US20150307552A1 - Novel d-enantiomeric peptides derived from d3 and use thereof - Google Patents

Novel d-enantiomeric peptides derived from d3 and use thereof Download PDF

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US20150307552A1
US20150307552A1 US14/426,797 US201314426797A US2015307552A1 US 20150307552 A1 US20150307552 A1 US 20150307552A1 US 201314426797 A US201314426797 A US 201314426797A US 2015307552 A1 US2015307552 A1 US 2015307552A1
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peptide
seq
beta
peptides
oligomers
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Dieter Willbold
Susanne Aileen Funke
Oleksander Brener
Luitgard Nagel-Steger
Dirk Bartnik
Antonia Nicole Klein
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Forschungszentrum Juelich GmbH
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4709—Amyloid plaque core protein
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2410/00—Assays, e.g. immunoassays or enzyme assays, involving peptides of less than 20 animo acids

Definitions

  • the present invention relates to novel D-enantiomeric A-beta-oligomer-binding peptides derived from D3 and use thereof, particularly in medicine.
  • AD Alzheimer's disease
  • latin Merbus Alzheimer
  • a feature of Alzheimer's disease are extracellular deposits of amyloid beta peptide (A-beta peptide). This deposition of A-beta peptide in plaques is typically determined post mortem in the brains of AD patients. Therefore, various forms of A-beta peptide—such as fibrils—are blamed for the emergence and progression of the disease. In addition, for some years the small, freely diffusible A-beta oligomers have been recognized as the major cause of the emergence and progress of AD.
  • A-beta monomers as building blocks of A-beta oligomers, form continuously in the human body and are probably not toxic per se. They may even have a neuroprotective function.
  • A-beta monomers can be randomly stored together depending on their concentration. The concentration is dependent on their rate of formation and rate of degradation in the body. An increase in the concentration of A-beta monomers occurs in the body with increasing age, such that spontaneous aggregation of the monomers to form A-beta oligomers is more likely. The A-beta oligomers thus formed could proliferate in analogy to prions and eventually lead to Alzheimer's disease.
  • AD Alzheimer's disease
  • A-beta oligomers from numerous monomers is a higher order reaction and is therefore dependent to a high degree on the A-beta monomer concentration. Therefore, a small reduction in the active A-beta monomer concentration already leads to prevention of the formation of the first A-beta oligomers.
  • the preventive therapeutic concepts and substances currently in development are based on this mechanism.
  • A-beta oligomers or possibly also even larger polymers or fibrils are present which proliferate by prion-like mechanisms.
  • this proliferation is a lower order reaction and is therefore scarcely dependent on the A-beta monomer concentration.
  • the substances known from the prior art reduce the concentration of A-beta monomers and/or oligomers in a variety of ways. For instance, gamma secretase modulators are known, which have been used for prevention in animal experiments.
  • Hybrid compounds consisting of aminopyrazoles and peptides which prevent A-beta oligomerization, are known from WO 2011/147797.
  • AD is mainly diagnosed by neuropsychological tests by examining people in whom symptoms of dementia have already occurred. It is known, however, that A-beta oligomers and the fibrils and plaques formed therefrom over time in the course of the disease occur up to 20 years before the onset of symptoms in the brain of patients, and may have already caused irreversible damage. However, in practice there is currently no possibility of diagnosing AD before the outbreak of symptoms.
  • novel compounds active ingredients which bind very specifically and with high affinity to A-beta oligomers, and thus prevent their proliferation.
  • Said compounds should have no undesirable side effects and in particular not provoke any immune response.
  • the compounds should also recognize toxic A-beta oligomers and therefore also the small, freely diffusible oligomers at low concentrations, and completely destroy and/or prevent the (prion-like) proliferation thereof.
  • a further object of the present invention was to provide substances which not only focus on extracellular A-beta peptides, like most compounds known from the prior art, but specifically bind soluble A-beta oligomers. Furthermore, the novel compounds should inhibit or prevent formation of fibrils of A-beta peptides.
  • D-enantiomeric D-peptide D3 SEQ ID NO: 11
  • the properties include, inter alia, binding affinity and specificity for A-beta species, inhibition of A-beta fibril formation, inhibition of A-beta cytotoxicity, precipitation of A-beta oligomers and conversion of A-beta fibrils to non-toxic, non-amyloidogenic species.
