Peptide therapeutics for treating alzheimer's disease and related conditions

Abstract

To provide peptide compounds in treatments of Alzheimer's disease, neuroinflammation, and cerebral amyloid angiopathy.SOLUTION: The present invention provides peptide compounds derived from cathepsin G (CG), neutrophil elastase, and cationic antimicrobial protein of 37 kDa (CAP37), and methods of their use in inhibiting and / or reversing polymerization of amyloid β peptide into oligomers and / or fibrils, and / or disaggregating amyloid β plaques, and methods of use of the peptide compounds in treatments of, for example, Alzheimer's disease, neuroinflammation, and cerebral amyloid angiopathy.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is expressly incorporated by reference in its entirety herein. This application claims priority to U.S. Provisional Patent Application No. 62 / 682,455, filed on the 6th of this application.

[0002] government support This invention was made possible by grant numbers R21EY026229 and R21EY026229 awarded by the National Institutes of Health. This invention was made with government support under grants P20GM103640-06 and P20GM103640-06. The Company has certain rights. [Background technology]

[0003] Alzheimer's disease (AD) is the most common cause of dementia among people aged 65 and over. It is the sixth leading cause of death in the United States. It currently affects approximately 5 million people. The number of people diagnosed with AD is estimated to double over the next 20 years. It cannot be cured, prevented or delayed, and therefore there is no efficient treatment approach. There is a significant need for development of novel approaches.

[0004] Cationic antimicrobial protein of 37 kDa kDa (CAP37) was associated with increased neuroprotection in patients with AD compared with age-matched controls. It has previously been shown that CAP37 is significantly upregulated in the retina and hippocampal vasculature. The protein amyloid beta (amyloid β, or Aβ) in the brains of people with AD It binds to and cleaves CAP37 protein (a major component of amyloid plaques found in It has also been shown that proteins interact with the proinflammatory receptor for advanced glycation end products (RAGE). Aβ accumulates in the brain early during the progression of AD and is a key hallmark of AD pathology. As its concentration increases, Aβ polymerizes, initially forming soluble oligomers and fibrils. They then precipitate to form insoluble amyloid plaques. The accumulation is due to (1) increased production of amyloid precursor protein (APP) through cleavage; (2) This may be the result of (3) increased influx from the blood and / or (4) decreased clearance.

[0005] RAGE is a member of the immunoglobulin superfamily of transmembrane receptors and is a pro-inflammatory cytokine. Aβ (in the form of monomers, oligomers, and fibrils) mediates the RAGE signal. Other known ligands for RAGE include advanced glycation end products (AGEs), S100 / calgranulin and high mobility group box 1 (HMGB1) proteins RAGE is expressed at low levels in adult tissues and is involved in the regulation of RAGE expression. When the concentration of the ligand increases, it is upregulated by a positive feedback loop. AGE expression is upregulated in the brains of patients with AD. RAGE mediates the synthesis of Aβ oligomers and fibrils in neurons and microglial cells. It is believed to play a role in mediating the toxic effects of RAGs. RAGE can mediate the induction of oxidative stress and hyperphosphorylation of tau. also mediate the activation of microglial cells and underlying chronic neuroinflammation associated with AD. At the blood-brain barrier, RAGE activation leads to the breakdown of barrier function, which is associated with AD. Finally, RAGE is thought to be involved in the transcytosis of inflammatory bowel disease and other pathological features. It mediates the entry of Aβ from the circulation into the brain via the endothelial cell membrane.

