The present invention relates to antibodies that bind N3pGlu Amyloid Beta peptide and their use in treating diseases related to Amyloid Beta (herein referred to as Αβ or Abeta) peptide.

The cleavage of the amyloid precursor protein results in Αβ peptides ranging in size from 38 and 43 amino acids. Conversion of Αβ from soluble to insoluble forms having high β-sheet content and the deposition of these insoluble forms as neuritic and cerebrovascular plaques in the brain has been associated with a number of conditions and diseases, including Alzheimer's disease (AD), Down's syndrome, and cerebral amyloid angiopathy (CAA) .

The deposits found in plaques are comprised of a heterogeneous mixture of Αβ peptides. N3pGlu Αβ, also referred to as N3pE, pE3-42, or Αβp3-42, is a truncated form of Αβ peptide and is found only in plaques. N3pGlu Αβ lacks the first two amino acid residues at the N-terminus of human Αβ and has a pyroglutamate which was derived from the glutamic acid at the third amino acid position. Although N3pGlu Αβ peptide is a minor component of the deposited Αβ in the brain, studies have demonstrated that N3pGlu Αβ peptide has aggressive aggregation properties and accumulates early in the deposition cascade.

Antibodies to N3pGlu Αβ are known in the art. For example, U.S. Patent Number 8,679,498 discloses human N3pG3u Αβ antibodies ( e.g. B12L) and methods of treating diseases, such as Alzheimer's disease, with said antibodies. However, there still remains a need for N3pGlu Αβ antibodies with higher affinity, improved thermostability, reduced non-specific binding, and lower levels of immunogenicity. Such N3pGlu Αβ antibodies would provide an improved safety profile for a potential human therapeutic with pharmacokinetics for a better dosing schedule.

The antibodies within the scope of the present invention possess these desirable characteristics. The present invention provides an antibody that binds human N3pG3u Αβ, wherein said antibody comprises a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 9 or 10, and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 8. In a particular embodiment, the present

invention provides an antibody that binds human N3pGlu Αβ, wherein said antibody comprises a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 9, and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 8. In another particular embodiment, the present invention provides an antibody that binds human N3pG3u Αβ, wherein said antibody comprises a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 10, and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 8.

In an embodiment, the present invention provides an antibody that binds human N3pGlu Αβ, comprising a light chain (LC) and a heavy chain (HC), wherein the amino acid sequence of the LC is the amino acid sequence of SEQ ID NO: 12 or 13, and the amino acid sequence of the HC is the amino acid sequence of SEQ ID NO: 11 , In a more particular embodiment, the present invention provides an antibody that binds human N3pGlu Αβ, comprising a light chain (LC) and a heavy chain (HC), wherein the amino acid sequence of the LC is the amino acid sequence of SEQ ID NO: 12, and the amino acid sequence of the HC is the amino acid sequence of SEQ ID NO: 1 1. In another particular embodiment, the present invention provides an antibody that binds human N3pGlu Αβ, comprising a light chain (LC) and a heavy chain (HC), wherein the amino acid sequence of the LC is the amino acid sequence of SEQ ID NO: 13, and the amino acid sequence of the HC is the amino acid sequence of SEQ ID NO: 1 1. In a further embodiment, the present invention provides an antibody comprising two LC and two HC, wherein the amino acid sequence of each LC is SEQ ID NO: 12 or 13, and the amino acid sequence of each HC is SEQ ID NO:l 1. In a more particular embodiment, the present invention provides an antibody comprising two LC and two HC, wherein the amino acid sequence of each LC is SEQ ID NO: 12, and the amino acid sequence of each HC is SEQ ID NO: 11 , In a more particular embodiment, the present invention provides an antibody comprising two LC and two HC, wherein the amino acid sequence of each LC is SEQ ID NO: 13, and the amino acid sequence of each HC is SEQ ID NO:l 1.