  • a peptide comprising an amino acid sequence according to SEQ ID NO: 1 (D3-Delta-HTH), SEQ ID NO: 2 (RD2), SEQ ID NO: 3 (RD 1), SEQ ID NO: 4 (RD3), SEQ ID NO: 5 (DB3), SEQ ID NO: 6 (NT-D3), SEQ ID NO: 7 (DB1), SEQ ID NO: 8 (DB2), SEQ ID NO: 9 (DB4) and/or SEQ ID NO. 10 (DB5) and/or homologs, fragments and parts thereof.
  • SEQ ID NO: 1 D3-Delta-HTH
  • SEQ ID NO: 2 RD2
  • SEQ ID NO: 3 RD 1
  • SEQ ID NO: 4 RD3
  • SEQ ID NO: 5 DB3
  • SEQ ID NO: 6 N-D3
  • SEQ ID NO: 7 DB1
  • SEQ ID NO: 8 DB2
  • SEQ ID NO: 9 DB4
  • SEQ ID NO. 10 DB5 and/or homologs thereof.
  • Fragments and parts show a similar or identical effect to the peptides according to the invention.
  • the peptides according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and/or SEQ ID NO. 10 and homologs thereof consist substantially and preferably of at least 60%, 75%, 80%, particularly preferably 85%, 90%, 95%, in particular 96%, 97%, 98%, 99%, 100% of D-amino acids.
  • a polymer in the context of the invention is formed from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more monomers selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and/or SEQ ID NO. 10 and homologs thereof, which for already A-beta oligomer.
  • the polymers according to the invention are composed of identical monomers or a combination of 2, 3, 4, 5, 6, 7, 8, 9 or 10 different monomers, different to the abovementioned monomers, as so-called combination polymers.
  • the monomers may also be, in part, identical.
  • the number of identical monomers in the combination polymers is arbitrary.
  • the combination polymers comprise at least one monomer selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO. 10 and/or SEQ ID NO: 11 and homologs thereof and also parts or fragments thereof and at least one monomer selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO. 10 and/or SEQ ID NO: 11 and homologs thereof and also parts or fragments thereof and at least one
  • A-beta-oligomer-binding peptide which differs from the monomers selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO. 10 and/or SEQ ID NO: 11 and homologs thereof and also parts or fragments thereof, preferably of substantially D-enantiomers, as a further monomer.
  • Polymers may be prepared, for example, via chemical synthesis or peptide synthesis.
  • the monomers are covalently linked to one another. In a further embodiment, the monomers are non-covalently bound to one another.
  • a covalent binding or linking of the monomer units is present in the context of the invention if the peptides are linearly linked head-to-head, tail-to-tail or head-to-head with one another without intervening linker or linker groups being used.
  • a non-covalent linking is present in the context of the invention if the monomers are linked to one another via biotin and streptavidin, particularly streptavidin tetramer.
  • the monomers may be linked together linearly, in particular as described above.
  • the monomers are linked together branched to give the polymer according to the invention.
  • a branched polymer according to the invention can be a dendrimer, in which the monomers are linked together covalently or non-covalently.
  • the monomers may also be linked to a platform molecule (such as PEG or sugar) and thus form a branched polymer.
  • a platform molecule such as PEG or sugar
  • Monomers and polymers are hereinafter referred to as peptides according to the invention.
  • One variant of the invention is a peptide having the amino acid sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and/or SEQ ID NO. 10 and/or homologs thereof with an identity of 50%.
  • homologous sequences or “homologs” signifies that an amino acid sequence has an identity with any of the abovementioned amino acid sequences of the monomers of at least 50, 55, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%.
  • identity the terms “homologous” or “homology” are used synonymously in the present description.
  • the identity between two nucleic acid sequences or polypeptide sequences is calculated by comparison with the BESTFIT program based on the algorithm of Smith, T. F. and Waterman, M. S (Adv. Appl. Math. 2: 482-489 (1981)) setting the following parameters for amino acids: Gap creation penalty: 8 and Gap extension penalty: 2; and the following parameters for nucleic acids: Gap creation penalty: 50 and Gap extension penalty: 3.
