Pd-1 targeted il-15/il-15ralpha fc fusion proteins with improved properties

A composition comprising an anti-PD-1 antigen binding domain (ABD) comprising:

• a) a variant variable heavy domain as compared to SEQ ID NO:5 comprising an W100F amino acid substitution and at least one additional amino acid substitution selected from the group consisting of F32L, S52aG, R97E, R97Y, R97W, L98R, S100aT, R97A, V99T, V99L, S100aA, L98Q, R97Q, V99F, V99L, S100aN, V99I, P100bS, G96H, L98V, V99A, V99Q, G96V, R97K, L98S, L98F, R97T, L98K, L98S, V99I, R97L, G96A, R97A, V99S, R97S, V99Y, R97H, L98R, according to Kabat numbering;
• and
• b) a variable light domain selected from the group consisting of:
• i) SEQ ID NO:168;
• and
• ii) a variant light domain as compared to SEQ ID NO:168 comprising an amino acid substitution selected from the group consisting of N27dH, N27dS, K30Y, S93T and Y94W, according to Kabat numbering;
• wherein said ABD binds to human PD-1.

A targeted IL-15/Rα heterodimeric Fc fusion protein comprising:

• a) a first monomer comprising, from N- to C-terminal:
• i) a IL-15 Rα sushi domain protein;
• ii) a first domain linker;
• iii) a variant IL-15 protein as compared to SEQ ID NO:2, comprising an amino acid substitution selected from the group consisting of N71Q, N79Q, N112Q, S114del and S114A;
• and
• iv) a first variant Fc domain;
• and
• b) a second monomer comprising a heavy chain comprising VH-CH1-hinge-CH2-CH3, wherein said CH2-CH3 is a second variant Fc domain;
• c) a third monomer comprising a light chain comprising VL-CL;
• wherein said VH and VL domains form an antigen binding domain that binds to human PD-1 and does not compete for binding to said human PD-1 with nivolumab and/or pembrolizumab.

A targeted IL-15/Rα heterodimeric Fc fusion protein comprising:

• a) a first monomer comprising, from N- to C-terminal:
• i) a IL-15 Rα sushi domain protein;
• ii) a first domain linker;
• iii) a variant IL-15 protein;
• and
• iv) a first variant Fc domain;
• and
• b) a second monomer comprising a heavy chain comprising VH-CH1-hinge-CH2-CH3, wherein said CH2-CH3 is a second variant Fc domain;
• c) a third monomer comprising a light chain comprising VL-CL;
• wherein said VH and VL domains form an antigen binding domain that binds to human PD-1;
• wherein said VH is a variant variable heavy domain as compared to SEQ ID NO:5 comprising an W100F amino acid substitution and at least one additional amino acid substitution selected from the group consisting of F32L, S52aG, R97E, R97Y, R97W, L98R, S100aT, R97A, V99T, V99L, S100aA, L98Q, R97Q, V99F, V99L, S100aN, V99I, P100bS, G96H, L98V, V99A, V99Q, G96V, R97K, L98S, L98F, R97T, L98K, L98S, V99I, R97L, G96A, R97A, V99S, R97S, V99Y, R97H, L98R, according to Kabat numbering;
• wherein said VL domain is selected from the group consisting of:
• i) SEQ ID NO:168;
• and
• ii) a variant light domain as compared to SEQ ID NO:168 comprising an amino acid substitution selected from the group consisting of N27dH, N27dS, K30Y, S93T and Y94W, according to Kabat numbering.

A heterodimeric protein comprising:

• a) a first fusion protein comprising:
• i) a variant IL-15 protein comprising the amino acid substitutions D30N/E64Q/N65D and N71Q/N79Q/N112Q as compared to SEQ ID NO:2;
• ii) a domain linker;
• and
• iii) a first variant Fc domain;
• and
• b) a second fusion protein comprising:
• i) an IL-15Rα sushi domain;
• ii) a domain linker;
• and
• iii) a second variant Fe domain.

A targeted IL-15/Rα heterodimeric Fc fusion protein comprising:

• a) a first monomer comprising, from N- to C-terminal:
• i) a IL-15 Rα sushi domain protein;
• ii) a first domain linker;
• iii) a variant IL-15 protein as compared to SEQ ID NO:2, comprising an amino acid substitution selected from the group consisting of N71Q, N79Q, N112Q, S114del and S114A;
• and
• iv) a first variant Fc domain;
• and
• b) a second monomer comprising a heavy chain comprising VH-CH1-hinge-CH2-CH3, wherein said CH2-CH3 is a second variant Fc domain;
• c) a third monomer comprising a light chain comprising VL-CL;
• wherein said VH and VL domains form an antigen binding domain that binds to human PD-1;
• wherein said VH is a variant variable heavy domain as compared to SEQ ID NO:5 comprising an W100F amino acid substitution and at least one additional amino acid substitution selected from the group consisting of F32L, S52aG, R97E, R97Y, R97W, L98R, S100aT, R97A, V99T, V99L, S100aA, L98Q, R97Q, V99F, V99L, S100aN, V99I, P100bS, G96H, L98V, V99A, V99Q, G96V, R97K, L98S, L98F, R97T, L98K, L98S, V99I, R97L, G96A, R97A, V99S, R97S, V99Y, R97H, L98R, according to Kabat numbering;
• wherein said VL domain is selected from the group consisting of:
• i) SEQ ID NO:168;
• and
• ii) a variant light domain as compared to SEQ ID NO:168 comprising an amino acid substitution selected from the group consisting of N27dH, N27dS, K30Y, S93T and Y94W, according to Kabat numbering.

A targeted IL-15/Rα heterodimeric Fc fusion protein comprising:

• a first monomer comprising the amino acid sequence of SEQ ID NO:
• 225;
• a second monomer comprising the amino acid sequence of SEQ ID NO:
• 244;
• and a third monomer comprising the amino acid sequence of SEQ ID NO:
• 196.