[0006] Previous findings have established that: (1) CAP37 expression is consistent with age However, more patients with Alzheimer's disease (AD) than those without a diagnosis of AD This suggests that the amount of CAP37 in the brain is increased in the brain cells of the affected individual. They established a positive correlation between two closely related proteins, neutrophil elastase and neutrophil β-actin. (2) CAP37, NE, and cathepsin G (CG) were not included. and regulation of CG expression is a ligation mechanism for the proinflammatory receptor for advanced glycation end products (RAGE). (3) CAP3, which generally correlates with the regulation of proteins that act as 7 binds to RAGE in a manner similar to the Aβ peptide, a known ligand of RAGE. NE and CG also bind to RAGE; (4) CAP37-RAGE and NE-RAGE interaction is inhibited by specific RAGE inhibitors. (5) CA P37, NE, and CG bind to Aβ, and (6) CAP37, NE, and and CG exert protease activity on Aβ, which is mediated by several identified cleavage sites. Cut at the cut site.

[0007] Thus, both Aβ and RAGE are identified as important pharmacological targets in the pathogenesis of AD. The present disclosure is directed to a method for targeting Aβ and RAGE, and can be used to treat patients with AD and / or neuroinflammation. Drug identification. [Brief description of the drawings]

[0008] [Figure 1] Figure 1 shows the binding of amyloid beta 1-42 (Aβ1-42) to full-length CAP37, NE, and CG proteins. Binding is stronger for CAP37 and CG and is not significantly altered by the addition of antiproteases (phenylmethylsulfonyl fluoride (PMSF) or chymostatin). Binding of Aβ1-42 to NE is almost undetectable but is significantly increased in the presence of PMSF, which inhibits cleavage of Aβ1-42 by NE. [Diagram 2] Figure 1 shows the levels of Aβ1-42 binding to wild-type CAP37-derived peptides 20-44 (20-44 CC, 95-122, and 120-146 QR) and mutant CAP37-derived peptides (20-44 CS, 20-44 SC, 120-146 WR, 120-146 QH, and 120-146 WH). Aβ1-42 did not bind to peptide 95-122. [Diagram 3] FIG. 3 shows the levels of Aβ1-42 binding to the “5RMP” modified CAP37-derived peptide of FIG. 2 (two AEEA mini-PEG molecules and four consecutive arginines linked to the N-terminus of the peptide, and one arginine linked to the C-terminus of the peptide). [Figure 4] Graph showing the level of fibrillation (aggregation) of Aβ1-42 in the presence of the indicated concentrations of full-length CAP37, NE, or CG. The time for complete fibrillization of Aβ1-42 (without added protein) is indicated by the vertical dotted line. [Diagram 5] 1 is a graph showing the level of Aβ1-42 fibrillization (aggregation) in the presence of full-length CAP37, NE, or CG at various concentrations of protein. NE has the highest inhibitory effect on Aβ1-42 fibrillization, followed by CG and then CAP37. [Figure 6]Figure 1 shows that the protease inhibitor PMSF impairs the inhibitory effect of CAP37 on Aβ1-42 fibrillogenesis. The IC50 of CAP37 is increased 3.5-fold in the presence of PMSF, indicating that the inhibitory effect of CAP37 on fibrillogenesis is at least in part due to the cleavage of Aβ1-42. [Figure 7] Figure 1 shows that the protease inhibitor PMSF abolishes the inhibitory effect of NE on Aβ1-42 fibrillogenesis. The IC50 of NE is increased 36-fold in the presence of PMSF, indicating that the inhibitory effect of NE on fibrillogenesis is due, at least in part, to cleavage of Aβ1-42. [Figure 8] Figure 1 shows a direct correlation between NE cleavage of Aβ1-42 and inhibition of fibrillization, supporting the idea that NE inhibition of fibrillization is primarily mediated by cleavage of Aβ. This experiment was performed in the absence of PMSF. [Figure 9] Figure 1 shows the direct correlation between cleavage of Aβ1-42 by NE and inhibition of fibrillization. This experiment was performed in the presence of PMSF. [Figure 10] Figure 1 shows that the protease inhibitor chymostatin abolishes the inhibitory effect of CG on Aβ1-42 fibrillogenesis. The IC50 of CG is increased 17-fold in the presence of chymostatin, indicating that the inhibitory effect of CG on fibrillogenesis is due, at least in part, to cleavage of Aβ1-42. [Figure 11] 1 shows a direct correlation between CG cleavage of Aβ1-42 and inhibition of fibrillization, supporting