The present invention also provides a N3pGlu Αβ antibody comprising an LCVR and an HCVR, wherein said LCVR comprises LCDR1, LCDR2, and LCDR3, and wherein said HCVR comprises HCDR1, HCDR2, and HCDR3, and wherein the amino acid sequences of said CDRs are SEQ ID NO:4 for LCDR1 , SEQ ID NO:6 for LCDR2,

SEQ ID NO:7 for LCDR3, SEQ ID NO: 1 for HCDR1, SEQ ID NO:2 for HCDR2, and SEQ ID NO:3 for HCDR3

The present invention further provides pharmaceutical compositions comprising an antibody of the present invention and one or more pharmaceutically acceptable carriers, diluents or excipients. Further, the present invention provides a method of treating a disease characterized by deposition of Αβ, comprising administering to a patient in need thereof a pharmaceutical composition of the present invention.

Particularly, the present invention provides a method of treating or preventing clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical CAA comprising administering to a patient in need thereof an effective amount of an antibody of the present invention. More particularly, the present invention provides a method of treating or preventing a condition selected from prodromal AD (sometimes also referred to as Αβ-related mild cognitive impairment, or MCI), mild AD, moderate AD and severe AD, comprising administering to a patient in need thereof a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of slowing cognitive decline in a patient diagnosed with pre-clinical Alzheimer's disease or clinical Alzheimer's disease, comprising administering a pharmaceutical composition of the present invention. More particularly, the present invention further provides a method of slowing cognitive decline in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD and severe AD, comprising administering a pharmaceutical composition of the present invention.

In another embodiment the presen t invention provides a method of slowing functional decline in a patient diagnosed with pre-clinical Alzheimer's disease or clinical Alzheimer's disease, comprising administering a pharmaceutical composition of the present invention. More particularly, the present invention provides a method of slowing functional decline in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD and severe AD, comprising administering a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of reducing brain

Αβ amyloid plaque load in a patient diagnosed with pre-clinical or clinical Alzheimer's disease, comprising administering a pharmaceutical composition of the present invention. More particularly, the present invention provides a method of reducing brain Αβ amyloid plaque load in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD, and severe AD, comprising administering a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of preventing memory loss or cognitive decline in asymptomatic patients with low levels of Αβ 1-42 in the cerebrospinal fluid (CSF) or Αβ plaques in the brain comprising administering to the said patient a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of treating asymptomatic patients known to have an Alzheimer's disease-causing genetic mutation, comprising administering to the said patient a pharmaceutical composition of the present invention. In a particular embodiment, the present in vention provides a method of treating asymptomatic patients known to have a PSEN1 E280A Alzheimer's disease-causing genetic mutation (Paisa mutation), comprising administering to the said patient a pharmaceutical composition of the present invention. In another particular embodiment, the present invention provides a method of treating asymptomatic patients with a genetic mutation that causes autosomal-dominant Alzheimer's disease, comprising administering to the said patient a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of preventing memory loss or cognitive decline in asymptomatic patients known to have an Alzheimer's disease-causing genetic mutation, comprising administering to the said patient a pharmaceutical composition of the present invention. In a particular embodiment, the present invention provides a method of preventing memory loss or cognitive decline in asymptomatic patients known to have a PSEN1 E280A Alzheimer's disease-causing genetic mutation (Paisa mutation), comprising administering to the said patient a pharmaceutical composition of the present invention. In another particular embodiment, the present invention provides a method of preventing memory loss or cognitive decline in asymptomatic patients with a genetic mutation that causes autosomal-dominant Alzheimer's disease, comprising administering to the said patient a pharmaceutical composition of the present invention.

In another embodiment the present invention provides a method of slowing cognitive decline in an asymptomatic patient known to have an Alzheimer's disease-causing genetic mutation, comprising administering to the said patient a pharmaceutical composition of the present invention. In a particular embodiment, the present invention provides a method of slowing cognitive decline in asymptomatic patients known to have a PSEN1. E280A Alzheimer's disease-causing genetic mutation (Paisa mutation), comprising administering to the said patient a pharmaceutical composition of the present invention. In another particular embodiment, the present invention provides a method of slowing cognitive decline in asymptomatic patients with a genetic mutation that causes autosomal -dominant Alzheimer's disease, comprising administering to the said patient a pharmaceutical composition of the present invention.