  • the identity between two nucleic acid sequences or polypeptide sequences is defined by the identity of the nucleic acid sequence/polypeptide sequence respectively over the entire sequence length, which is calculated by comparison using the GAP program based on the algorithm of Needleman, S. B. and Wunsch, C. D. (J. Mol. Biol. 48: 443-453) setting the following parameters for amino acids: Gap creation penalty: 8 and Gap extension penalty: 2; and the following parameters for nucleic acids: Gap creation penalty: 50 and Gap extension penalty: 3.
  • two amino acid sequences are identical if they have the same amino acid sequence.
  • homologs are understood to mean the corresponding retro-inverso sequences of the monomers mentioned above.
  • the peptides according to the invention bind to parts of the amyloid beta peptide.
  • the peptides according to the invention have sequences which differ from the specified sequences by up to three amino acids.
  • sequences are also used as peptides which comprise the sequences mentioned above.
  • the peptides have fragments of the abovementioned sequences or have sequences homologous to the abovementioned sequences.
  • the invention relates to a peptide for use in medicine, preferably for the treatment of Alzheimer's disease.
  • the peptide is largely composed of D-amino acids.
  • the term “largely composed of D-amino acids” signifies that the monomers to be used according to the invention are made up of at least 50%, 60%, preferably 75%, 80%, particularly preferably 85%, 90%, 95%, in particular 96%, 97%, 98%, 99% or 100% of D-amino acids.
  • the peptides according to the invention are derivatives of the D-enantiomeric D-peptide D3.
  • Derivatives in the context of the invention are peptide sequences derived from D3 which are achieved by one of the following three methods:
  • the invention relates to a peptide for the inhibition of formation of fibrils of amyloid beta peptides.
  • the polymers according to the invention detoxify the A-beta oligomers or polymers formed therefrom, and also fibrils, by binding thereto and thus converting them into non-toxic compounds. Accordingly, the present invention also provides a method for detoxifying A-beta oligomers, polymers formed therefrom or fibrils.
  • the invention also relates in one embodiment to peptides in accordance with the invention which are linked to a further substance.
  • the linking takes the form of a chemical bond as defined in Römpp Chemie Lexikon, 9th Edition, Volume 1, page 650 if, Georg Thieme Verlag Stuttgart, preferably a main valency bond, particularly a covalent bond.
  • the substances are medicaments or active ingredients, defined according to the Medicines Act ⁇ 2 or ⁇ 4 (19), as of September 2012.
  • active ingredients are therapeutically active substances which are used as medicinally active substances.
  • Anti-inflammatories are preferably used.
  • the substances are compounds which enhance the effect of the peptides.
  • such compounds are aminopyrazole and/or aminopyrazole derivatives.
  • Aminopyrazole derivatives in the context of the invention are 3-aminopyrazole-5-carboxylic acid or 3-nitropyrazole-5-carboxylic acid and also all derivatives thereof in which the heterocyclic CH group has been replaced by —CR— or —N— or —O— or —S—, and also all peptidic dimers, trimers or tetramers, preferably aminopyrazole trimer, derived therefrom.
  • said compounds are compounds which improve the solubility of the peptides and/or passage of the blood-brain barrier.
  • the peptides according to the invention have any desired combination of at least two or more features of the variants, embodiments and/or alternatives described above.
  • the invention also relates to a peptide in accordance with the invention for binding to aggregated A-beta peptides.
  • the invention further relates to a method for preparing the peptide according to the invention by peptide synthesis, as is known to those skilled in the art, for example, organic synthetic methods for any compounds with low molecular weights (low-molecular weight compounds) and/or mutagenesis and recombinant preparation.
  • the invention also relates to a composition comprising the peptide according to the invention, particularly for the treatment of Alzheimer's disease.
  • the present invention also relates to a composition comprising the peptide according to the invention, particularly for preventing toxic A-beta oligomers, or for destroying polymers or fibrils formed therefrom.
  • composition may be, for example, a vaccine, a medicament (e.g. in tablet form), an injectable solution, a food or food supplement, comprising the peptide according to the invention in a formulation being prepared due to technical expertise.
  • the invention also relates to a kit comprising the peptide according to the invention.
