US 12,226,481 B2
Cytokine fusion proteins
Yiyou Chen, San Jose, CA (US); and Dagang Guo, Beijing (CN)
Assigned to Beijing Percans Oncology Co., Ltd., Beijing (CN)
Appl. No. 16/964,316
Filed by Beijing Percans Oncology Co., Ltd., Beijing (CN)
PCT Filed Jan. 24, 2019, PCT No. PCT/US2019/014985
§ 371(c)(1), (2) Date Oct. 30, 2020,
PCT Pub. No. WO2019/147837, PCT Pub. Date Aug. 1, 2019.
Claims priority of application No. PCT/CN2018/073940 (WO), filed on Jan. 24, 2018; and application No. PCT/CN2018/119071 (WO), filed on Dec. 4, 2018.
Prior Publication US 2021/0052727 A1, Feb. 25, 2021
Int. Cl. A61K 39/395 (2006.01); A61K 39/00 (2006.01); A61K 45/06 (2006.01); A61P 35/00 (2006.01); C07K 14/525 (2006.01); C07K 14/535 (2006.01); C07K 14/54 (2006.01); C07K 14/55 (2006.01); C07K 14/56 (2006.01); C07K 14/565 (2006.01); C07K 14/57 (2006.01); C07K 16/28 (2006.01)
CPC A61K 39/3955 (2013.01) [A61P 35/00 (2018.01); C07K 14/525 (2013.01); C07K 14/535 (2013.01); C07K 14/5418 (2013.01); C07K 14/5434 (2013.01); C07K 14/55 (2013.01); C07K 14/56 (2013.01); C07K 14/565 (2013.01); C07K 14/57 (2013.01); C07K 16/2818 (2013.01); A61K 2039/585 (2013.01); A61K 2039/80 (2018.08); A61K 45/06 (2013.01)] 14 Claims
 
1. A cytokine fusion protein, comprising a first human cytokine fused to the N-terminus of a second human cytokine, wherein the first cytokine is IL-2 and wherein the second cytokine is TNF-α, wherein the human IL-2 comprises a C125S or C125A mutation and the human TNF-α has increased binding to TNFR1 relative to TNFR2 and B comprises at least one mutation selected from S86T and R32W, including combinations thereof, wherein the first and second cytokines are separated by a stable peptide linker, and wherein the cytokine fusion protein comprises:
 
 (SEQ ID NO: 19)
 
 IL-2(C125S)-(GGGGS)3-TNF-α (S86T);
 
 or
 
 
 
 (SEQ ID NO: 20)
 
 IL2(C125A)-(GGGGS)2-TNF-α (S86T);
 
 
 
 (SEQ ID NO: 21)
 
 IL2(C125A)-(GGGGS)2-TNF-α (R32W);
 
 
 
 (SEQ ID NO: 22)
 
 IL2(C125A)-PAPAP-TNF-α (S86T);
 
 
 
 (SEQ ID NO: 23)
 
 IL2(C125A)-PAPAP-TNF-α (R32W);
 
 
 
 (SEQ ID NO: 24)
 
 IL2(C125A)-PAEAAAKEAAAKA-TNF-α (S86T);
 
 
 
 (SEQ ID NO: 25)
 
 IL2(C125A)-PAEAAAKEAAAKA-TNF-α (R32W);
 
 
 
 IL-2(C125S)-(GGGGS)3-TNF-α (R31E, S86T);
 
 or
 
 
 
 (SEQ ID NO: 26)
 
 IFN-β-(GGGGS)2-TNF-α.
an amino acid sequence that is at least 95% identical to a sequence selected from SEQ ID NOs: 19-25 and which has a cytokine signaling activity and anti-tumor activity.