The present invention also provides an antibody of the present invention for use in therapy. In an embodiment, the present invention provides an antibody of the present invention for use in the treatment of a disease characterized by deposition of Αβ. In another embodiment, the present invention provides an antibody of the present invention for use in treatment of clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical cerebral amyloid angiopathy. In another embodiment, the present invention provides an antibody of the present invention for u se in treatment of a condition selected from prodromal AD, mild AD, moderate AD and severe AD. In another embodiment, the present invention provides an antibody of the present invention for use in slowing cognitive decline in a patient diagnosed with a condition selected from clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical cerebral amyloid angiopathy. In another embodiment, the present invention provides an antibody of the present invention for use in slowing cognitive decline in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD and severe AD.

Further, the present invention provides a pharmaceutical composition comprising an antibody of the present invention for use in therapy. In an embodiment, the present invention provides a pharmaceutical composition comprising an antibody for use in the treatment of a disease characterized by deposition of Αβ.

The present invention also provides the use of an antibody of the present invention in the manufacture of a medicament for the treatment of a disease characterized by deposition of Αβ. In an embodiment, the present invention provides the use of an antibody of the present invention in the manu facture of a medicament for the treatment of clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical cerebral amyloid angiopathy. In an embodiment, the present invention provides the use of an antibody of the present invention in the manufacture of a medicament for the treatment of prodromal AD, mild AD, moderate AD or severe AD. in another embodiment, the present invention pro vides the use of an antibody of the present mvention in the manufacture of a medicament for slowing cognitive decline in a patient diagnosed with a condition selected from clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical cerebral amyloid angiopathy. In another embodiment, the present invention provides the use of an antibody of the present invention in the manufacture of a medicament for slowing cognitive decline in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD and severe AD.

In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 8. In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:9. In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 10. In a further embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:8, and comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:9. In another embodiment, the present invention provides a DNA molecule comprising a

polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:8, and comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 10.

In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ

ID NO: 11 , In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 12. In an embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 13. In a further embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 , and comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 12, In another embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:l 1, and comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 13. In another embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 , and a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 12. In another embodiment, the present invention provides a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 , and a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 13. In a particular embodiment the polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 is SEQ ID NO: 14 and the polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 12 is SEQ ID NO: 15. In a particular embodiment the polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 1 1 is SEQ ID NO: 14 and the polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 13 is SEQ ID NO: 16.

Further, the present invention provides a mammalian cell comprising a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 and a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 12, Preferably the mammalian cell comprises a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 and a polypeptide having the amino acid sequence SEQ ID NO: 12. The present invention also provides a mammalian cell comprising a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 and a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 13. Preferably the mammalian cell comprises a DNA molecule comprising a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 11 and a polypeptide having the amino acid sequence of SEQ ID NO: 13. In an embodiment the mammalian cell line is a Chinese Hamster Ovary (CHO) or Hamster embryonic kidney (HEK) cell line.

WE CLAIM:

An antibody that binds human N3pGlu Αβ, comprising a light chain

variable region (LCVR) having the amino acid sequence of SEQ ID NO:9 or SEQ ID NO: 10, and a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 8.

2. The antibody of Claim 1, comprising a LCVR having the amino acid

sequence of SEQ ID NO: 9, and a HCVR having the amino acid sequence of SEQ ID NO:8.

3. The antibody of Claim 1, comprising a LCVR having the amino acid

sequence of SEQ ID NO: 10, and a HCVR having the amino acid sequence of SEQ ID NO:8.

4. The antibody of Claim 1 , comprising a light chain (LC) having the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 13 and a heavy chain (HC) having the amino acid sequence of SEQ ID NO: 1 1.

5. The antibody of Claim 1 , comprising a LC having the amino acid sequence of SEQ ID NO: 12 and a HC having the amino acid sequence of SEQ ID NO: 1 1.

6. The antibody of Claim 1, comprising a LC having the amino acid sequence of SEQ ID NO: 13 and a HC having the amino acid sequence of SEQ ID NO: 11.

7. The antibody of Claim 1 , comprising two LCs and two HCs, wherein the amino acid sequence of each LC is SEQ ID NO: 12 or SEQ ID NO: 13, and the amino acid sequence of each HC is SEQ ID NO: l l .