the idea that CG inhibition of fibrillization is primarily mediated by Aβ cleavage. This experiment was performed in the absence of chymostatin. [Figure 12] Figure 1 shows the direct correlation between cleavage of Aβ1-42 by CG and inhibition of fibrillization. The experiment was performed in the presence of chymostatin. [Figure 13] Figure 1 shows that CAP37-derived peptides 120-146 WR and 120-146 WR 5RMP inhibit, but not peptide 95-122 5RMP, fibrosis. [Figure 14] FIG. 1 shows that Aβ1-42 oligomers are more toxic to neurons than fibrils, with the monomeric form of Aβ1-42 being the least toxic. [Figure 15] FIG. 1 shows that NE and CG, but not CAP37, inhibit Aβ1-42 toxicity when incubated under conditions that favor fibril formation. [Figure 16] Figure 1 shows the effect of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of NE. NE significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 17] Figure 1 shows the effect of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CG. CG significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 18] Figure 1 shows the effect of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CAP37 protein.Full-length CAP37 does not inhibit Aβ1-42 oligomer toxicity. [Figure 19] Figure 1 shows the effect of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CAP37-derived peptide 120-146 WR. The modified CAP37 peptide significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 20] Figure 1 shows the effect of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of the CAP37-derived peptide 120-146 WR-5RMP. The 5RMP modified CAP37 peptide variant significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Detailed Description In at least certain embodiments, the present disclosure provides a method for the detection of three neutrophil granule proteins: CG, neutrophil elastase, and a 37 kDa cationic antibody Contains peptide sequences obtained or derived from biological proteins (CAP37). A peptide compound that inhibits the fibril formation of amyloid beta (Aβ) ) and methods of their use in inhibiting oligomerization, as well as, for example, AD, cerebral amyloid angiopathy, dementia, and / or neuroinflammation, and / or their To treat, alleviate, and / or inhibit Aβ-related diseases and conditions, including symptoms. The present invention is directed to methods of their use as therapeutic agents for the treatment of Aβ 1-42 (the 42 amino acid form of Aβ) at different sites and with different kinetics. It was discovered that CAP37 is involved in the regulation of the oxidative stress response in the brains of patients with Alzheimer's disease (AD). It was also observed that IL-1 was upregulated in IL-1, but not in NE or CG. CAP37 binds to Aβ peptides, which are known ligands of RAGE. , have been found to bind to RAGE. NE and CG also bind to RAGE. CAP37-RAGE and NE-RAGE interactions are specific RAGE inhibitors It is inhibited by Aβ 1-42 Binding and cleavage of toxic oligomers and fibrils It is hypothesized that the present disclosure would inhibit the ability of CAP3 to form 7, NE, and CG, as well as their peptide derivatives, inhibit Aβ aggregation (fibril formation). The results of this study will describe whether Aβ oligomers and polymers can be inhibited. However, it is believed that the monomeric form is more toxic than the monomeric form. This would reduce β toxicity and slow the progression of AD.

[0010] Without wishing to be bound by theory, the peptide compounds of the present disclosure inhibit RA at the blood-brain barrier. It can bind to GE, thereby inhibiting the binding of systemically produced Aβ, and Therefore, it is thought that the peptide, not Aβ, is then transported to the brain. It will be transported to the brain through the blood-brain barrier by RAGE-mediated transcytosis. Once in the brain, the peptides inhibit Aβ oligomerization and / or polymerization, preventing its aggregation. This causes disaggregation of Aβ (depolymerizing Aβ oligomers and fibrils), thereby It reduces the toxic effects of Aβ and its deposition in amyloid plaques. The peptide inhibits the neuronal inflammatory response mediated by aggregated Aβ and other agonists. and bind RAGE and TLR4 on microglial cells and their pro-inflammatory effects , thereby reducing neuroinflammation.

