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KR-102964291-B1 - Amyloid beta epitopes and antibodies against them

KR102964291B1KR 102964291 B1KR102964291 B1KR 102964291B1KR-102964291-B1

Abstract

The present disclosure relates to an epitope identified in A-beta comprising a morphological epitope, an antibody against it, and an immunogen and antibody specific thereto, and a method for manufacturing and using the antibody.

Inventors

  • 캐쉬맨, 닐 알.
  • 플로트킨, 스티븐 에스.

Assignees

  • 더 유니버시티 오브 브리티쉬 콜롬비아

Dates

Publication Date
20260513
Application Date
20161109
Priority Date
20151109

Claims (20)

  1. A cyclic compound composed of an amino acid sequence, an A-beta peptide consisting of QKLV (sequence identification number: 1), and a linker consisting of the amino acid sequence GCG, A cyclic compound in which the above linker is covalently coupled to the above A-beta peptide N-terminal residue and the above A-beta peptide C-terminal residue.
  2. A cyclic compound according to claim 1, wherein the cyclic compound further comprises a detectable label.
  3. Immunogen comprising the cyclic compound of claim 1.
  4. Immunogen according to claim 3, wherein the cyclic compound is coupled to a carrier protein.
  5. Immunogen according to claim 4, wherein the carrier protein is bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH).
  6. A composition for the treatment of Alzheimer's disease and/or other A-beta amyloid-related diseases, comprising a cyclic compound according to claim 1 or an immunogen according to any one of claims 3 to 5, wherein the other A-beta amyloid-related diseases are selected from the group consisting of Lewy body dementia, inclusion body myositis, and cerebral amyloid angiopathy.
  7. A composition according to claim 6, further comprising an adjuvant.
  8. A composition according to claim 7, wherein the adjuvant is aluminum phosphate, aluminum hydroxide, QS21, polyphosphazene, or CpG oligonucleotide.
  9. An isolated antibody that specifically binds to an A-beta peptide composed of QKLV (sequence identification number: 1), wherein the antibody comprises a light chain variable region and a heavy chain variable region, wherein the heavy chain variable region comprises complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, and the light chain variable region comprises complementarity determining regions CDR-L1, CDR-L2, and CDR-L3, and the amino acid sequence of the CDR is the following sequence: CDR-H1 GYTFTDYE (Sequence identification number: 11) CDR-H2 IDPETGDT (Sequence identification number: 12) CDR-H3 TSPIYYDYDWFAY(Sequence identification number: 13) CDR-L1 QSLLNNRTRKNY(Sequence identification number: 14) CDR-L2 WAS (Sequence identification number: 15) CDR-L3 KQSYNLRT(sequence identification number: 16); or CDR-H1 GFSLSTSGMG (Sequence identification number: 21) CDR-H2 IWWDDDK (Sequence identification number: 22) CDR-H3 ARSITTVVATPFDY (Sequence identification number: 23) CDR-L1 QNVRSA (Sequence identification number: 24) CDR-L2 LAS (Sequence identification number: 25) CDR-L3 LQHWNSPFT (Sequence identification number: 26).
  10. An isolated antibody according to claim 9, wherein the antibody is a conformation-specific and/or selective antibody that specifically or selectively binds to QKLV present in a cyclic compound.
  11. An isolated antibody according to claim 10, wherein the antibody selectively binds to an A-beta oligomer rather than an A-beta monomer and/or A-beta fibril.
  12. An isolated antibody according to claim 11, wherein the selectivity is at least 3 times, at least 5 times, at least 10 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 100 times, at least 500 times, or at least 1000 times more selective for A-beta oligomers than for A-beta monomers and/or A-beta fibrils.
  13. An isolated antibody according to any one of claims 9 to 12, wherein the antibody does not specifically and/or selectively bind to a linear A-beta peptide comprising sequence QKLV (sequence identification number: 1).
  14. In any one of claims 9 to 12, the antibody is an antibody that lacks binding to an A-beta monomer and/or an A-beta fibril plaque within the same system, or has a minute amount of said binding.
  15. An isolated antibody according to any one of claims 9 to 12, wherein the antibody is a monoclonal antibody or a polyclonal antibody.
  16. An isolated antibody according to any one of claims 9 to 12, wherein the antibody is a humanized antibody.
  17. An isolated antibody according to any one of claims 9 to 12, wherein the antibody is an antibody-binding fragment selected from Fab, Fab', F(ab')2, scFv, dsFv, ds-scFv, dimers, minibodies, diabodies, and multimers thereof.
  18. An isolated antibody according to any one of claims 9 to 12, wherein the antibody comprises a heavy chain variable region comprising the following: i) Sequence identification number: Amino acid sequence presented in 18 or 28; ii) an amino acid sequence having at least 80% or at least 90% sequence identity with sequence identification number: 18 or 28, wherein the CDR sequence is the amino acid sequence presented in sequence identification numbers: 11, 12, 13, 21, 22 and 23.
  19. An antibody according to any one of claims 9 to 12, wherein the isolated antibody comprises a light chain variable region comprising the following: i) Sequence identification number: Amino acid sequence presented in 20 or 30; ii) an amino acid sequence having at least 80% or at least 90% sequence identity with sequence identification number: 20 or 30, wherein the CDR sequence is the amino acid sequence presented in sequence identification numbers: 14, 15, 16, 24, 25 and 26.
  20. An isolated antibody according to any one of claims 9 to 12, wherein the heavy chain variable region amino acid sequence is encoded by the nucleotide sequence presented in sequence identification number: 17 or 27 and/or; the antibody comprises a light chain variable region amino acid sequence encoded by the nucleotide sequence presented in sequence identification number: 19 or 29.