8. The antibody of Claim 1, comprising two LCs and two HCs, wherein the amino acid sequence of each LC is SEQ ID NO: 12, and the amino acid sequence of each HC is SEQ ID NO: l 1.

9. The antibody of Claim 1 , comprising two LCs and two HCs, wherein the amino acid sequence of each LC is SEQ ID NO: 13, and the amino acid sequence of each HC is SEQ ID NO: l 1.

10. A pharmaceutical composition comprising the antibody of any one of Claims 1 to 9 and one or more pharmaceutically acceptable carriers, diluents or excipients.

11. A method of treating a patient having a disease characterized by deposition of Αβ, comprising administering to said patient an effective amount of the antibody of any one of Claims 1 to 9.

12. A method of treating a patient having a condition selected from clinical or pre-clinicai Alzheimer's disease, Down's syndrome, and clinical or preclinical cerebral amyloid angiopathy, comprising administering to said patient an effective amount of an antibody of any one of Claims 1 to 9. 13. A method of treating a patient having a condition selected from prodromal AD, mild AD, moderate AD, and severe AD, comprising administering an effective amount of an antibody of any one of Claims 1 to 9.

14. A method of treating a patient having a condition selected from clinical or pre-clinicai Alzheimer's disease, Down's syndrome, and clinical or preclinical cerebral amyloid angiopathy, comprising administering to said patient an effective amount of a pharmaceutical composition of claim 10. 15. A method of treating a patient having a condition selected from prodromal AD, mild AD, moderate AD, and severe AD, comprising administering to said patient an effective amount of a pharmaceutical composition of claim 10.

16. A method of slowing cognitive decline in a patient diagnosed with a

condition selected from clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-cltnical cerebral amyloid angiopathy, comprising administering to said patient an effective amount of an antibody of any one of Claims 1 to 9.

17. A method of slowing cognitive decline in a patient diagnosed with a

condition selected from prodromal AD, mild AD, moderate AD and severe AD, comprising administering to said patient an effective amount of an antibody of any one of Claims 1 to 9.

18. A method of slowing cognitive decline in a patient diagnosed with a condition selected from clinical or pre-clinical Alzheimer's disease,

Down's syndrome, and clinical or pre-clinical cerebral amyloid angiopathy, comprising administering to said patient an effective amount of a pharmaceutical composition of claim 10.

19. A method of slowing cognitive decline in a patient diagnosed with a

condition selected from prodromal AD, mild AD, moderate AD and severe

AD, comprising administering to said patient an effective amount of a pharmaceutical composition of claim 10.

20. The antibody of any one of Claims 1 to 9 for use in therapy.

21. The antibody of any one of Claims 1 to 9 for use in the treatment of a disease characterized by deposition of Αβ.

22. The antibody of any one of Claims 1 to 9 for use in the treatment of a condition selected from clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or pre-clinical cerebral amyloid

23. The antibody of any one of Claims 1 to 9 for use in the treatment of a condition selected from prodromal AD, mild AD, moderate AD, and severe AD.

24. The antibody of any one of Claims 1 to 9 for use in slowing cognitive decline in a patient diagnosed with a condition selected from clinical or pre-clinical Alzheimer's disease, Down's syndrome, and clinical or preclinical cerebral amyloid angiopathy.

25. The antibody of any one of Claims 1 to 9 for use in slowing cognitive decline in a patient diagnosed with a condition selected from prodromal AD, mild AD, moderate AD and severe AD.

26. Use of the antibody of any one of Claims 1 to 9 in the manufacture of a medicament for the treatment of a disease characterized by deposition of Αβ.

27. Use of the antibody of any one of Claims 1 to 9 in the manufacture of a medicament for the treatment of a condition selected from clinical or pre- clinical Alzheimer's disease, Down's syndrome, and clinical or preclinical cerebral amyloid angiopathy.

28. Use of the antibody of any one of Claims 1 to 9 in the manufacture of a medicament for the treatment of a condition selected from prodromal AD, mild AD, moderate AD, and severe AD.