[0011] Before describing in further detail various embodiments of the compositions of the present disclosure and methods of their use, In particular, the disclosure applies to the details of the methods and compositions as set forth in the following description. It should be understood that the present invention is not limited to the above-mentioned embodiments. The description provided herein is by way of example only. It is intended for the purposes of illustration only and is not intended to be construed in a limiting sense. , other embodiments are possible or the invention may be practiced or carried out in various ways. As such, the language used herein is to be construed in the broadest possible scope and meaning. The following examples are intended to give a taste and the embodiments are intended to be exemplary and not comprehensive. Additionally, the phraseology and terminology used herein is for purposes of description and not to be construed as limiting the scope of the invention. It should be understood that unless otherwise stated, the disclosures contained herein should not be construed as limiting. Furthermore, in the following detailed description, numerous specific details are set forth in order to provide a more complete understanding of the present disclosure. These are presented for the purposes of understanding. However, various embodiments of the present disclosure may It will be apparent to one skilled in the art that the present invention may be practiced without the specific details. Well-known features have not been described in detail to avoid unnecessarily complicating the description. All alternatives, substitutions, modifications, and equivalents apparent to those skilled in the art are as defined herein. It is intended to be within the scope of the present disclosure, including specific embodiments thereof. The following examples serve to illustrate the practice of the disclosure, the specifics given being presented by way of example and not limitation. For illustrative purposes only, specific embodiments are discussed and the procedures and principles of the inventive concept are not necessarily intended to be limiting. Provide what is believed to be a useful and easily understood explanation of the theoretical and conceptual aspects of It is understood that the disclosed compositions and methods are presented for the purpose of Although the method has been described with reference to specific embodiments, the concepts of the inventive concepts disclosed herein are not intended to be limiting. without departing from the spirit and scope of the compositions and / or Variations may be applied to the method and in the steps or sequence of steps of the method. It will be apparent to one skilled in the art that

[0012] All patents, published patent applications, and non-patent publications mentioned herein are incorporated by reference in their entirety. The following examples are indicative of the level of skill of those skilled in the art to which they pertain. Each patent, published patent application, and non-patent publication referenced herein is treated as if it were an individual patent, or published patent application. Each such patent application or non-patent publication is specifically and individually indicated to be incorporated by reference. No. 6,393,945, filed on Oct. 13, 2003, and is expressly incorporated herein by reference in its entirety to the same extent as if incorporated herein by reference in its entirety. In particular, International Patent Publication Nos. 2014 / 089088 and 2014 / 09089 are incorporated herein by reference. / 172747 pamphlet, and U.S. Patent No. 9096679, No. 960 3896, 9624283, and 9862748 are all No. 6,393,945, which is expressly incorporated herein by reference in its entirety.

[0013] Unless otherwise defined herein, the scientific and technical terms used in connection with this disclosure All technical terms shall have the meaning commonly understood by one of ordinary skill in the art. Unless otherwise required by , singular terms include plurals and plural terms shall refer to the singular. This includes:

[0014] As utilized in accordance with the methods and compositions of the present disclosure, the following terms are used herein, unless otherwise indicated: and shall be understood to have the following meanings:

[0015] The term "comprising" in the claims and / or specification When used with "one," the words "a" or "an" mean "one can mean "one or more," "at least one," and The meaning of the term "one or more than one" in the claims is consistent with that of the term "one or more than one" in the claims. The use of "or" does not necessarily imply a substitute, unless expressly indicated to refer only to the alternative. Unless the terms are mutually exclusive, even if the disclosure refers only to alternatives and "and / or," The term is used to mean "and / or" even if the definition supports that definition. The use of "at least one" means one, as well as two, three, four, five, six, seven, eight, nine, ten, Contains 15, 20, 30, 40, 50, 100, or any integer therein It is understood that the term "at least one" includes any amount greater than one, including but not limited to. "One more" can mean up to 100 or 1000 or more depending on the term to which it is attached. in addition, the amount of 100 / 1000 may be expanded beyond this limit, even if the higher limit is also satisfactory. In addition, the term "X The use of "at least one of X, Y, and Z" includes X alone, Y alone, and Z alone, as well as It is understood that any combination of X, Y, and Z is included.

