IP Library › Granted Patent US 12,540,189
Granted Patent B2
US 12,540,189 · App. 18/844,732 · Granted Feb 3, 2026

Method of treating cancer by administering an inhibitor of erythropoietin receptor activity comprising either erythropoietin receptor or erythropoietin protein

Inventors: Bo Yu (Mountain View, CA); Edgar G. Engleman (Atherton, CA); Xiangyue Zhang (Sunnyvale, CA); David Kung-Chun Chiu (Mountain View, CA)
Assignees: ImmunEdge, Inc.; The Board of Trustees of the Leland Stanford Junior University
C07K16/2866A61P37/02A61P37/06C07K14/505C07K16/22C12N15/113A61K2039/505C07K2317/31C07K2317/565C07K2317/70C07K2317/76C07K2319/30C07K2319/31C12N2310/14
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Quick Facts
Patent No.
US 12,540,189
App. No.
18/844,732
Granted
Feb 3, 2026
Kind
B2
Abstract

The present disclosure provides EPO analogs, anti-EPOR antibodies, anti-EPO antibodies, and fragments thereof that specifically bind to the hetero-EPOR or homo-EPOR or EPO with high affinity. Also provided herein are engineered EPOs. The EPO analogs, anti-EPOR antibodies, anti-EPO antibodies, and/or engineered EPOs can be used to treat patients.

Claims (63)

1 . A method of treating a cancer in a patient in need thereof, wherein said method comprises administering a composition to the patient, wherein said composition comprises a therapeutically effective amount of an inhibitor of erythropoietin receptor activity, thereby treating the cancer in the patient in need thereof,

wherein said inhibitor of erythropoietin receptor activity comprises:

(i) an engineered erythropoietin receptor comprising a first subunit comprising a soluble fragment of an erythropoietin receptor, the soluble fragment comprising an extracellular domain from a human erythropoietin receptor, wherein said engineered erythropoietin receptor binds to erythropoietin and inhibits said erythropoietin receptor activity; or

(ii) an engineered human erythropoietin protein, wherein said engineered human erythropoietin protein binds to an erythropoietin receptor to form a complex and inhibits said erythropoietin receptor activity.

2 . The method of claim 1 , wherein said erythropoietin receptor is expressed on a myeloid cell.

3 . The method of claim 1 , wherein said engineered erythropoietin receptor further comprises:

(a) a second subunit comprising a soluble fragment of an erythropoietin receptor comprising an extracellular domain from a human erythropoietin receptor; or

(b) a second subunit comprising a soluble fragment of CD131 comprising an extracellular domain from a human CD131 protein.

4 . The method of claim 1 , wherein said engineered erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor of said first subunit.

5 . The method of claim 1 , wherein said engineered erythropoietin receptor comprises a single extracellular domain from a human erythropoietin, and wherein said single extracellular domain comprises domains D1 and D2 of said human erythropoietin receptor.

6 . The method of claim 3 , wherein said engineered erythropoietin receptor comprises a homo-erythropoietin receptor comprising:

(a) said first subunit comprising said soluble fragment of said erythropoietin receptor comprising said extracellular domain from said human erythropoietin receptor, and

(b) said second subunit comprising said soluble fragment of said erythropoietin receptor comprising said extracellular domain from said human erythropoietin receptor.

7 . The method of claim 3 , wherein said engineered erythropoietin receptor comprises a hetero-erythropoietin receptor comprising:

(a) said first subunit comprising said soluble fragment of said erythropoietin receptor comprising said extracellular domain from said human erythropoietin receptor, and

(b) said second subunit comprising said soluble fragment of CD131 comprising said extracellular domain from said human CD131 protein, or a soluble fragment of an erythropoietin receptor comprising an extracellular domain from a human erythropoietin receptor, wherein said extracellular domain from said human erythropoietin receptor of said second subunit comprises an amino acid variation compared to said extracellular domain from said human erythropoietin receptor of said first subunit.

8 . The method of claim 6 , wherein:

(a) said first subunit of said homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and

(b) said second subunit of said homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor.

9 . The method of claim 7 , wherein:

(a) said first subunit of said hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and

(b) said second subunit of said hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human CD131 protein, or said extracellular domain from said human erythropoietin receptor of said second subunit.

10 . The method of claim 7 , wherein said extracellular domain from said human erythropoietin receptor of said first subunit comprises an F93A amino acid substitution as compared to SEQ ID NO: 3897 without the signal peptide sequence corresponding to residues 1-24 of SEQ ID NO: 3897.

11 . The method of claim 7 , wherein said extracellular domain from said human CD131 protein comprises D3 and D4 domains of human CD131 protein.

12 . The method of claim 4 , wherein said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3897.

13 . The method of claim 6 , wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human erythropoietin receptor of said second subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3897.

14 . The method of claim 7 , wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human CD131 protein of said second subunit comprises an amino acid sequence having at least 85% identity to amino acids 20-221 of SEQ ID NO: 3901.

15 . The method of claim 1 , wherein said cancer is a cancer that is resistant to immune checkpoint inhibitor therapy.

16 . The method of claim 1 , further comprising administering one or more immune checkpoint inhibitors to said patient.

17 . The method of claim 16 , wherein said one or more immune checkpoint inhibitors comprises an inhibitor of Cytotoxic T-Lymphocyte Associated Protein 4 (CTLA-4), Programmed Death 1 (PD-1), Programmed Death Ligand 1 (PD-L1), Killer-Cell Immunoglobulin-Like Receptors (KIRs), or Lymphocyte-Activation Gene 3 (LAG3).

