Delivering biological drugs to tissues

Patent metadata

Field	Value
Publication number	US-2020405805-A1
Application number	US-201916979143-A
Country	US
Kind code	A1
Filing date	Mar 11, 2019
Priority date	Mar 9, 2018
Publication date	Dec 31, 2020
Grant date	—

Abstract

Disclosed herein are fusion proteins for use in treating an inflammatory or immune disorder and methods of use. In some examples, the fusion proteins include an anchor domain and a therapeutic polypeptide. In some examples, the fusion proteins and methods herein can be used to treat inflammatory or immune disorders.

First claim

1 . A fusion protein comprising an anchor domain and a therapeutic polypeptide, wherein:
(a) the anchor domain comprises a lectin carbohydrate-binding anchor domain, a von Willebrand factor (vWF) collagen-binding anchor domain, a Clostridium collagenase (ColH) collagen-binding anchor domain, or a heparin-binding (HS) anchor domain, and the therapeutic polypeptide comprises:
(i) an antibody that specifically binds to, or an antagonist of, interleukin (IL)-17A, IL-6, IL-1β, IL-1α, IL-18, interferon (IFN)γ, IL-12, granulocyte-macrophage colony-stimulating factor (GM-CSF), transforming growth factor beta (TGFβ), IL-21, IL-22, IL-23, IL-4, IL-5, IL-13, IL-17F, IL-9, or VEGF-A, or
(ii) an immunoglobulin-binding (Ig) polypeptide or an antagonist of tumor necrosis factor alpha (TNFα); or
(b) the anchor domain comprises a lectin carbohydrate-binding anchor domain, a von Willebrand factor (vWF) collagen-binding anchor domain, or a Clostridium collagenase (ColH) collagen-binding anchor domain, and the therapeutic polypeptide comprises:
(i) an anti-inflammatory cytokine; or
(ii) an antagonist of, or antibody that specifically binds to, a pro-inflammatory cytokine.

2 . The fusion protein of claim 1 , wherein:
(a) the anti-inflammatory cytokine comprises IL-10; (b) the antagonist of the pro-inflammatory cytokine comprises interleukin-1 receptor antagonist (IL-1Ra); or (c) the antibody that specifically binds to a pro-inflammatory cytokine comprises an antibody that specifically binds to tumor necrosis factor alpha (TNFα).

3 . The fusion protein of claim 1 , wherein the lectin carbohydrate-binding anchor domain is a wheat germ agglutinin (WGA), jacalin (Jac), or concanavalin A (conA) carbohydrate-binding anchor domain.

4 . The fusion protein of claim 1 , wherein:
the antagonist of TGFβ is P17 or P144; the antagonist of TNFα comprises a TNF receptor 1 polypeptide; and/or the Ig-binding polypeptide comprises protein A, protein G, or protein L or Ig-binding fragment thereof.

5 . The fusion protein of claim 1 , wherein:
(a) the anchor domain comprises a lectin carbohydrate-binding anchor domain, and the therapeutic polypeptide comprises the inhibitor of TNFα polypeptide from the TNF receptor 1, IL-10, IL-1Ra, or an antibody that specifically binds TGFβ, TNFα, IL-1β, IL-6, IFNγ, IL-23, or IL-17A; (b) the anchor domain comprises the vWF collagen-binding anchor domain or the ColH collagen-binding anchor domain, and the therapeutic polypeptide comprises the inhibitor of TNFα polypeptide from the TNF receptor 1, IL-1Ra, IL-10, P17, P144, or the Ig-binding polypeptide; or (c) the anchor domain comprises the HS anchor domain, and the therapeutic polypeptide comprises the inhibitor of TNFα polypeptide from the TNF receptor 1.

6 . The fusion protein of claim 5 wherein the lectin carbohydrate-binding anchor domain comprises a WGA.

7 . The fusion protein of claim 1 , wherein:
the lectin carbohydrate-binding anchor domain comprises the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8; the vWF collagen-binding anchor domain comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 26; the ColH collagen-binding anchor domain comprises the amino acid sequence of SEQ ID NO: 3; and/or the HS anchor domain comprises the amino acid sequence of SEQ ID NO: 5.

8 . The fusion protein of claim 1 , further comprising a linker between the anchor binding domain and the therapeutic domain.

9 . The fusion protein of claim 8 , wherein linker has the amino acid sequence of SEQ ID NO:9 or SEQ ID NO: 19.

10 . The fusion protein of claim 1 , further comprising a label, solubility enhancing sequence, purification tag, or combinations thereof.

11 . The fusion protein of claim 1 , wherein the anchor domain, therapeutic peptide or combination thereof are benzylated, glycosylated, acetylated, phosphorylated, amidated, pegylated, and combinations thereof.

12 - 14 . (canceled)

15 . An isolated nucleic acid molecule encoding the fusion protein of claim 1 for use in treating an inflammatory or immune disorder.

16 . The isolated nucleic acid molecule of claim 15 , operably linked to a heterologous promoter.

17 . An expression vector comprising the isolated nucleic acid molecule of claim 16 , for use in treating an inflammatory disorder.

18 . A method for treating an inflammation of a tissue in a subject, comprising administering to the tissue of the subject a therapeutically effective amount of the fusion protein of claim 1 thereby treating the inflammation of the tissue in the subject.

19 . The method of claim 18 , wherein the therapeutic protein comprises an immunoglobulin-binding (Ig) polypeptide or fragment thereof, further comprising administering an antibody that specifically binds the Ig-binding polypeptide or fragment thereof.

20 . The method of claim 19 , wherein the tissue is in a mouth, a bladder, a nasal cavity, a gastrointestinal tract, a lung, a joint, an eye, or the reproductive system of a subject.

21 . The method of claim 19 , wherein the subject has chronic obstructive pulmonary disease, asthma, bronchitis, rhinosinusitis, mucositis, left-sided ulcerative colitis, inflammatory bowel disease, periodontal disease, peri-implantitis, interstitial cystitis, atrophic vaginitis, or arthritis.

22 . The method of claim 19 , wherein tissue is the eye, the therapeutic polypeptide comprises an antibody that specifically binds IL-17A or IL-23, and method treats the inflammation in the eye of the subject.

23 . The method of claim 22 , wherein the subject has dry eye.

24 . The method of claim 22 , wherein the subject has Sjögren's syndrome.

25 . A pharmaceutical composition comprising a therapeutically effective amount of the fusion protein of claim 1 and a pharmaceutically acceptable carrier formulated for external and/or topical administration to a tissue.

26 . A unit dose dispenser for dispensing the pharmaceutical composition claim 25 locally to an inflamed tissue.

27 . The pharmaceutical composition of claim 25 , wherein the pharmaceutical composition is formulated as a gel, cream, lotion, or ointment.

28 . (canceled)

Assignees

• Univ Pittsburgh Commonwealth Sys Higher Education

Inventors

• Shanks Robert Michael Queen
• Klarlund Jes K

Classifications

• C07K14/4726 Primary

  Lectins · CPC title

• C07K14/42

  Lectins, e.g. concanavalin, phytohaemagglutinin · CPC title

• C07K16/247

  IL-4 · CPC title

• C07K16/249

  Interferons · CPC title

• C07K16/22

  against growth factors {; against growth regulators} · CPC title