[0016] As used in this specification and claims, the word "comprising" means ng)" (and "comprise" and "comprises" any form of "comprising" such as "having" (and "have" and "has" "including" (any form of "including") "es" and "include" "containing" (and "contains"), or "containing" (and "contains") "contains" and "contain" Any form of aining) is inclusive or open-ended and may include additional, enumerated does not exclude any element or method step not included in the disclosure.

[0017] As used herein, the term "or combinations thereof" refers to the list preceding the term. refers to all permutations and combinations of the items listed. For example, "A, B, C, or any combination thereof." "A" is intended to include at least one of the following: A, B, C, AB, AC, BC , or ABC, and also, if the order is important in the particular context, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, BB, AA A, AAB, BBC, AAABCCCC, CBBAAA, CABABB, etc. The phrase explicitly includes combinations containing multiple items or repetitions of terms. Unless otherwise apparent from the context, there is no limit to the number of items or terms in any combination. Those skilled in the art will understand that.

[0018] Throughout this application, the terms "about" or "approximately" refer to values ​​that are consistent with the time of administration of a composition, a composition the inherent variability of error for the method used to determine the As used herein, the modifiers "about" or " "Approximately" does not include an exact value, amount, degree, orientation, or other qualified characteristic or value. The invention is not intended to be limited to the above-mentioned technical aspects, but rather to take into account, for example, measurement errors, manufacturing tolerances, observer errors, and the like. It is also intended to include some slight variations in the amount, duration, and time of the combination of the above. The terms "about" or "approximately" when used herein to refer to measurable values ​​such as "Otherwise" means that such variations are reasonable and appropriate for practicing the disclosed methods. For example, ±20% or ±10% from the specified value as understood by the trader. or ±5%, or ±1%, or ±0.1% variation. When used in this document, the term "substantially" means that the events or circumstances described thereafter occur completely. or the events or circumstances described therein will change to a greater or lesser extent. For example, the term "substantially" means that the The event or situation is at least 80% of the time, at least 90% of the time, or at least 90% of the time. 1% of the time, at least 92% of the time, at least 93% of the time, at least 94% of the time % of the time, at least 95% of the time, at least 96% of the time, at least 97% occurs at least 98% of the time, or at least 99% of the time. This means:

[0019] As used herein, "one embodiment" or "one embodiment" refers to Any reference to the embodiments or "an embodiment" refers to the embodiments. A particular element, feature, composition, structure, or characteristic described in the series may be present in at least one embodiment. In various places in this specification, the phrase "in one embodiment" means The appearances of "in" do not necessarily all refer to the same embodiment.

[0020] As used herein, the terms "AEEA," "AEEEA," "AEEP," and and "AEEEP" are, in certain embodiments, part of the compounds disclosed herein. The short polyethylene glycol molecules ("miniPE") that make up the G").

[0021] The term AEEA refers to [2-(2-amino-ethoxy)-ethoxy]-acetic acid (8-amino The N- or C-terminal group is derived from arylamino-3,6-dioxaoctanoic acid (also known as arylamino-3,6-dioxaoctanoic acid), -NH(CH2CH2O)2CH2CO- or NH2(CH2CH2O)2CH2CO - represents.

[0022] The term AEEEA stands for {2-[2-(2-amino-ethoxy)-ethoxy]-ethoxy 11-amino-3,6,9-trioxaundecanoic acid (also known as 11-amino-3,6,9-trioxaundecanoic acid) The N- or C-terminal groups, -NH(CH2CH2O)3CH2CO- or NH2 (CH2CH2O)3CH2CO-.