18 . The method of claim 16 , wherein said one or more immune checkpoint inhibitors comprises nivolumab, pembrolizumab, cemiplimab, atezolizumab, avelumab, durvalumab, ipilimumab, lirilumab, or BMS-986016.

19 . The method of claim 1 , wherein said cancer is a cancer selected from the group consisting of a lung cancer, a breast cancer, a colon cancer, a brain cancer, a skin cancer, a colorectal cancer, a liver cancer, a gastric cancer, a renal cancer, and a lymph node metastasis and a liver metastasis.

20 . The method of claim 19 , wherein said cancer is a solid tumor.

21 . The method of claim 1 , wherein said erythropoietin receptor activity comprises phosphorylation of an intracellular domain of said erythropoietin receptor or activation of Janus tyrosine kinase 2 (Jak2), Signal transducer and activator of transcription 5 (Stat5), mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), phosphatidylinositol 3-kinase (PI3K), v-Akt Murine Thymoma Viral Oncogene/Protein Kinase-B (Akt/PKB), or Mammalian target of rapamycin (mTOR); wherein said erythropoietin receptor activity is measured by western blotting, an enzyme-linked immunosorbant assay (ELISA), a flow cytometry assay, a cell proliferation assay, an apoptosis assay, or a combination thereof.

22 . The method of claim 1 , wherein said inhibitor of erythropoietin receptor activity is an engineered erythropoietin protein comprising at least one amino acid substitution comprising: K20A, N24Q, N24A, N24S, N38Q, N38A, N38S, K45A, K52A, Q58A, E62R, E62A, Q65A, L69A, E72A, R76E, R76A, L80A, N83Q, N83A, N83S, S84A, S85A, K97A, K116A, G151A, R103A, K45D, N147K, R150E, Q65A, E72R, N83A, K140A, K152A, or K154A, wherein the amino acid position is determined by alignment with SEQ ID NO: 1.

23 . The method of claim 7 , wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3903 with the proviso that residue 93 as compared to SEQ ID NO: 3903 without the signal peptide sequence corresponding to residues 1-24 of SEQ ID NO: 3903 is an alanine, and

(b) said extracellular domain from said human CD131 protein of said second subunit comprises an amino acid sequence having at least 85% identity to amino acids 20-221 of SEQ ID NO: 3901, or said extracellular domain from said human erythropoietin receptor of said second subunit comprising said amino acid variation compared to said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 85% identity to amino acids 25-250 of SEQ ID NO: 3897.

24 . The method of claim 6 , wherein said first subunit of the homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and said second subunit of the homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor; wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 95% identity to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human erythropoietin receptor of said second subunit comprises an amino acid sequence having at least 95% identity to amino acids 25-250 of SEQ ID NO: 3897.

25 . The method of claim 7 , wherein said first subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and said second subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human CD131 protein or said extracellular domain from said human erythropoietin receptor of said second subunit; wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 95% identity to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human CD131 protein of said second subunit comprises an amino acid sequence having at least 95% identity to amino acids 20-221 of SEQ ID NO: 3901, or said extracellular domain from said human erythropoietin receptor of said second subunit comprising said amino acid variation compared to said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence having at least 95% identity to amino acids 25-250 of SEQ ID NO: 3897.

26 . The method of claim 25 , wherein residue 93 of said extracellular domain from said human erythropoietin receptor of said first subunit as compared to SEQ ID NO: 3903 without the signal peptide sequence corresponding to residues 1-24 of SEQ ID NO: 3903 is an alanine.

27 . The method of claim 6 , wherein said first subunit of the homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and wherein said second subunit of the homo-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor; wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human erythropoietin receptor of said second subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3897.

28 . The method of claim 7 , wherein said first subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and wherein said second subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human CD131 protein or said extracellular domain from said human erythropoietin receptor of said second subunit; wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3897, and

(b) said extracellular domain from said human CD131 protein of said second subunit comprises an amino acid sequence according to amino acids 20-221 of SEQ ID NO: 3901, or said extracellular domain from said human erythropoietin receptor of said second subunit comprising said amino acid variation compared to said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3897.

29 . The method of claim 7 , wherein said first subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human erythropoietin receptor, and wherein said second subunit of the hetero-erythropoietin receptor further comprises an Fc portion of an immunoglobulin molecule fused to said extracellular domain from said human CD131 protein or said extracellular domain from said human erythropoietin receptor of said second subunit; wherein:

(a) said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3903, and

(b) said extracellular domain from said human CD131 protein of said second subunit comprises an amino acid sequence according to amino acids 20-221 of SEQ ID NO: 3901, or said extracellular domain from said human erythropoietin receptor of said second subunit comprising said amino acid variation compared to said extracellular domain from said human erythropoietin receptor of said first subunit comprises an amino acid sequence according to amino acids 25-250 of SEQ ID NO: 3897.

30 . The method of claim 1 , wherein said treating comprises increasing an immune response to said cancer.

31 . The method of claim 1 , wherein said treating promotes differentiation of naive T cells into effector T cells.

32 . The method of claim 1 , wherein said treating inhibits differentiation of naive T cells into regulatory T cells.

33 . The method of claim 1 , wherein said cancer is associated with immunosuppressive cells.

34 . The method of claim 1 , wherein said cancer is associated with effector T cells.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 6, 2024
From: YU, BO
To: IMMUNEDGE, INC.
Reel/Frame 068517/0690 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 6, 2024
From: ENGLEMAN, EDGAR G.; ZHANG, XIANGYUE; CHIU, DAVID KUNG-CHUN
To: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Reel/Frame 068517/0792 →
Continuity (2)
Provisional Application 63317943 · Mar 8, 2022
Related Publication 20250129170A1 · Apr 24, 2025
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