Amyloid beta(1-42) oligomers, derivatives thereof and antibodies thereto, methods of preparation thereof and use thereof

• US 9,176,150 B2
• Filed: 08/01/2011
• Issued: 11/03/2015
• Est. Priority Date: 01/31/2003
• Status: Expired due to Term

• Current Assignee: Abbvie Deutschland GmbH & Company KG (Abbvie Incorporated)
• Original Assignee: Abbvie Deutschland GmbH & Company KG (Abbvie Incorporated)
• Inventors: Hillen, Heinz, Striebinger, Andreas, Krantz, Carsten, Moeller, Achim, Mueller, Reinhold
• Primary Examiner(s): BALLARD, KIMBERLY
• Application Number:
  US13/195,533
• Publication Number:
  US 20120034166A1
• Time in Patent Office: 1,555 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 39/0007 Nervous system antigens; Prions
• 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 37/02 Immunomodulators
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 16/005 constructed by phage libraries
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: G01N 2333/4709 Amyloid plaque core protein
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The invention relates to neuromodulatory oligomers of the amyloid-β(1-42) protein, a particular production method, by means of which the oligomer can be obtained in a reproducible manner at high yield, the use of the oligomers as diagnostic and therapeutics agents, for the generation of oligomer-specific antibodies and for the discovery of substances which can interact with the oligomers and in the formation thereof. Corresponding methods for the production of the antibodies and for discovery of the substances are also disclosed as are the antibodies themselves and the use of the antibodies or substances as diagnostic and therapeutic agents. The invention further relates to derivatives of the oligomers and oligomers based on abbreviated forms of the amyloid-β(1-42) proteins, the production and use thereof.

Claims

1. An isolated antibody which binds an N-terminal truncated amyloid-β

(“

Aβ

”

)(1-42) oligomer comprising an Aβ

(X-42) fragment, wherein X is 10 to 24, the isolated antibody obtainable by a method comprising;

a) immunizing a host with the N-terminal truncated Aβ

(1-42) oligomer; and

b) obtaining an antibody-containing host serum produced as a response to said immunization,

wherein the isolated antibody binds the N-terminal truncated Aβ

(1-42) oligomer with 10-fold greater affinity than an amyloid derived, diffusible and dementing ligand (ADDL),

wherein the N-terminal truncated Aβ

(1-42) oligomer is produced by a process comprising;

i) exposing a monomeric Aβ

(1-42) protein to a detergent to form an intermediate oligomer;

ii) reducing the detergent action and further incubating the intermediate oligomer to form an Aβ

(1-42) oligomer; and

iii) treating the Aβ

(1-42) oligomer with a protease to form the N-terminal truncated Aβ

(1-42) oligomer,

wherein the detergent is a compound of formula R—

X, wherein the radical R is unbranched or branched alkyl having from 6 to 20 carbon atoms or unbranched or branched alkenyl having from 6 to 20 carbon atoms, and the radical X is an acidic group or salt thereof.

2. The isolated antibody of claim 1, which binds the N-terminal truncated Aβ

(1-42) oligomer with an affinity greater than K_d=10<sup>−

8</sup> M.

3. The isolated antibody of claim 1, which binds monomeric Aβ

(1-42) protein and/or monomeric Aβ

(1-40) protein with an affinity of less than K_d=10<sup>−

8</sup> M.

4. The isolated antibody of claim 1, wherein the affinity of said antibody to the N-terminal truncated Aβ

(1-42) oligomer is at least 10 times greater than to monomeric Aβ

(1-42) protein and/or monomeric Aβ

(1-40) protein.

5. The isolated antibody of claim 1, which binds the N-terminal truncated Aβ

(1-42) oligomer with a rate constant k_off of 0.1 S<sup>−

1</sup> or lower, as determined by means of surface plasmon resonance.

6. A pharmaceutical agent, comprising the isolated antibody of claim 1 and, optionally, a pharmaceutically suitable carrier.

7. The isolated antibody of claim 1, wherein the fragment is selected from Aβ

(12-42), Aβ

(13-42), Aβ

(14-42), Aβ

(15-42), Aβ

(16-42), Aβ

(17-42), Aβ

(18-42), Aβ

(19-42), and Aβ

(20-42).

8. The isolated antibody of claim 1, wherein the fragment is an Aβ

(20-42) fragment.

9. The isolated antibody of claim 8, wherein the protease is thermolysin.

10. The isolated antibody of claim 1, wherein the Aβ

(1-42) oligomer has an apparent molecular weight of about 38 kDa and/or about 48 kDa on SDS gel electrophoresis.

11. The isolated antibody of claim 1, wherein the radical X is selected from among —

COO⁻

M⁺, —

SO³⁻

M⁺, and —

OSO³⁻

M⁺, wherein M⁺ is a hydrogen cation or an inorganic or organic cation selected from alkali metal and alkaline earth metal cations and ammonium cations.

12. The isolated antibody of claim 1, wherein the detergent is sodium dodecyl sulfate.