[0023] The term AEEP refers to [3-(2-amino-ethoxy)-ethoxy]-propanoic acid ( 9-amino-4,7-dioxanonanoic acid) -NH(CH2CH2O)2CH2CH2CO- or NH2(CH2CH2O)2 It represents CH2CH2CO-.

[0024] The term AEEEP stands for [3-[2-(2-amino-ethoxy)-ethoxy]-ethoxy 12-amino-4,7,10-trioxadodecanoic acid (also known as ), an N- or C-terminal group derived from -NH(CH2CH2O)3CH2CH2CO- Or NH2(CH2CH2O)3CH2CH2CO-.

[0025] Below are other abbreviations used in this specification, as well as the whole words or abbreviations they represent: or a list of phrases: AGE: Advanced glycation end products Aβ: Amyloid beta or amyloid beta ANOVA: Analysis of variance APP: Amyloid precursor protein CAP37: A cationic antimicrobial protein with a molecular weight of 37 KDa CG: Cathepsin G BCA: Bicinchoninic acid BSA: bovine serum albumin DAB: diaminobenzidine tetrahydrochloride BPI: Bactericidal Permeability Enhancer DAG: diacylglycerol EDTA: Ethylenediaminetetraacetic acid EGF: Epidermal growth factor EGFR: epidermal growth factor receptor ERK: Extracellular signal-regulated kinase FBS: Fetal bovine serum 5RMP: 5Arginine-miniPEG GCSF: Granulocyte colony-stimulating factor GM-CSF: Granulocyte-macrophage colony-stimulating factor GPCR: G protein-coupled receptor HB-EGF: Heparin-binding epidermal growth factor HBSS: Hanks' balanced salt solution HCEC: human corneal epithelial cells H&E: Hematoxylin and Eosin HGF: Hepatocyte growth factor HBD-1: Human beta-defensin-1 HMGB1: high mobility group box 1 IL-6: Interleukin-6 IL-8: Interleukin-8 IP-10: Interferon-inducible protein-10 KC: keratinocyte-derived chemokine KSFM: Keratinocyte serum-free medium LPS: lipopolysaccharide MCP-1: Monocyte chemotactic protein-1 NADP: Nicotinamide adenine dinucleotide phosphate NE: neutrophil elastase PBS: Phosphate-buffered saline PDGF-BB: Platelet-derived growth factor-BB PKC: Protein kinase C PMA: phorbol 12-myristate 13-acetate PMSF: Phenylmethylsulfonyl fluoride RAGE: Receptor for advanced glycation end products RIPA: Radioimmunoprecipitation ROS: reactive oxygen species SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis SFM: Serum-free medium TBS: Tris-buffered saline TBST: Tris-buffered saline TWEEN® 20 (Thermo Fisher Scientific) R Scientific, Pittsburgh, PA) TGF-β: Transforming growth factor beta TNF-α: Tumor necrosis factor alpha CG 119-144 : Amino acids 119-144 of cathepsin G protein NE 119-145 : Amino acids 119-145 of neutrophil elastase CAP37 120-146 : Amino acids 120 to 146 of the CAP37 protein CG 20-47 : Amino acids 20-47 of cathepsin G protein NE 20-44 : Amino acids 20-44 of neutrophil elastase CAP37 20-44 : Amino acids 20 to 44 of the CAP37 protein

[0026] The following standard amino acid abbreviations are used herein: Alanine: ala: A, Cysteine: cys: C, Aspartic acid: asp: D, Gluta Glu:E, Phenylalanine:Phe:F, Glycine:Gly:G, Histidine: his:H, isoleucine:ile:I, lysine:lys:K, leucine:leu: L, methionine: met: M, asparagine: asn: N, glutamine: gln: Q, Proline: P, Arginine: R, Serine: S, Threonine: th r:T, valine:val:V, tryptophan:trp:W, and tyrosine:tyr: T.

