim 1 , wherein the human interleukin-2 (IL2) or a functional variant thereof is substituted with a lysine at glutamic acid 62.
8 . The polypeptide of claim 1 , wherein the human IL2 or functional variant thereof further comprises one or more amino acid substitutions which enhance the affinity for IL2Rβγ.
9 . A polypeptide comprising a mutein of human interleukin-2 (IL2) or of a functional variant of human IL2, wherein the human IL2 or functional variant thereof comprises at least (i) one or more amino acid substitutions which reduce the affinity for the alpha subunit of IL2Rαβγ and (ii) one or more amino acid substitutions which enhance the affinity for IL2Rβγ, wherein the human IL2 or functional variant thereof is substituted with at least a glutamic acid at lysine 43 and a lysine at glutamic acid 61.
10 . The polypeptide of claim 9 , wherein the one or more amino acid substitutions which reduce the affinity for the alpha subunit of IL2Rαβγ comprise substitutions at one or more positions of IL2 or a functional variant thereof selected from the group consisting of K35, T37, R38, T41, F42, K43, F44, Y45, E61, E62, K64, P65, E68, L72, and Y107, and wherein the one or more amino acid substitutions which enhance the affinity for IL2Rβγ comprise substitutions at one or more positions of IL2 selected from the group consisting of K9, L12, Q13, E15, H16, D20, Q74, L80, R81, D84, L85, 186, N88, I92, L94, and E95.
11 . The polypeptide of claim 8 , wherein the one or more amino acid substitutions which enhance the affinity for IL2Rβγ comprise substitutions at one or more positions of IL2 selected from the group consisting of K9, L12, Q13, E15, H16, D20, Q74, L80, R81, D84, L85, 186, N88, I92, L94, and E95.
12 . The polypeptide of claim 9 , wherein the one or more amino acid substitutions which enhance the affinity for IL2Rβγ comprise the following set of substitutions: 80F, 81D, 85V, 86V, 92F.
13 . The polypeptide of claim 1 , which is an extended pharmacokinetic (PK) IL2 further comprising an amino acid sequence which is heterologous to the IL2 or functional variant thereof fused to the substituted IL2 or functional variant thereof.
14 . The polypeptide of claim 13 , wherein the amino acid sequence which is heterologous to the IL2 or functional variant thereof is selected from the group consisting of serum albumin, an immunoglobulin fragment, transferrin, and Fn3, or variants thereof.
15 . The polypeptide of claim 14 , wherein the serum albumin comprises mouse serum albumin or human serum albumin.
16 . A polynucleotide encoding the polypeptide of claim 1 .
17 . The polynucleotide of claim 16 , which is RNA.
18 . A host cell comprising the polynucleotide of claim 16 .
19 . A pharmaceutical composition comprising the polypeptide of claim 1 , a polynucleotide encoding the polypeptide of claim 1 , or a host cell comprising the polynucleotide.
20 . A medical preparation comprising: a. the polypeptide of claim 1 , a polynucleotide encoding the polypeptide of claim 1 , a host cell comprising the polynucleotide, or a pharmaceutical composition comprising the polypeptide or the polynucleotide; and b. a peptide or polypeptide comprising an epitope for inducing an immune response against an antigen in a subject or a polynucleotide encoding the peptide or polypeptide.
21 . The medical preparation of claim 20 , which comprises each component a. and b. in a separate container.
22 . The medical preparation of claim 20 , further comprising instructions for use of the medical preparation for treating or preventing cancer wherein the antigen is a tumor-associated antigen.
23 . The polypeptide of claim 11 , wherein the one or more amino acid substitutions which enhance the affinity for IL2Rβγ comprise the following set of substitutions: 80F, 81D, 85V, 86V, 92F.
24 . The polypeptide of claim 9 , wherein (i) the one or more amino acid substitutions which reduce the affinity for the alpha subunit of IL2Rαβγ comprise a substitution at position 43 (lysine) with glutamic acid and a substitution at position 61 (glutamic acid) with lysine.
25 . The polypeptide of claim 9 , wherein the human interleukin-2 (IL2) or a functional variant thereof is substituted with a lysine at glutamic acid 62.