Modulation of Tau Using Protein-Like Polymers and Their Use

Abstract

A polymer comprising a first repeating unit comprising a first polymer main chain subunit directly or indirectly covalently linked to a first functional side chain comprising a peptide that (i) inhibits the aggregation of at least a part of the tau protein, (ii) promotes the aggregation of the tau protein, (iii) binds to the tau protein, and / or (iv) mimics the tau protein.

Description

Technical Field

[0001] Cross - Reference to Related Applications

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 389,616, filed on July 15, 2022, which is hereby incorporated by reference in its entirety. Reference to Electronic Sequence Listing

[0002] The content of the electronic sequence listing (339677_93 - 21_WO_ST1.xml; size: 14,746 bytes; and creation date: July 11, 2023) is hereby incorporated by reference in its entirety. Description of Research or Development Sponsored by the Federal Government

[0003] This invention was made with government support under grant number 1F31AG076334 - 01 awarded by the Department of Health and Human Services, National Institute of Health. The government has certain rights in this invention.

Background Art

[0002]

[0004] Hundreds of human diseases that are collectively known as protein conformational diseases or protein folding disorders occur due to protein misfolding resulting from intrinsic and extrinsic errors amplified by exposure to environmental and physiological stress conditions. Zhao J-H et al., Chemical Chaperone and Inhibitor Discovery: Potential Treatments for Protein Conformational Diseases. Perspect Medicin Chem 2007;1:PMC.S212; Voisine C et al., Chaperone networks: Tipping the balance in protein folding diseases. Neurobiol Dis 2010;40:12-20; Ciryam P et al., Widespread Aggregation and Neurodegenerative Diseases Are Associated with Supersaturated Proteins. Cell Rep 2013;5:781-90. Such events can attack proteome integrity, causing acute clearance, mislocalization, and protein dysfunction or aggregation, thus affecting cellular robustness, health, and lifespan. These diseases include type II diabetes, cystic fibrosis, cancer, and neurodegenerative diseases, exemplified by, for example, Alzheimer's disease (AD), frontotemporal dementia, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and Huntington's disease.

[0003]

[0005] In each case, it has been demonstrated that inhibiting, controlling, and even investigating the progression of these processes is difficult with conventional approaches such as small molecule and antibody-type systems. As described herein, the inventors have focused on neurodegenerative diseases, specifically on the dysregulation and aberrant behavior of the microtubule-associated protein tau (MAPT) that cause tauopathies, including AD, frontotemporal dementia, and traumatic brain injury. It is thought that mediating tau dysregulation should reduce the etiology of dementia in tauopathies. Here, the inventors disclose an approach for investigating tau in the transient, early, and toxic situations of amyloid formation and for adding to drugs by engaging the cellular machinery with the phase separation state of tau (FIGS. 1A, 1B). This approach is thought to provide a basis for bringing about the treatment of a larger class of age-related degenerative diseases of protein misfolding.

[0004]

[0006] The inventors provide the use of proteomimetic materials called protein-like polymers (PLPs) that engage proteins with the intracellular quality control machinery (FIGS. 1A, 1B). Callmann CE et al., Poly(peptide): Synthesis, Structure, and Function of Peptide-Polymer Amphiphiles and Protein-like Polymers. Acc Chem Res 2020;53:400-13; Gianneschi NC et al., Biomolecular Densely Grafted Brush Polymers: Oligonucleotides, Oligosaccharides and Oligopeptides. Angew Chemie Int Ed 2020; Blum AP, Kammeyer JK, Gianneschi NC. Activating peptides for cellular uptake via polymerization into high density brushes. Chem Sci 2016;7:989-94; Sun H et al., Peptide-Brush Polymers as Globular Proteomimetics. The proposed PLPs are modular, scalable, rapidly formulated using advanced polymerization strategies, and provide cell permeability, binding to specific targets, and multivalency. The inventors believe that the successful integration of PLPs into native protein assemblies will have broad implications in understanding and treating many diseases associated with the misfolding of major proteins such as TDP43, huntingtin, and SOD1.

[0005]

[0007] As a proof of concept, a robust methodological framework is disclosed herein for creating a single protein / disease (tau / Alzheimer's) to convert these proteomimetic polymers into a therapeutic platform that can ultimately be widely adopted for protein misfolding disorders. PLP is thought to be optimizable to alter the biological consequences of the phase transition of tau protein into amyloid fibrils. This strategy is based on evidence that misfolding and aggregation are fundamental problems related to tauopathy in Alzheimer's disease, but current efforts to develop treatments with small molecules or antibodies have proven to be at best challenging and at worst unsuccessful.

[0006]

[0008] Accordingly, there remains a need for therapeutic agents and methods that target age-related degenerative diseases of protein misfolding. The present invention provides such therapeutic agents and methods. This and other advantages of the present invention will become apparent from the detailed description provided herein.

SUMMARY OF THE INVENTION

[0007]

[0009] In one aspect, the present invention provides a polymer comprising a first repeating unit comprising a first polymer backbone subunit directly or indirectly covalently linked to a first functional side chain comprising a peptide that (i) inhibits the aggregation of at least a portion of the tau protein, (ii) promotes the aggregation of at least a portion of the tau protein, (iii) binds to at least a portion of the tau protein, and / or (iv) mimics at least a portion of the tau protein.

[0008]

[0010] The present invention further includes a polymer comprising a first repeating unit comprising a first polymer backbone subunit directly or indirectly covalently linked to a first functional side chain comprising a peptide that (i) inhibits the aggregation of at least a portion of the microtubulin protein, (ii) promotes the aggregation of at least a portion of the microtubulin protein, (iii) binds to at least a portion of the microtubulin protein, and / or (iv) mimics at least a portion of the microtubulin protein.

[0009]

[0011] A polymer characterized by formula (FX1): [Chemical formula] (In the formula, each P 1 independently contains a peptide; each P 2 independently contains a peptide, and each instance of P 2 is different from each instance of P 1 ; at least one P 1 independently or in combination with other instances of P 1 either (a) inhibits the aggregation of, and / or (b) promotes the aggregation of, and / or (c) binds to, and / or (d) mimics at least a part of the tau protein and / or at least a part of the microtubulin protein; T 1 and T 2 are each independently polymer backbone end groups which may be the same or different; B 1 , B 2 , and B 3 are each independently polymer backbone subunits; L 1 and L 2 are each independently linking groups; R 1 is independently a substituent; m is an integer from 2 to 1000; n is an integer from 0 to 1000; o is an integer from 0 to 1000; each connecting line in formula (FX1) represents a connection by a covalent bond including a single bond, a double bond, at least one of one or more atoms, or any combination thereof, and optionally, one or more atoms include carbon, nitrogen, and / or oxygen atoms; each instance of B 1 , B 2 , B 3 , L 1 , L 2 , R 1 , P 1 , and P 2 is respectively B 1 , B 2 , B 3 , L 1 , L 2 , R 1 , P 1 , and P2 is the same as or different from any other case; (i) n is an integer from 1 to 1000, o is an integer from 1 to 1000, P 1 at least one case of 1 is different from another case of P 2 and / or at least one case of P 2 is different from another case of P, (ii) the polymer is a block copolymer or a statistical copolymer) is further disclosed herein.

[0010]

[0012] In other aspects, the present invention provides a method for preventing, disrupting, promoting, or detecting tau and / or microtubulin protein aggregation, the method comprising contacting oligomers, protofibrils, amyloid fibers, and / or cross-β sheet amyloid species of tau and / or microtubulin with any of the polymers disclosed herein in a therapeutically effective amount, or a composition thereof.

[0011]

[0013] The present invention further provides a method for preventing, treating, or detecting a tauopathy-related disease or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of any of the polymers disclosed herein, or a composition thereof.

[0012]

[0014] While not wishing to be bound by any particular theory, discussions may be presented herein regarding ideas or understandings of the underlying principles related to the devices and methods disclosed herein. Regardless of the fundamental accuracy of any mechanistic explanations or hypotheses, it is recognized that embodiments of the present invention may nevertheless function and be useful.

Brief Description of the Drawings

[0013]

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Mode for Carrying Out the Invention

[0014] Description Regarding Chemical Substances and Academic Names

[0051] The following abbreviations are used in this specification: RP-HPLC refers to reverse-phase high-performance liquid chromatography; ESI-MS refers to electrospray ionization mass spectrometry; SEC-MALS refers to size exclusion chromatography combined with multi-angle light scattering; PLP refers to protein-like polymer; SPPS refers to solid-phase peptide synthesis; TEM refers to transmission electron microscopy; STEM refers to scanning TEM; SE or SEM refers to scanning electron microscope; CD refers to circular dichroism; FPLC refers to high-performance liquid chromatography for proteins; DP refers to degree of polymerization.

[0015]

[0052] In one embodiment, the peptide, polymer, or composition (e.g., formulation) of the present invention is isolated or purified. In one embodiment, the isolated or purified peptide, polymer, or composition (e.g., formulation) is at least partially isolated or purified as understood in the art. In one embodiment, the peptide, polymer, or composition (e.g., formulation) of the present invention has a chemical purity of at least 95%, optionally at least 99% for some uses, optionally at least 99.9% for some uses, optionally at least 99.99% for some uses, optionally at least 99.999% pure for some uses. The present invention includes any isolated and purified composition of the brush block polymers described herein, including peptide brushes and block copolymers, and brushes and brush block copolymers having one or more side chains including peptide analogs, derivatives, variants or fragments.

[0016]

[0053] As used herein, the term "polymer" refers to a molecule composed of repeating structural units connected by covalent chemical bonds, often characterized by a significant number of repeating units (e.g., repeating units equal to or greater than 3, optionally, in some embodiments, repeating units equal to or greater than 5, in some embodiments, repeating units greater than or equal to 10) and a high molecular weight (e.g., greater than or equal to 1 kDa, in some embodiments, greater than or equal to 5 kDa, or greater than or equal to 50 kDa). Polymers are generally the polymerization products of one or more monomer precursors. The term "polymer" includes homopolymers, or polymers consisting essentially of a single repeating monomer subunit. The term "polymer" also includes copolymers formed when two or more different types of monomers are linked within the same polymer. Copolymers may include two or more monomer subunits (e.g., three or more monomer subunits, four or more monomer subunits, five or more monomer subunits, or six or more monomer subunits) and may include random, block, brush, brush-block, alternating, segmented, grafted, tapered, and other architectures. In some embodiments, the copolymers of the present invention include 2 to 10 different monomer subunits. Useful polymers include organic polymers that can be in an amorphous, semi-amorphous, crystalline, or semi-crystalline state. For some applications, cross-linked polymers having linked monomer chains are useful, for example, cross-linked polymers having monomer chains linked by one or more disulfide linkages. The present invention provides polymers comprising therapeutic agents, such as brush polymers having at least a portion of repeating units comprising polymer side chains, such as peptide side chains.

[0017]

[0054] An "oligomer" is a molecule composed of repeating structural units connected by covalent chemical bonds, often characterized by a number of repeating units less than that of the repeating units of a polymer (e.g., repeating units equal to or less than 3) and a molecular weight lower than that of a polymer (e.g., less than or equal to 1,000 Da). An oligomer may be a polymerization product of one or more monomer precursors.

[0018]

[0055] As used interchangeably herein, "peptide" or "oligopeptide" refers to a polymer having repeating structural units connected by peptide bonds. Typically, the repeating structural units of a peptide are amino acids such as naturally occurring amino acids, non-naturally occurring amino acids, amino acid analogs, or any combination thereof. The number of repeating structural units of a peptide is typically less than that of a "protein" as understood in the art, and thus peptides often have a lower molecular weight than proteins. In some embodiments, the peptide has a chain length of 3 to 150 amino acids, optionally 3 to 100 amino acids, optionally 5 to 50 amino acids, optionally 5 to 30 amino acids.

[0019]

[0056] A "block copolymer" is a type of copolymer that contains blocks or spatially separated domains, where different domains contain different polymerized monomers, for example, at least two chemically distinguishable blocks. A block copolymer may further contain one or more other structural domains, such as hydrophobic groups, hydrophilic groups, etc. In a block copolymer, adjacent blocks are structurally different, that is, adjacent blocks contain structural units derived from different species of monomers or contain structural units of the same species of monomers but with different compositions or sequence distributions of the structural units. Different blocks (or domains) of a block copolymer may be present at different ends or within the polymer (e.g., [A][B]), or may be provided in a selected sequence ([A][B][A][B]). A "diblock copolymer" refers to a block copolymer having two different polymer blocks. A "triblock copolymer" refers to a block copolymer having three different polymer blocks, where two non-adjacent blocks have the same or similar composition. A "pentablock" copolymer refers to a copolymer having five different polymers, where two or more non-adjacent blocks have the same or similar composition.

[0020]

[0057] A "statistical copolymer", also generally known in the art as a "random copolymer", is a copolymer in which the order of the main-chain groups is determined by the reaction kinetics. Statistical copolymers are generally contrasted with block copolymers.

[0021]

[0058] "Polymer main chain group" or "polymer main chain subunit" refers to a group that is covalently linked to form the main chain of a polymer such as a block copolymer. The polymer main chain group may be linked to a side chain group, for example, a polymer side chain group. Some polymer main chain groups useful in the compositions of the present invention are derived from the polymerization of monomers selected from the group consisting of substituted or unsubstituted norbornene, olefin, cyclic olefin, norbornene anhydride, cyclooctene, cyclopentadiene, styrene, acrylamide, and acrylate. Some polymer main chain groups useful in the compositions of the present invention are obtained from ring-opening metathesis polymerization (ROMP) reactions. The polymer main chain may be terminated with various main chain end groups including hydrogen, C1-C 10 alkyl, C3-C 10 cycloalkyl, C5-C 10 aryl, C5-C 10 heteroaryl, C1-C 10 acyl, C1-C 10 hydroxyl, C1-C 10 alkoxy, C2-C 10 alkenyl, C2-C 10 alkynyl, C5-C 10 alkylaryl, -CO2R 30 , -CONR 31 R 32 , -COR 33 , -SOR 34 , -OSR 35 , -SO2R 36 , -OR 37 , -SR 38 , -NR 39 R 40 , -NR 41 COR 42 , C1-C 10 alkyl halide, phosphonate, phosphonic acid, silane, siloxane, acrylamide, acrylate, or catechol, and each of R 30 ~R 42 is independently hydrogen, C1-C 10 alkyl or C5-C 10 aryl.

[0022]

[0059] "Polymer side chain group" (and sometimes, in this specification, for example, R 1 is also referred to as a "substituent") refers to a group that is covalently linked (either directly or indirectly) to a polymer backbone group containing a polymer side chain, and optionally imparts steric properties to the polymer. In one embodiment, for example, a polymer side chain group is characterized by a plurality of repeating units having the same or similar chemical composition. The polymer side chain group may be linked directly or indirectly to the polymer backbone group. In some embodiments, the polymer side chain group provides a steric bulk and / or interaction that results in an extended polymer backbone and / or a rigid polymer backbone. Some polymer side chain groups useful in the compositions of the present invention include unsubstituted or substituted peptide groups. Some polymer side chain groups useful in the compositions of the present invention contain repeating units obtained via anionic polymerization, cationic polymerization, free radical polymerization, group transfer polymerization, or ring-opening polymerization. The polymer side chain may be terminated with a variety of polymer side chain end groups including hydrogen, C1-C 10 alkyl, C3-C 10 cycloalkyl, C5-C 10 aryl, C5-C 10 heteroaryl, C1-C 10 acyl, C1-C 10 hydroxyl, C1-C 10 alkoxy, C2-C 10 alkenyl, C2-C 10 alkynyl, C5-C 10 alkylaryl, -CO2R 30 , -CONR 31 R 32 , -COR 33 , -SOR 34 , -OSR 35 , -SO2R 36 , -OR 37 , -SR 38 , -NR 39 R 40 , -NR 41 COR 42 , C1-C 10 alkyl halide, phosphonate, phosphonic acid, silane, siloxane, acrylamide, acrylate, or catechol, and R30 ~R 42 Each of them is independently hydrogen or C1-C5 alkyl.

[0023]

[0060] The term "polymer segment" (e.g., the first polymer segment, the second polymer segment, etc.), as used herein, refers to a section (e.g., a portion) of a polymer that includes a particular monomer or arrangement of monomers. A polymer segment may be a homopolymer or a copolymer. In embodiments where the polymer segment is a copolymer, the copolymer may be present in any suitable arrangement of monomers (e.g., random, block, brush, brush-block, alternating, segmented, grafted, tapered, statistical, and other architectures). In some embodiments, the polymer segment is a homopolymer, random copolymer, statistical copolymer, or block copolymer. Any polymer described herein (e.g., a brush polymer) may have a single polymer segment or multiple polymer segments. In embodiments where the polymer has multiple polymer segments, the polymer segments may be present in any suitable arrangement (random, block, brush, brush-block, alternating, segmented, grafted, tapered, statistical, and other architectures).

[0024]

[0061] The term "degree of polymerization", as used herein, refers to the average number of monomer units per polymer chain. For example, for a particular polymer described herein that includes B 1 , B 2 , and / or B 3 main chain units, the degree of polymerization would be represented by the total of the B 1 , B 2 , and B 3 main chain units. Since the degree of polymerization can vary from polymer to polymer, the degree of polymerization is generally represented by an average.

[0025]

[0062] As used herein, the term "brush polymer" refers to a polymer comprising repeating units each independently containing a polymer backbone group covalently linked to at least one polymer side chain group. Brush polymers can be characterized by a brush density, which refers to the percentage of repeating units containing polymer side chain groups. Brush polymers of certain embodiments are characterized by a brush density greater than or equal to 50% (e.g., greater than or equal to 60%, greater than or equal to 65%, greater than or equal to 70%, greater than or equal to 75%, greater than or equal to 80%, greater than or equal to 85%, or greater than or equal to 90%), and optionally for some embodiments, by a density greater than or equal to 70%, or optionally for some embodiments, by a density greater than or equal to 90%. Brush polymers of certain embodiments are characterized by a brush density selected from the range of 50% to 100%, and optionally for some embodiments, by a density selected from the range of 75% to 100%, or optionally for some embodiments, by a density selected from the range of 90% to 100%. Brush polymers, such as the polymers disclosed herein (e.g., the polymer of formula (1)), can be prepared by any suitable method including a "grafting from" method, a "grafting onto" method, a "grafting through" method, or any combination thereof. Such suitable methods can include, for example, ring-opening metathesis polymerization (ROMP) synthetic routes and / or non-ROMP synthetic routes, such as, by way of example, reversible addition-fragmentation chain transfer (RAFT) polymerization, stable free radical-mediated polymerization, and atom transfer radical polymerization (ATRP).

[0026]

[0063] As used herein, the term "peptide density" refers to the percentage of monomer units in a polymer chain having peptides covalently linked to the polymer chain, and such "peptide density" may generally be calculated for all peptides or for a specific peptide. The percentage is based on the overall total of monomer units in the polymer chain. For example, in the case of a particular polymer described herein, peptide P 1 with m repeating units, B 2 -R 1 with n repeating units, and peptide P 2 with o repeating units, the density of peptide P 1 (or the percentage of monomer units containing peptide P 1 ) is given by the formula:

Number

[0027]

[0064] In one aspect, the polymer side chain groups (also referred to herein as substituents) may have any suitable spacing in the polymer backbone. Typically, the spacing between adjacent polymer side chain groups is from 3 angstroms to 30 angstroms, optionally from 5 to 20 angstroms, and optionally from 5 to 10 angstroms. By way of illustration, in certain embodiments having a 100% brush density, the polymer side chain groups are typically separated in the polymer backbone by a spacing of 6 ± 5 angstroms. In some embodiments, the brush polymer has a high brush density (e.g., greater than 70%), in which case the polymer side chain groups are separated in the polymer backbone by a spacing of 5 to 20 angstroms.

[0028]

[0065] The term "sequence homology" or "sequence identity", as used herein, means the ratio of amino acids that match between two different peptides, taking into account the order of the amino acids, between two amino acid sequences of interest. A match occurs when, in one peptide, the amino acids are in the same order as compared to the other peptide. When sequence homology is expressed as a percentage, such as 50%, the percentage means the proportion of matches over the length of the sequence being compared to some other sequence, taking into account the order of the amino acids. Gaps (in either of the two sequences) are allowed to maximize the match, and for example, a gap length of 5 amino acids or less, optionally 3 amino acids or less, is typically used. In other words, a sequence having at least 75% sequence identity to an amino acid sequence having 9 amino acids can be shown to have the 9 amino acid sequence with one or two point mutations (i.e., amino acid changes), one or two amino acid deletions, one or two amino acid additions, one point mutation and one amino acid deletion, or one point mutation and one amino acid addition. Even if two such amino acids are different, 7 out of 9 amino acids still match in the correct order, and thus there is sequence identity greater than 75%. For clarity, the analysis of whether there is sequence homology between two amino acid sequences of interest is performed with respect to a particular portion of one peptide or protein (i.e., the first amino acid sequence of interest) compared to a particular portion of another peptide or protein (i.e., the second amino acid sequence of interest), and is not performed with respect to all amino acids present in the peptide or protein (i.e., this analysis does not include amino acids outside the particular amino acid sequences of interest).

[0029]

[0066] As used herein, the terms "amino acid composition similarity" or "amino acid similarity" mean the ratio of amino acids that match between two different peptide amino acid sequences, regardless of the order of the amino acids. A match occurs when an amino acid is present in both amino acid sequences, regardless of order. When amino acid composition similarity is expressed as a percentage, such as 50%, the percentage means the proportion of matches over the length of the sequence being compared to some other sequence(s), regardless of the order of the amino acids. Gaps (in either of the two sequences) are allowed to maximize the matches, and for example, a gap length of 5 amino acids or less, optionally 3 amino acids or less, is commonly used. As an example, if two amino acid sequences each containing 10 amino acids have 3 amino acids in common in any order, there is 30% amino acid composition similarity between the sequences. For clarity, the analysis of whether there is amino acid composition similarity between two target amino acid sequences is performed with respect to a particular portion of one peptide or protein (i.e., the first target amino acid sequence) compared to a particular portion of another peptide or protein (i.e., the second target amino acid sequence), and is not performed with respect to all amino acids present in the peptide or protein (i.e., this analysis does not include amino acids outside of the particular target amino acid sequences of interest).

[0030]

[0067] The term "fragment" refers to a portion, but not all, of a composition or material, such as a peptide composition or material. In one embodiment, a fragment of a peptide refers to 50% or more of the amino acid sequence, optionally 70% or more of the amino acid sequence, optionally 90% or more of the amino acid sequence.