Description

Amyloid beta epitopes and antibodies against them Related applications This is a PCT application claiming the advantage of priority of U.S. patent application serial number 62/253044, filed November 9, 2015; U.S. patent application serial number 62/289,893, filed Monday, February 1, 2016; U.S. patent application serial number 62/365,634, filed July 22, 2016; and U.S. patent application serial number 62/393,615, filed September 12, 2016, each of which is incorporated herein by reference. field The present disclosure relates to amyloid beta (A-beta or Aβ) epitopes and antibodies against them, and more specifically to morphological A-beta epitopes predicted to be selectively accessible within A-beta oligomers, and related antibody compositions and uses thereof. Amyloid-beta (A-beta), existing as a 36–43 amino acid peptide, is a product released from amyloid precursor proteins (APP) by the enzymes β and γ secretases. In patients with Alzheimer's disease (AD), A-beta can exist as soluble monomers, insoluble fibrils, and soluble oligomers. In monomeric form, A-beta usually exists as unstructured polypeptide chains. In fibrilic form, A-beta can aggregate into distinct morphologies, often referred to as strains. Several of these forms have been identified by solid-state NMR. For example, several strain forms of fibrils are available in the Protein Data Bank (PDB), a crystallographic database of atomic-resolution three-dimensional morphological data, including the 3-fold symmetric Aβ form (PDB entry, 2M4J); the 2-fold symmetric form of the Aβ-40 monomer (PDB entry 2LMN); and the single-chain, parallel register-intraform form of the Aβ-42 monomer (PDB entry 2MXU). The form of 2M4J is J. CELL Vol. Reported in 154 p.1257 (2013), the form of 2MXU is Y. ALZHEIMER'S DISEASE. NAT.STRUCT.MOL.BIOL. Vol. It is reported in 22 p.499 (2015). It has been revealed that A-beta oligomers block key synaptic activity supporting memory, referred to as long-term potentiation (LTP), in brain slice cultures and living animals, and cause cell lines and neurons to die in the culture. The morphology of the oligomers has not been measured to date. Furthermore, NMR and other evidence indicate that oligomers do not exist in a single distinct form, but rather as a morphologically plastic, malleable morphological ensemble with limited regularity. Additionally, the concentration of toxic oligomer species is significantly lower than that of either the monomer or the fibril (estimates vary, but are approximately 1,000-fold higher or lower), making it difficult to achieve the target. U.S. Patents No. 5,766,846; No. 5,837,672; and No. 5,593,846 (which is incorporated herein by reference) describe the production of mouse monoclonal antibodies against the central domain of an Aβ peptide. No. WO 01/62801 describes an antibody that binds to A-beta between amino acids 13 to 28. No. WO2004071408 discloses a humanized antibody. WO2008088983A1 describes an antibody fragment that binds to an amyloid beta (A-beta) peptide and is covalently attached to one or more polyethylene glycol (PEG) molecules, and said antibody fragment specifically binds to the human A-beta peptide between amino acids at positions 13 to 28. Solanezumab and crenezumab bind to amino acids 16 to 26 in A-beta. Crenezumab interacts with monomers, oligomers, and fibrils. Midregion antibodies, including solanezumab (picomolar affinity) and crenezumab (nanomolar affinity), are believed to preferentially bind to monomeric A-beta[1]. An antibody that binds preferentially or selectively to an A-beta oligomer is preferred. Summary In A-beta consisting of residue QKLV (sequence identification number: 1) or a part thereof, an epitope, optionally a morphological epitope, and an antibody against it are described herein. The epitope is identified as being selectively exposed to an oligomeric species of A-beta in a form that distinguishes it from A-beta within the monomer. One aspect comprises an A-beta peptide comprising QKL and 8, 7, or 6 or fewer A-beta residues and a linker, wherein the linker comprises a compound, preferably a cyclic compound, that is covalently coupled to the A-beta peptide N-terminal residue and the A-beta C-terminal residue. In one embodiment, the A-beta peptide is selected from any one of the sequences of sequence identification numbers: 1-10, and optionally selected from: QKLV (sequence identification number: 1), HQKLV (sequence identification number: 2), HQKLVF (sequence identification number: 9) and QKLVF (sequence identification number: 10). In another embodiment, the cyclic compound is a cyclic peptide. In another embodiment, the cyclic compound comprises at least Q and/or ii) a form for Q, and/or K, and/or L, measured by an entropy limited to at least 10%, at least 20%, at least 25%, at least 30%, at least 35%, and at least 40% compared to the corresponding straight-chain compound. In another embodiment, the A-beta peptide is as follows: QKLV (sequence identification number