  • the peptides according to the invention may be packaged in containers, optionally with/in buffers or solutions. All components of the kit can be packaged in the same container or separately. Furthermore, the kit may include instructions for use.
  • a kit may comprise, for example, in accordance with the invention, in an injection vial with a stopper and/or septum. Furthermore, a disposable syringe can also be contained therein.
  • the present invention further relates to the use of the peptide according to the invention as a probe for the identification and qualitative and/or quantitative determination of amyloid beta oligomers or fibrils.
  • the present invention also relates to a probe comprising the peptide according to the invention for the identification and qualitative and/or quantitative determination of amyloid beta oligomers.
  • Such probes are of major significance, since an early diagnosis of AD is thereby made possible. With early diagnosis, the disease may already be counteracted at a very early stage.
  • Such molecular probes comprise the polymer according to the invention and optionally dyes, fluorescent dyes, radioactive isotopes, (PET etc.), gadolinium (MRI), and also alternative substances suitable for imaging the probes and the patients, for example, may be injected intravenously. After passage through the blood-brain barrier, the probes can bind to A-beta oligomers and/or plaques. The A-beta oligomers and/or plaques thus labeled can be visualized by imaging methods such as SPECT, PET, CT, MRT, proton MR spectroscopy etc.
  • imaging methods such as SPECT, PET, CT, MRT, proton MR spectroscopy etc.
  • the invention also relates to the use of the peptide for the prevention of amyloid beta oligomers and/or amyloid beta peptide aggregates and/or amyloid beta fibrils.
  • the peptide according to the invention is also used for the detoxification of toxic amyloid beta oligomers and/or aggregates. Said peptide is particularly used to bind to amyloid beta oligomers and/or aggregates and thus to form amorphous, non-toxic aggregates.
  • the invention further relates to the use of the peptide according to the invention as a therapeutic agent for Alzheimer's disease.
  • the peptides according to the invention bind particularly effectively to A-beta oligomers, particularly to soluble A-beta oligomers.
  • a particularly strong binding of the inventive peptides to the target molecules is caused by high specificity and/or affinity of the inventive peptides for the target molecule.
  • the complexes formed have a low dissociation constant (KD).
  • the peptides according to the invention very efficiently inhibit the formation of fibrils of A-beta peptides, particularly SEQ ID NO: 1-5, in particular SEQ ID NO: 1 and/or SEQ ID NO: 5.
  • the invention further relates to the use of the peptides according to the invention in a method for treating (in vitro, ex vivo) blood, blood products and/or organs, characterized in that the blood, the blood products and/or organs are taken from a human or animal body and A(amyloid)-beta oligomers are removed and/or detoxified.
  • the D3 variants listed in Table 1 were chemically synthesized.
  • A-beta was mixed with the respective peptide and the particles formed are separated according to size.
  • the sample to be investigated was placed on the surface of the centrifuge tube in which a density gradient was charged, which consisted of layers of different iodixanol concentrations in this example. During the several hours of separation, the molecules sediment at different rates in the solvent, namely, more rapidly the larger the particles. The centrifugation is terminated at a suitable time point and different constituents of the sample are obtained in the various layers which are analyzed by SDS-PAGE.
  • A-beta without addition of peptide served as control. For example, the evaluation of runs with A-beta and A-beta with RD2 is shown in FIG. 1 .
  • A-beta could be detected in all fractions in the A-beta sample. This changes on addition of RD2.
  • fewer A-beta oligomers could be detected in the fractions 3-9 (A-beta oligomer fraction). Large aggregates are formed which are detectable in fractions 10-15.
  • D3 showed no effect when the concentration of peptide used was 20 ⁇ M. In earlier studies, it was found that D3 in higher concentrations precipitates A-beta oligomers and converts them into large, amorphous and non-toxic aggregates (Funke at al., ACS Chem. Neurosci. 2010).
  • the peptides were also tested with respect to their in vitro binding to various A-beta conformers (A-beta monomers, oligomers, fibrils) using ELISA. All peptides showed weak binding to A-beta monomers but relatively high affinity to oligomers and fibrils ( FIG. 2 ). Interestingly, biotinylated monomers were not detected at the amino terminus, whereas the “seedless” monomers (aggregation seed-free preparation) were weakly bound to the carboxy terminus biotinylation. This may be interpreted as a potential indication of an epitope binding site of the D3 derivatives localized to the amino terminus.