[0031]

[0068] "Polymer blend" refers to a mixture comprising at least one polymer, such as a brush polymer, such as a brush block copolymer, and at least one additional component, optionally more than one additional component. In some embodiments, for example, the polymer blends of the present invention comprise a first brush copolymer and one or more additional brush polymers having a composition different from that of the first brush copolymer. In some embodiments, for example, the polymer blends of the present invention further comprise one or more additional brush block copolymers, homopolymers, copolymers, block copolymers, brush block copolymers, oligomers, solvents, small molecules (e.g., having a molecular weight of less than 500 Da, optionally less than 100 Da), or any combination thereof. Polymer blends useful for some applications comprise a first brush polymer and one or more additional components comprising a polymer, block copolymer, brush polymer, linear block copolymer, random copolymer, homopolymer, or any combination thereof. Polymer blends of the present invention include mixtures of 2, 3, 4, 5 and more polymer components.

[0032]

[0069] The term "compound" as used herein can be used to refer to either a peptide or a polymer described herein. Alternatively, or in addition, the term compound may refer to any of the synthetic precursors, reagents, additives, excipients, etc. used in the preparation or formulation of a peptide or polymer described herein.

[0033]

[0070] The term "group" as used herein may refer to a functional group of a chemical substance. A group of a compound of the present invention refers to an atom or a collection of atoms that is part of the compound. A group of the present invention may be attached to other atoms of the compound via one or more covalent bonds. A group can also be characterized with respect to its valence state. The present invention includes groups characterized as having valence states such as monovalent, divalent, trivalent, etc.

[0034]

[0071] The term "substituted", as used herein, generally refers to a compound in which hydrogen is replaced by another functional group, unless there is a contradiction in the context.

[0035]

[0072] The term "average molecular weight" or "molecular weight", unless otherwise specified, refers to the number average molecular weight. The number average molecular weight is defined as the value obtained by dividing the total weight of the sample volume by the number of molecules in the sample. As is customary and well-known in the art, the peak average molecular weight and the weight average molecular weight may also be used to characterize the molecular weight of the polymer distribution in the sample.

[0036]

[0073] The term "K18", as used herein, refers to a fragment of a recombinant tau protein having four domains involved in microtubule binding and amyloid fibril formation. The 129-residue long chain is regarded as the core peptide of tau, or the active tau monomer.

[0037]

[0074] References to compounds, oligomers, and / or polymers (e.g., “proteomimetic”) that mimic a given species, e.g., one or more oligo- or polypeptides (e.g., proteins), the terms “mimic,” “mimicking,” “mimetic,” and grammatically equivalent variations thereof, as used herein, mean that the compound, oligomer, and / or polymer has a portion that has an amino acid sequence similar to and / or corresponding to a portion of the given species. In some embodiments, the similar and / or corresponding portions typically relate to the presence of a certain level of sequence homology and / or similarity in amino acid composition between the given species and one or more oligo- or polypeptides (e.g., proteins). In embodiments, a mimetic refers to a material capable of mimicking the primary structure and / or function of a peptide or protein. A mimetic can be synthetically produced and modified to include specific properties depending on desired outcomes such as variable size, greater stability, greater affinity, protease resistance, and improved solubility. In embodiments, a mimetic refers to a protein-like polymer (PLP) designed to engage a protein with the intracellular quality control machinery. Callmann CE et al., Poly(peptide): Synthesis, Structure, and Function of Peptide-Polymer Amphiphiles and Protein-like Polymers. Acc Chem Res 2020;53:400-13; Gianneschi NC et al., Biomolecular Densely Grafted Brush Polymers: Oligonucleotides, Oligosaccharides and Oligopeptides. Angew Chemie Int Ed, 2020; Blum AP, Kammeyer JK, Gianneschi NC, each of these is incorporated herein by reference in its entirety, to the extent that it does not conflict with the description herein, more specifically to facilitate the understanding of PLP.

[0038]

[0075] In aspects of the invention, the mimetic may be modified to include residue-specific modifications, peptide backbone modifications, N-terminal modifications, C-terminal modifications, or any combination thereof. In embodiments, the modifications can improve peptide stability, alter peptide structure, incorporate imaging and / or detection agents, improve solubility, enhance non-specific enzyme resistance, reduce steric hindrance, increase cell permeability, improve binding affinity to a target, enhance safety, or any combination thereof. For example, modifications can include biotinylation, labeling with a contrast agent such as labeling with Gd-DOTA, labeling with a fluorescent dye such as labeling with a cyanine, labeling with fluorescein and 7-methoxycoumarin acetic acid, labeling with dansyl and / or 2,4-dinitrophenyl, labeling with EDANS, labeling with coumarin, and / or labeling with rhodamine, one or more point mutations, introduction of one or more spacers, labeling with an isotope, introduction of one or more chelating agents, acetylation, amidation, methylation, palmitoylation, hydroxylation, glycosylation, sulfation and sulfonation, esterification, phosphorylation, peptide stapling, lipidation, cyclization, or one or more of any combination thereof.

[0039]

[0076] The phrase "charge modulating domain" as used herein refers to one or more amino acids added to the peptide sequences described herein to modulate the charge of a peptide. For example, the charge modulating domain may be a TAT sequence, a glycine - serine domain, a cationic residue domain, or a combination thereof, or optionally a glycine - serine domain, a cationic residue domain, or a combination thereof. In certain embodiments, the charge modulating domain has 2 - 7 amino acid residues. The 2 - 7 amino acids may be added in a single block containing 2 - 7 amino acid residues or in more than one block containing 1 - 6 amino acid residues. In some embodiments, the charge modulating domain is a cationic residue domain having 2 - 7 amino acid residues selected from lysine, arginine, histidine, or a combination thereof. In some embodiments, the charge modulating domain includes aspartic acid residues. Generally, the charge modulating domain modulates the charge of the peptide to have a net positive charge. Without wishing to be bound by any particular theory, the net positive charge is thought to increase the intracellular uptake of the peptide or the polymer containing the peptide. The overall charge of the peptide or the copolymer containing the peptide can be determined by any suitable means. For example, the overall charge can be determined by (i) a structural analysis of the functional residues on the peptide sequence and their respective pKa's, (ii) physical characterization by measuring the zeta potential, and / or (iii) materials that migrate towards the cathode in an electrophoretic polymer gel. In certain embodiments, the overall charge of the peptide or the copolymer containing the peptide is determined by measuring the zeta potential.

[0040]

[0077] A "degrader agent" or "degron", as used herein, refers to a class of agents capable of facilitating the regulation of proteolysis, either directly or indirectly. For example, the regulation may include promoting proteolysis of a protein, inhibiting proteolysis, increasing the rate of proteolysis, and / or decreasing the rate of proteolysis. For example, in embodiments where the degrader agent is incorporated into PLP, the degrader agent facilitates specific degradation of the targeted protein. In some aspects of the invention, the degrader agent facilitates ubiquitin mobilization. Without wishing to be bound by a particular theory, in aspects where the degrader agent facilitates ubiquitin mobilization, the degrader agent is thought to participate in the polyubiquitination process for targeting proteins, or fragments thereof, for degradation by the proteasome. In these aspects, the degrader agent may be referred to herein as a "proteasome recruiter". In embodiments, the degrader agent includes a proteasome-targeting chimera ("PROTAC"). In aspects, the degrader agent may comprise any suitable number of amino acid units, provided that the peptide comprises a sequence having at least 75% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to SEQ ID NO: 11 (ALAPYIP) or SEQ ID NO: 12 (ALAPYIPR).

[0041]

[0078] In some embodiments, the degrader agent is a degrader peptide or component or a fragment thereof. In embodiments, the degrader agent may be a degrader peptide having a chain length of 3 to 150 amino acids, optionally 3 to 100 amino acids, optionally 5 to 50 amino acids, optionally 5 to 20 amino acids, or optionally 4 to 10 amino acids. In some embodiments, the degrader agent may include a small molecule degrader. In an example, the small molecule degrader includes a low molecular weight organic compound having a molecular weight less than or equal to 2 kDa, optionally less than or equal to 1.5 kDa, or optionally less than or equal to 1 kDa. In some embodiments, the degrader agent is characterized by a molecular weight of 100 Da to 2000 Da. In some embodiments, the degrader agent is characterized by a molecular weight of 250 Da to 1500 Da.

[0042]

[0079] As used herein, "HYDRAC" refers to Heterofunctional polYmeric DegRading Chimeras. In an aspect, HYDRACs are a subclass of PLPs containing heterologous side chains with distinct functionalities, where one domain binds to the protein of interest and a second domain targets it for degradation.

[0043]

[0080] The terms "alkylene" and "alkylene group" are used interchangeably herein and refer to a divalent group derived from an alkyl group as defined herein. The present invention includes compounds having one or more alkylene groups. The alkylene groups in some compounds function as linking groups and / or spacer groups. The compounds of the present invention may have, for example, one or more linking groups (e.g., L 1 L 2 ) as substituted and / or unsubstituted C1-C 20 alkylene, C1-C 10It may have an alkylene and a C1-C5 alkylene group.

[0044]

[0081] The terms "cycloalkylene" and "cycloalkylene group" are used synonymously herein and refer to a divalent group derived from a cycloalkyl group as defined herein. The present invention includes compounds having one or more cycloalkylene groups. In some compounds, the cycloalkyl group functions as a linking group and / or a spacer group. The compounds of the present invention may have, for example, one or more linking groups (e.g., L 1 , L 2 ) as a substituted and / or unsubstituted C3-C 20 cycloalkylene, C3-C 10 cycloalkylene, and may have a C3-C5 cycloalkylene group.

[0045]

[0082] The terms "arylene" and "arylene group" are used synonymously herein and refer to a divalent group derived from an aryl group as defined herein. The present invention includes compounds having one or more arylene groups. In some embodiments, arylene is a divalent group obtained from an aryl group by removal of hydrogen atoms from two in-ring carbon atoms of the aromatic ring of the aryl group. In some compounds, the arylene group functions as a linking group and / or a spacer group. In some compounds, the arylene group functions as a chromophore, fluorophore, aromatic antenna, dye and / or imaging group. The compounds of the present invention may have, for example, one or more linking groups (e.g., L 1 , L 2 ) as a substituted and / or unsubstituted C3-C 30 arylene, C3-C 20 arylene, C3-C 10 arylene and a C1-C5 arylene group.

[0046]

[0083] The terms "heteroarylene" and "heteroarylene group" are used interchangeably herein and refer to a divalent group derived from a heteroaryl group as defined herein. The present invention includes compounds having one or more heteroarylene groups. In some embodiments, heteroarylene is a divalent group obtained from a heteroaryl group by removal of a hydrogen atom from two carbon atoms within the aromatic ring of a heterocyclic aromatic compound or heteroaryl group or from a nitrogen atom within the ring. The heteroarylene groups in some compounds function as linking groups and / or spacer groups. The heteroarylene groups in some compounds function as chromophores, aromatic antennas, fluorophores, dyes and / or imaging groups. The compounds of the present invention include, for example, one or more linking groups (e.g., L 1 , L 2 ) and substituted and / or unsubstituted C3-C 30 heteroarylene, C3-C 20 heteroarylene, C1-C 10 heteroarylene and C3-C5 heteroarylene groups.

[0047]

[0084] The terms "alkenylene" and "alkenylene group" are used interchangeably herein and refer to a divalent group derived from an alkenyl group as defined herein. The present invention includes compounds having one or more alkenylene groups. The alkenylene groups in some compounds function as linking groups and / or spacer groups. The compounds of the present invention include, for example, one or more linking groups (e.g., L1, L2) and substituted and / or unsubstituted C2-C 20 alkenylene, C2-C 10 alkenylene and C2-C5 alkenylene groups.

[0048]

[0085] The terms "cycloalkenylene" and "cycloalkenylene group" are used interchangeably herein and refer to a divalent group derived from a cycloalkenyl group as defined herein. The present invention includes compounds having one or more cycloalkenylene groups. The cycloalkenylene groups in some compounds function as linking groups and / or spacer groups. The compounds of the present invention include, for example, one or more linking groups (e.g., L 1 、L 2 ), and substituted and / or unsubstituted C3-C 20 cycloalkenylene, C3-C 10 cycloalkenylene, and C3-C5 cycloalkenylene groups.

[0049]

[0086] The terms "alkynylene" and "alkynylene group" are used interchangeably herein and refer to a divalent group derived from an alkynyl group as defined herein. The present invention includes compounds having one or more alkynylene groups. The alkynylene groups in some compounds function as linking groups and / or spacer groups. The compounds of the present invention include, for example, one or more linking groups (e.g., L 1 、L 2 ), and substituted and / or unsubstituted C2-C 20 alkynylene, C2-C 10 alkynylene, and C2-C5 alkynylene groups.

[0050]

[0087] The term "halo" as used herein refers to a halogen group such as fluoro (-F), chloro (-Cl), bromo (-Br), iodo (-I), or astatine (-At).

[0051]

[0088] The term "heterocyclic" refers to a ring structure that contains, in addition to carbon, at least one other type of atom in the ring. Examples of such heteroatoms include nitrogen, oxygen, and sulfur. Heterocycles include heterocyclic alicyclic rings and heterocyclic aromatic rings. Examples of heterocycles include, but are not limited to, pyrrolidinyl, piperidyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, triazolyl, and tetrazolyl groups. The atoms of the heterocycle may be bonded to various other atoms and functional groups and may be provided, for example, as substituents.

[0052]

[0089] The term "carbocyclic" refers to a ring structure that contains only carbon atoms in the ring. The carbocyclic carbon atoms may be bonded to various other atoms and functional groups and may be provided, for example, as substituents.

[0053]

[0090] The term "alicyclic ring" refers to a ring or a plurality of fused rings that are not aromatic rings. Alicyclic rings include both carbocyclic and heterocyclic rings.

[0054]

[0091] The term "aromatic ring" refers to a ring or a plurality of fused rings that contain at least one aromatic ring group. The term aromatic ring includes aromatic rings that contain carbon, hydrogen, and heteroatoms. Aromatic rings include carbocyclic and heterocyclic aromatic rings. An aromatic ring is a component of an aryl group.

[0055]

[0092] The term "fused ring" or "fused ring structure" refers to a plurality of alicyclic and / or aromatic rings provided in a stereoconfiguration of a fused ring, for example, in a fused ring that shares at least two carbon atoms and / or heteroatoms within the ring.

[0056]

[0093] The term "alkoxyalkyl", as used herein, refers to a substituent of the formula alkyl-O-alkyl.

[0057]

[0094] As used herein, the term "polyhydroxyalkyl" refers to a substituent having 2 to 12 carbon atoms and 2 to 5 hydroxyl groups, for example, a 2,3-dihydroxypropyl, 2,3,4-trihydroxybutyl or 2,3,4,5-tetrahydroxypentyl residue.

[0058]

[0095] As used herein, the term "polyalkoxyalkyl" refers to a substituent having the formula alkyl-(alkoxy) n -alkoxy, where n is an integer from 1 to 10, preferably from 1 to 4, and more preferably, for some embodiments, from 1 to 3.

[0059]

[0096] Examples of amino acids include glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, asparagine, glutamine, glycine, serine, threonine, serine, threonine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid and glutamic acid. References to "side chain residues of natural α-amino acids", as used herein, specifically include the side chains of the amino acids referenced above. A peptide is composed of two or more amino acids connected via peptide bonds.

[0060]

[0097] Examples of the alkyl group include linear, branched, and cyclic alkyl groups. The alkyl group may have 1 to 30 carbon atoms. Examples of the alkyl group include small alkyl groups having 1 to 3 carbon atoms. Examples of the alkyl group include medium-length alkyl groups having 4 to 10 carbon atoms. Examples of the alkyl group include long alkyl groups having more than 10 carbon atoms, particularly those having 10 to 30 carbon atoms. The term cycloalkyl specifically refers to an alkyl(alky) group having a ring structure, for example, an alkyl group having a ring structure containing 3 to 30 carbon atoms, optionally 3 to 20 carbon atoms, and optionally 2 to 10 carbon atoms. Examples include alkyl groups having one or more rings. Examples of the cycloalkyl group include those having a 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, or 10-membered carbon ring(s), particularly those having a 3-membered, 4-membered, 5-membered, 6-membered, or 7-membered ring(s). The carbon ring in the cycloalkyl group may have an alkyl group. Examples of the cycloalkyl group include bicyclic and tricyclic alkyl groups. The alkyl group may be optionally substituted. Examples of the substituted alkyl group include those substituted with an aryl group, which may be optionally substituted in turn. Specific examples of the alkyl group include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, s-butyl, t-butyl, cyclobutyl, n-pentyl, branched pentyl, cyclopentyl, n-hexyl, branched hexyl, and cyclohexyl groups, all of which may be optionally substituted. Examples of the substituted alkyl group include fully halogenated or partially halogenated alkyl groups, for example, alkyl groups in which one or more hydrogens are replaced by one or more fluorine atoms, chlorine atoms, bromine atoms, and / or iodine atoms. Examples of the substituted alkyl group include fully fluorinated or partially fluorinated alkyl groups, for example, alkyl groups in which one or more hydrogens are replaced by one or more fluorine atoms.An alkoxy group is an alkyl group modified by linkage to oxygen, which can be represented by the formula R-O and is sometimes referred to as an alkyl ether group. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, and heptoxy. As the alkoxy group, there may be mentioned a substituted alkoxy group in which the alkyl part of the group is substituted as provided in connection with the description of the alkyl group herein. MeO- refers to CH3O- when used herein. The compositions of some embodiments of the present invention include an alkyl group as a terminal group, for example, as a polymer main chain terminal group and / or a polymer side chain terminal group.

[0061]

[0098] Examples of the alkenyl group include linear, branched, and cyclic alkenyl groups. The alkenyl group may have one, two or more double bonds, and two or more of the double bonds may be conjugated double bonds. The alkenyl group may have 2 to 20 carbon atoms. The alkenyl group may include small alkenyl groups having 2 to 3 carbon atoms. The alkenyl group may include medium-length alkenyl groups having 4 to 10 carbon atoms. The alkenyl group may include long alkenyl groups having more than 10 carbon atoms, particularly those having 10 to 20 carbon atoms. Examples of the cycloalkenyl group include those in which the double bond is in the ring or in the alkenyl group attached to the ring. The term cycloalkenyl specifically refers to an alkenyl group having a ring structure, and examples thereof include alkenyl groups having a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-membered carbon ring(s), particularly those having a 3-, 4-, 5-, 6- or 7-membered ring(s). The carbon ring in the cycloalkenyl group may have an alkyl group. The cycloalkenyl group may include bicyclic and tricyclic alkenyl groups. The alkenyl group may be optionally substituted. Examples of the substituted alkenyl group include those substituted with an alkyl or aryl group, and these groups may be optionally substituted in turn. Specific examples of the alkenyl group include ethenyl, prop-1-enyl, prop-2-enyl, cycloprop-1-enyl, but-1-enyl, but-2-enyl, cyclobut-1-enyl, cyclobut-2-enyl, pent-1-enyl, pent-2-enyl, branched pentenyl, cyclopent-1-enyl, hex-1-enyl, branched hexenyl, cyclohexenyl, and all of these may be optionally substituted. Examples of the substituted alkenyl group include fully halogenated or partially halogenated alkenyl groups, for example, alkenyl groups in which one or more hydrogens are replaced by one or more fluorine atoms, chlorine atoms, bromine atoms and / or iodine atoms.Examples of the substituted alkenyl group include a fully fluorinated or partially fluorinated alkenyl group, for example, an alkenyl group in which one or more hydrogen atoms are replaced by one or more fluorine atoms. The compositions of some embodiments of the present invention include an alkenyl group as a terminal group, for example, as a polymer main chain terminal group and / or a polymer side chain terminal group.

[0062]

[0099] Examples of the aryl group include a group having one or more 5-membered, 6-membered or 7-membered aromatic rings including a heterocyclic aromatic ring. The term heteroaryl specifically refers to an aryl group having at least one 5-membered, 6-membered or 7-membered heterocyclic aromatic ring. The aryl group may contain one or more fused aromatic rings, for example, one or more fused heteroaromatic rings, and / or a combination of one or more aromatic rings and one or more non-aromatic rings which may be fused or may be linked via a covalent bond. The heterocyclic aromatic ring may contain one or more N, O, or S atoms in the ring. Examples of the heterocyclic aromatic ring include those having 1, 2 or 3 N atoms, those having 1 or 2 O atoms, those having 1 or 2 S atoms, or those having a combination of 1, 2 or 3 N, O or S atoms. The aryl group may be optionally substituted. Examples of the substituted aryl group include those substituted with an alkyl or alkenyl group, and these groups may be optionally substituted in turn. Specific examples of the aryl group include phenyl, biphenyl group, pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, and naphthyl group, and all of these may be optionally substituted. Examples of the substituted aryl group include a fully halogenated or partially halogenated aryl group, for example, an aryl group in which one or more hydrogens are replaced by one or more fluorine atoms, chlorine atoms, bromine atoms and / or iodine atoms. Examples of the substituted aryl group include a fully fluorinated or partially fluorinated aryl group, for example, an aryl group in which one or more hydrogens are replaced by one or more fluorine atoms.Examples of aryl groups include, but are not limited to, the following: benzene, naphthalene, naphthoquinone, diphenylmethane, fluorene, anthracene, anthraquinone, phenanthrene, tetracene, tetracenedione, pyridine, quinoline, isoquinoline, indole, isoindole, pyrrole, imidazole, oxazole, thiazole, pyrazole, pyrazine, pyrimidine, purine, benzimidazole, furan, benzofuran, dibenzofuran, carbazole, acridine, acridone, phenanthridine, thiophene, benzothiophene, dibenzothiophene, xanthene, xanthone, flavone, coumarin, azulene, or anthracycline; or a group containing a heterocyclic aromatic group corresponding to any one of these. As used herein, a group corresponding to the groups explicitly listed above that include an aromatic or heterocyclic aromatic group, for example, monovalent, divalent, and polyvalent groups of the aromatic and heterocyclic aromatic groups listed herein, are provided in a configuration covalently bonded to the compounds of the present invention at any suitable point of attachment. In an embodiment, the aryl group contains 5 to 30 carbon atoms. In an embodiment, the aryl group contains a 6-membered ring of one aromatic or heterocyclic aromatic compound and one or more additional 5- or 6-membered aromatic or heteroaromatic rings. In an embodiment, the aryl group contains 5 to 18 carbon atoms in the ring. The aryl group optionally has one or more aromatic rings or heterocyclic aromatic rings having one or more electron-donating groups, electron-withdrawing groups, and / or targeting ligands provided as substituents. Compositions of some embodiments of the present invention include an aryl group as a terminal group, for example, as a polymer main chain terminal group and / or a polymer side chain terminal group.

