Alzheimer abeta peptide binding polypeptide

Abstract

The present disclosure relates to a class of engineered polypeptides having a binding affinity for amyloid β (Aβ) peptides (in the following referred to as Aβ), comprising the amino acid sequence EX 2 X 3 YX 5 X 6 NLX 9 AX 11 QLCAX 16 IX 18 X 19 X 20 ED. The present disclosure also relates to the use of such Aβ peptide binding polypeptides as therapeutic, prognostic and/or diagnostic agents.

1 . An Aβ peptide binding polypeptide, which comprises
 a first moiety comprising a first Aβ peptide binding motif BM1, 
 a second moiety comprising a second Aβ peptide binding motif BM2, 
 which motifs may be the same or different, and 
 a linker,
 wherein each one of said binding motifs BM1 and BM2 consists of an amino acid sequence selected from 
 i) EX 2 X 3 YX 5 X 6 NLX 9 A X 11 QLCAX 16 IX 18 X 19 X 20  ED 
 
 wherein, independently from each other,
 X 2  is selected from I, M, Q, R, T and Y; 
 X 3  is selected from H and V; 
 X 5  is selected from F, I and L; 
 X 6  is selected from P and T; 
 X 9  is selected from N and T; 
 X 11  is selected from D and H; 
 X 16  is selected from F and I; 
 X 18  is selected from N, Q and R; 
 X 19  is selected from K and S; and 
 X 20  is selected from F, I, L and R; 
 and 
 ii) an amino acid sequence which has at least 95% identity to the sequence defined in i). 
 
 
     
     
         2 . The Aβ peptide binding polypeptide according to  claim 1 , wherein at least one of first and second moieties comprises a binding module amino acid sequence, Bmod, selected from
 iii) [BM]-DXaSQXbXcXdLLXe EAKKLXfXgXhQA PXi 
 wherein BM is BM1 or BM2 as defined in  claim 1 , and, independently of each other, 
 X a  is selected from P, Q, R and S; 
 X b  is selected from N, Q, R and S; 
 X c  is selected from A and S; 
 X d  is selected from K, N and E; 
 X e  is selected from A, S and C; 
 X f  is selected from E, N and S; 
 X g  is selected from D, E and S; 
 X h  is selected from A and S; and 
 X i  is selected from no amino acid, A and K, 
 and 
 iv) an amino acid sequence which has at least 97% identity to a sequence defined in iii). 
 
     
     
         3 . The Aβ peptide binding polypeptide according to  claim 2 , wherein sequence iii) consists of the amino acid sequence
 [BM]-DXaSQXbAXdLLA EAKKLNDAQA PXi 
 wherein BM is BM1 or BM2 as defined in  claim 1 , and, independently of each other, 
 X a  is selected from P, Q, R and S; 
 X b  is selected from N, Q, R and S; 
 X d  is selected from K and N; and 
 X i  is selected from no amino acid, A and K. 
 
     
     
         4 . The Aβ peptide binding polypeptide according to  claim 1 , wherein
 sequence i) of BM1 corresponds to amino acid residues 4-25 in any one of SEQ ID NO:1-106 and 118-553, and 
 sequence i) of BM2 corresponds to amino acid residues 64-85 in any one of SEQ ID NO:1-17, 19-106 and 118-553 or to amino acid residues 69-90 in SEQ ID NO:18. 
 
     
     
         5 . The Aβ peptide binding polypeptide according to  claim 4 , wherein
 sequence i) of BM1 corresponds to amino acid residues 4-25 in SEQ ID NO:95 and 
 sequence i) of BM2 corresponds to amino acid residues 64-85 in SEQ ID NO:95. 
 
     
     
         6 . The Aβ peptide binding polypeptide according to  claim 2 , wherein
 sequence iii) in Bmod of the first moiety corresponds to amino acid residues 4-47 in any one of SEQ ID NO:1-106 and 118-553, and 
 sequence iii) in Bmod of the second moiety corresponds to amino acid residues 64-107 in any one of SEQ ID NO:1-17, 19-106 and 118-553 or amino acid residues 69-112 in SEQ ID NO:18. 
 
     
     
         7 . The Aβ peptide binding polypeptide according to  claim 6 , wherein
 sequence iii) in Bmod of the first moiety corresponds to amino acid residues 4-47 in SEQ ID NO:95, and 
 sequence iii) in Bmod of the second moiety corresponds to amino acid residues 64-107 in SEQ ID NO:95. 
 
