Methods for diagnosing CNS disorders with fusion antibodies that cross the blood-brain barrier in both directions

• US 8,753,610 B2
• Filed: 04/07/2010
• Issued: 06/17/2014
• Est. Priority Date: 08/18/2006
• Status: Active Grant

• Current Assignee: JCR Pharmaceuticals Co., Ltd.
• Original Assignee: ArmaGen Technologies, Inc.
• Inventors: Pardridge, William M., Boado, Ruben J.
• Primary Examiner(s): Emch, Gregory S
• Application Number:
  US12/756,093
• Publication Number:
  US 20100290985A1
• Time in Patent Office: 1,532 Days
• Field of Search: 435/7.1, 530/387.3, 424/135.1, 424/9.1, 424/136.1, 424/1.49, 424/9.4, 424/9.341
• US Class Current: 424/9.341
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/1081 Togaviridae, e.g. flavivirus, rubella virus, hog cholera virus
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 16/2881 against CD71
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides diagnostic and therapeutic macromolecular compositions that cross the blood-brain barrier, in some embodiments in both directions, while allowing their activity to remain substantially intact once across the barrier. Also provided are methods for using such compositions in the diagnosis or treatment of CNS disorders such as Alzheimer'"'"'s disease.

Claims

1. A method of diagnosing a brain disorder comprising detecting a signal emitted by a composition in the central nervous system (CNS) of an individual, wherein the composition comprises a fusion antibody that:

(i) comprises a first antibody and a second antibody, wherein the first antibody is an ScFv antibody that is linked to the carboxy terminus of the second antibody;

(ii) is capable of crossing the blood-brain barrier (BBB) from the blood to the brain by binding an endogenous receptor of the BBB;

(iii) comprises an Fc region that enables crossing the BBB from the brain to the blood by binding to an Fc receptor; and

(iv) interacts with a pathological substance associated with the brain disorder, wherein the ScFv retains at least 10% of its affinity compared to the affinity of the tetrameric immunoglobulin from which the ScFv can be derived.

2. The method of claim 1, wherein the pathological substance is a protein.

3. The method of claim 2, wherein the protein is Aβ

amyloid, α

-synuclein, huntingtin protein, PrP prion protein, West Nile envelope protein, tumor necrosis factor (TNF) related apoptosis inducing ligand (TRAIL), Nogo A, HER2, epidermal growth factor receptor (EGFR), hepatocyte growth factor (HGF), or oligodendrocyte surface antigen.

4. The method of claim 3, wherein the protein is Aβ

amyloid.

5. The method of claim 1, wherein the composition is labeled with a substance that emits the signal.

6. The method of claim 5, wherein the substance that emits the signal is selected from the group consisting of positron emitters, radionuclide, and magnetic substances.

7. The method of claim 1, further comprising systemically administering the composition to the individual prior to the detecting step.

8. The method of claim 1, wherein the detecting step comprises using positron emission tomography (PET), single photon emission computed tomography (SPECT), or magnetic resonance imaging (MRI).

9. The method of claim 1, wherein the detecting step comprises quantitation by a sandwich enzyme-linked immunosorbent assay (ELISA).

10. The method of claim 1, wherein the pathological substance is selected from the group consisting of a protein, a nucleic acid, a carbohydrate, a carbohydrate polymer, a lipid, a glycolipid, a small molecule, and a combination thereof.

11. The method of claim 1, wherein the endogenous receptor of the BBB is an insulin receptor, transferrin receptor, leptin receptor, lipoprotein receptor, or IGF receptor.

12. The method of claim 1, wherein the fusion antibody is able to cross the BBB by binding an insulin receptor.

13. The method of claim 1, wherein the ScFv is capable of binding the endogenous receptor of the BBB.

14. The method of claim 1, wherein the ScFv is fused at its amino terminus to the carboxy terminus of the heavy chain of the second antibody.

15. The method of claim 1, wherein the ScFv is fused at its amino terminus to the carboxy terminus of the light chain of the second antibody.

16. The method of claim 1, wherein the ScFv interacts with the pathological substance associated with the brain disorder.

17. The method of claim 1, wherein the ScFv is able to bind to amyloid β

(Aβ

) peptide.

18. The method of claim 1, wherein the ScFv dissociation constant of the ScFv for its antigen is less than 100 nM.

19. The method of claim 1, wherein the second antibody interacts with the pathological substance associated with the brain disorder.

20. The method of claim 1, wherein the second antibody is able to bind to amyloid β

(Aβ

) peptide.

21. The method of claim 1, wherein the second antibody is a monoclonal antibody.

22. The method of claim 1, wherein the first antibody is covalently linked to the carboxy terminus of the second antibody.

23. The method of claim 1, wherein the brain disorder is Alzheimer'"'"'s disease, Parkinson'"'"'s disease, Huntington'"'"'s disease, bovine spongiform encephalopathy, West Nile virus encephalitis, Neuro-AIDS, brain injury, spinal cord injury, metastatic cancer of the brain, metastatic breast cancer of the brain, primary cancer of the brain, or multiple sclerosis.