[0063]

[0100] An arylalkyl group is an alkyl group substituted with one or more aryl groups, the alkyl group optionally having additional substituents, and the aryl group optionally being substituted. Specific alkylaryl groups are alkyl groups substituted with phenyl, for example, phenylmethyl groups. The alkylaryl group is alternatively described as an aryl group substituted with one or more alkyl groups, in which case the alkyl group optionally has additional substituents and the aryl group is optionally substituted. Specific alkylaryl groups are phenyl groups substituted with alkyl, for example, methylphenyl. Substituted arylalkyl groups include fully halogenated or partially halogenated arylalkyl groups, for example, arylalkyl groups having one or more alkyl and / or aryl groups in which one or more hydrogens are replaced with one or more fluorine, chlorine, bromine, and / or iodine atoms. The compositions of some embodiments of the present invention include an arylalkyl group as a terminal group, for example, as a polymer main chain terminal group and / or a polymer side chain terminal group.

[0064]

[0101] With respect to any of the groups described herein that contain one or more substituents, it is understood that such groups do not contain any substitutions or substitution patterns that are sterically unrealistic and / or synthetically infeasible. Optional substitution of the alkyl group includes substitution with one or more alkenyl groups, aryl groups, or both, and the alkenyl group or aryl group is optionally substituted. Optional substitution of the alkenyl group includes substitution with one or more alkyl groups, aryl groups, or both, and the alkyl group or aryl group is optionally substituted. Optional substitution of the aryl group includes substitution of the aryl ring with one or more alkyl groups, alkenyl groups, or both, and the alkyl group or alkenyl group is optionally substituted.

[0102] Optional substituents for any alkyl, alkenyl, and aryl groups include substitution with one or more of the following substituents, among which substitution with a halogen such as fluorine, chlorine, bromine, or iodine; pseudohalides such as -CN;

[0103] -COOR (wherein R is hydrogen, an alkyl group, or an aryl group, more specifically, R is a methyl, ethyl, propyl, butyl, or phenyl group, and all of these groups may optionally be substituted);

[0104] -COR (wherein R is hydrogen, an alkyl group, or an aryl group, more specifically, R is a methyl, ethyl, propyl, butyl, or phenyl group, and all of these groups may optionally be substituted);

[0105] -CON(R)2 (wherein each R is independently hydrogen, an alkyl group, or an aryl group, more specifically, R is a methyl, ethyl, propyl, butyl, or phenyl group, and all of these groups may optionally be substituted; R and R may form a ring containing one or more double bonds and may contain one or more additional carbon atoms);

[0106] -OCON(R)2 (wherein each R is independently hydrogen, an alkyl group, or an aryl group, more specifically, R is a methyl, ethyl, propyl, butyl, or phenyl group, and all of these groups may optionally be substituted; R and R may form a ring containing one or more double bonds and may contain one or more additional carbon atoms);

[0107] -N(R)2 (wherein each R is independently hydrogen, an alkyl group, an acyl group, or an aryl group, more specifically, R is a methyl, ethyl, propyl, butyl, phenyl, or acetyl group, and all of these may optionally be substituted; R and R may form a ring containing one or more double bonds and may contain one or more additional carbon atoms);

[0108] -SR (wherein R is hydrogen, an alkyl group or an aryl group, more specifically, R is hydrogen, methyl, ethyl, propyl, butyl, or a phenyl group, and these may be optionally substituted);

[0109] -SO2R, or -SOR (wherein R is an alkyl group or an aryl group, more specifically, R is methyl, ethyl, propyl, butyl, or a phenyl group, and all of these may be optionally substituted);

[0110] -OCOOR (wherein R is an alkyl group or an aryl group);

[0111] -SO2N(R)2 (wherein each R is independently hydrogen, an alkyl group, or an aryl group, and all of these may be optionally substituted, and R and R may form a ring containing one or more double bonds and may contain one or more additional carbon atoms);

[0112] -OR (wherein R is H, an alkyl group, an aryl group, or an acyl group, and all of these may be optionally substituted. In certain examples, R may be an acyl group that gives rise to -OCOR”, and R” is hydrogen, an alkyl group, or an aryl group, more specifically, R” is methyl, ethyl, propyl, butyl, or a phenyl group, and all of these groups may be optionally substituted) may be mentioned.

[0065]

[0113] Specific substituted alkyl groups include haloalkyl groups, especially trihalomethyl groups, specifically trifluoromethyl groups. Specific substituted aryl groups include mono-, di-, tri-, tetra- and pentahalophenyl groups; mono-, di-, tri-, tetra-, penta-, hexa- and hepta-halogenated naphthalene groups; 3- or 4-halophenyl groups, 3- or 4-alkylphenyl groups, 3- or 4-alkoxyphenyl groups, 3- or 4-RCO-substituted phenyl, 5- or 6-halogenated naphthalene groups. More specifically, substituted aryl groups include acetylphenyl groups, especially 4-acetylphenyl groups; fluorophenyl groups, especially 3-fluorophenyl and 4-fluorophenyl groups; chlorophenyl groups, especially 3-chlorophenyl and 4-chlorophenyl groups; methylphenyl groups, especially 4-methylphenyl groups; and methoxyphenyl groups, especially 4-methoxyphenyl groups.

[0066]

[0114] Regarding any of the above groups containing one or more substituents, it is understood that such groups do not contain any substitution or substitution pattern that is sterically unrealistic and / or synthetically infeasible.

[0067]

[0115] The term "pharmaceutically acceptable salt" means salts of the active compounds prepared with relatively non-toxic acids or bases depending on certain substituents found on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, the base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of any desired base, either in pure or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of any desired acid, either in pure or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonic acid, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, monohydrogen sulfate, hydroiodic acid, or phosphorous acid, and in addition, salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginate, and salts of organic acids such as glucuronic acid or galactunoric acid (see, for example, Berge et al., Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functional groups that allow the compound to be converted into either a base or an acid addition salt. Other pharmaceutically acceptable carriers known to those skilled in the art are suitable for the present invention. The salts tend to be more soluble in aqueous solvents or other protic solvents in the corresponding free base form. In other cases, the preparation may be a lyophilized powder in 1 mM to 50 mM histidine, 0.1% to 2% sucrose, 2% to 7% mannitol within a pH range of 4.5 to 5.5, which is combined with a buffer before use.

[0068]

[0116] Thus, the compounds, oligomers, or polymers disclosed herein may exist as salts, e.g., salts with pharmaceutically acceptable acids. Examples of such salts include hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate (e.g., a mixture including (+)-tartrate, (-)-tartrate, or a racemic mixture thereof), succinate, benzoate, and salts with amino acids such as glutamate. These salts can be prepared by methods known to those skilled in the art.

[0069]

[0117] The neutral form of the compound is preferably regenerated by contacting the salt with a base or an acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties such as solubility in polar solvents.

[0070]

[0118] In addition to the salt form, the present invention provides compounds in prodrug form. A prodrug of a compound described herein is a compound that undergoes an easy chemical change under physiological conditions to provide the compound of the present invention. In addition, a prodrug can be converted to the compound of the present invention by chemical or biochemical methods in an ex vivo environment. For example, a prodrug can be slowly converted to the compound of the present invention when placed in a transdermal patch reservoir together with a suitable enzyme or chemical reagent.

[0071]

[0119] Certain compounds, oligomers, or polymers disclosed herein may exist in an unsolvated form and, in addition, in a solvated form including a hydrate form. Generally, the solvated form is equivalent to the unsolvated form and is encompassed within the scope of the present invention. Certain compounds disclosed herein may exist in multiple crystalline or amorphous forms. Generally, all physical forms are equivalent for the intended uses herein and are intended to be within the scope of the disclosed compounds, oligomers, or polymers.

[0072]

[0120] As used herein, the term "salt" refers to the acid or base salt of a compound used in the method of the present invention. Examples useful in the description of acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, etc.) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid, etc.) salts, quaternary ammonium (methyl iodide, ethyl iodide, etc.) salts.

[0073]

[0121] Certain compounds of the present invention have an asymmetric carbon atom (optical center or chiral center) or a double bond; they can be defined as (R)- or (S)- from the perspective of absolute stereochemistry, or as D- or L- in the case of amino acids, and have enantiomers, racemic compounds, diastereomers, tautomers, geometric isomers, stereoisometric forms. Each individual isomer is included within the scope of the present invention. The compounds of the present invention do not include those that are known in the art to be too unstable to synthesize and / or isolate. The present invention means that it includes compounds in racemic form and optically pure form. Optically active (R)- and (S)-, or D- or L-isomers may be prepared using a chiral synthon or chiral reagent, or may be resolved using conventional techniques. If the compounds described herein contain an olefinic bond or other center with geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers.

[0074]

[0122] As used herein, the term "isomer" refers to compounds that have the same number and type of atoms, and thus the same molecular weight, but differ in the structural arrangement or stereostructure of the atoms. Isomers include structural isomers and stereoisomers, such as enantiomers.

[0075]

[0123] As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily convertible from one isomeric form to another.

[0076]

[0124] One skilled in the art will understand that a particular compound of the present invention may exist in tautomeric forms, and that all tautomeric forms of such compounds are within the scope of the present invention.

[0077]

[0125] Unless otherwise specified, the structures depicted herein also mean all stereochemical forms of the structure; that is, including R and S configurations for each chiral center. Therefore, in addition to single stereochemical isomers of the compounds of the present invention, enantiomers and mixtures of diastereomers are also within the scope of the present invention.

[0078]

[0126] Unless otherwise specified, the structures depicted herein also mean compounds that differ only in the presence of one or more isotope-enriched atoms. For example, replacement of hydrogen by deuterium or tritium, or replacement of carbon by 13 C- or 14 Compounds having the structure of the present invention except for replacement of carbon by C-enriched carbon are within the scope of the present invention.

[0079]

[0127] As is customary and well known in the art, the hydrogen atoms in formulas (FX1) and (S1a)-(S2c) are not necessarily explicitly shown. For example, hydrogen atoms bonded to carbon atoms of aromatic, heteroaromatic compounds, and alicyclic rings are not necessarily explicitly shown in formulas (FX1) and (S1a)-(S2c). The structures provided herein are intended to show the chemical composition of the compounds of the methods and compositions of the present invention to those of ordinary skill in the art in the context of, for example, the description of formulas (FX1) and (S1a)-(S2c) and the schematic diagrams and structures in the drawings. As understood by those skilled in the art, the structures provided do not specify the specific positions and / or orientations of the atoms of these compounds and the corresponding bond angles between the atoms.

[0080]

[0128] The compounds of the present invention may also contain unnatural ratios of atomic isotopes in one or more of the atoms constituting such compounds. For example, the compound may be, for example, tritium ( 3H), iodine-125( 125 I), or carbon-14( 14 C), etc., may be radiolabeled with a radioisotope. All isotope variations of the compounds of the present invention are included within the scope of the present invention, whether radioactive or not.

[0081]

[0129] The symbol "

Chemical Formula

[0082]

[0130] The term "treating" or "treatment" refers to any indication of success in the treatment or amelioration of an injury, disease, medical condition or state, e.g., alleviation; remission; reducing symptoms or making an injury, medical condition or state more tolerable to a subject such as a patient in need of treatment; slowing the rate of degeneration or decline; making the debility at the end point of degeneration less; improving the physical or mental health state of the subject, etc., including any objective or subjective parameters. Treatment or amelioration of symptoms may be based on objective or subjective parameters including the results of a physical examination, neuropsychiatric examination, and / or psychiatric evaluation.

[0083]

[0131] "Effective amount" means an amount sufficient to achieve the stated purpose (e.g., achieve the intended effect of its administration, treat a disease, reduce enzyme activity, increase enzyme activity, reduce transcriptional activity, increase transcriptional activity, reduce one or more symptoms of a disease or condition). Examples of "effective amount" are amounts sufficient to contribute to the treatment, prevention, or reduction of one or more symptoms of a disease, and an effective amount may also be referred to as a "therapeutically effective amount". "Reduction" (and grammatical equivalents of this phrase) of one or more symptoms means decreasing the severity or frequency of the symptom(s) or eliminating the symptom(s). A "prophylactically effective amount" of a drug is the amount of the drug that, when administered to a subject, results in the intended prophylactic effect, e.g., preventing or delaying the onset (or recurrence) of an injury, disease, medical condition or condition, or reducing the likelihood of the onset (or recurrence) of an injury, disease, medical condition, or condition, or their symptoms. A complete prophylactic effect does not necessarily occur upon administration of a single dose and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations. "Amount to decrease activity", as used herein, refers to the amount of an antagonist (inhibitor) required to decrease the activity of an enzyme or protein (e.g., a transcription factor) compared to the absence of the antagonist. "Amount to increase activity", as used herein, refers to the amount of an agonist (activator) required to increase the activity of an enzyme or protein (e.g., a transcription factor) compared to the absence of the agonist. "Amount to disrupt function", as used herein, refers to the amount of an antagonist (inhibitor) required to disrupt the function of an enzyme or protein (e.g., a transcription factor) compared to the absence of the antagonist. "Amount to increase function", as used herein, refers to the amount of an agonist (activator) required to increase the function of an enzyme or protein (e.g., a transcription factor) compared to the absence of the agonist.The correct amount is determined by the purpose of the treatment and can be ascertained by one of ordinary skill in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1 - 3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th ed., 2003, ed. Gennaro, Lippincott, Williams & Wilkins).

[0084]

[0132] As used herein, the terms "inhibit", "inhibiting", "inhibition", and grammatically equivalent variations in reference to a compound, oligomer, and / or polymer that inhibits aggregation mean to disrupt, prevent, or otherwise negatively affect (e.g., reduce) the ability of a given species, such as one or more oligo- or polypeptides (e.g., proteins), to bind together (e.g., non-covalently) or otherwise associate, compared to the level of such species association in the absence of the compound, oligomer, and / or polymer. In reference to a protein inhibitor (e.g., antagonist) interaction, the terms "inhibit", "inhibiting", "inhibition", etc. mean to negatively affect (e.g., reduce) the activity or function of a protein, compared to the activity or function of the protein in the absence of the inhibitor, as defined herein. In some embodiments, inhibition refers to a reduction in a disease or a symptom of a disease. In some embodiments, inhibition refers to a reduction in the activity of a signaling or signal transduction pathway. Thus, inhibition includes, at least in part, partially or completely blocking a stimulus, reducing activation, preventing or delaying it, or inactivating a protein, reducing its sensitivity, or downregulating its signal transduction or enzyme activity or amount.

[0085]

[0133] In reference to protein activator (e.g., agonist) interaction, terms such as "activation", "activate", "activating", etc. mean to have a positive effect (e.g., increase) on the activity or function of a protein, as defined herein.

[0086]

[0134] The term "modulator" refers to a composition that increases or decreases the level or function of a target molecule.

[0087]

[0135] "Patient", "subject", or "subject in need thereof" refers to an organism that has or is susceptible to a disease or condition that can be treated by administration of a compound or pharmaceutical composition provided herein. Non-limiting examples include humans, other mammals, cows, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human. In some embodiments, the patient is a mammal. In some embodiments, the patient is a mouse. In some embodiments, the patient is a laboratory animal. In some embodiments, the patient is a rat. In some embodiments, the patient is a test animal.

[0088]

[0136] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" are substances that assist in the administration of the active agent to the subject and its absorption by the subject, and that may be included in the compositions of the present invention without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer's solutions, normal sucrose, normal glucose, binders, diluents, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (e.g., Ringer's solutions), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and dyes, among others. Such preparations may be sterilized and, if necessary, may be mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts for influencing osmotic pressure, buffer solutions, coloring agents, and / or flavoring agents, for example, which do not react detrimentally with the compounds of the present invention. Those skilled in the art will recognize that other pharmaceutical excipients may be useful in the present invention.

[0089]

[0137] The term "preparation" is intended to include formulations of active compounds containing encapsulating materials as carriers for providing capsules, in which case, within the capsule, the active ingredient is surrounded by the carrier, with or without other carriers, and thus the carrier is in an associated state with the active ingredient. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0090]

[0138] As used herein, the term "administering" means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, nasal or subcutaneous administration, or implantation of a delayed release device, e.g., a mini osmotic pump, into a subject. Administration may be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). In embodiments, administration includes direct administration to a tumor. Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intracardiac, and intracranial. Other delivery modes include, but are not limited to, use of liposomal formulations, intravenous infusions, transdermal patches, etc. "Co-administering" means that the compositions described herein are administered simultaneously with, immediately before, or immediately after the administration of one or more additional therapeutic agents (e.g., an anti-cancer agent or chemotherapeutic agent). The compounds of the invention may be administered alone or co-administered to a patient. Co-administration means administering the compounds individually or in combination (more than one compound or agent) simultaneously or sequentially. Thus, the preparations may also be combined with other active substances (e.g., for reducing metabolic degradation) if desired. The compositions of the invention can be delivered transdermally or by a topical route and can be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols. Oral preparations include tablets, pills, powders, dragees, capsules, solutions, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for ingestion by a patient. Solid formulations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. Liquid formulations include solutions, suspensions, and emulsions, e.g., water or water / propylene glycol solutions. The compositions of the invention may additionally contain components for providing sustained release and / or comfort. Such components include high molecular weight, anionic mucomimetic polymers, gelling polysaccharides, and granulated drug carrier substrates.These components are discussed in more detail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212,162; and 4,861,760. The entire content of these patents is hereby incorporated by reference in its entirety for all purposes. The compositions of the present invention can also be delivered as microspheres for delayed release in the body. For example, the microspheres can be delivered via intradermal injection of microspheres containing a drug that is slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. 7:623-645, 1995); as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12:857-863, 1995); or as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). In another embodiment, the formulations of the compositions of the present invention may be delivered by the use of liposomes that fuse with the cell membrane, or may be incorporated by endocytosis, i.e., by employing receptor ligands attached to the liposomes that bind to surface membrane protein receptors of the cells and cause endocytosis. By using liposomes, the delivery of the compositions of the present invention to target cells in vivo can be focused on, particularly when the liposome surface has receptor ligands specific for the target cells or is otherwise preferentially directed to specific organs. (See Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587, 1989).

[0091]

[0139] As used herein, the term "conjugated", when stated with respect to two moieties, means that the two moieties are joined, where one or more linkages connecting the two moieties may be covalent or non-covalent. In embodiments, the two moieties are covalently bonded to each other (e.g., directly or through a covalently bonded mediator). In embodiments, the two moieties are bonded by non-covalent bonds (e.g., via ionic bond(s), van der Waals bond(s) / interaction, hydrogen bond(s), polar bond(s), or combinations or mixtures thereof).

[0092]

[0140] The term "about", as used herein, means a range of values that includes the specified value that would be considered by one of ordinary skill in the art to be reasonably similar to the specified value. In some aspects, about means within one standard deviation using measurements generally acceptable in the art. In some aspects, about means a range extended + / −10% from the specified value. In embodiments, about means the specified value.

[0093]

[0141] The various polymers disclosed herein are characterized in part by the relative amounts of discrete functional side chains present in the polymer. In aspects, the relative amounts of discrete functional side chains are expressed as an average ratio defined herein as the "peptide ratio". Since the degree of polymerization can vary between polymers, the composition of the monomers (and the functional side chains of said monomers) can also vary. Therefore, the peptide ratio should be understood as an average. The polymerization method and subsequent analytical methods are under the influence of random experimental error, and thus it will be understood by one of ordinary skill in the art that the peptide ratio should be read to encompass reasonable variations from the stated values. Specifically, in some aspects, the peptide ratio associated with the functional side chains of the polymer includes variations of ±20% of the stated ratio. According to this aspect, a 2:1 P 1 :P 2The ratio is 1.6:1 to 2.4:1 (e.g., 1.6:1, 1.8:1, 2:1, 2.2:1, 2.4:1) and 2:0.8 to 2:1.2 (e.g., 2:0.8, 2:0.9, 2:1, 2:1.1, 2:1.2) of P 1 :P 2 including variations in the ratio. In some embodiments, the peptide ratio related to the functional side chains of the polymer includes variations of ±10% of the stated ratio. In some embodiments, the peptide ratio related to the functional side chains of the polymer includes variations of ±5% of the stated ratio. In some embodiments, the peptide ratio related to the functional side chains of the polymer includes variations of ±1% of the stated ratio.

[0094]

[0142] In reference to a compound, oligomer, and / or polymer that promotes aggregation, the terms "promote", "promotes", "promoting", and grammatically equivalent variations, as used herein, mean to promote, facilitate, or otherwise have a positive effect (e.g., increase or speed up) the ability of one or more oligo or polypeptides (e.g., proteins) and the like of a given species to bind together (e.g., non-covalently) or otherwise associate, compared to the level of such species association in the absence of the compound, oligomer, and / or polymer. The promotion of aggregation is relevant when the aggregation is sped up to higher order aggregates, for example, past toxic species such as nanofibiril, so that the overall toxicity of the system is reduced.

[0095]

[0143] In reference to a compound, oligomer, and / or polymer that binds to one or more oligo or polypeptides (e.g., proteins) and the like of a given species, the terms "binds", "binding", and grammatically equivalent variations, as used herein, mean that the compound, oligomer, and / or polymer makes multiple non-covalent bonds to the given species. In some embodiments, the compound, oligomer, and / or polymer is capable of multiple non-covalent bonds because it has a portion with an amino acid sequence similar to and / or corresponding to a portion of the given species.

[0096]

[0144] The statement "at least a portion of each instance of P 1 independently includes at least (a specified percentage of amino acid composition similarity and / or sequence homology)", and similar language, when used herein, refers to separate segments of P 1 having the indicated amino acid composition similarity and / or sequence homology. For clarity, the specified percentage is not determined by reference to a single amino acid taken from non-related segments of P 1 .

[0097]

[0145] The statement "at least a portion of each instance of P 1 is, independently, at least one or includes (a specified amino acid sequence)", and similar language, when used herein, means that each P 1 can be either the specified amino acid sequence or any combination of the specified amino acid sequences.

[0098]

[0146] "Inducing the ubiquitination cellular mechanism of an organism", or similar language, when used herein, means activating a molecular component of the organism, such as an enzyme, to cause ubiquitination of a target oligo or polypeptide (e.g., a protein), such as tau and / or microtubulin.

[0099]

[0147] "Liquid-liquid phase separation" (LLPS), when used herein, means that initially a homogeneous solution is concentrated into droplets when it exceeds a critical saturation concentration, forming a dense (protein-rich) phase of droplets within a dilute (protein-poor) phase.

[0100]

[0148] The "aggregation region" of an oligo or polypeptide (e.g., a protein), as used herein, is that portion thereof which readily aggregates with the same or similar oligos or polypeptides to form aggregated species, including, for example, species that aggregate to amyloid or aggregated species that include amyloid (e.g., of tau and / or microtubulin protein).

[0101]

[0149] "Metaphilic", as used herein, means a compound, oligomer, or polymer that is temporarily amphiphilic, e.g., by having a hydrophobic backbone with hydrophilic side chains. In some embodiments, metaphilicity results in a globular but fluid structure, which can be useful for making cell walls permeable.