  • ThT thioflavin T
  • ThT tests are used for measuring the fibrillation of A-beta and are used especially in ligands to detect potential inhibitory effects of these on A-beta aggregation.
  • the peptides were used in a ratio of 1:10 (A-beta: peptide) with a 10 ⁇ M A-beta solution.
  • D3 variants were identified which have an increased binding strength for A-beta, particularly for A-beta oligomers, with at least the same effect on the aggregation thereof, compared to D3. Stronger binding may suggest a more efficient therapeutic effect.
  • the binding signals were evaluated as signal intensity/area.
  • the signal intensity of the original D3 (mean value) was compared with that of the derivatives.
  • the amino acid exchanges which resulted in stronger binding of A-beta oligomers are summarized in table 3.
  • FIG. 1 Modulation of the A-beta1-42 aggregation behavior by RD2, analyzed by an iodixanol density gradient.
  • the iodixanol gradient was overlayed with 100 ⁇ l of a 80 ⁇ M A-beta1-42 solution and 100 ⁇ l of a 80 ⁇ M A-beta and 20 ⁇ M RD2 mixture.
  • the mixture was then centrifuged at 259 000 ⁇ g for 3 h at 4° C.
  • the 15 fractions fraction 15 is the pellet boiled with the loading buffer
  • FIG. 2 ELISA for the relative quantification of the binding of the peptides to various A-beta1-42 conformers. Seedless monomers of carboxy terminal biotinylated A-beta (light grey bars) and oligomers (dark grey bars) and fibrils (black bars) of amino terminal biotinylated A-beta monomers were each immobilized at 5 ⁇ g/ml and the D-peptide applied at a concentration of 10 ⁇ g/ml. The relative quantification of the binding of the peptides in a duplicate determination is shown as absorption at 450 nm after subtraction of the background absorption.
  • FIG. 3 ThT aggregation test of A-beta1-42 for the quantification of the relative fibril content in the presence of various peptides.
  • concentration of A-beta was 10 ⁇ M and the peptides were added in a ratio of 1:10 (A-beta: peptide).
  • the fluorescence of 10 ⁇ M A-beta was set to 100% and the values and standard deviations of the other incubations are given as percentages of this maximum value. None of the peptides showed a significant ThT fluorescence with no A-beta.
  • FIG. 4 Results of a PepChip experiment. Different D3 variants were immobilized on the PepChip. The PepChip was incubated with FITC labeled A-beta (5 ⁇ M). The FITC fluorescence intensity was read as a measure of the binding strength of the respective D3 derivatives. Values of the mean and standard deviation were calculated from the 11 values obtained for the D3 controls likewise applied at 11 different positions on the chip. Mean and standard deviation were added and the result was defined as the limit which a D3 derivative must attain in order to clearly have higher affinity for A-beta compared to D3. The integrals of the binding signals of all peptides are shown which exceed this limit. The individual bars are based on the means of three exactly identical peptide spots. a.u.: arbitrary units, relative fluorescence units. Due to the observed variance in the results, this entire experiment was carried out six times.

