Dual cytokine fusion proteins comprising IL-10
The application relates to a dual cytokine fusion protein composition, pharmaceutical composition, and/or formulation thereof comprising IL-10 or IL-10 variant molecules fused to a single chain variable fragment scaffolding system and a second cytokine, where the second cytokine is linked in the hinge region of the scFv. The application also relates to methods of using the dual cytokine fusion protein composition for treating cancer, inflammatory diseases or disorders, and immune and immune mediated diseases or disorders.
1. A dual cytokine fusion protein of formula (I)
NH2-(IL-10)-(X 1 )—(Z n )—(X 2 )—(IL-10)-COOH (Formula I);
wherein
“IL-10” is a monomer of SEQ ID No: 16;
“X 1 ” is a variable light (VL) or variable heavy (VH) chain region comprising three complementarity determining regions (CDRs) that bind to epidermal growth factor receptor (EGFR);
“X 2 ” is a VH or VL region comprising three CDRs that bind to EGFR;
wherein when X 1 is a VL, X 2 is a VH and when X 1 is a VH, X 2 is a VL, and wherein the CDRs are obtained from an anti-EGFR monoclonal antibody and wherein X 1 and X 2 together form a single chain variable fragment (scFv);
“Z” is IL 2; and
“n” is an integer of 1.
2. The dual cytokine fusion protein according to claim 1 , wherein the fusion protein is SEQ ID No: 35.
3. A pharmaceutical composition comprising the dual cytokine fusion protein according to claim 1 , pharmaceutically acceptable buffers, and pharmaceutically acceptable excipients.
4. The fusion protein according to claim 1 , wherein the VH and VL regions comprise a framework region obtained from a human anti-Ebola antibody.
5. The fusion protein according to claim 4 , wherein the frame region from the anti-Ebola antibody is engrafted with the three VH CDRs and three VL CDRs from an anti-EGFR monoclonal antibody.
6. The fusion protein according to claim 2 , wherein the fusion protein is at least 95% identical to SEQ ID No: 35.