[0102] Detailed Description

[0150] In the following description, numerous specific details of the devices, device elements, and methods of the present invention are set forth in order to provide a thorough explanation of the exact nature of the invention. However, it will be apparent to those skilled in the art that the invention may be practiced without these specific details.

[0103]

[0151] Microtubule-associated protein tau has been strongly implicated in Alzheimer's disease (AD) and related dementias. When hyperphosphorylated, tau aggregates into insoluble neurofibrillary tangles (NFTs). Tau aggregation causes dysfunction of numerous cellular properties, such as aggregation-related toxicity, impairment of the cell quality control machinery, and spread of pathological "seeds" of the aggregate template to adjacent neurons across synapses, thereby causing widespread impairment of brain function. Regardless of the molecular mechanisms underlying tau neurotoxicity, promoting, reducing, or preventing tau aggregation should mitigate the etiology of dementia in AD and other tauopathies.

[0104]

[0152] Numerous hypotheses regarding the etiology and thus potential therapeutic targets in AD have been proposed regarding β-amyloid (Aβ) plaques, tau aggregation, chronic inflammation, oxidative stress, and acetylcholine abnormalities. Immunotherapy approaches targeting tau are attractive, but one concern regarding treatment in the preclinical phase is the unwanted immune response. Similarly, small molecule approaches have typically shown poor pharmacokinetics and off-target binding side effects. Here the inventors propose that PLP can be modularly designed to directly incorporate major peptide fragments involved in the etiology of AD, followed by a systematic investigation to determine the specific mechanisms driving tau aggregation in AD.

[0105]

[0153] The dynamic, unorganized peptide side chains of the PLP described herein serve as a contrast to the common view that the exact conformation of an antibody is necessary to target disordered tau, while the brush architecture provides polyvalency in binding that mimics the polyvalency of disordered proteins and is not seen in small molecules. Furthermore, the disclosed PLP has an essentially amphiphilic nature (a combination of a hydrophobic backbone with hydrophilic side chains and a globular but fluid structure). In embodiments, this is a feature that can be utilized for cell permeation or to drive materials into a phase-separated state.

[0106]

[0154] The PLP described in this specification is considered to be a therapeutic agent that targets ideal disordered proteins by having one or more, or preferably all, of the following characteristics: 1) showing binding that is tolerant of the conformation to the target protein while having antibody-like specificity, 2) the ability to alter the phase separation properties of the target protein, and 3) interaction energy with the target protein sufficient to induce a change in the energy landscape in the cellular environment by accelerating aggregation, capping aggregation, dissolving condensates, or degrading aggregates (FIG. 1A, FIG. 1B). In some aspects of the invention, polymers capable of assembly into multiple types of phase separation states are disclosed. In some aspects, the ability of the polymer to bind to the target is conformation-independent. According to this aspect, this conformation-independent feature is thought to facilitate binding to non-aggregated tau protein.

[0107]

[0155] In some embodiments, the present invention provides a polymer comprising a first polymer segment comprising at least two first repeating units, wherein each of the first repeating units of the first polymer is directly or indirectly covalently linked to a first functional side chain group comprising a peptide that (i) inhibits the aggregation of, (ii) promotes the aggregation of, (iii) binds to, and / or (iv) mimics at least a portion of a tau protein. In some embodiments, the present invention provides a polymer comprising a first polymer segment comprising at least two first repeating units, wherein each of the first repeating units of the first polymer is directly or indirectly covalently linked to a first functional side chain group comprising a peptide that (i) inhibits the aggregation of, (ii) promotes the aggregation of, (iii) binds to, and / or (iv) mimics at least a portion of a microtubule tubulin protein. In some embodiments, the present invention provides a polymer comprising a first polymer segment comprising at least two first repeating units, wherein each of the first repeating units of the first polymer comprises a first polymer backbone subunit directly or indirectly covalently linked to a first functional side chain group comprising a peptide; the peptide comprises a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to one or more of SEQ ID NOs: 1-16. The polymers of the present invention may be of any suitable type of polymer described herein, may contain any suitable number of monomers, or may be derived therefrom. For example, in some embodiments, the polymer is a homopolymer (i.e., derived from / incorporating one type of monomer). Alternatively, in some embodiments, the polymer may be a copolymer comprising more than one type of monomer (e.g., 2-10 types of monomers) (e.g., derived from / incorporating them). It will be understood that the polymers of the present invention may have any suitable configuration with the attached polymer side chains.For example, in some embodiments where the polymer is a homopolymer, the polymer may be a brush polymer. In other embodiments where the polymer is a copolymer, the polymer may be a brush block copolymer or a brush random / statistical copolymer.

[0108]

[0156] In some aspects, the polymer is P selected from the range of 50% to 100% 1 characterized by peptide density and, optionally for some embodiments, by a density selected from the range of 75% to 100%, or, optionally for some embodiments, by a density selected from the range of 90% to 100%. In an aspect, the polymer is P selected from the range of 50% to 100% 2 characterized by peptide density and, optionally for some embodiments, by a density selected from the range of 75% to 100%, or, optionally for some embodiments, by a density selected from the range of 90% to 100%.

[0109]

[0157] In some embodiments, a peptide having a functional side chain group includes any suitable number of amino acid units as long as the peptide includes a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to a sequence found in tau protein or microtubulin protein. In some embodiments, a peptide having a functional side chain group includes any suitable number of amino acid units as long as the peptide includes a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to one or more of SEQ ID NOs: 1-5 or SEQ ID NO: 13. In some embodiments, a peptide having a functional side chain group includes any suitable number of amino acid units as long as the peptide includes a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to one or more of SEQ ID NOs: 6-7 or SEQ ID NO: 16. In some embodiments, a peptide having a functional side chain group includes any suitable number of amino acid units as long as the peptide includes a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to one or more of SEQ ID NOs: 8-10 or SEQ ID NOs: 14-15. In some embodiments, a peptide having a functional side chain group includes any suitable number of amino acid units as long as the peptide includes a sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or 100%) sequence identity to one or more of SEQ ID NOs: 11-12. In some embodiments, the peptide includes at least 5 amino acid residues. For example, the peptide includes 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, 24 or more, 25 or more, 26 or more, 27 or more, 28 or more, 29 or more, 30 or more, 31 or more, 32 or more, or 33 or more amino acid units.Alternatively, or in addition, the peptide may comprise 100 or fewer amino acid units, such as, for example, 90 or fewer, 80 or fewer, 70 or fewer, 60 or fewer, 59 or fewer, 58 or fewer, 57 or fewer, 56 or fewer, 55 or fewer, 54 or fewer, 53 or fewer, 52 or fewer, 51 or fewer, 50 or fewer, 49 or fewer, 48 or fewer, 47 or fewer, 46 or fewer, 45 or fewer, 44 or fewer, 43 or fewer, 42 or fewer, 41 or fewer, 40 or fewer amino acid units. Thus, the peptide may comprise a number of amino acid units bounded by any two of the aforementioned upper and lower limits. For example, the peptide may comprise 5 to 100 amino acid units, such as, for example, 5 to 100, 5 to 90, 5 to 80, 5 to 70, 5 to 60, 5 to 50, 5 to 40, 5 to 30, 5 to 20, 5 to 16, 5 to 15, 5 to 14, 6 to 100, 6 to 90, 6 to 80, 6 to 70, 6 to 60, 6 to 50, 6 to 40, 6 to 30, 6 to 20, 6 to 16, 6 to 15, 6 to 14, 7 to 100, 7 to 90, 7 to 80, 7 to 70, 7 to 60, 7 to 50, 7 to 40, 7 to 30, 7 to 20, 7 to 16, 7 to 15, 7 to 14, 8 to 100, 10 to 100, 10 to 90, 10 to 80, 10 to 70, 10 to 60, 10 to 50, 10 to 40, 10 to 30, 10 to 20, 10 to 16, 10 to 15, 10 to 14, 11 to 16, 11 to 15, 11 to 14, 12 to 20, 12 to 16, 12 to 15, or 12 to 14 amino acid units. In some embodiments, the peptide comprises 6 to 20 amino acid residues. In certain embodiments, the peptide comprises 6 to 16 amino acid residues.

[0110]

[0158] The functional side groups that make up the peptide may have any suitable structure described herein (e.g., primary, secondary, tertiary, or quaternary structure). The peptide may be a branched peptide, a linear peptide, a cyclic peptide, or a cross-linked peptide. In some embodiments, the polymer is characterized by a structure in which at least a portion of the peptide is linked to the polymer backbone group via an enzymatically degradable linker, such as a matrix metalloproteinase (MMP) cleavage sequence, a cathepsin B cleavage sequence, an ester bond, a disulfide bond that is a reduction-sensitive bond, an imine bond that is a pH-sensitive bond, or any combination thereof. In other embodiments, the polymer is characterized by a structure in which at least a portion of the peptide side chain is linked to the polymer backbone or consists of a degradable or inducible linker.

[0111]

[0159] In some aspects, the peptide includes a sequence having a hydrophobic region, such as a leucine-rich region. In an aspect, the hydrophobic region may be modified to substitute a hydrophilic amino acid residue or a non-hydrophobic amino acid residue. Such substitutions are thought to facilitate improvement of the solubility of the polymer, if necessary for a particular application. Even in embodiments where excellent stability is desired, the polymer may be modified. Acceptable polymer modifications include asparagine β-hydroxylation, a higher degree of polymerization (e.g., greater than 10 DP, greater than 15 DP, greater than 30 DP, or greater than 45 DP), single point mutations, and other suitable modifications.

[0112]

[0160] In addition, the peptide may include one or more gaps in its sequence. For example, the peptide may include five consecutive amino acid residues that do not affect or contribute to the properties of the peptide sequence. In an aspect, the one or more gaps are spacer molecules, five or fewer amino acid residues, three or fewer amino acid residues, or a combination thereof.

[0113]

[0161] In some specific embodiments, the polymer comprises tags for imaging and / or analysis. In an aspect, the polymer comprises a fluorescein-based, biotin-based, or rhodamine-based tag that results in a fluorescently labeled PLP. For example, each polymer segment B of formula (FX1) 1 , B 2 , or B 3 may independently comprise a tag for imaging and / or analysis. Similarly, each P of formula (FX1) 1 or P 2 may independently comprise a tag for imaging and / or analysis. Additionally, each T of any of the formulas described herein 1 or T 2 may independently comprise a tag for imaging and / or analysis. For example, the polymer may comprise one or more of a dye, a radiolabeling agent, a contrast agent, a titrant, etc.

[0114]

[0162] The polymer of the present invention may have any suitable degree of polymerization. If the degree of polymerization is too low, the molecular weight of the polymer is lower than the clearance threshold through the kidney, so the polymer may not have resistance to adhesion, or may not have resistance to enzymatic cleavage by proteases, or may be eliminated from the body too rapidly. In addition, if it is too low, the polymer may exhibit poor solubility and structural instability. Alternatively, if the degree of polymerization is too high, the peptide side chain groups presented on the polymer may be too dense to engage its biological targets such as cell receptors, enzymes, PPIs, etc. In addition, a high degree of polymerization may result in a polymer that is too large to penetrate cells. Typically, the polymer has a degree of polymerization of 2 to 1000 (e.g., 2 to 500, 2 to 250, 2 to 100, 2 to 60, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 60, 5 to 50, 5 to 45, 5 to 30, 7 to 45, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30). In certain embodiments, the polymer has a degree of polymerization of 5 to 100. In preferred embodiments, the polymer has a degree of polymerization of 7 to 30. For example, the polymer may have a degree of polymerization of 2 or about 2, 5 or about 5, 10 or about 10 (e.g., 11), 15 or about 15 (e.g., 17), 20 or about 20, 30 or about 30, 50 or about 50, 60 or about 60, 100 or about 100, 150 or about 150, or 200 or about 200. In some embodiments, the polymer has a degree of polymerization of 2 to 50. In certain embodiments, the polymer has a degree of polymerization of at least 5. In other certain embodiments, the polymer has a degree of polymerization of at least 7.

[0115]

[0163] In addition, in an aspect, the polymer comprises a brush density greater than or equal to 50% (e.g., greater than or equal to 60%, greater than or equal to 65%, greater than or equal to 70%, greater than or equal to 75%, greater than or equal to 80%, greater than or equal to 85%, or greater than or equal to 90%), optionally for some embodiments, a density greater than or equal to 70%, or optionally for some embodiments, a density greater than or equal to 90%. The brush polymer of a particular aspect has a brush density selected from the range of 50% to 100%, optionally for some embodiments, a density selected from the range of 75% to 100%, or optionally for some embodiments, a density selected from the range of 90% to 100%. The brush polymer of a preferred aspect has a brush density selected from the range of 75% to 100%. The brush polymer of a particular aspect has a "high brush density" selected from the range of 90% to 100%, optionally for some embodiments, a density selected from the range of 95% to 100%, or optionally for some embodiments, a density selected from the range of 99% to 100%. For example, in an aspect of the present invention, the polymer can be characterized by a formulation in which 90% of its polymer segments comprise a polymer backbone group covalently linked to a functional side chain group, where each polymer segment may contain the same or different functional side chain groups. In an aspect, the functional side chain group comprises a peptide comprising a sequence having at least 75% sequence identity to a sequence found in a tau protein such as a portion of K18 tau. In an aspect, the functional side chain group comprises a peptide comprising a sequence having at least 75% sequence identity to a sequence found in a microtubulin protein such as SEQ ID NO: 6. In some embodiments, the brush density of the polymer is equal to the peptide density of a particular peptide (e.g., all polymer segments of the polymer are P 1(comprising a polymer backbone covalently linked to a polymer side chain containing). In other embodiments, the brush density of the polymer is different from the peptide density of a particular peptide (e.g., at least one polymer segment is P 1 comprising a polymer backbone covalently linked to a polymer side chain containing, and at least one other polymer segment is P 2 comprising a polymer backbone covalently linked to a polymer side chain containing, P 1 and P 2 are characterized by different sequences).

[0116]

[0164] In addition, for each of the polymers characterized by formula (FX1) described herein, the first repeating unit (i.e.,

Chemical formula

Chemical formula

Chemical formula

[0117]

[0165] The polymer main chain group units (e.g., B of the polymers characterized by formula (FX1) and / or substructures (S1a) to (S2c)) 1 , B 2 , and B 3 each can be independently selected from any suitable polymer main chain sub-units. In an aspect, each of the first polymer main chain sub-unit, the second polymer main chain sub-unit, and the third polymer main chain sub-unit (subusit) may be a monomer capable of undergoing ring-opening metathesis. For example, each of B 1 , B 2 , and B 3 can be independently a substituted or unsubstituted norbornene, oxanorbornene, olefin, cyclic olefin, cyclooctene, or cyclopentadiene. In some aspects, each of the first polymer main chain group sub-unit, the second polymer main chain group sub-unit, and / or the third polymer main chain group sub-unit is a polymerized norbornene dicarboximide monomer. In some embodiments, each polymer main chain sub-unit of the polymer is a polymerized norbornene dicarboximide monomer. In an aspect where the polymer has poor solubility, one or more of the polymer main chain sub-units may be replaced with an oxanorbornene-based sub-unit (if not already used) or other suitable hydrophilic main chain sub-units.

[0118]

[0166] Preferably, in any embodiment of the polymers, methods, uses, compositions, or medicaments disclosed herein, the polymer is stable to enzymatic digestion. Optionally, in any embodiment of the polymers, methods, uses, compositions, or medicaments disclosed herein, the polymer is stable to enzymatic digestion by metalloproteinases. Optionally, in any embodiment of the polymers, methods, uses, compositions, or medicaments disclosed herein, the polymer is stable to enzymatic digestion by matrix metalloproteinases and thermolysin. Preferably, in any embodiment of the polymers, methods, uses, compositions, or medicaments disclosed herein, the polymer is stable to enzymatic digestion for at least 450 minutes. Optionally, in any arbitrary embodiment of the polymers, methods, uses, compositions, or medicaments disclosed herein, when a plurality of said polymers are dispersed in water, each polymer is individually solvated by water.

[0119]

[0167] In another aspect, the present invention provides a pharmaceutical composition comprising one or more of the peptides and / or one or more of the polymers described herein. In some embodiments, the composition comprises one or more pharmaceutically acceptable excipients. For example, the peptides and / or polymers of the present invention may be formulated for parenteral administration, for example, for intravenous (IV) administration or for administration into the body cavity or lumen of an organ. Alternatively, the peptides and / or polymers may be injected into the tumor. Formulations for injection generally comprise a solution of the peptide and / or polymer dissolved in a pharmaceutically acceptable carrier. Acceptable vehicles and solvents that may be employed include, among others, water and isotonic sodium chloride. In addition, sterile, fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectable compositions. These solutions are sterile and generally free of undesirable matter. These formulations may be sterilized by conventional, well-known sterilization techniques. The formulations may contain pharmaceutically acceptable adjuncts as required to approximate physiological conditions, such as pH adjusters and buffers, tonicity adjusters, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, and the like. The concentrations of the peptides and / or polymers in these formulations can be widely varied and are selected primarily based on the volume of fluid, viscosity, body weight, etc., in accordance with the particular mode of administration selected and the needs of the patient. In certain embodiments, the concentration of the peptide and / or polymer in the solution formulation for injection ranges from 0.1% (w / w) to 10% (w / w) or from about 0.1% (w / w) to about 10% (w / w).

[0120]

[0168] The present invention further provides a method for producing the polymers of the present invention disclosed herein. In an aspect of the method for producing the polymers of the present invention, at least one peptide is capped at its terminus with any suitable polymerizable monomer. In an aspect, the polymerizable monomer may comprise an ethylenically unsaturated monomer. In an aspect, the polymerizable monomer may comprise an olefin-based functional group, norbornene amide hexanoic acid, (meth)acrylate, or norbornene dicarboxamide. The polymerizable monomer may be polymerized by ROMP, RAFT, or ATRP, and this aspect is further described in Kammeyer et al., Polymerization of Protecting-Group-Free Peptides via ROMP, Polym. Chem. 2013, 4(14), 3929-3933 and Nomura et al., Precise Synthesis of Polymers Containing Functional End Groups by Living Ring-Opening Metathesis Polymerization (ROMP): Efficient Tools for Synthesis of Block / Graft Copolymers, Polym. 2010, 51(9), 1861-1881, each of which is incorporated herein by reference in its entirety to the extent that it does not conflict with the description set forth herein, more specifically with respect to the techniques for making polymers.

[0121]

[0169] After polymerization, the polymers of the present invention can be characterized using any suitable technique(s). Typically, the polymers of the present invention are characterized by SDS-PAGE or size exclusion chromatography combined with multi-angle light scattering (SEC-MALS), sometimes also referred to as gel permeation chromatography (GPC), to confirm the degree of polymerization (DP) and the molecular weight distribution (dispersity or Mw / Mn). Alternatively, or in addition, the polymers of the present invention can be characterized by SDS-PAGE to confirm the degree of polymerization (DP) and the molecular weight. Preferably, there is a match between the obtained DP and the theoretical DP based on the initial monomer to initiator ratio ([M]0 / [I]0).

[0122]

[0170] The present invention further provides a method for using the polymers of the present invention disclosed herein. In an aspect, the polymers of the present invention can be used as a therapeutic agent, a PPI disintegrant, an agent for detecting tau or microtubulin aggregation, an agent for promoting tau or microtubulin aggregation, or any combination thereof. In some embodiments, the methods described herein can be used to treat or manage a neurodegenerative disease or condition, such as Alzheimer's disease (AD). In some embodiments, the methods described herein can be used to treat or manage a tauopathy-related disease or condition. The method includes administering a therapeutically effective amount of the polymers described herein and a pharmaceutically acceptable excipient to a subject, cell, or tissue in need thereof. In some aspects, a suitable dosage of the pharmaceutical composition is determined in a conventional manner based on factors such as the subject's condition, immune status, weight, and age. For example, the amount of polymer or peptide required to be administered to treat or manage a neurodegenerative disease or condition in a subject varies depending on factors such as the risk and severity of the underlying condition(s), any other medical condition or disease, age, the form of the composition, and any other drug therapies being administered. Further, such amounts can vary depending on whether the polymer or peptide is being used for treatment (where higher dosages may be possible) or whether the polymer or peptide is being used as secondary prevention / maintenance (where lower dosages may be possible). However, the amount required can be readily set by a medical practitioner. For example, the method may include administering the polymer such that a dosage in the range of 10 ng / kg to 50 mg / kg is provided to the subject. For example, the dosage of the polymer may be in the range of 5 mg / kg to 50 mg / kg, 10 μg / kg to 5 mg / kg, or 100 μg / kg to 1 mg / kg. The dosage of the polymer may also be outside of these ranges depending on the particular polymer and, in addition, the type of disease being treated. The frequency of administration may range from a single dose per week to multiple doses per week, or may be more frequent. In some embodiments, the polymer is administered from about once per month to about five times per week.In some embodiments, the polymer is administered once per week.

[0123]

[0171] The polymer may be administered to the subject by oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, nasal or subcutaneous administration, or implantation of a sustained release device, such as a mini-osmotic pump. In some embodiments, the polymer is administered intravenously, subcutaneously, intramuscularly, topically, orally, or in combination thereof. In some embodiments, the polymer is administered to the subject's brain, spinal cord, cerebrospinal fluid, or any combination thereof. The methods described herein may include contacting a target tissue of the subject with the polymer or a metabolite or product thereof, contacting a target cell of the subject with the polymer or a metabolite or product thereof, and / or contacting a target receptor of the subject with the polymer or a metabolite or product thereof, and / or contacting a target peptide of the subject with the polymer or a metabolite or product thereof. In embodiments, the polymers described herein cross cell membranes and contact intracellular targets. Without wishing to be bound by any particular theory, the structure and charge of the PLP described herein are thought to play an essential role in providing cell permeability.

[0124]

[0172] Aspects of the present invention

[0173] Various aspects are contemplated herein, some of which are described in the following paragraphs. It is expressly contemplated that any aspect or combination of some of them can form an aspect. Further, although the following aspects are divided into aspects A, B, C, D, etc., it is expressly contemplated that the aspects in each of the sub-categories A, B, C, D, etc. can be combined in any form. Further, the term "any of the foregoing aspects" means any aspect that appears before the aspect containing such a phrase (in other words, the sentence "Aspect B13: the method described in any one of Aspects B1~B12 or any of the foregoing aspects, ~" means that any aspect before Aspect B13 including all of Aspects B1~B12 and Aspect "A" is referred to). For example, optionally, any method or composition of any of the following aspects can be useful in combination with or in combination with any other aspect provided below. Further, for example, it is contemplated that any embodiment described elsewhere in this specification including before this paragraph can optionally be combined with any of the aspects listed below. In some cases of the following aspects or elsewhere in this specification, two open-ended ranges are disclosed and these can be combined into one range. For example, "at least X" is disclosed and this can be combined with "less than Y" to form a range, where X and Y are numerical values. For the purpose of forming a range herein, the combination of "at least X" and "less than Y" is expressly contemplated to form a range of X~Y including the X value and the Y value even if "less than Y" alone does not include Y.