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10239923B2 (en) 2015-03-20 2019-03-26 Forschungszentrum Juelich Gmbh Specific amyloid beta binding peptides and the use of same for treating and diagnosing alzheimer's dementia
WO2019070645A1 (en) * 2017-10-02 2019-04-11 Cedars-Sinai Medical Center METHODS AND COMPOSITIONS FOR EFFECTIVE ADMINISTRATION THROUGH MULTIPLE BIOLOGICAL BARRIERS

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102012108598A1 (de) * 2012-09-14 2014-04-10 Forschungszentrum Jülich GmbH Neue, von D3 abgeleitete D-enantiomere Peptide und deren Verwendung
DE102015003676A1 (de) 2015-03-20 2016-09-22 Forschungszentrum Jülich GmbH Fachbereich Patente Spezifisch A-Beta Spezies bindende Peptide für die Therapie und/oder die Diagnose der Alzheimerschen Demenz
DE102016125645A1 (de) * 2016-12-23 2018-06-28 Forschungszentrum Jülich GmbH D-enantiomere Peptide zur Therapie von chronischem und neuropathischem Schmerz
DE102021121435A1 (de) * 2021-08-18 2023-02-23 Priavoid Gmbh Aß-BINDENDE SUBSTANZEN

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK1346041T3 (da) * 2000-11-27 2007-07-02 Praecis Pharm Inc Terapeutiske midler og fremgangsmåder til anvendelse deraf til behandling af en amyloidogen sygdom
WO2002081515A1 (fr) * 2001-04-03 2002-10-17 Kyowa Hakko Kogyo Co., Ltd. Proteine de liaison de facteur de croissance de type insuline
DE10117281A1 (de) 2001-04-06 2002-10-24 Inst Molekulare Biotechnologie Peptid zur Diagnose und Therapie der Alzheimer-Demenz
US20050158306A1 (en) * 2004-01-16 2005-07-21 Halikas James A. Treatment for neuro-degenerative disease by a blood cleansing system using monoclonal antibodies
AT500483B1 (de) * 2004-07-13 2006-01-15 Mattner Frank Dr Set zur vorbeugung oder behandlung der alzheimer'schen erkrankung
US20070092508A1 (en) 2005-10-21 2007-04-26 Recombiant Technologies, Llc Detoxification depot for Alzheimer's disease
US8445438B2 (en) 2008-10-27 2013-05-21 Recombinant Technologies Llc Compositions comprising capture peptides for A β-amyloid peptide
DE102008037564A1 (de) * 2008-11-19 2010-05-20 Forschungszentrum Jülich GmbH Zusammensetzung zur Herstellung von anti-Amyloid beta-Peptid-Antikörpern mit D-Peptiden
DE102010017130B4 (de) 2010-05-28 2018-11-29 Forschungszentrum Jülich GmbH Hybrid-Verbindung, deren Verwendung und Verfahren zu deren Herstellung
DK2834643T3 (da) * 2012-04-05 2020-02-03 Forschungszentrum Juelich Gmbh Fremgangsmåde til behandling af blod, blodprodukter og organer
DE102012108598A1 (de) * 2012-09-14 2014-04-10 Forschungszentrum Jülich GmbH Neue, von D3 abgeleitete D-enantiomere Peptide und deren Verwendung
US9464118B2 (en) 2012-04-05 2016-10-11 Forschungszentrum Juelich Gmbh Polymers containing multivalent amyloid-beta-binding D-peptides and their use

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Hampel et al. The future of Alzheimer's disease: the next 10 years. Prog Neurobiol. 2011 Dec;95(4):718-28. Epub 2011 Nov 22. *
Perrin et al. Multimodal techniques for diagnosis and prognosis of Alzheimer's disease. Nature 2009 Oct 15;461(7266):916-22, Epub 2009 Oct 14. *
Vickers. A vaccine against Alzheimer's disease: developments to date. Drugs Aging 2002; 19(7):487-94. *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10239923B2 (en) 2015-03-20 2019-03-26 Forschungszentrum Juelich Gmbh Specific amyloid beta binding peptides and the use of same for treating and diagnosing alzheimer's dementia
WO2019070645A1 (en) * 2017-10-02 2019-04-11 Cedars-Sinai Medical Center METHODS AND COMPOSITIONS FOR EFFECTIVE ADMINISTRATION THROUGH MULTIPLE BIOLOGICAL BARRIERS

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US20190085030A1 (en) 2019-03-21
JP2015531348A (ja) 2015-11-02
JP6345670B2 (ja) 2018-06-20
US20170313744A1 (en) 2017-11-02
EP3346273A8 (de) 2018-08-29
EP2895867A2 (de) 2015-07-22
WO2014041115A2 (de) 2014-03-20
CN107556375A (zh) 2018-01-09
EP2895867B1 (de) 2019-10-23
WO2014041115A3 (de) 2014-06-26
US10889618B2 (en) 2021-01-12
JP6611858B2 (ja) 2019-11-27
EP3346273B1 (de) 2023-01-04
DE102012108598A1 (de) 2014-04-10
US10167318B2 (en) 2019-01-01
ES2938773T3 (es) 2023-04-14
JP2018141008A (ja) 2018-09-13
PL3346273T3 (pl) 2023-04-11
DK2895867T3 (da) 2020-02-03
EP3346273A1 (de) 2018-07-11

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