     
     
         8 . An Aβ peptide binding polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-106 and 118-553. 
     
     
         9 . The Aβ peptide binding polypeptide according to  claim 1 , which is capable of binding to Aβ peptide such that the KD value of the interaction is at most 1×10 −7  M. 
     
     
         10 . A Fusion protein or conjugate, comprising
 a first part consisting of an Aβ peptide binding polypeptide according to  claim 1 ; and   a second part consisting of a polypeptide having a desired biological activity.   
     
     
         11 . The Fusion protein or conjugate according to  claim 10 , wherein said desired biological activity is a binding activity. 
     
     
         12 . The Fusion protein or conjugate according to  claim 10 , wherein the second part is selected from the group consisting of antibodies and antigen binding fragments thereof. 
     
     
         13 . A Composition comprising an Aβ peptide binding polypeptide, fusion protein or conjugate according to  claim 1  and at least one pharmaceutically acceptable excipient or carrier. 
     
     
         14 .- 15 . (canceled) 
     
     
         16 . The Aβ binding polypeptide of  claim 8 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, 5, 9, 13, 14, 16, 18, 20, 21, 25, 26, 28, 33, 35, 37, 40, 42, 48, 49, 50, 51, 53, 54, 55, 59, 60, 61, 62, 64, 65, 70, 72, 75, 78, 84, 89, 90, 95, 96, 97, 100, 104 and 118-159. 
     
     
         17 . The Aβ binding polypeptide of  claim 8 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO:18, 28, 35, 49, 50, 51, 55, 59, 64, 65, 70, 72, 78, 84, 95 and 118-159. 
     
     
         18 . The Aβ binding polypeptide of  claim 8 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO 18, 70 and 95. 
     
     
         19 . The Aβ binding polypeptide of  claim 8 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO:95. 
     
     
         20 . The fusion protein or conjugate according to  claim 11 , wherein said binding activity is binding to a target selected from the group consisting of Tau, apolipoprotein E (ApoE), beta-secretase 1 (BACE1), gamma-secretase, transferrin receptor (TfR), complement factor C1s, presenilin 1, presenilin 2, nicastrin, alpha-synuclein and Bri. 
     
     
         21 . The composition of  claim 13 , further comprising at least one active agent, said active agent selected from the group consisting of inhibitors of neurotransmitter degradation, neurotransmitters, acetylcholinesterase inhibitors, NMDA receptor antagonists, TNF inhibitors, antihistamines, anti-viral agents, alpha-secretase activators, inhibitors of beta- or gamma-secretase, inhibitors of α-synuclein aggregation, inhibitors of tau aggregation, calcium channel blockers, compounds effective against oxidative stress, anti-apoptotic compounds, metal chelators, pirenzepin and metabolites, attractants for Aβ clearing/depleting cellular components, inhibitors of N-terminal truncated Aβ including pyroglutamated Aβ3-42, anti-inflammatory molecules, clozapine, ziprasidone, risperidone, aripiprazole, olanzapine, tacrine, rivastigmine, donepezil, galantamine, amyloid or tau modifying drugs, vitamin B12, cysteine, acetylcholine precursor, lecithin, choline,  Ginkgo biloba , acetyl-L-carnitine, idebenone, propentofylline, xanthine derivatives, and combinations thereof. 
     
     
         22 . A method for the treatment of an Aβ peptide associated condition, comprising administering to a subject in need thereof, an effective amount of an Aβ peptide binding polypeptide according to  claim 1 , a fusion protein according to  claim 10 , or a composition according to  claim 13 . 
     
     
         23 . The method of  claim 22 , wherein the Aβ associated condition is selected from the group consisting of amyloidosis, secondary amyloidosis, age-related amyloidosis, Alzheimer's disease (AD), mild cognitive impairment (MCI), Lewybody dementia, Down's syndrome, hereditary cerebral hemorrhage with amyloidosis (Dutch type), the Guam Parkinson-dementia complex; cerebral amyloid angiopathy, primary and secondary systemic amyloidosis, familial amyloid polyneuropathy 1, progressive supranuclear palsy, multiple sclerosis, Creutzfeld Jacob disease, Parkinson's disease, HIV-related dementia, ALS (amyotropic lateral sclerosis), inclusion-body myositis (IBM), type II diabetes, senile cardiac amyloidosis, glaucoma, macular degeneration, drusen-related optic neuropathy, and cataract due to beta-amyloid deposition.