Genetically encoded multifunctional compositions bidirectionally transported between peripheral blood and the cns

• US 8,759,297 B2
• Filed: 08/20/2007
• Issued: 06/24/2014
• 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): Stucker, Jeffrey
• Assistant Examiner(s): Fontainhas, Aurora M
• Application Number:
  US11/841,541
• Publication Number:
  US 20080152645A1
• Time in Patent Office: 2,500 Days
• Field of Search: None
• US Class Current: 514/17.7
• 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

Provided herein are compositions for increasing transport of agents across the blood-brain barrier, in some embodiments in both directions, while allowing their activity once across the barrier to remain substantially intact. The agents are transported across the blood-brain barrier via one or more endogenous receptor-mediated transport systems. In some embodiments the agents are therapeutic, diagnostic, or research agent. Also provided herein are nucleic acids encoding proteins contained in the compositions.

Claims

1. A nucleic acid encoding a fusion antibody that:

(i) comprises a first antibody and a second antibody, wherein the first antibody is an ScFv that is covalently linked to the carboxy terminus of either the heavy or light chain of the second antibody;

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

(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 a brain disorder, wherein the ScFv retains an average of at least about 20% of its activity compared to its activity as a separate entity.

2. The nucleic acid of claim 1, wherein the second antibody comprises a CH₂ region, a CH₃ region, and a VH region.

3. The nucleic acid of claim 2, wherein the ScFv is covalently linked to the carboxyl terminus of the CH₃ region.

4. The nucleic acid of claim 2, wherein the VH region is a VH region derived from an antibody that binds to an endogenous receptor of the BBB.

5. The nucleic acid of claim 4, wherein the VH region is derived from an antibody that is generated in vivo or by a recombinant method.

6. The nucleic acid of claim 5, wherein the recombinant method comprises selection of libraries of recombinant antibodies in phage or vectors.

7. The nucleic acid of claim 1, wherein the ScFv is generated in vivo or by a recombinant method.

8. The nucleic acid of claim 7, wherein the recombinant method comprises selection of libraries of recombinant antibodies in phage or vectors.

9. The nucleic acid of claim 1, wherein the ScFv interacts with a pathological substance associated with a brain disorder.

10. The nucleic acid of claim 1, wherein the second antibody interacts with a pathological substance associated with a brain disorder.

11. The nucleic acid of claim 1, wherein the endogenous BBB receptor-mediated transport system is selected from the group consisting of an insulin receptor, transferrin receptor, leptin receptor, lipoprotein receptor, and an insulin-like growth factor (IGF) receptor.

12. The nucleic acid of claim 11, wherein the endogenous BBB receptor-mediated transport system is the insulin receptor.

13. The nucleic acid of claim 1, wherein the brain disorder is a disorder selected from the group consisting of 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, and multiple sclerosis.

14. The nucleic acid of claim 1, wherein the pathological substance is a protein.

15. The nucleic acid of claim 14, wherein the protein is selected from the group consisting of 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), and oligodendrocyte surface antigen.

16. The nucleic acid of claim 14, wherein the protein is Aβ

amyloid.

17. The nucleic acid of claim 1, comprising the nucleotide sequence of SEQ ID NO:

11.

18. The nucleic acid of claim 1, comprising the nucleotide sequence of SEQ ID NO:

13.

19. The nucleic acid of claim 1, comprising the nucleotide sequence of SEQ ID NO:

15 or SEQ ID NO;

17.

20. The nucleic acid of claim 1, comprising the nucleotide sequence of SEQ ID NO:

19, SEQ ID NO;

21, SEQ ID NO;

23, or SEQ ID NO;

25.

21. The nucleic acid of claim 1, comprising the nucleotide sequence of SEQ ID NO:

27.

22. A tandem expression vector comprising (i) a first promoter sequence operably linked to an open reading frame corresponding to the nucleic acid sequence of claim 2;

and (ii) a second promoter operably linked to a second open reading frame encoding an immunoglobulin light chain amino acid sequence comprising a CL region and a VL region.

23. A cell comprising the tandem expression vector of claim 22.