Anti-ADDL antibodies and uses thereof
View Patent ↗The present invention relates to antibodies that differentially recognize multi-dimensional conformations of Aβ-derived diffusible ligands, also known as ADDLs. The antibodies of the invention can distinguish between Alzheimer's Disease and control human brain extracts and are useful in methods of detecting ADDLs and diagnosing Alzheimer's Disease. The present antibodies also block binding of ADDLs to neurons, assembly of ADDLS, and tau phosphorylation and are there useful in methods for the preventing and treating diseases associated with soluble oligomers of amyloid β 1-42.
1. An isolated antibody, or an antigen binding fragment of the antibody, that binds amyloid β-derived diffusible ligands comprising:
(a) a light chain variable region comprising,
(i) a CDR1 having the sequence Arg-Ser-Ser-Gln-Ser-Xaa 1 -Xaa 2 -His-Ser-Asn-Gly-Asn-Thr-Tyr-Leu-Xaa 3 (SEQ ID NO:52), wherein Xaa 1 and Xaa 2 are independently Ala, Val, Leu, Ile or P, and Xaa 3 is Asp, Glu, Arg, His or Lys,
(ii) a CDR2 having the sequence Lys-Xaa 1 -Ser-Asn-Arg-Phe-Xaa 2 (SEQ ID NO:57), wherein Xaa 1 is Ala, Val, Leu, Ile, or Pro, and Xaa 2 is Ser or Phe, and
(iii) a CDR3 having the sequence Xaa 1 -Gln-Xaa 2 -Xaa 3 -Xaa 4 -Val-Pro-Xaa 5 -Thr (SEQ ID NO:65), wherein Xaa 1 is Ser or Phe, Xaa 2 is Gly, Ser or Thr, Xaa 3 is Ser or Thr, Xaa 4 is His, Tyr or Leu, and Xaa 5 is Ala, Val, Leu, Ile, or Pro; or Xaa 1 -Gln-Xaa 2 -Thr-Arg-Val-Pro-Leu-Thr, wherein Xaa 1 is Phe or Leu, and Xaa 2 is Ala or Thr, and
(b) a heavy chain variable region comprising,
(i) a CDR1 having the sequence Thr-Ser-Gly-Met-Gly-Val-Xaa (SEQ ID NO:27), wherein Xaa is Ser, Gly or Ala,
(ii) a CDR2 having the sequence His-Ile-Xaa 1 -Trp-Asp-Asp-Asp-Lys-Xaa 2 -Tyr-Asn-Pro-Ser-Leu-Lys-Ser (SEQ ID NO:32), wherein Xaa 1 is Phe, Tyr or Trp and Xaa 2 is Ser, Arg, or Tyr, and
(iii) a CDR3 of SEQ ID NO:43.
2. The isolated antibody of claim 1 , further comprising a heavy chain constant region of SEQ ID NO:254.
3. The isolated antibody of claim 1 , wherein the antibody is a monoclonal antibody.
4. A pharmaceutical composition comprising the antibody or antigen binding fragment of claim 1 in admixture with a pharmaceutically acceptable carrier.
5. A kit for detecting Aβ-derived diffusible ligands comprising the antibody or antigen binding fragment of claim 1 .
6. A method for attenuating binding of Aβ-derived diffusible ligands to a neuron comprising contacting the neuron with the antibody or antigen binding fragment of claim 1 so that binding of Aβ-derived diffusible ligands to the neuron is attenuated.
7. A method for inhibiting assembly of Aβ-derived diffusible ligands comprising contacting a sample containing amyloid β 1-42 peptides with the antibody or antigen binding fragment of claim 1 thereby inhibiting assembly of Aβ-derived diffusible ligands.
8. A method for inhibiting the phosphorylation of tau protein at Ser202/Thr205 comprising contacting a sample containing a tau protein with the antibody or antigen binding fragment of claim 1 thereby inhibiting the phosphorylation of tau protein at Ser202/Thr205.
9. A method for attenuating the symptoms of a disease associated with Aβ-derived diffusible ligands comprising administering an effective amount of the pharmaceutical composition of claim 4 .
10. A method for identifying a putative therapeutic agent that attenuates the binding of Aβ-derived diffusible ligands to neurons comprising
(a) contacting a composition comprising a neuron with Aβ-derived diffusible ligands in the presence of an agent;
(b) contacting the composition with the antibody or antigen binding fragment of claim 1 ; and
(c) detecting the amount of antibody or antigen binding fragment bound in the presence of the agent,
wherein a decrease in the amount of antibody or antigen binding fragment bound in the presence of the agent as compared to the amount of antibody bound in the absence of the agent indicates that the agent is a putative therapeutic agent for attenuating binding of Aβ-derived diffusible ligands to neurons.
11. A method for detecting Aβ-derived diffusible ligands in a sample comprising contacting a sample with the antibody or antigen binding fragment of claim 1 and determining the presence of a complex comprising the Aβ-derived diffusible ligands and antibody or antigen binding fragment.
12. A method for diagnosing a disease associated with Aβ-derived diffusible ligands comprising contacting a sample the antibody or antigen binding fragment of claim 1 and determining the presence of a complex comprising the Aβ-derived diffusible ligands and antibody or antigen binding fragment wherein the complex is diagnostic of a disease associated with Aβ-derived diffusible ligands.