[0125]

[0174] Aspect A1: A polymer comprising a first repeating unit comprising a first polymer main chain subunit directly or indirectly covalently linked to a first functional side chain containing a peptide that (i) inhibits the aggregation of at least a part of the tau protein, (ii) promotes the aggregation of the tau protein, (iii) binds to the tau protein, and / or (iv) mimics the tau protein.

[0126]

[0175] Aspect B1: A polymer comprising a first repeating unit comprising a first polymer main chain subunit directly or indirectly covalently linked to a first functional side chain containing a peptide that (i) inhibits the aggregation of at least a portion of the microtubule protein, (ii) promotes the aggregation of at least a portion of the microtubule protein, (iii) binds to at least a portion of the microtubule protein, and / or (iv) mimics at least a portion of the microtubule protein.

[0127]

[0176] Aspect C1: A polymer characterized by formula (FX1):

Chemical formula

[0128]

[0177] Aspect C2: Each instance of P 1 independently comprises 5 to 50 amino acids (e.g., 5 to 50, 5 to 40, 5 to 30, 5 to 20, 6 to 50, 6 to 40, 6 to 30, or 6 to 20 amino acids), the polymer according to Aspect C1 or any of the preceding aspects.

[0129]

[0178] Aspect C3: At least one P 1 is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) and comprises a sequence having at least 75% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) sequence identity to any of the above, the polymer according to Aspect C1 or C2 or any of the preceding aspects.

[0130]

[0179] Aspect C4: At least one P 1 is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) and has a sequence identity of 85% or more (e.g., 85% or more, 90% or more, 95% or more, or 99% or more) with any one of the sequences described in Aspect C1 - C3 or any of the foregoing aspects, and a polymer comprising the sequence.

[0131]

[0180] Aspect C5: At least one P 1 is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) The polymer according to any one of Aspects C1 to C4 or any of the aforementioned aspects, comprising:

[0132]

[0181] Aspect C6: P 1 At least a part of each example of SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); SEQ ID NO: 16 (YQQYQDATADEQGRR); or Any combination thereof The polymer according to any one of Aspects C1 to C5 or any of the aforementioned aspects, comprising:

[0133]

[0182] Aspect C7: P 1 At least 75% (e.g., 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, or 99% or more) of all cases of SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); Sequence number 6 (YQQYQDATADEQG); Sequence number 7 (YQQYQDATADEQGRRR); Sequence number 8 (WMINK); Sequence number 9 (WMINKRR); Sequence number 10 (VQPINK); Sequence number 13 (VQIVYKRR); Sequence number 14 (DRWMINKRR); Sequence number 15 (VQPINKR); Sequence number 16 (YQQYQDATADEQGRR); or any combination thereof A polymer according to any one of Aspects C1 to C6 or any of the foregoing aspects, comprising:

[0134]

[0183] Aspect C8: At least one P 1 is Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR) A polymer according to any one of Aspects C1 to C7 or any of the foregoing aspects, comprising a sequence having at least 75% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) sequence identity to any of the sequences of Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR).

[0135]

[0184] Aspect C9: At least one P 1 is Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR) A polymer according to any one of Aspects C1 to C8 or any of the foregoing aspects, comprising a sequence having at least 85% (e.g., at least 85%, at least 90%, at least 95%, or at least 99%) sequence identity to any of the sequences of Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR).

[0136]

[0185] Aspect C10: At least one P 1 is SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 10 (VQPINK); or SEQ ID NO: 15 (VQPINKR) and is a polymer according to any one of Aspects C1 to C9 or any of the preceding aspects.

[0137]

[0186] Aspect C11: At least a part of each instance of P 1 is independently SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 15 (VQPINKR); or any combination thereof and is a polymer according to any one of Aspects C1 to C10 or any of the preceding aspects.

[0138]

[0187] Aspect C12: At least 75% (for example, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, or 99% or more) of all instances of P 1 is, on a numerical basis SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 15 (VQPINKR); or any combination thereof and is a polymer according to any one of Aspects C1 to C11 or any of the preceding aspects.

[0139]

[0188] Aspect C13: At least one P 1 includes SEQ ID NO: 1 (VQIVYK) or SEQ ID NO: 2 (DRVQIVYKRR); and at least one other P 1 includes SEQ ID NO: 8 (WMINK) or SEQ ID NO: 9 (WMINKRR) A polymer according to any one of embodiments C1 to C12 or any of the preceding embodiments, comprising

[0140]

[0189] Embodiment C14: A polymer according to embodiment C13 or any of the preceding embodiments, characterized by a proportional amount of at least one P 1 and at least one other P 1

[0141]

[0190] Embodiment C15: A polymer according to embodiment C13 or any of the preceding embodiments, characterized by a peptide ratio of at least one P of 1:14 1 : at least one other P 1 For example, a peptide ratio in the range of 0.8:14 to 1.2:14 or 1:11.2 to 1:16.8.

[0142]

[0191] Embodiment C16: A polymer according to embodiment C14 or any of the preceding embodiments, characterized by a peptide ratio of at least one P of 7:8 1 : at least one other P 1 For example, a peptide ratio in the range of 5.6:8 to 8.4:8 or 7:6.4 to 7:9.6.

[0143]

[0192] Embodiment C17: A polymer according to embodiment C14 or any of the preceding embodiments, characterized by a peptide ratio of at least one P of 14:1 1 : at least one other P 1 For example, a peptide ratio in the range of 11.2:1 to 16.8:1 or 14:0.8 to 14:1.2.

[0144]

[0193] Embodiment C18: A polymer according to any one of embodiments C1 to C17 or any of the preceding embodiments, wherein at least one P 1 comprises SEQ ID NO: 4 (VQIINKRR) or SEQ ID NO: 3 (VQIINK); and at least one other P 1 comprises SEQ ID NO: 8 (WMINK) or SEQ ID NO: 9 (WMINKRR)

[0145]

[0194] Aspect C19: At least one P 1 and at least one other P 1 The polymer according to Aspect C18 or any of the preceding aspects, characterized by a proportional amount of.

[0146]

[0195] Aspect C20: At least one P of 1:14 1 : At least one other P 1 The polymer according to Aspect C18 or any of the preceding aspects, characterized by a peptide ratio of. For example, a peptide ratio in the range of 0.8:14 to 1.2:14 or 1:11.2 to 1:16.8.

[0147]

[0196] Aspect C21: At least one P of 7:8 1 : At least one other P 1 The polymer according to Aspect C18 or any of the preceding aspects, characterized by a peptide ratio of. For example, a peptide ratio in the range of 5.6:8 to 8.4:8 or 7:6.4 to 7:9.6.

[0148]

[0197] Aspect C22: At least one P of 14:1 1 : At least one other P 1 The polymer according to Aspect C18 or any of the preceding aspects, characterized by a peptide ratio of. For example, a peptide ratio in the range of 11.2:1 to 16.8:1 or 14:0.8 to 14:1.2.

[0149]

[0198] Aspect C23: At least one P 1 is comprising SEQ ID NO: 10 (VQPINK) or SEQ ID NO: 15 (VQPINKR); at least one other P 1 is SEQ ID NO: 3 (VQIINK), SEQ ID NO: 4 (VQIINKRR), SEQ ID NO: 5 (KVQIINKKLDRR), SEQ ID NO: 1 (VQIVYK), or SEQ ID NO: 2 (DRVQIVYKRR) The polymer according to any one of Aspects C1 to C22 or any of the preceding aspects, comprising.

[0150]

[0199] Aspect C24: at least one P 1 and at least one other P 1 The polymer according to aspect C23 or any of the preceding aspects, characterized by a proportional amount of

[0151]

[0200] Aspect C25: at least one P of 1:14 1 : at least one other P 1 The polymer according to aspect C23 or any of the preceding aspects, characterized by a peptide ratio of

[0152]

[0201] Aspect C26: at least one P of 8:7 1 : at least one other P 1 The polymer according to aspect C23 or any of the preceding aspects, characterized by a peptide ratio of

[0153]

[0202] Aspect C27: at least one P of 14:1 1 : at least one other P 1 The polymer according to aspect C23 or any of the preceding aspects, characterized by a peptide ratio of

[0154]

[0203] Aspect C28: P 2 In each case, when present, independently contains 5 to 50 amino acids (for example, 5 to 50, 5 to 40, 5 to 30, 5 to 20, 6 to 50, 6 to 40, 6 to 30, or 6 to 20 amino acids), the polymer according to any one of aspects C1 to C27 or any of the preceding aspects.

[0155]

[0204] Aspect C29: o is an integer from 1 to 1000 (for example, 1 to 500, 1 to 250, 1 to 100, 1 to 50, 1 to 30, 2 to 1000, 2 to 500, 2 to 250, 2 to 100, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 50, 5 to 30, 7 to 1000, 7 to 500, 7 to 250, 7 to 100, 7 to 50, 7 to 30, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30); At least one P 2 comprises a sequence capable of inducing the ubiquitination cellular mechanism of an organism, and optionally, the amino acid sequence binds to an E3 ubiquitin ligase, the polymer according to any one of Aspects C1 to C28 or any of the preceding aspects.

[0156]

[0205] Aspect C30: The sequence capable of inducing the ubiquitination cellular mechanism of an organism is SEQ ID NO: 11 (ALAPYIP) or SEQ ID NO: 12 (ALAPYIPRR) for which (i) it has at least 75% (for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) or at least 90% (for example, at least 90%, at least 95%, or at least 99%) amino acid composition similarity, (ii) it has at least 60% (for example, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) or at least 80% (for example, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) sequence identity, or (iii) it contains, the polymer according to Aspect C29 or any of the preceding aspects.

[0157]

[0206] Aspect C31: At least one P 1comprises SEQ ID NO:6 (YQQYQDATADEQG), SEQ ID NO:7 (YQQYQDATADEQGRRR), SEQ ID NO:16 (YQQYQDATADEQGRR), SEQ ID NO:3 (VQIINK), SEQ ID NO:4 (VQIINKRR), SEQ ID NO:5 (KVQIINKKLDRR), SEQ ID NO:1 (VQIVYK), or SEQ ID NO:2 (DRVQIVYKRR); at least one P 2 is a polymer according to embodiment C30 or any of the preceding embodiments, comprising SEQ ID NO:11 (ALAPYIP) or SEQ ID NO:12 (ALAPYIPRR).

[0158]

[0207] Embodiment C32: o is an integer from 1 to 1000 (such as 1 to 500, 1 to 250, 1 to 100, 1 to 50, 1 to 30, 2 to 1000, 2 to 500, 2 to 250, 2 to 100, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 50, 5 to 30, 7 to 1000, 7 to 500, 7 to 250, 7 to 100, 7 to 50, 7 to 30, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30), and the polymer has a P 1 :P 2 peptide ratio characterized by the peptide ratio of, for example, in the range of 11.2:1 to 16.8:1 or 14:0.8 to 14:1.2. A polymer according to any one of embodiments C1 to C31 or any of the preceding embodiments.

[0159]

[0208] Embodiment C33: o is an integer from 1 to 1000 (such as 1 to 500, 1 to 250, 1 to 100, 1 to 50, 1 to 30, 2 to 1000, 2 to 500, 2 to 250, 2 to 100, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 50, 5 to 30, 7 to 1000, 7 to 500, 7 to 250, 7 to 100, 7 to 50, 7 to 30, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30), and the polymer has a P 1 :P 2A polymer according to any one of embodiments C1 - C31 or any of the preceding embodiments, characterized by a peptide ratio. For example, a peptide ratio in the range of 10.4:2 to 15.6:2 or 13:1.6 to 13:2.4.

[0160]

[0209] Embodiment C34: o is an integer from 1 to 1000 (for example, 1 to 500, 1 to 250, 1 to 100, 1 to 50, 1 to 30, 2 to 1000, 2 to 500, 2 to 250, 2 to 100, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 50, 5 to 30, 7 to 1000, 7 to 500, 7 to 250, 7 to 100, 7 to 50, 7 to 30, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30), and the polymer has a P of 5:1 1 :P 2 A polymer according to any one of embodiments C1 - C31 or any of the preceding embodiments, characterized by a peptide of P

[0161]

[0210] Embodiment C35: o is an integer from 1 to 1000 (for example, 1 to 500, 1 to 250, 1 to 100, 1 to 50, 1 to 30, 2 to 1000, 2 to 500, 2 to 250, 2 to 100, 2 to 50, 2 to 30, 5 to 1000, 5 to 500, 5 to 250, 5 to 100, 5 to 50, 5 to 30, 7 to 1000, 7 to 500, 7 to 250, 7 to 100, 7 to 50, 7 to 30, 20 to 500, 20 to 250, 20 to 100, 20 to 50, or 20 to 30), and the polymer has a P of 3:1 1 :P 2 A polymer according to any one of embodiments C1 - C31 or any of the preceding embodiments, characterized by a peptide ratio. For example, a peptide ratio in the range of 2.4:1 to 3.6:1 or 3:0.8 to 3:1.2.

[0162]

[0211] Embodiment C36: A polymer according to any one of embodiments A1 - C35, wherein the tau protein and / or microtubulin protein is in the form of an oligomer, protofibril, amyloid fiber, cross-β sheet amyloid species, or any combination thereof.

[0163]

[0212] Aspect C37: When the tau protein is in a liquid-liquid phase separation (LLPS) state, at least one P 1 is, independently, or in combination with other instances of P 1 a polymer according to any one of Aspects C1 - C36 or any of the preceding aspects, which (a) for at least a part of the tau protein, (i) inhibits its aggregation, (ii) promotes its aggregation, (iii) binds to it, and / or (iv) mimics it.

[0164]

[0213] Aspect C38: A polymer according to Aspect C37 or any of the preceding aspects that disrupts and / or prevents further growth of the LLPS state of the tau protein.

[0165]

[0214] Aspect C39: A polymer according to any one of Aspects A1 - C38 that is capable of being in an LLPS state under physiological conditions.

[0166]

[0215] Aspect C40: (a) Whether at least a part of the tau protein contains an aggregating region of the tau protein, (b) Whether at least a part of the microtubulin tau protein contains an aggregating region of the microtubulin tau protein; or (c) A combination thereof A polymer according to any one of Aspects A1 - C39.

[0167]

[0216] Aspect C41: At least one P 1 and / or at least one P 2 is characterized by a net positive charge, a polymer according to any one of Aspects C1 - C40 or any of the preceding aspects.

[0168]

[0217] Aspect C42: At least one P 1 and / or at least one P 2The polymer according to any one of aspects C1 to C41 or any of the preceding aspects, characterized by a net positive charge of 1 to 5 (for example, a net positive charge of 1, 2, 3, 4, or 5).

[0169]

[0218] Aspect C43: P 1 The polymer according to any one of aspects C1 to C42 or any of the preceding aspects, wherein at least 75% (for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) of all cases of are characterized by a net positive charge on a number basis.

[0170]

[0219] Aspect C44: P 2 The polymer according to any one of aspects C1 to C43 or any of the preceding aspects, wherein at least 75% (for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) of all cases of are characterized by a net positive charge on a number basis.

[0171]

[0220] Aspect C45: At least one P 1 And / or at least one P 2 The polymer according to any one of aspects C1 to C44 or any of the preceding aspects, wherein at least one P and / or at least one P contains or further contains at least one arginine.

[0172]

[0221] Aspect C46: At least one P 1 And / or at least one P 2 The polymer according to any one of aspects C1 to C45 or any of the preceding aspects, wherein at least one P and / or at least one P contains or further contains at least one aspartic acid.

[0173]

[0222] Aspect C47: At least one P 1 And / or at least one P 2 The polymer according to any one of aspects C1 to C46 or any of the preceding aspects, wherein at least one P and / or at least one P contains or further contains at least one proline.

[0174]

[0223] Aspect C48: A polymer according to any one of Aspects A1 to C47, which is metaphilic.

[0175]

[0224] Aspect C49: A polymer according to any one of Aspects A1 to C48, characterized by an average degree of polymerization of 2 to 100 (e.g., 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 30, 2 to 20, 7 to 100, 7 to 80, 7 to 60, 7 to 40, 7 to 30, 7 to 20, 15 to 50, 15 to 40, or 15 to 30).

[0176]

[0225] Aspect C50: A polymer according to any one of Aspects A1 to C49, characterized by an average degree of polymerization of 2 to 50 (e.g., 2 to 50, 2 to 30, 2 to 20, 7 to 50, 4 to 40, 4 to 30, 4 to 20, 15 to 50, 15 to 40, or 15 to 30).

[0177]

[0226] Aspect C51: A polymer according to any one of Aspects A1 to C50, characterized by an average degree of polymerization of 2 to 30 (e.g., 2 to 30, 2 to 20, 5 to 30, 5 to 20, 7 to 30, 7 to 20, 15 to 40, or 15 to 30).

[0178]

[0227] Aspect C52: A polymer according to any one of Aspects A1 to C51, characterized by an average degree of polymerization of 7 to 30 (e.g., 7 to 30, 7 to 25, 7 to 20, 7 to 15).

[0179]

[0228] Aspect C53: A polymer according to any one of Aspects A1 to C52, characterized by a number average molecular weight of 1 kDa to 50 kDa (e.g., 1 kDa to 50 kDa, 1 kDa to 30 kDa, 5 kDa to 50 kDa, 5 kDa to 30 kDa, 7 kDa to 50 kDa, 7 kDa to 30 kDa, 10 kDa to 50 kDa, or 10 kDa to 30 kDa).

[0180]

[0229] Aspect C54: A polymer according to any one of Aspects A1 to C53, characterized by a number average molecular weight of 1 kDa to 30 kDa (for example, 1 kDa to 30 kDa, 5 kDa to 30 kDa, 7 kDa to 30 kDa, or 10 kDa to 30 kDa).

[0181]

[0230] Aspect C55: A polymer according to any one of Aspects A1 to C54, characterized by a brush density of at least 75% (for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%).

[0182]

[0231] Aspect C56: A polymer according to any one of Aspects A1 to C55, characterized by a brush density of at least 80% (for example, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%).

[0183]

[0232] Aspect C57: The following characteristics: (a) P 1 contains 5 to 100 amino acids (for example, 5 to 100, 5 to 50, 5 to 40, 5 to 30, 5 to 20, 6 to 50, 6 to 100, 6 to 40, 6 to 30, or 6 to 20 amino acids); (b) m is an integer from 2 to 100 (for example, 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 30, 2 to 20, 2 to 10, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30); (c) n is an integer from 0 to 100 (for example, 0 to 100, 0 to 80, 0 to 60, 0 to 50, 0 to 30, 0 to 20, 0 to 10, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30); (d) o is an integer from 0 to 100 (e.g., 0 to 100, 0 to 80, 0 to 60, 0 to 50, 0 to 30, 0 to 20, 0 to 10, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30); (d) m is an integer from 2 to 100 (e.g., 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 30, 2 to 20, 2 to 10, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30), n is 0, p is 0, and at least one instance of P 1 is different from another instance of P 1 ; (e) The degree of polymerization (m + n + o) is an integer from 2 to 200 (e.g., 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 30, 2 to 20, 2 to 10, 4 to 200, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 200, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30), or an integer from 2 to 50 (e.g., 2 to 50, 2 to 30, 2 to 20, 2 to 10, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30); (f) A molecular weight of 1 kDa to 1,000 kDa (e.g., 1 kDa to 1000 kDa, 1 kDa to 500 kDa, 1 kDa to 100 kDa, 1 kDa to 50 kDa, 1 kDa to 30 kDa, 5 kDa to 50 kDa, 5 kDa to 30 kDa, 7 kDa to 1000 kDa, 7 kDa to 50 kDa, 7 kDa to 30 kDa, 10 kDa to 1000 kDa, 10 kDa to 50 kDa, or 10 kDa to 30 kDa); (g) When the polymer is defined by the equation m / (m + n + o) × 100, it has a density of at least 50% (e.g., at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) of P 1 peptide; (h) Combinations thereof; (i) Any combination thereof A polymer according to any one of Aspects C1 to C56 or any of the preceding aspects, having at least one of the following.

[0184]

[0233] Aspect C58: m is an integer from 2 to 100 (for example, 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 30, 2 to 20, 2 to 10, 4 to 100, 4 to 80, 4 to 60, 4 to 50, 4 to 30, 4 to 20, 4 to 10, 10 to 50, 10 to 40, 10 to 30, 20 to 50, 20 to 40, or 20 to 30), and the polymer is polyvalently bonded via at least two P to at least a part of a tau protein and / or at least a part of a microtubulin protein. A polymer according to any one of Aspects C1 to C57 or any of the preceding aspects. 1 A polymer according to any one of Aspects C1 to C57 or any of the preceding aspects, polyvalently bonded via at least two P to at least a part of a tau protein and / or at least a part of a microtubulin protein.

[0185]

[0234] Aspect C59: B 1 , B 2 , or B 3 A polymer according to any one of Aspects C1 to C57 or any of the preceding aspects, wherein at least one of them, or all three of them, are polymerized monomers containing an unsaturated monomer.

[0186]

[0235] Aspect C60: A polymer according to Aspect C59 or any of the preceding aspects, wherein the unsaturated monomer contains an ethylenically unsaturated monomer, a norbornene monomer, or a norbornene dicarboximide.

[0187]

[0236] Aspect C61: A polymer according to any one of Aspects A1 to C60, prepared by a living polymerization method optionally selected from ring-opening metathesis polymerization (ROMP), reversible addition-fragmentation chain transfer polymerization (RAFT), or atom transfer radical polymerization (ATRP).

[0188]

[0237] Aspect C62: Each instance of the partial structure (S1a) in formula (FX1):

Chemical formula

[0189]

[0238] Embodiment C63: Each case of the substructure (S2a) in formula (FX1): [Chemical formula] is, independently, a substructure (S2b) or a substructure (S2c): [Chemical formula] [Chemical formula] comprising, wherein R 16 is H or C1-C3 alkyl, a polymer according to embodiment C1-C62 or any one of any of the foregoing embodiments.

[0190]

[0239] Embodiment C64: Each case of L 1 and L 2 , if present, is, independently, a single bond, -O-, -(CH2CH2O) x -, C1-C 10 alkyl, C1-C 10 acyl, C2-C 10 alkenyl, C3-C 10 aryl, C1-C 10 alkoxyl, or any combination thereof, wherein x is an integer from 1 to 20, and each L 1 and L 2If present, B 1 is attached to P 1 by a covalent bond, and B 3 is attached to P 2 by a covalent bond and is composed of one or more suitable functional groups, a polymer according to any one of aspects C1 to C63 or any of the preceding aspects.

