Epha3 and multi-valent targeting of tumors

Patent metadata

Field	Value
Publication number	US-2016297863-A1
Application number	US-201415035560-A
Country	US
Kind code	A1
Filing date	Nov 11, 2014
Priority date	Nov 11, 2013
Publication date	Oct 13, 2016
Grant date	—

Abstract

Provided herein is a construct comprising, in combination: an EphA3, EphA2 and/or EphB2 binding ligand; and at least one effector molecule. In some embodiments, the at least one effector molecule comprises a therapeutic agent, a nanoparticle, a detectable group, a lipid, or a liposome. In some embodiments, the construct is a fusion protein and/or a covalent conjugate. Further provided is a construct comprising in combination: a ligand that binds to EphA2, EphA3 and/or EphB2; a ligand that binds to IL-13Rα2; and at least one effector molecule. Also provided are methods of use thereof for treating cancer.

First claim

1 . A construct comprising, in combination:
a ligand that binds to EphA2, EphA3, and EphB2; and at least one effector molecule.

2 . (canceled)

3 . The construct of claim 1 , wherein said construct is a fusion protein and/or a covalent conjugate.

4 . The construct of claim 1 , wherein said ligand is eA5, a mutant of eA5, or an EphA2, EphA3 and EphB2 binding fragment thereof.

5 . The construct of claim 4 , wherein said eA5, mutant of eA5, or EphA2, EphA3 and EphB2 binding fragment thereof is glycosylated.

6 . The construct of claim 1 , wherein said at least one effector molecule comprises a diphtheria toxin or a Pseudomonas exotoxin A.

7 . (canceled)

8 . The construct of claim 1 , wherein said construct further comprises:
a cytosol localization element covalently coupled between said ligand and said at least one effector molecule; and a subcellular compartment localization signal element covalently coupled between said ligand and said at least one effector molecule.

9 .- 10 . (canceled)

11 . The construct of claim 8 , wherein said subcellular compartment localization signal element is a nuclear localization element or a lysosomal localization element.

12 . (canceled)

13 . A construct of claim 1 , wherein said construct has the formula, from N terminus to C terminus, selected from the group consisting of:
A-B-C-D-E, E-D-C-B-A, A-B-D-C-E; and E-C-D-B-A,
wherein:
A is said ligand that binds to EphA2, EphA3 and EphB2;
B is a linker;
C is a cytosol localization element;
D is optionally present and is a subcellular compartment localization signal element; and
E is said at least one effector molecule.

14 . The construct of claim 13 , wherein each of A, C, and D, and optionally B and E, is a protein or peptide.

15 . The construct of claim 13 , wherein E is a toxin or an amphipathic antimicrobial peptide.

16 . The construct of claim 15 , wherein E is an amphipathic antimicrobial peptide comprising a sequence selected from the group consisting of: (KLAKLAK) 2 ; (KLAKKLA) 2 ; (KAAKKAA) 2 and (KLGKKLG) 2 .

17 . The construction of claim 13 , wherein D is present and is a nuclear localization element and wherein E is a radiopharmaceutical or a chemotherapeutic agent.

18 .- 19 . (canceled)

20 . A construct comprising, in combination:
a ligand that binds to EphA2, EphA3 and EphB2; a ligand that binds to IL-13Rα2; and at least one effector molecule.

21 . (canceled)

22 . The construct of claim 20 , wherein said construct is a fusion protein and/or a covalent conjugate.

23 . The construct of claim 22 , wherein said construct is a covalent conjugate of two or more fusion proteins.

24 . The construct of claim 20 , wherein said ligand that binds to EphA2, EphA3 and EphB2 is a monomer or dimer of:
eA5; a mutant of eA5; or an EphA2, EphA3 and EphB2 binding fragment thereof.

25 . The construct of claim 24 , wherein said eA5, mutant of eA5, or EphA2, EphA3 and EphB2 binding fragment thereof is glycosylated.

26 . The construct of claim 20 , wherein said ligand that binds to IL-13Rα2 is:
IL-13;
a mutant of IL-13; or
an IL-13Rα2 binding fragment thereof.

27 . The construct of claim 20 , wherein said construct has a first end and a second end, said first end comprising said ligand that binds to EphA2, EphA3 and EphB2; and said second end comprising said ligand that binds to IL-13Rα2.

28 . The construct of claim 27 , wherein said construct comprises a protein linker between said first end and said second end.

29 . The construct of claim 28 , wherein said protein linker comprises an ADCC and/or CDC activating domain.

30 . The construct of claim 28 , wherein said protein linker comprises an Fc fragment of human IgG 1 .

31 . A nucleic acid that encodes a construct of claim 1 , or a protein or peptide portion thereof.

32 . A host cell that contains a nucleic acid of claim 31 and is configured to express the encoded peptide.

33 . (canceled)

34 . A method of treating cancer in a subject in need thereof, comprising administering to said subject a construct of claim 1 in a treatment effective amount.

35 . The method of claim 34 , wherein said cancer is breast cancer, bladder cancer, pancreatic cancer, colorectal cancer, head and neck cancer, thyroid cancer, prostate cancer, melanoma, or glioma.

36 . The method of claim 34 , wherein said cancer is glioblastoma, prostate cancer or melanoma.

37 . (canceled)

38 . A method of detecting EphA2, EphA3 and/or EphB2 expressing cells, comprising administering said construct of claim 1 to a cell or group of cells, wherein said construct comprises a detectable group, and detecting said detectable group.

39 . A method of delivering at least one effector molecule to a cell of interest, comprising:
contacting the construct of claim 1 to a cell of interest under conditions in which said construct is internalized therein.

40 . The method of claim 39 , wherein said effector molecule is delivered to a subcellular compartment.

41 . The method of claim 39 , wherein said subcellular compartment is the nucleus.

42 . (canceled)

43 . A polypeptide comprising a mutant eA5, or an EphA2, EphA3 and/or EphB2 binding fragment thereof, or a nucleic acid encoding a mutant eA5, or an EphA2, EphA3 and/or EphB2 binding fragment thereof.

44 . The polypeptide or nucleic acid of claim 43 , wherein the mutant eA5 comprises a P123A, S125A and/or G127A mutation.

45 . (canceled)

Assignees

• Univ Wake Forest Health Sciences

Inventors

• Debinski Waldemar
• Tome Carla Lema
• Ferluga Sara
• Sonawane Poonam S

Classifications

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• C07K2319/55

  containing a fusion with a toxin, e.g. diphteria toxin · CPC title

• C07K14/52

  Cytokines; Lymphokines; Interferons · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K14/5437 Primary

  IL-13 · CPC title