Monoclonal antibodies against pathological alpha-synuclein, and methods using same
The present disclosure provides monoclonal antibodies that bind α-Synuclein. In certain aspects, the antibodies preferentially bind to α-Synuclein fibrils over α-Synuclein monomer. In other aspects, the invention comprises a method of treating α-Synucleopathic disease in a subject, comprising administering any of the antibodies of the invention to the subject. In yet other aspects, the invention comprises methods of detecting α-Synuclein fibrils using any of the antibodies of the invention.
1. An isolated monoclonal antibody that preferentially binds pathological alpha-synuclein (α-Syn) over native α-Syn, the antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.
2. The monoclonal antibody of claim 1 , wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1 or 11;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2 or 12; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3 or 13, and wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.
3. The monoclonal antibody of claim 1 , wherein one of the following applies:
(a) the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 1;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 2; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 3; and
the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8;
(b) the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 11;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 12; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 13, and
the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8;
(c) the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 23, and
the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.
4. The monoclonal antibody of claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NOs: 4 or 14, and wherein the VH comprises the amino acid sequence of SEQ ID NO: 9.
5. The monoclonal antibody of claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NOs: 4, and wherein the VH comprises the amino acid sequence of SEQ ID NO: 9.
6. The monoclonal antibody of claim 1 , wherein
the VL comprises the amino acid sequence of SEQ ID NO: 14, and the VH comprises SEQ ID NO: 9.
7. The monoclonal antibody of claim 1 , which is humanized or labeled.
8. A pharmaceutical composition comprising the monoclonal antibody of claim 1 and at least one pharmaceutical excipient.
9. An isolated polynucleotide, comprising:
(a) the nucleic acid sequences of SEQ ID NOs: 5 and 10;
(b) the nucleic acid sequences of SEQ ID NOs: 15 and 20; or
(c) the nucleic acid sequences of SEQ ID NOs: 25 and 30.
10. An autonomously replicating or an integrative mammalian cell vector comprising a recombinant nucleic acid encoding an antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH),
wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23;
and wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.
11. The vector of claim 10 , which comprises at least one of the following: a plasmid, a virus, a mammalian cell expression vector, at least one nucleic acid sequence that directs or controls expression of the antibody.
12. An isolated host cell comprising the vector of claim 10 .
13. The cell of claim 12 , which is at least one of the following:
a non-human cell; a mammalian cell.
14. A method of treating a synucleopathic disease in a subject, the method comprising administering to the subject a therapeutically effective amount of an isolated monoclonal antibody that preferentially binds to pathological α-synuclein over native α-synuclein, comprising a light chain variable region (VL) and a heavy chain variable region (VH),
wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and
wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8.
15. The method of claim 14 , wherein the synucleopathic disease is at least one from the group consisting of Parkinson's disease, Parkinson's disease with dementia, dementia with Lewy bodies, Alzheimer's disease, Down's syndrome, multiple-system atrophy, prion diseases, and other α-Syn related neurodegenerative disorders.
16. The method of claim 14 , wherein at least one applies:
the antibody is provided to the subject as a pharmaceutical composition;
the antibody is administered parenterally to the subject.
17. A method of detecting a synucleopathic disease in a subject, the method comprising
administering to the subject a labeled isolated monoclonal antibody that preferentially binds to pathological α-synuclein, comprising a light chain variable region (VL) and a heavy chain variable region (VH),
wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and
wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8; and
detecting presence or absence of a complex of the labeled isolated monoclonal antibody with any α-Syn fibrils present in the subject,
wherein, if the complex is detected, the subject has a synucleopathic disease.
18. A method of detecting α-Syn fibrils in a sample, the method comprising
contacting the sample with a labeled isolated monoclonal antibody that preferentially binds to pathological α-synuclein over native α-synuclein, comprising a light chain variable region (VL) and a heavy chain variable region (VH),
wherein the VL comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NOs: 1, 11, or 21;
a CDR2 region comprising the amino acid sequence of SEQ ID NOs: 2, 12, or 22; and
a CDR3 region comprising the amino acid sequence of SEQ ID NOs: 3, 13, or 23, and
wherein the VH comprises
a CDR1 region comprising the amino acid sequence of SEQ ID NO: 6;
a CDR2 region comprising the amino acid sequence of SEQ ID NO: 7; and
a CDR3 region comprising the amino acid sequence of SEQ ID NO: 8; and
detecting presence or absence of a complex of the labeled isolated monoclonal antibody with any α-Syn fibrils present in the sample,
wherein, if the complex is detected, α-Syn fibrils are present in the sample.