[0191]

[0240] Aspect C65: R 1 , T 1 , and T 2 each independently is hydrogen, C1 - C 30 alkyl, C3 - C 30 cycloalkyl, C5 - C 30 aryl, C5 - C 30 heteroaryl, C1 - C 30 acyl, C1 - C 30 hydroxyl, C1 - C 30 alkoxy, C2 - C 30 alkenyl, C2 - C 10 alkynyl, C5 - C 30 alkylaryl, -CO2R 3 , -CONR 4 R 5 , -COR 6 , -SOR 7 , -OSR 8 , -SO2R 9 , -OR 10 , -SR 11 , -NR 12 R 13 , -NR 14 COR 15 , C1 - C 30 alkyl halide, phosphonate, phosphonic acid, silane, siloxane, silsesquioxane, C2 - C 30 halocarbon chain, C2 - C 30 perfluorocarbon, C2 - C 30 polyethylene glycol, metal, metal complex, a moiety containing a fluorophore, or a moiety containing a contrast agent, and each of R 3 ~R 15 is independently H, C5 - C 10 aryl, or C1 - C 10The polymer according to embodiment C1 - C64 which is alkyl or any one of the above - mentioned embodiments.

[0192]

[0241] Embodiment C66: At least one P 1 , P 2 , R 1 , T 1 , or T 2 contains a fluorophore - containing moiety, or a contrast - agent - containing moiety, or further contains the same, the polymer according to embodiment C1 - C65 or any one of the above - mentioned embodiments.

[0193]

[0242] Embodiment C67: At least one P 1 , P 2 , R 1 , T 1 , or T 2 contains rhodamine, fluorescein, Cy5.5, gadoteric acid, or a combination thereof, or further contains the same, the polymer according to embodiment C1 - C66 or any one of the above - mentioned embodiments.

[0194]

[0243] Embodiment D1: A composition comprising the polymer according to any one of embodiments A1 - C67 and a pharmaceutically acceptable carrier.

[0195]

[0244] Embodiment E1: A medicament for use in preventing, disintegrating, promoting, or detecting tau and / or microtubulin protein aggregation, comprising a composition having a therapeutically effective amount of the polymer according to any one of embodiments A1 - C67.

[0196]

[0245] Embodiment E2: A medicament for use in preventing, treating, or detecting tauopathy - related diseases or conditions in a subject, comprising a composition having a therapeutically effective amount of the polymer according to any one of embodiments A1 - C67.

[0197]

[0246] Embodiment F1: A method for preventing, disintegrating, promoting, or detecting tau and / or microtubulin protein aggregation, comprising Contacting oligomers, protofibrils, amyloid fibrils, and / or cross-β sheet amyloid species of tau and / or microtubulin with a polymer as described in any one of therapeutic effective amounts of aspects A1 - C67 or a composition as described in aspect D1 or any of the foregoing aspects A method comprising

[0198]

[0247] Aspect F2: The method according to aspect F1 or any of the foregoing aspects, wherein oligomers, protofibrils, amyloid fibrils, and / or cross-β sheet amyloid species of tau and / or microtubulin are in a patient or in a fluid derived from a subject

[0199]

[0248] Aspect F3: At least one R 1 is a moiety containing a fluorophore or a moiety containing a contrast agent, and the method further comprises an imaging step after the contacting step, the method according to aspect F1 or aspect F2 or any of the foregoing aspects

[0200]

[0249] Aspect G1: A method for preventing, treating, or detecting a tauopathy-related disease or condition in a subject, comprising administering to the subject a polymer as described in any one of therapeutic effective amounts of aspects A1 - C67 or a composition as described in aspect D1 or any of the foregoing aspects A method comprising

[0201]

[0250] Aspect G2: The tauopathy-related disease or condition optionally includes a neurodegenerative disease, which is selected from or related to Alzheimer's disease (AD), primary age-related tauopathy, chronic traumatic encephalopathy, traumatic brain injury, progressive supranuclear palsy, corticobasal degeneration, dementia, frontotemporal dementia, argyrophilic grain dementia, frontotemporal dementia linked to chromosome 17, and parkinsonism, Parkinson's disease, parkinsonism, postencephalitic parkinsonism, amyotrophic lateral sclerosis (ALS), Huntington's disease, vacuolar tauopathy, corticobasal body disease, glioma, gangliocytoma, meningovascular amyloidosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, lipofuscinosis, Pick's disease, Pick complex, or any combination thereof, in the method according to Aspect G1 or any of the preceding aspects.

[0202]

[0251] Aspect G3: The method according to Aspect G1 or Aspect G2 or any of the preceding aspects, wherein the polymer is administered to the subject's brain, spinal cord, cerebrospinal fluid, or any combination thereof.

[0203]

[0252] Aspect G4: At least one R 1 is a moiety containing a fluorophore or a moiety containing a contrast agent, and the method further includes an imaging step after the administering step, in the method according to any one of Aspects G1 - G3 or any of the preceding aspects.

[0204]

[0253] Aspect H1: Use of a composition for preventing, disrupting, promoting, or detecting tau and / or microtubulin protein aggregation, wherein the composition includes the polymer according to any one of Aspects A1 - C67 or any of the preceding aspects.

[0205]

[0254] Aspect I1: Use of a composition for preventing, treating, or detecting a tauopathy-related disease or condition in a subject, wherein the composition includes the polymer according to any one of Aspects A1 - C67 or any of the preceding aspects.

[0206]

[0255] Aspect J1: A method for producing a polymer according to any one of Aspects A1 to C67 or any of the foregoing aspects, comprising: at least one P 1 synthesizing a peptide; at least one P 1 capping the peptide at the terminus with a polymerizable monomer that becomes polymer main chain subunit B once polymerized, thereby forming a polymerizable P 1 monomer; 1 forming a polymerizable P polymerizable P 1 polymerizing the monomer A method comprising.

[0207]

[0256] Aspect J2: The method according to Aspect J1 or any of the foregoing aspects, wherein the synthesizing step comprises solid-phase synthesis using protected amino acids; the polymerizable monomer comprises an ethylenically unsaturated monomer optionally containing norbornene or (meth)acrylate; and the polymerizing step comprises ROMP, RAFT, or ATRP.

[0208]

[0257] Aspect K1: A method for producing a medicament for use in preventing, disrupting, promoting, or detecting tau and / or microtubulin protein aggregation, comprising combining a composition having a therapeutically effective amount of a polymer according to any one of Aspects A1 to C67 or any of the foregoing aspects, and an optional carrier A method comprising.

[0209]

[0258] Aspect L1: A method for producing a medicament for use in preventing, treating, or detecting a tauopathy-related disease or condition in a subject, comprising combining a composition having a therapeutically effective amount of a polymer according to any one of Aspects A1 to C67 or any of the foregoing aspects, and an optional carrier A method comprising.

Example

[0210]

[0259] The present invention can be further understood by the following non-limiting examples. The examples are provided to illustrate a part of the concepts described within this disclosure. Each example is considered to provide specific individual embodiments of the compositions and methods of preparation and use, but no example should be considered to limit the more general embodiments described herein.

[0211]

[0260] In the following examples, efforts have been made to confirm the accuracy regarding the numerical values (e.g., amounts, temperatures, etc.) used, but some experimental errors and deviations should be taken into account.

[0212]

[0261] Materials and methods : The following description of materials and methods applies to one or more of the following examples. In case of any degree of contradiction between the materials and methods of these descriptions and those provided in the examples, the examples shall prevail.

[0213]

[0262] PLP: The PLP tested in the following examples is sometimes referred to as "TPX", where "X" means a specific peptide sequence. For example, TP1 refers to the PLP generated from the peptide monomer characterized by SEQ ID NO: 16; TP4 refers to the PLP generated from the peptide monomer characterized by SEQ ID NO: 4; TP7 refers to the PLP generated from the peptide monomer characterized by SEQ ID NO: 2; TP12 refers to the PLP generated from the peptide monomer characterized by SEQ ID NO: 10. Similarly, the free peptides (i.e., peptides not incorporated into the PLP platform) in the following examples are sometimes referred to as "PX", where "X" means a specific peptide sequence.

[0214]

[0263] Peptide Synthesis: Peptide monomers are synthesized on Rink resin (0.67 mmol / g) using standard Fmoc SPPS procedures on a Liberty Blue automated microwave synthesizer. Peptide monomers are prepared via amide coupling to N-(hexanoic acid)-cis-5-norbornene-exo-dicarboximide (e.g., 3.0 equivalents) in the presence of HBTU (e.g., 2.9 equivalents) and DIPEA (e.g., 6.0 equivalents). Aspects of this process are described in more detail, specifically the synthetic methodology, in Blum et al., Peptides Displayed as High Density Brush Polymers Resist Proteolysis and Retain Bioactivity, J. Am. Chem. Soc. 2014, 136(43), 15422 - 15437, which is incorporated herein by reference in its entirety to the extent not inconsistent with the description herein. Peptide monomers are cleaved from the resin by treating the resin with trifluoroacetic acid (TFA):H2O:triisopropylsilane (95:2.5:2.5 v / v) for 4 hours. The crude product is obtained by precipitation in cold diethyl ether.

[0215]

[0264] Peptides are further purified by Armen Glider CPC prep-HPLC on a Jupiter Proteo 90Å Phenomenex column (2050×25.0 mm) to obtain a purity of 90 - 95% as confirmed by analytical HPLC. For all RP-HPLC purifications, the gradient solvent system utilizes buffer A (water containing 0.1% TFA) and buffer B (acetonitrile containing 0.1% TFA). All peptides and monomers are purified using a gradient of 15 - 45% buffer B over 30 minutes. The pure product is then analyzed by electrospray ionization mass spectrometry (ESI-MS) on a Bruker amaZon SL to confirm the molecular weight.

[0216]

[0265] Coincidence: PLP is achieved by ring-opening metathesis polymerization (ROMP) in a glove box under nitrogen gas. A norbornene-conjugated peptide monomer (e.g., 15.0 equivalents, 30 mM) is dissolved in degassed DMF containing 1 M LiCl. Next, an olefin metathesis initiator (IMesH2)(C5H5N)2(Cl)2Ru=CHPh stock solution (e.g., 1.0 equivalent, 20 mg / mL in DMF) is rapidly added to the monomer solution. This solution is kept stirring until the monomer is completely consumed. In the case of the polymer having a rhodamine tag, this is achieved by the addition of 1 equivalent of rhodamine linked to norbornene via a 6-carbon chain linker containing an amide bond. After polymerization, the polymer solution is precipitated in diethyl ether and further purified via dialysis against deionized water. Finally, the polymer product is obtained by lyophilization. Aspects of this process are described in more detail in Kammeyer et al., Polymerization of Protecting-Group-Free Peptides via ROMP, Polym. Chem. 2013, 4(14), 3929 - 3933 and Nomura et al., Precise Synthesis of Polymers Containing Functional End Groups by Living Ring-Opening Metathesis Polymerization (ROMP): Efficient Tools for Synthesis of Block / Graft Copolymers, Polym. 2010, 51(9), 1861 - 1881, the entireties of which are incorporated herein by reference to the extent not inconsistent with the description herein, more specifically with respect to the polymerization techniques.

[0217]

[0266] 1H NMR is used to confirm the completion of peptide monomer consumption and determine the period required to reach completion. The percent conversion from monomer to polymer is tracked over time by comparing monomer peaks that decrease by NMR relative to an internal standard. A linear plot of NMR elucidates pseudo-first order kinetics (see, e.g., FIG. 7C showing polymerization kinetics using SEQ ID NO: 16 as the peptide monomer; FIG. 8 using SEQ ID NO: 1; FIG. 9B using SEQ ID NO: 4 (“TP4”); FIG. 10B using SEQ ID NO: 2 (“TP7”); FIG. 32 using SEQ ID NO: 10 (“TP12”)). The polymer is terminated with ethyl vinyl ether (10 equivalents) for 1 hour at room temperature, precipitated, washed three times with cold diethyl ether, and collected by centrifugation. The molecular weight and polydispersity of the polymer are determined by SEC MALS (Phenomenex Phenogel 5μ 103Å, 1K-75K, 300×7.8 mm and in series with Phenomenex Phenogel 5μ 103Å, 10K-100K, 300×7.8 mm) in 0.05 M LiBr in DMF at 65° C. using a ChromTech series 1500 pump equipped with a multi-angle light scattering detector (DAWN-HELIOS II, Wyatt Technology) and a refractive index detector (Wyatt Optilab TrEX) normalized to a 30,000 MW polystyrene standard (see, e.g., FIG. 33 showing SEC-MALS results for a PLP homopolymer incorporating SEQ ID NO: 10 (“TP12”)). Molecular weight can also be determined via SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). The polymer is run at 2 mg / ml on a pre-cast Mini-PROTEAN TGX gel 4-10% and visualized using Instant Blue or Coomasie blue staining (see, e.g., FIG. 7A showing molecular weight results for a PLP homopolymer (DP10) using SEQ ID NO: 16; FIG. 9A using SEQ ID NO: 4 (“TP4”) and SEQ ID NO: 2 (“TP7”) at DP12; FIG. 10A using SEQ ID NO: 4 (“TP4”) and SEQ ID NO: 2 (“TP7”) at DP14).

[0218] Example 1

[0267] This example describes exemplary PLP properties, design, and performance for the investigation of protein phase separation.

[0219]

[0268] PLP platform:In PLP, the target peptide sequences are synthesized via SPPS followed by peptide functionalization at the reactive monomer moiety. The resulting monomers thus have peptide side-chain pendants to polymerizable motifs, which leads to the formation of brush polymers with high-density peptide side-chains formed via grafting polymerization (Figures 35C, D). The choice of polymerizable units, and thus the choice of polymer backbone, has been shown to be flexible, but norbornene imide-based monomers are particularly effective due to their rapid polymerization kinetics by ring-opening metathesis polymerization (ROMP) (Figures 35C, D). In solution, the hydrophobic backbone of PLP collapses to form spherical single chains with surfaces decorated with high-density peptides. The spherical structure and the densely packed peptide corona minimize the solvent-exposed surface area of the peptides and inhibit proteolysis even in the case of L-amino acids (Figures 35A, B). Callmann CE et al., Poly(peptide): Synthesis, Structure, and Function of Peptide-Polymer Amphiphiles and Protein-like Polymers. Acc Chem Res 2020;53:400-413; Gianneschi NC et al., Biomolecular Densely Grafted Brush Polymers: Oligonucleotides, Oligosaccharides and Oligopeptides. Angew Chemie Int Ed, 2020; Blum AP, Kammeyer JK, Gianneschi NC. Furthermore, as demonstrated by a 100- to 500-fold increase in the binding equilibrium constant compared to individual peptides, the high-density peptides in PLP are thought to confer a high degree of multivalency (Figures 35A, B). Additionally, PLP can be designed to penetrate cells and has been shown to be distributed in the cytosol, where they exhibit bioactivity in multiple cell types ranging from cancer cells to neurons (Figures 35A, B) and stem cells.

[0220]

[0269] Peptide side chainHere, the peptide side chains are selected to represent the intrinsically disordered regions of proteins of high clinical interest. For example, the microtubule-binding region (MBTR) of tau (SEQ ID NO: 1 (VQIVYK), SEQ ID NO: 3 (VQIINK)). The peptides can be strategically modified by adding or deleting certain amino acids in a bottom-up approach, enabling precise control over the strength and solubility of peptide-peptide interactions. The resulting monomers can then be polymerized to effectively use the synthetic toolkit of polymer chemistry to incorporate a high degree of multivalency into the protein mimics and form graft polymers with high-density peptide side chains. Furthermore, the peptide monomers can also be copolymerized with non-peptide monomers to create statistical and block copolymers with adjustable peptide graft densities. The ability to adjust the graft density of peptide side chains, essentially the distance between each peptide side chain along the backbone, is predicted to modulate multivalency and ultimately the strength of protein-PLP interactions. The inventors predict that the insights obtained by conducting such systematic studies within the framework of well-established theories will provide a paradigm shift in the perspective regarding the major factors driving LLPS of proteins.

[0221] Example 2

[0270] Table 1. List of peptide sequences for integration into the PLP embodiments described herein. Charges are related to the average pH in the human body (i.e., 7.35 - 7.45).

[0222]

Table 1

[0223] Example 3

[0271] This example provides an exemplary cell permeability assay for a particular PLP.

[0272] Table 2. Characterization of the PLP used in the cell permeability assay.

[0224]

Table 2

[0273] The results in Table 2 were partially obtained from FIG. 34 showing the results of SDS-PAGE for rhodamine-labeled PLP incorporating either Array No. 4 (i.e., "TP4Rho" in FIG. 34) or Array No. 2 (i.e., "TP7Rho" in FIG. 34).

[0225]

[0274] Mouse striatal cells HdhQ7 / 7 from the Coriell Institute were plated at a density of 50,000 per well in a 4-chamber 35 mm round-bottom glass culture dish. The cells were incubated at 33 °C for 24 hours in a 5% CO2 atmosphere. Rhodamine-labeled PLP, rhodamine dye, and vehicle control in 10% FBS DMEM medium without phenol red were each incubated with the cells for 24 hours. A 405 nm laser at 15% laser power was used to reach the cell nucleus (stained with Hoechst). A 488 nm laser at 12% laser power was used to reach the cell membrane (stained with wheat germ agglutinin, Alexa Fluor 488 conjugate). A 581 nm laser at 8% laser power was used to achieve cell imaging for the fluorescence of rhodamine.

[0226]

[0275] Each of the polymers listed in Table 2 demonstrated the ability to penetrate striatal cells, as shown in FIG. 7B (Array No. 16) and FIG. 13 (left: Array No. 4; right: Array No. 2). A hypothesis is proposed that the penetration is facilitated in part by additional residues, particularly arginine, which modify the known structures of tau and microtubulin to promote solubility.

[0227] Example 4

[0276] A hypothesis is proposed that PLPs capable of liquid-liquid phase separation (LLPS) may be more suitable for engaging phase-separated tau. Thus, this example demonstrates the LLPS of a particular rhodamine-labeled PLP.

[0228]

[0277] The solution was dissolved in HEPES buffer: 10 mM HEPES buffer (pH 7.4) containing 0.1 mM EDTA and 2 mM DTT, medium buffer: Dulbecco's modified Eagle's medium (high glucose) containing 2 mM L-glutamine with 10% fetal bovine serum and 1% penicillin / streptomycin, or water. Various crowding agents were also tested using HEPES buffer - PEG and heparin. In the case of heparin, a stock solution of PLP (using either SEQ ID NO: 4 or SEQ ID NO: 7 as the peptide monomer) labeled with 10 μM of each rhodamine and 30 μM of sodium heparin was prepared in buffer and maintained refrigerated. Before the experiment, the solution was left undisturbed for at least 30 minutes to come to room temperature, and the experiment was conducted at room temperature. In the microscopy room, 20 μL of PLP was added to 5 μL of heparin and gently mixed in a microcentrifuge tube. 5 μL of the solution was immediately pipetted onto a Fisherbrand glass microscope slide and covered with a coverslip. All reactions were visualized using an Olympus BX53 fluorescence microscope with a 581 nm laser, monitoring the bright field in addition to rhodamine. LLPS occurred within 60 minutes from the start.

[0229]

[0278] Figures 11A - 11B demonstrate the effect of various solvents on the results of LLPS for TP4 (Figure 11A) and TP7 (Figure 11B), in which an increase in salt concentration appears to induce salting-out effects seen in proteins, and HEPES buffer appears to promote aggregation. In the case of samples tested with PEG, the crowding agent appeared to induce LLPS phase condensates of PLP homopolymers incorporating SEQ ID NO: 4 as the peptide side chain, as shown in Figure 12. The LLPS results for solutions in HEPES buffer containing heparin are shown in Figures 14 (SEQ ID NO: 4) and 15 (SEQ ID NO: 2), suggesting that both of the PLP homopolymers tested are capable of liquid - liquid phase separation.

[0230] Example 5

[0279] Based on the results obtained in Example 4, in this Example 5, it was evaluated whether the same PLP reacted in the same or substantially similar manner as tau, including thioflavin T.

[0231]

[0280] All solutions were dissolved in 10 mM HEPES buffer (pH 7.4) containing HEPES buffer: 0.1 mM EDTA and 2 mM DTT. Stock solutions of 10 μM PLP labeled with each rhodamine (using either SEQ ID NO: 4 or SEQ ID NO: 7 as the peptide monomer), 30 μM sodium heparin, and 100 μM thioflavin T were prepared in HEPES buffer and maintained refrigerated. Before the experiment, the solutions were left undisturbed for at least 30 minutes to reach room temperature, and the experiment was conducted at room temperature. 20 μL of PLP was added to 5 μL of heparin and 1 μL of thioflavin T and gently mixed in a microcentrifuge tube. The centrifuge tube was left standing at room temperature for 24 hours and then 5 μL of the solution was pipetted onto a Fisherbrand glass microscope slide and covered with a coverslip. The results were visualized using a 581 nm laser to monitor rhodamine and a 488 nm laser to visualize the bright field in addition to thioflavin T. Since the Olympus microscope was removed from the facility, images were taken with a Leica DM6B fluorescence microscope.

[0232]

[0281] As depicted in Figure 16, these results suggest that the PLP tested is capable of reacting with thioflavin T in the same manner as tau through binding to residues that form beta-sheets.

[0233] Example 6

[0282] Following the results of Examples 4 and 5, PLP homopolymers were subjected to additional tests to evaluate their material properties.

[0234]

[0283] Using standard STEM protocols, PLPs having SEQ ID NO:2 or SEQ ID NO:4 as peptide monomers were prepared and visualized in H2O. The first STEM results in Figure 3 appear to demonstrate beta-sheet conformations (SEQ ID NO:4 (top), SEQ ID NO:2 (bottom)). Further, from the results depicted in Figure 17, the property of forming beta-sheets of the homopolymers was confirmed. After placing the PLP in HEPES LLPS buffer and heating to 37°C (i.e., body temperature), over time, polymerization fibrillation was established in dry-state TEM. Images were captured at 10 minutes, 1 hour, and 7 hours as shown in Figure 18. As depicted in Figure 19, the same fibrillation was seen in purified K18 tau protein after incubating with heparin in PBS for 24 hours. The strikingly similar features were evidence of the mimicking performance of the PLP.

