IL-10 variant molecules conjugated with antibody variable domain regions

1. A fusion protein of formula (I, II, III, IV, V, VI, or VII)

IL10-L1-X1-L1-X2-L1-IL10  (Formula I);

(Z) n -X1-L2-Y2-L1-IL10  (Formula II);

IL10-L1-Y1-L2-X2-(Z) n   (Formula III);

X1-L2-X2-L1-IL10  (Formula IV);

IL10-L1-X1-L2-X2  (Formula V);

X1-L1-IL10  (Formula VI);

IL10-L1-X2  (Formula VII),

wherein

“IL-10” is a monomer sequence selected from SEQ ID NOs: 15, 19, or 57;

“L1” is a linker of SEQ ID NO: 31 or 54;

“L2” is a linker of SEQ ID NO: 30;

“X1” is a VH region obtained from a first antibody specific for epidermal growth factor receptor (EGFR), CD3α, MAdCAM, α4β7 integrin, HIV, or Ebola;

“X2” is a VL region obtained from the same antibody as X1;

“Y1” is a VH region obtained from a second antibody specific for EGFR, CD3α, MAdCAM, α4β7 integrin, HIV, or Ebola;

“Y2” is a VL region obtained from the same second antibody as Y1; and

“Z” is a cytokine other than IL-10, wherein “n” is an integer of zero.

2. The fusion protein according to claim 1 , wherein formula IV and V are capable of forming a fusion protein complex where the IL-10 monomer from each of formula IV and V are capable of forming a functional homodimer.

3. The fusion protein according to claim 2 , wherein formula IV is SEQ ID NO: 48.

4. The fusion protein according to claim 2 , wherein formula V is SEQ ID NO: 49.

5. The fusion protein according to claim 2 , wherein the fusion protein complex is formed between SEQ ID NOs: 48 and 49.

6. The fusion protein according to claim 1 , wherein formula VI and VII are capable of forming a fusion protein complex where the IL-10 monomer from each of formula VI and VII are capable of forming a functional homodimer; and wherein the VH region and VL region pair to form a scFv.

7. The fusion protein according to claim 1 , wherein formula I is SEQ ID NOs: 37, 40, 43.

8. The fusion protein according to claim 1 , wherein the VH region and the VL region are from an anti-Ebola antibody; and wherein the VH region and VL region comprise a framework region with 6 CDRs engrafted with 6 CDRs from a second antibody selected from EGFR; CD52; PD-L1; PD-1; TIM3; BTLA; LAG3; CTLA4; CD20; CD47; GD-2; HER2; EpCAM; ICAM-1; ICAM-2; ICAM-3; ICAM-4; ICAM-5; VCAM, FAPα; 5T4; Trop2; EDB-FN; TGFβ Trap; MAdCAM, β7 integrin subunit; α4β7 integrin; α4 integrin SR-A1; SR-A3; SR-A4; SR-A5; SR-A6; SR-B; dSR-C1; SR-D1; SR-E1; SR-F1; SR-F2; SR-G; SR-H1; SR-H2; SR-I1; or SR-J1.

9. The fusion protein according to claim 8 , wherein the framework region with 6 CDRs is engrafted with 6 CDRs from an anti-MAdCAM antibody.

10. The fusion protein according to claim 1 , wherein the VH region and the VL region comprise modifications that reduce the antigenicity in a subject.

11. A pharmaceutical composition comprising a therapeutically effective amount of the fusion protein according to claim 1 , and a pharmaceutically acceptable carrier, excipient, preservative, stabilizers, or any combination thereof.

12. The fusion protein according to claim 8 , wherein the VH region and VL region comprise the framework regions from an anti-Ebola antibody.

13. The fusion protein according to claim 1 , wherein formula II and III are capable of forming a fusion protein complex where the IL-10 monomer from each of formula II and III are capable of forming a functional homodimer.