[0235]

[0284] The proteomimetic similarity was further investigated using circular dichroism. The samples tested included K18 tau, PLPs having SEQ ID NO:4, SEQ ID NO:2, and SEQ ID NO:10, and the PLPs containing K18 tau. Each of these samples was dissolved in Tris buffer (10 mM Tris, 50 mM NaF, 0.5 mM dithiothreitol) and the pH was adjusted to 7.4. The samples were tested with and without the addition of heparin. For samples containing heparin, the samples were incubated for 24 hours in the presence of 10 μM sodium heparin salt. The polymers and peptides were run at a concentration of 3 mg / 10 ml at 37°C, and K18 tau was run at 10 μM. The parameters were as follows: 1 mm quartz cuvette, continuous scan at 10 nm / min over 260 - 200 nm, and an average of 3 acquisitions per trace.

[0236]

[0285] Samples with added heparin showed CD results consistent with the properties of a random coil (Figure 20, top). When combined with K18, PLP appeared to maintain the characteristics of a random coil (Figure 20, top). However, for samples containing heparin, the CD results depicted a beta-sheet conformation (Figure 20, bottom). As demonstrated by circular dichroism having a peak near 220 nm for PLP-sequence number 4 in Figure 20, heparin is thought to have induced aggregation and beta-sheet formation. Interestingly, there appears to be an interaction between K18 and the polymer, in which case the degree of beta-sheet characteristics decreases. This was initially ignored as it was enigmatic and further investigated in subsequent experiments (discussed in Example 7 below).

[0237]

[0286] Free peptides (i.e., peptides characterized by sequence number 4, sequence number 2, or sequence number 10) were synthesized as controls and run on circular dichroism. As shown in Figure 21, each of the free peptides yielded CD characterizations consistent with beta-sheet formation regardless of the addition of heparin. These results suggested that the polymer inhibits beta-sheet formation unless heparin is introduced as an aggregating agent. These results contradicted the inventors' hypothesis and were to be further examined in a thioflavin T kinetics assay (discussed in Example 7 below).

[0238] Example 7

[0287] Based on the unexpected results obtained from Example 6, a thioflavin T kinetics assay was performed to establish whether free peptides had the same or substantially similar effects when incorporated into the PLP platform.

[0239]

[0288] Expression and purification of K18 tau:Before the K18 tau dynamics became executable, K18 tau was first expressed, collected, and purified. Through the expression and collection of K18 tau, in combination with several days of purification, a large amount of K18 was obtained (see Figures 22A - 22B regarding the results of purification and quantification (previous iterations are shown in Figure 4)). However, this K18 was tagged with a His tag for collecting / purifying K18 using a nickel bead column. Therefore, TEV protease was used to cleave the His tag from K18. Once cleaved, as shown in Figure 23, K18 was electrophoresed through an FPLC column in two iterative applications due to the column size limitation. Then the two iterative applications were combined and subjected to dialysis following standard dialysis procedures. The pooled fractions were quantified using BSA quantification (Figure 6), and 28 mg of purified K18 tau was obtained. Figure 5 demonstrates silver staining regarding the purity of K18, in which uncleaved K18 refers to K18 with a His tag.

[0240]

[0289] Tau kinetics thioflavin T plate reader assay protocol :First, the effects of control and various concentrations of K18 tau were examined (Figure 24). In Figure 24, 1× tau refers to a final concentration of 10 μM K18 tau in a well with a total volume of 100 μl (i.e., 0.5× tau refers to 5 μM K18 tau and 2× tau refers to 20 μM K18 tau). Before adding PLP to K18 tau, the concentration at which the mimetic aggregating polymer does not activate thioflavin T was evaluated. Based on the results depicted in Figure 25, a PLP concentration of 1 μM was selected for further experiments. Notably, for TP4, 1 μM still produced a significant signal (Figure 25, top) when compared to 1 μM of TP7, which showed only a very weak signal (Figure 25, bottom). Considering the purpose of investigating the effect of PLP, it was determined that concentrations that are too high can be easily adjusted, while concentrations that are too low may produce false negatives regarding the effect.

[0241]

[0290] PLP incorporating the peptide side chains of SEQ ID NO: 4 ("TP4"), SEQ ID NO: 2 ("TP7"), or SEQ ID NO: 10 ("TP12") was tested. PLP, K18 tau (10 μM), and Thio T (Sigma) were added at a final concentration of 10 μM, and aggregation was induced by the addition of freshly prepared sodium heparin solution (Santa Cruz) at a final concentration of 44 μg / ml. For the non-induced control, assay buffer was added instead of the heparin solution until a final volume of 100 μl per well was reached. The aggregation reaction was carried out with continuous shaking at 37 °C and monitored via ThT fluorescence (excitation, 444 nm; emission, 485 nm; cutoff, 480 nm) in a Spectramax M5 microplate reader (Molecular Devices). Readings were taken for at least 24 hours.

[0242]

[0291] The results depicted in FIG. 26 suggest that the mimetic aggregating polymer promotes or amplifies tau fibril formation.

[0243]

[0292] The mimetic polymer was subjected to additional experiments to determine whether it acts as a seeding agent and induces K18 tau fibrillation in the absence of heparin. Surprisingly, the mimetic polymer did not induce fibrillation (FIG. 27), which indicates that seeding is not necessarily relevant to biological systems where seeding is a natural phenomenon. Furthermore, based on the beta-sheet forming peptide control (FIG. 24), the effect of PLP on tau fibrillation was evaluated at two different concentrations: 1 μM, which matches the polymer, and 10 μM, which partially matches the actual concentration of the peptide on the polymer. Unfortunately, the amplitudes were "unstable" and the plotting software GraphPad Prism was unable to compare whether the obtained values shown in FIG. 28 were significant. More replicates are expected to smooth the amplitudes and enable comparison.

[0244]

[0293] The tau kinetic thioflavin T assay was performed using TP12 (i.e., PLP incorporating SEQ ID NO: 10, a known inhibitor). Assays were performed at two concentrations of TP12, 1 μM and 10 μM, and the results (Figure 29) suggest that the inhibitor polymer significantly prevented fibril formation of K18 tau. However, recent attempts to perform a concentration gradient of TP12 to confirm the lowest possible dose for inhibition were interrupted by the lag time of control K18 tau (Figures 30 and 31). In fact, the lag time of control K18 tau was 5 hours, which is significantly higher than the standard 2.5 hours.

[0245]

[0294] Table 3. Summary of the results depicted in Figures 26, 28, and 29 for events where graphical representation is difficult to confirm. Values are rounded to the nearest integer at the first decimal place. Tau 10 hours refers to the average amplitude (fluorescent intensity readout) of the trace of control tau (meaning no peptide or polymer added, only heparin, thioflavin T, and PBS) with n = 6 at 10 hours. The same applies to tau 15 hours and tau 20 hours. Average tau amplitude refers to the average of the amplitudes at 10, 15, and 20 hours in the previous column. Tau is the control for each individual plate, and since these readings are taken on three separate plates, there are three different sets of average amplitudes. Similarly, PLP or Pep 10 hours refers to the average amplitude of the tau trace treated with that compound at 10 hours (n = 6 for PLP and n = 3 for the peptide). Average P amplitude refers to the average of the amplitudes over three times, or the average of the three previous columns. PLP or Pep / tau % is the result of dividing the average P amplitude by the average tau amplitude and multiplying by 100. % Amplitude change refers to the change in amplitude obtained by subtracting the tau average amplitude from the P average amplitude, dividing by the tau average amplitude, and multiplying by 100.

[0246]

Table 3

[0247] Example 8

[0295] In this example, toxicity and off-target effects in cell culture are evaluated.

[0248]

[0296] To measure the biological effects of tau-PLP introduction into the cellular environment, monitoring assays can be used that include global transcriptional profiling by RT-PCR and RNA-seq. Each of these enables monitoring of the acute and chronic effects of PLP on cytotoxicity, effects on the stress response and protein homeostasis. Since the proposed therapeutic pathway is a sequence strand rather than a single protein-protein interaction in a single cell line, it is important to understand the biological effects. It will be verified by, for example, a monitoring assay of cell processes via cell viability that autophagy is not induced and that important cell processes are not disrupted.

[0249]

[0297] Tau is highly expressed in neurons and has an important role in maintaining neuronal function. Therefore, it is important to titrate tau-PLP and tau-HYDRAC to confirm that the health of neurons is not affected according to concentration. The selection of tau-PLP and tau-HYDRAC that specifically target pathological tau species is predicted to reduce toxic effects. Tau-binding PLPs have different water solubilities and may cause off-target effects. At high local concentrations of PLP in the cytosol, the seeding and amplification properties inherent in endogenous MAPT wild-type or P301S mutant proteins should be monitored. Depending on the physico-chemical dynamics of the event, the effect on cellular protein homeostasis can be observed. In cases of promoted tau aggregation, the sequestration of functional cellular proteins such as chaperones, as well as autophagy and the proteasome machinery, are monitored.

[0250] Example 9

[0298] In this example, a biosensor for detecting leaky tau prior to symptom onset was developed using tau-binding PLP.

[0251]

[0299] As a biosensor that specifically binds to free tau collected from a blood sample, PLP with a fluorophore tag is used. Various forms of tau are doped into pooled blood, and the detection conditions for tau are optimized. When determined by BLI, the tau-mimicking PLP with the lowest k off rate may be most suitable in capturing tau in this application, which is thought to either not bind as strongly or is in contrast to inhibitor-based PLP. A screening assay containing tau-binding PLP as a biosensor can be used in parallel with other techniques to improve brain scans and early identification of AD.

[0252] Example 10

[0300] In this example, the aggregation and protofibril formation of tau in cell culture are regulated.

[0253]

[0301] In a cell-free assay, 1N4R tau that conditionally expresses and forms amyloid when seeded with MAPT P301S fibrils formed in the brain is screened in human 293T cells using a polymer tagged at the fluorophore end. Cell permeability, intracellular localization, and stability are evaluated using the polymer. Intracellular tau is visualized by immunostaining with anti-total tau antibodies (DAKO total tau pAb and HT7 mAb). Co-localization image analysis and biochemical pull-down experiments detect the interaction between fluorescent tau PLP and intracellular tau. The performance of tau PLP is evaluated to determine whether tau PLP shows higher affinity for the MAPT P301S mutant protein than wild-type tau. The appearance of tau inclusions occurs after exposing HEK293 tau P301S cells to sarcosyl-insoluble tau seeds from the brains of TgP301S mice, and tau amyloid is monitored using thioflavin T staining and antibody immunostaining for oligomeric and / or aggregated forms of tau (MC1 and PG5), and these results are predicted to be mirror images of each other. Inhibitor PLP and HYDRAC are predicted to reduce aggregation compared to tau-mimicking homopolymer PLP. Tau-binding PLP is delivered before, during, and after seeding to understand how it affects tau aggregation at various stages of the tau seeding / amplification process. For example, addition of tau PLP before seeding is predicted to delay tau amyloid formation by inhibiting the dynamics of seed amplification, or alternatively, to stimulate seed amplification and amyloid formation. This could lead to the development of tau PLP that either inhibits or enhances amyloid formation. Similarly, another class of tau PLP causes dissociation of the tau protofibril or amyloid state, is detected when added to each of these states, and either blocks further growth or causes its dissolution. What is important here is that PLP is predicted to engage with tau, the predicted target, with various binding activities.Studies are performed to develop design rules and interaction mappings and how PLP engages tau to prevent either seeding or propagation of tau amyloid in a complex liquid-liquid phase separated species environment.

[0254] Example 11

[0302] In this example, the ability of the PLP described herein to disassemble pre-formed aggregates during cell culture is evaluated.

[0255]

[0303] The overall purpose of tau HYDRAC is to disrupt and disassemble tau aggregates before they become neurofibrillary tangles. PROTACs generally function by reducing the proximity of the protein of interest to an E3 ligase that facilitates polyubiquitination (in this case via an E3 ligase recruiter, via the von Hippel-Lindau protein (VHL)), thereby resulting in degradation of that protein. HYDRAC applies the PLP system in this manner and has shown success for peptide-based PROTACs to date. The inventors have made HYDRAC novel by having superior resistance to degradation, allowing it to be delivered at lower concentrations for the same therapeutic effect compared to established peptide PROTACs.

[0256]

[0304] A peptide that binds to tau is copolymerized with a peptide motif that binds to an E3 ubiquitin ligase to create tau HYDRAC. Tau-targeted HYDRAC is synthesized in the same manner as the polymers described previously, where in this case, a block of the targeted peptide sequence is polymerized first, and upon completion, an E3 ligase recruiter motif (e.g., SEQ ID NO: 11 (ALAPYIP) or SEQ ID NO: 12 (ALAPYIPRR)) is added to form a second block. The block copolymer will be characterized via both SEC MALS and SDS PAGE in cases where SEC MALS is not performed, taking into account the aggregability of the targeted motif. Since much of the pathogenic tau is sequestered in the LLPS condensed state (Figure 2), HYDRAC is evaluated with respect to its ability to phase separate, which can have a large impact on its targeting motif. The absence of LLPS is not necessarily detrimental since some free monomeric tau and aggregates are present outside of these condensates (Figure 2).

[0257]

[0305] The turnover rate of tau is evaluated by using the pulse-chase method in HEK293 MAPT WT and P301S cells and then treating with tau HYDRAC. By tracking Ub G76V-GFP, a proteasomal degradation reporter co-expressed in HEK293 cells, it is examined whether the increase in proteasomal degradation is specific to the tau protein. A tau seeding / aggregation assay is used to quantify the results of tau HYDRAC. A positive result is considered to be any substantial change in the aggregation rate measured by a plate reader, predicting that HYDRAC significantly disrupts and destroys aggregates. The hypothesis is put forward that HYDRAC provides a multivalent degradation strategy that is a proof of concept for the idea of extending to many other desired protein targets in neurodegenerative diseases and beyond.

[0258]

[0306] Using tau binding and HYDRAC PLP, which were first characterized in a conditional system of human 293T cells, candidates (i.e., one or more of SEQ ID NOs: 1-10, 13-16; and a PLP having a peptide side chain comprising one or more of SEQ ID NOs: 11-12, or any other suitable degrader agent) are evaluated using cortical neurons directly differentiated and induced from patient-derived MAPT. Cortical neurons induced from fibroblasts of tauopathy patients are similar to the Alzheimer's brain and express high levels of the 4R isoform of tau, which is different from normal brains that express all six tau isoforms at an approximate 4R / 3R ratio. The directly induced neurons retain an epigenetic aging-related signature, and thus, the induced neurons are an ideal cell line for studying human tauopathy. See, for example, Huh CJ et al., Maintenance of age in human neurons generated by microRNA-based neuronal conversion of fibroblasts. Elife 2016;5:1-14. Total tau isoform expression and the 4R / 3R tau ratio are determined biochemically after PLP treatment. Similarly, thioflavin T staining is used to monitor amyloid formation. The effect of PLP on neuronal survival rate and restoration of the protein homeostasis pathway is evaluated. This allows for an assessment of the broad efficiency of tau-targeting PLP in neurons containing various disease-related tau mutations. As a result, PLP is established as a generalizable chemical biology tool in understanding how to inhibit aggregation and, further, in simulating a new treatment modality in diseases that cannot be prevented by other means and enabling it, providing a new polymer-based platform technology for this purpose.

[0259]

[0307] Success is partially dependent on the ability of HYDRACS to penetrate cells expressing tau, regardless of whether they are HEK293 or SH-SY5Y cells with increased expression. The addition of cationic residues (e.g., arginine residues) to the peptide side chains is predicted to facilitate this. PLP modified with arginine is predicted to enter cells mainly through direct membrane penetration and enable efficient cytosolic targeting of tau. However, the cell surface interaction of arginine-rich peptides may activate the small GTPase Rac, leading to actin organization, formation of lamellipodia, and uptake by macropinocytosis. If this membrane-bound uptake pathway is adopted, tau-binding PLP may be mislocalized to the vesicular trafficking pathway by endocytosis, affecting the efficiency of tau targeting. The entry mechanism of PLP targeting tau is examined, and additional peptide modifications (e.g., using endosomal escape peptides or acid-sensitive functional groups) may be made to increase its vesicular escape if necessary.

[0260]

[0308] Once inside the cell, HYDRACS is predicted to co-localize with tau. The experiments discussed in Examples 6 and 7 were evaluated using fluorophore-labeled polymers and visualized via confocal microscopy. Specifically, the signal of labeled HYDRAC was examined to be within the boundaries of the cell membrane, and in another experiment, it was examined to overlap with tau staining. Co-localization with tau also indicates whether the polymer induces degradation when the tau signal is quantified in the cell via antibody staining or thioflavin T staining.

[0261]

[0309] The mechanism of action of HYDRAC is further investigated by probing this pathway by inhibiting the proteasome (e.g., via MG132) and autophagy (e.g., via bafilomycin A1) to determine whether polyubiquitination guided by HYDRAC is actually the major origin of tau degradation. Western blots probed for tau aggregated with specific antibodies clarify tau levels not only in cell samples but also in tissue samples for in vivo studies. Total tau aggregate concentration can be quantified by Western blot by collecting and lysing the brains of treated and untreated transgenic mice and compared to untreated and treated wild-type controls.

[0262] Example 12

[0310] In this example, the pharmacokinetics, biodistribution, and efficacy of the disease model are evaluated in vivo.

[0263]

[0311] The PLP platform itself enables facile labeling with fluorophores, biotin, and rare earth metal complexes for sensitivity detection. To date, such labels have been used to aid in quantifying PLP in blood and organs for pharmacokinetic (PK) and biodistribution (BD) analysis. Furthermore, an elimination half-life of 152 h for PLP was observed, with the compound detectable in blood and brain 1 week after a single injection. See, for example, Blum AP et al., Peptides displayed as high density brush polymers resist proteolysis and retain bioactivity. J Am Chem Soc 2014;136:15422–37; Blum AP et al., Activating peptides for cellular uptake via polymerization into high density brushes. Chem Sci 2016;7:989–94; Thompson MP et al., Labelling polymers and micellar nanoparticles via initiation, propagation and termination with ROMP. Polym Chem 2014;5:1954–64. Each of these is incorporated herein by reference to the extent not inconsistent with the description herein, and more specifically, is incorporated to demonstrate the facile labeling and detection performance of PLP. After weekly tail vein (IV) injection over an 8-week treatment in Balb / c mice, a biotin tag is added to the most successful tau PLP and tau HYDRAC to examine PK and BD profiles. The successful tau PLP and tau HYDRAC are selected by the utility of each PLP in rescuing cells by disassembling aggregates (HYDRAC) or in collapsing / facilitating tau lesions to improve tauopathy.

[0264]

[0312] The elimination half-lives of tau PLP and tau HYDRAC leads with biotin tags are measured separately after lethality and bleeding and preparation for biotin quantification. This is expected to provide quantification via a streptavidin-biotin detection complex in a commercially available kit. Standard curves are generated from whole blood doped with each PLP to ensure accurate quantification is reliably achieved from precision assays. Additional organs, such as the brain, liver, and kidneys, are collected for maceration and biotin quantification and pathology. Eighteen time points over an 8-week treatment period allow for complete characterization of the PK / BD profile and are expected to confirm that tau PLP does not reach the brain and has no toxic effects in mice. To address scientific rigor, both female and male mice will be evaluated in these studies.

[0265]

[0313] For validation after human neuron experiments, corresponding MAPT P301S tau mutant transgenic mice are treated with lead tau PLP and tau HYDRAC with respect to behavioral and neuropathological efficacy. The motor skills and memory of the animals are evaluated by rotarod and open field tests and weekly body weight measurements. Neuropathology is evaluated using western blot and immunohistochemical detection of tau in the brain when lethality occurs. In these studies, both female and male mice, including 55 males and 55 females distributed across various wild-type and disease groups, will be evaluated.

[0266] Description of Incorporation by Reference and Variations

[0314] All patent documents, such as all references cited throughout this application, e.g., issued or granted patents or equivalents thereof; published patent applications; and non-patent literature documents or other sources, are incorporated herein by reference in their entirety as if each reference were individually incorporated by reference to the extent that each reference does not conflict at least partially with the disclosure in this application (e.g., references that do not fully match are incorporated by reference except for the non-matching portions of the references).

[0267]

[0315] The terms and expressions employed in this specification are used as terms for explanation and not for limitation. In the use of such terms and expressions, it is not intended to exclude any equivalents or portions thereof of the features shown and described. However, it is recognized that various modifications are possible within the scope of the claimed invention. Accordingly, although the present invention has been specifically disclosed by preferred embodiments, exemplary embodiments and optional features, modifications and variations of the concepts disclosed herein can be reselected by those skilled in the art, and it is understood that such modifications and variations are considered to be within the scope of the present invention as defined by the appended claims. The specific embodiments provided herein are examples of useful embodiments of the present invention, and those skilled in the art will understand that the present invention may be practiced using numerous variations of the devices, components of the devices, and method steps described in the description of the present invention. As will be apparent to those skilled in the art, the methods and devices useful for the method of the present invention may include numerous optional compositions, processing elements, and steps.

[0268]

[0316] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and their equivalents known to those skilled in the art. Similarly, the terms "a" (or "an"), "one or more", and "at least one" can be used interchangeably herein. It should also be noted that the terms "comprising", "including", and "having" can be used interchangeably. The expression "of any of claims XX-YY" (where XX and YY refer to claim numbers) is intended to provide a plurality of dependent claims in alternative form, and in some embodiments, can be replaced by the expression "as in any one of claims XX-YY".

[0269]

[0317] When a group of substituents is disclosed herein, it is understood that all individual members of that group and all subgroups thereof, including any isomers, enantiomers, and diastereomers of the members of the group, are disclosed separately. When Markush groups or other groupings are used herein, it is intended that all individual members of the group, as well as all possible combinations and subcombinations of the group, are individually included in the present disclosure. When a compound is described herein such that a particular isomer, enantiomer, or diastereomer of the compound is not specified, for example, in the form of a formula or a chemical name, the description is intended to include each isomer and enantiomer of the compound described, individually or in any combination. In addition, unless otherwise specified, all isotopic variants of the compounds disclosed herein are intended to be encompassed by the present disclosure. For example, it will be understood that any one or more of the hydrogens in the disclosed molecule may be replaced with deuterium or tritium. Isotopic variants of a molecule are generally useful as standards in assays for the molecule, as well as in chemical and biological investigations related to the molecule or its use. Methods for making such isotopic variants are known in the art. Because it is known to those of ordinary skill in the art that the same compound may be named differently, the specific names of the compounds are intended to be exemplary.

[0270]

[0318] Certain molecules disclosed herein may contain one or more ionizable groups [protons from such groups may be removed (e.g., -COOH), added (e.g., amines), or quaternized (e.g., amines)]. All possible ionic forms of such molecules and their salts are intended to be individually included in the disclosure herein. With respect to the salts of the compounds herein, one of ordinary skill in the art can select, among the various available counterions, those appropriate for the preparation of the salts of this invention for a given application. In a particular application, the selection of a given anion or cation for the preparation of a salt may result in an increase or decrease in the solubility of that salt.

[0271]

[0319] All devices, systems, formulations, combinations of components, or methods described or exemplified in this specification can be used to practice the invention, unless otherwise specified.

[0272]

[0320] Whenever ranges are disclosed in this specification, such as ranges of temperature, time, or composition or concentration, all intermediate ranges and sub-ranges, as well as all individual values included in the disclosed ranges, are intended to be included in this disclosure. It will be understood that any sub-range or individual value within a range or sub-range included in the description in this specification may be excluded from the claims in this specification.

[0273]

[0321] All patents and publications referred to in this specification are indicative of the level of skill of those of ordinary skill in the art to which the invention pertains. The references cited herein are hereby incorporated by reference in their entirety to show that they were state of the art at the time of their publication or filing date, and this information is intended to be used in this specification, if necessary, to exclude specific prior art embodiments. For example, when a composition of matter is claimed, it is to be understood that compounds known in the art and available prior to the applicant's invention, such as compounds for which enabling disclosures are provided in the references cited herein, are not intended to be included in the claims for the composition of matter in this specification.

[0274]

[0322] As used herein, "comprising" is synonymous with "including", "containing", or "characterized by", is inclusive or open-ended, and does not exclude additional, unrecited elements or method steps. "Consisting of" excludes any element, step, or ingredient not specified in the claims of the patent. As used herein, "consisting essentially of" does not exclude materials or steps that do not substantially affect the novel features on which the claims of the patent are based. In any case herein, any of the terms "comprising", "consisting essentially of", and "consisting of" can be replaced by either of the other two terms. The inventions illustratively described herein can be suitably practiced in the absence of any one or more elements or limitations not specifically disclosed herein.

[0275]

[0323] Those skilled in the art will recognize that starting materials, biological materials, reagents, synthetic methods, purification methods, analytical methods, assay methods, and biological methods other than those specifically exemplified can be employed in the practice of the invention without undue experimentation. All functional equivalents known in the art of all such materials and methods are intended to be included in this invention. The terms and expressions employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions to exclude any equivalents or portions thereof of the features shown and described, but it is recognized that various modifications are possible within the scope of the claimed invention. Accordingly, although the invention has been specifically disclosed by preferred embodiments and optional features, modifications and variations of the concepts disclosed herein can be resorted to by those skilled in the art, and such modifications and variations are considered to be within the scope of the invention as defined by the appended claims.

Claims

1. A polymer comprising a first repeating unit comprising a first polymer backbone subunit directly or indirectly covalently linked to a first functional side chain containing a peptide that (i) inhibits the aggregation of at least a portion of a tau protein, (ii) promotes the aggregation of at least a portion of a tau protein, (iii) binds to at least a portion of a tau protein, and / or (iv) mimics at least a portion of a tau protein.

2. A polymer comprising a first repeating unit comprising a first polymer backbone subunit directly or indirectly covalently linked to a first functional side chain containing a peptide that (i) inhibits the aggregation of at least a portion of a microtubulin protein, (ii) promotes the aggregation of at least a portion of a microtubulin protein, (iii) binds to at least a portion of a microtubulin protein, and / or (iv) mimics at least a portion of a microtubulin protein.

3. A polymer characterized by formula (FX1): 【Chemical 1】 (wherein, Each P １ independently contains a peptide; Each P ２ independently contains a peptide, and each case of P ２ differs from each case of P １ ; At least one P １ is, independently, or in combination with other instances of P １ in combination with other instances of P (a) at least a portion of a tau protein, and / or (b) at least a portion of a microtubulin protein (i) inhibits the aggregation of at least a portion of a tau protein, (ii) promotes the aggregation of at least a portion of a tau protein, (iii) binds to at least a portion of a tau protein, and / or (iv) mimics at least a portion of a tau protein; T １ and T ２ are each independently a polymer main chain end group which may be the same or different; B １ , B ２ , and B ３ are each independently polymer main chain subunits; L １ and L ２ are each independently a linking group; R １ is, independently, a substituent; m is an integer from 2 to 1000; n is an integer from 0 to 1000; o is an integer from 0 to 1000; Each connecting line in formula (FX1) represents a connection by a covalent bond including a single bond, a double bond, at least one of one or more atoms, or any combination thereof, and optionally, one or more atoms include carbon, nitrogen, and / or oxygen atoms; B １ 、 B ２ 、 B ３ 、 L １ 、 L ２ 、 R １ 、 P １ 、 and P ２ Each case of, and P １ 、 B ２ 、 B ３ 、 L １ 、 L ２ 、 R １ 、 P １ 、 and P ２ is either the same as or different from any other case of, and P; (i) n is an integer from 1 to 1000, o is an integer from 1 to 1000, and at least one instance of P １ is different from another instance of P １ and / or at least one instance of P ２ is different from another instance of P ２ if so, (ii) the polymer is a block copolymer or a statistical copolymer).

4. P １ The polymer according to claim 3, wherein each case of P independently comprises 5 to 50 amino acids.

5. At least one P １ is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) The polymer according to claim 3 or 4, comprising a sequence having at least 75% sequence identity with the above sequences.

6. At least one P １ is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) The polymer according to any one of claims 3 to 5, comprising a sequence having at least 85% sequence identity to any of the foregoing. **Claim 7** At least one P １ is SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); or SEQ ID NO: 16 (YQQYQDATADEQGRR) The polymer according to any one of claims 3 to 6, comprising any of the foregoing. **Claim 8** P １ at least a part of each case of SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); SEQ ID NO: 14 (DRWMINKRR); SEQ ID NO: 15 (VQPINKR); SEQ ID NO: 16 (YQQYQDATADEQGRR); or Any combination thereof, the polymer according to any one of claims 3 to 7. **Claim 9** P １ At least 75% of all cases of SEQ ID NO: 1 (VQIVYK); SEQ ID NO: 2 (DRVQIVYKRR); SEQ ID NO: 3 (VQIINK); SEQ ID NO: 4 (VQIINKRR); SEQ ID NO: 5 (KVQIINKKLDRR); SEQ ID NO: 6 (YQQYQDATADEQG); SEQ ID NO: 7 (YQQYQDATADEQGRRR); SEQ ID NO: 8 (WMINK); SEQ ID NO: 9 (WMINKRR); SEQ ID NO: 10 (VQPINK); SEQ ID NO: 13 (VQIVYKRR); Sequence number 14 (DRWMINKRR); Sequence number 15 (VQPINKR); Sequence number 16 (YQQYQDATADEQGRR); or any combination thereof The polymer according to any one of claims 3 to 8, comprising

10. At least one P １ is Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR) The polymer according to any one of claims 3 to 9, comprising a sequence having at least 75% sequence identity to

11. At least one P １ is Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR) The polymer according to any one of claims 3 to 10, comprising a sequence having at least 85% sequence identity to

12. At least one P １ is Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); or Sequence number 15 (VQPINKR) The polymer according to any one of claims 3 to 11, comprising

13. P １ at least a portion of each of the cases of Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); Sequence number 15 (VQPINKR); or any combination thereof The polymer according to any one of claims 3 to 12, comprising

14. P １ In at least 75% of all cases of, on a numerical basis, Sequence number 2 (DRVQIVYKRR); Sequence number 4 (VQIINKRR); Sequence number 10 (VQPINK); Sequence number 15 (VQPINKR); or any combination thereof The polymer according to any one of claims 3 to 13, comprising

15. At least one P １ is comprising sequence number 1 (VQIVYK) or sequence number 2 (DRVQIVYKRR); At least another P １ is sequence number 8 (WMINK) or sequence number 9 (WMINKRR) The polymer according to any one of claims 3 to 14, comprising

16. At least one P １ and at least another P １ The polymer according to claim 15, characterized by a proportional amount of

17. At least one P of 1:14 １ : At least other P １ The polymer according to claim 15, characterized by the peptide ratio of

18. At least one P of 7:8 １ : At least other P １ The polymer according to claim 15, characterized by the peptide ratio of

19. At least one P of 14:1 １ : At least other P １ The polymer according to claim 15, characterized by the peptide ratio of

20. At least one P １ is comprising sequence number 4 (VQIINKRR) or sequence number 3 (VQIINK); At least another P １ is sequence number 8 (WMINK) or sequence number 9 (WMINKRR) The polymer according to any one of claims 3 to 19, comprising

21. At least one P １ and at least another P １ The polymer according to claim 20, characterized by a proportional amount of

22. At least one P of 1:14 １ : At least other P １ The polymer according to claim 20, characterized by the peptide ratio of

23. At least one P of 7:8 １ : At least other P １ The polymer according to claim 20, characterized by the peptide ratio of

24. At least one P of 14:1 １ : At least other P １ The polymer according to claim 20, characterized by the peptide ratio of

25. At least one P １ is comprising sequence number 10 (VQPINK) or sequence number 15 (VQPINKR); At least another P １ is SEQ ID NO: 3 (VQIINK), SEQ ID NO: 4 (VQIINKRR), SEQ ID NO: 5 (KVQIINKKLDRR), SEQ ID NO: 1 (VQIVYK), or SEQ ID NO: 2 (DRVQIVYKRR) The polymer according to any one of claims 3 to 24, comprising the same. **Claim 26** At least one P １ and at least another P １ The polymer according to claim 25, characterized by proportional amounts thereof. **Claim 27** At least one P of 1:14 １ : At least other P １ The polymer according to claim 25, characterized by the peptide ratio of **Claim 28** At least one P of 8:7 １ : At least other P １ The polymer according to claim 25, characterized by the peptide ratio of **Claim 29** At least one P of 14:1 １ : At least other P １ The polymer according to claim 25, characterized by the peptide ratio of **Claim 30** P ２ each instance of which, when present, independently contains 5 to 50 amino acids The polymer according to any one of claims 3 to 29. **Claim 31** o is an integer from 1 to 1000, At least one P ２ The polymer according to any one of claims 3 to 30, wherein the polymer contains a sequence capable of inducing the ubiquitination cell mechanism of an organism, and optionally, the amino acid sequence thereof binds to an E3 ubiquitin ligase. **Claim 32** A sequence capable of inducing the ubiquitination cellular mechanism of an organism is SEQ ID NO: 11 (ALAPYIP) or SEQ ID NO: 12 (ALAPYIPRR) For which, (i) having at least 75% or at least 90% amino acid composition similarity thereto, (ii) having at least 60% or at least 80% sequence identity thereto, or (iii) containing the same, the polymer according to claim 31. **Claim 33** At least one P １ comprises SEQ ID NO: 6 (YQQYQDATADEQG), SEQ ID NO: 7 (YQQYQDATADEQGRRR), SEQ ID NO: 16 (YQQYQDATADEQGRR), SEQ ID NO: 3 (VQIINK), SEQ ID NO: 4 (VQIINKRR), SEQ ID NO: 5 (KVQIINKKLDRR), SEQ ID NO: 1 (VQIVYK), or SEQ ID NO: 2 (DRVQIVYKRR); At least one P ２ The polymer according to claim 31, wherein the polymer contains SEQ ID NO: 11 (ALAPYIP) or SEQ ID NO: 12 (ALAPYIPRR). **Claim 34** o is an integer from 1 to 1000, and the polymer is P with a peptide ratio of 14:1 １ :P ２ The polymer according to any one of claims 3 to 33, characterized by a peptide ratio of **Claim 35** o is an integer from 1 to 1000, and the polymer is P with a peptide ratio of 13:2 １ :P ２ The polymer according to any one of claims 3 to 34, characterized by the peptide ratio of **Claim 36** o is an integer from 1 to 1000, and the polymer is P with a ratio of 5:1 １ :P ２ The polymer according to any one of claims 3 to 35, characterized by a peptide ratio of **Claim 37** o is an integer from 1 to 1000, and the polymer is P with a peptide ratio of 3:1 １ :P ２ The polymer according to any one of claims 3 to 36, characterized by a peptide ratio of **Claim 38** The polymer according to any one of claims 3 to 37, wherein the tau protein and / or the microtubulin protein is in the form of an oligomer, a protofibril, an amyloid fiber, a cross-β sheet amyloid species, or any combination thereof. **Claim 39** When the tau protein is in a liquid-liquid phase separation (LLPS) state, at least one P １ independently, or in combination with other instances of P １ for at least a portion of the tau protein, (a) inhibits its aggregation, (b) promotes its aggregation, (c) binds thereto, and / or (d) mimics it, a polymer according to any one of claims 3 to 38. **Claim 40** The polymer according to claim 39, which disrupts and / or prevents further growth of the LLPS state of the tau protein. **Claim 41** The polymer according to any one of claims 3 to 40, which can be in the LLPS state under physiological conditions. **Claim 42** (a) at least a part of the tau protein contains the aggregating region of the tau protein, or (b) at least a part of the microtubulin protein contains the aggregating region of the microtubulin protein; or (c) a combination thereof The polymer according to any one of claims 3 to 41. **Claim 43** At least one P １ and / or at least one P ２ The polymer according to any one of claims 3 to 42, wherein the polymer is characterized by a net positive charge. **Claim 44** At least one P １ and / or at least one P ２ The polymer according to any one of claims 3 to 43, characterized by a net positive charge of 1 to 5. **Claim 45** P １ The polymer according to any one of claims 3 to 44, wherein at least 75% of all cases of P are characterized by a net positive charge on a number basis. **Claim 46** P ２ The polymer according to any one of claims 3 to 45, wherein at least 75% of all cases of P are characterized by a net positive charge on a number basis. **Claim 47** At least one P １ and / or at least one P ２ The polymer according to any one of claims 3 to 46, wherein the polymer contains or further contains at least one arginine. **Claim 48** At least one P １ and / or at least one P ２ The polymer according to any one of claims 3 to 47, wherein it contains or further contains at least one aspartic acid. **Claim 49** At least one P １ and / or at least one P ２ The polymer according to any one of claims 3 to 48, wherein it contains or further contains at least one proline. **Claim 50** The polymer according to any one of claims 1 to 49, which is metaphilic. **Claim 51** The polymer according to any one of claims 1 to 50, characterized by an average degree of polymerization of 2 to 100. **Claim 52** The polymer according to any one of claims 1 to 51, characterized by an average degree of polymerization of 2 to 50. **Claim 53** The polymer according to any one of claims 1 to 52, characterized by an average degree of polymerization of 2 to 30.

54. The polymer according to any one of claims 1 to 53, characterized by an average degree of polymerization of 5 to 30.

55. The polymer according to any one of claims 1 to 54, characterized by a number average molecular weight of 1 kDa to 50 kDa.

56. The polymer according to any one of claims 1 to 55, characterized by a number average molecular weight of 1 kDa to 30 kDa.

57. The polymer according to any one of claims 1 to 56, characterized by a brush density of at least 75%.

58. The polymer according to any one of claims 1 to 57, characterized by a brush density of at least 80%.

59. The following characteristics: (a) P １ contains 5 to 100 amino acids; (b) m is an integer from 2 to 100; (c) n is an integer from 0 to 100; (d) o is an integer from 0 to 100; (d) m is an integer from 2 to 100, n is 0, p is 0, and at least one case of P １ is different from another case of P １ ; (e) The degree of polymerization (m + n + o) is an integer from 2 to 200, or 2 to 50; (f) A molecular weight of 1 kDa to 1,000 kDa; When the polymer has a density of at least 50% of P as defined by the equation m / (m + n + o) × 100; １ having a density of peptide; (h) Combinations thereof; (i) Any combination thereof The polymer according to any one of claims 3 to 58, having at least one of them.

60. m is an integer from 2 to 100, and the polymer is polyvalently bound to at least a part of the tau protein and / or at least a part of the microtubulin protein via at least two P １ The polymer according to any one of claims 3 to 59, which is polyvalently bound through.

61. B １ , B ２ , or B ３ The polymer according to any one of claims 3 to 60, wherein at least one of them, or all three of them, is a polymerized monomer containing an unsaturated monomer.

62. The polymer according to claim 61, wherein the unsaturated monomer includes an ethylenically unsaturated monomer, a norbornene monomer, or a norbornene dicarboximide.

63. The polymer according to any one of claims 1 to 62, wherein the polymer is prepared by a living polymerization method optionally selected from ring-opening metathesis polymerization (ROMP), reversible addition-fragmentation chain transfer polymerization (RAFT), or atom transfer radical polymerization (ATRP).

64. Each case of the partial structure (S1a) in formula (FX1): 【Chemical 2】 is independently a partial structure (S1b) or a partial structure (S1c): 【Chemical Formula 3】 [Chemical Formula 4] comprising, wherein R ２ is H or C １ to C ３ alkyl, the polymer according to any one of claims 3 to 63.

65. Each case of the partial structure (S2a) in formula (FX1): 【Chemical Formula 5】 is independently a partial structure (S2b) or a partial structure (S2c): [Chemical Formula 6] 【Chemical Formula 7】 comprising, wherein R １６ is H or C １ to C ３ alkyl, the polymer according to any one of claims 3 to 64.

66. L １ and L ２ In each case where they exist, independently, a single bond, -O-, -(CH ２ CH ２ O) ｘ -, C １ ~C １０ alkyl, C １ ~C １０ acyl, C ２ ~C １０ alkenyl, C ３ ~C １０ aryl, C １ ~C １０ alkoxyl, or any combination thereof, wherein x is an integer from 1 to 20, and each L １ and L ２ , when they exist, attaches B １ to P １ by a covalent bond, and attaches B ３ to P ２ by a covalent bond, and is composed of one or more suitable functional groups, the polymer according to any one of claims 3 to 65.

67. R １ 、 T １ 、 and T ２ each of which is independently hydrogen, C １ ~C ３０ alkyl, C ３ ~C ３０ cycloalkyl, C ５ ~C ３０ aryl, C ５ ~C ３０ heteroaryl, C １ ~C ３０ acyl, C １ ~C ３０ hydroxyl, C １ ~C ３０ alkoxy, C ２ ~C ３０ alkenyl, C ２ ~C １０ alkynyl, C ５ ~C ３０ alkylaryl, -CO ２ R ３ , -CONR ４ R ５ , -COR ６ , -SOR ７ , -OSR ８ , -SO ２ R ９ , -OR １０ , -SR １１ , -NR １２ R １３ , -NR １４ COR １５ , C １ ~C ３０ alkyl halide, phosphonate, phosphonic acid, silane, siloxane, silsesquioxane, C ２ ~C ３０ halocarbon chain, C ２ ~C ３０ perfluorocarbon, C ２ ~C ３０ a polyethylene glycol, a metal, a metal complex, a moiety containing a fluorophore, or a moiety containing a contrast agent, and each of R ３ ~R １５ is independently H, C ５ ~C １０ aryl, or C １ ~C １０ The polymer according to any one of claims 3 to 66, which is alkyl.

68. At least one P １ , P ２ , R １ , T １ , or T ２ contains a fluorophore-containing moiety, or a contrast agent-containing moiety, or further contains the same, the polymer according to any one of claims 3 to 67.

69. At least one P １ , P ２ , R １ , T １ , or T ２ is included in or further includes rhodamine, fluorescein, Cy5.5, gadoteric acid, or a combination thereof, the polymer according to any one of claims 3 to 68.

70. A composition comprising the polymer according to any one of claims 1 to 69 and a pharmaceutically acceptable carrier.

71. A medicament for use in preventing, disintegrating, promoting, or detecting tau and / or microtubulin protein aggregation, comprising a composition having a polymer according to any one of claims 1 to 69 in a therapeutically effective amount.

72. A medicament for use in preventing, treating, or detecting a tauopathy-related disease or condition in a subject, comprising a composition having a polymer according to any one of claims 1 to 69 in a therapeutically effective amount.

73. A method for preventing, disintegrating, promoting, or detecting tau and / or microtubulin protein aggregation, comprising: contacting oligomers, protofibrils, amyloid fibrils, and / or cross-β sheet amyloid species of tau and / or microtubulin with a polymer according to any one of claims 1 to 69 in a therapeutically effective amount or a composition according to claim 70.

74. The method according to claim 73, wherein the oligomers, protofibrils, amyloid fibrils, and / or cross-β sheet amyloid species of tau and / or microtubulin are in a patient or in a fluid derived from a subject.

75. at least one R １ is a fluorophore-containing moiety or a contrast agent-containing moiety, and the method further comprises an imaging step after the contacting step, the method according to claim 73.

76. A method for preventing, treating, or detecting a tauopathy-related disease or condition in a subject, comprising: administering to the subject a polymer according to any one of claims 0 to 69 in a therapeutically effective amount or a composition according to claim 70.

77. The method according to claim 76, wherein the tauopathy-related disease or condition is optionally selected from Alzheimer's disease (AD), primary age-related tauopathy, chronic traumatic encephalopathy, traumatic brain injury, progressive supranuclear palsy, corticobasal degeneration, dementia, frontotemporal dementia, argyrophilic grain dementia, frontotemporal dementia linked to chromosome 17, and parkinsonism, Parkinson's disease, parkinsonism, postencephalitic parkinsonism, amyotrophic lateral sclerosis (ALS), Huntington's disease, vacuolar tauopathy, Lytico-Bodig disease, ganglioglioma, gangliocytoma, meningeal hemangiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, lipofuscinosis, Pick's disease, Pick complex, or a neurodegenerative disease associated with any combination thereof.

78. The method according to claim 76, wherein the polymer is administered to the target brain, spinal cord, cerebrospinal fluid, or any combination thereof.

79. At least one R １ is a part containing a fluorophore or a part containing a contrast agent, and the method according to any one of claims 76 to 78, further comprising an imaging step after the administering step.

80. Use of a composition for preventing, disrupting, promoting, or detecting tau and / or microtubulin protein aggregation, the composition comprising a polymer according to any one of claims 1 to 69.

81. Use of a composition for preventing, treating, or detecting a tauopathy-related disease or condition in a subject, the composition comprising a polymer according to any one of claims 1 to 69.

82. A method for producing a polymer according to any one of claims 0 to 69, At least one P １ A step of synthesizing a peptide; At least one P １ The peptide is capped at the end with a polymerizable monomer that becomes the polymer main chain subunit B once polymerized １ thereby forming a polymerizable P １ monomer; Polymerizable P １ Step of polymerizing monomers comprising.

83. The synthesizing step comprises solid-phase synthesis using protected amino acids; The polymerizable monomer comprises an ethylenically unsaturated monomer optionally containing norbornene or (meth)acrylate; The method according to claim 82, wherein the polymerizing step comprises ROMP, RAFT, or ATRP.

84. A method for producing a medicament for use in preventing, disrupting, promoting, or detecting tau and / or microtubulin protein aggregation, a composition having a therapeutically effective amount of a polymer according to any one of claims 1 to 69, and an optional carrier comprising the step of combining.

85. A method for producing a medicament for use in preventing, treating, or detecting a tauopathy-related disease or condition in a subject, a composition having a therapeutically effective amount of a polymer according to any one of claims 1 to 69, and an optional carrier comprising the step of combining.

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