[0027] The term "mutant" or "variant" refers to a protein, peptide, nucleic acid, or organism. A gene having at least one amino acid or nucleotide that differs from the wild-type version of is intended to refer to a protein, peptide, nucleic acid, or organism, and includes point substitutions, multiple consecutive Contiguous or non-contiguous substitutions, insertions, chimeric or fusion proteins, and Examples of conservative amino acid substitutions include, but are not limited to, Basic amino acids (e.g., arginine, lysine, histidine), acidic amino acids (e.g., glutamic acid and aspartic acid), polar amino acids (e.g., glutamine and aspartic acid), argin), hydrophobic amino acids (e.g., leucine, isoleucine, and valine), aromatic Amino acids (e.g., phenylalanine, tryptophan, tyrosine) and small amino acids Within the same group of acids (e.g., glycine, alanine, serine, threonine, methionine) Substitutions that occur within a group include, but are not limited to, those listed below. It is listed.

[0028] As used herein, the terms "peptide," "peptide analog," "peptide derivative," "Peptide" or "peptide compound" may refer to a molecule that contains only amino acids, or comprises an amino acid and one or more non-amino acid structures (e.g., PEG units). The terms "peptide", "peptide analogue", "peptide derivative", Alternatively, a "peptide compound" may be a variant ("mutant") of a "wild-type" peptide, or Molecular weight compounds that contain amino acids and one or more non-amino acid structures (e.g., PEG units). It may refer to a child.

[0029] The term "amyloid beta" (Aβ) refers to 1 to 40 Aβ, unless otherwise specified. It refers to Aβ peptides containing any of 1 to 42 amino acids, or other forms thereof.

[0030] The term "pharmaceutical acceptable" means a substance that is free of toxicity, irritation, or other adverse effects, consistent with a reasonable benefit / risk ratio. and / or without undue adverse side effects, such as allergic responses, in humans and / or It refers to compounds and compositions that are suitable for administration to an animal.

[0031] "Bioactive" means how an active agent produces its physiological effect. By "modification," we mean the ability to modify the physiological system of an organism, regardless of whether it is induced by a chemical reaction or not.

[0032] As used herein, "pure" or "substantially pure" means that the object species is present in a substantially pure form. It is the predominant species present (i.e., on a molar basis, how much of the composition it is present in) More specifically, the substantially purified fraction is more abundant than the total amount of any other target species present. A composition in which at least about 50 percent (on a molar basis) of the species is the target species. Generally, a substantially pure composition means that all of the macromolecular species present in the composition are pure. More than about 80%, more particularly, more than about 85%, more than about 90%, or more than about 95% or greater than about 99%. "Pure" also means that the species of interest (e.g., a peptide compound) is at least 60% (w / w) pure, or at least 70% (w / w) pure, or at least 75% (w / w) pure, or is at least 80% (w / w) pure, or at least 85% (w / w) pure, or At least 90% (w / w) pure, or at least 92% (w / w) pure, or At least 95% (w / w) pure, or at least 96% (w / w) pure, or at least 97% (w / w) pure, or at least 98% (w / w) pure, or at least 99 % (w / w) pure, or 100% (w / w) pure.

[0033] The terms "subject" and "patient" are used interchangeably herein and refer to warm-blooded animals, particularly mammals. It is understood that this term refers to animals. Non-limiting examples of animals within the scope and meaning of this term include dogs, Cats, rabbits, rats, mice, guinea pigs, chinchillas, hamsters, ferrets, horses , pigs, goats, cows, sheep, zoo animals, camels, llamas; Old World and New World monkeys, etc. and non-human primates (e.g., cynomolgus monkeys, chimpanzees, rhesus monkeys, orangutans, non-human primates, including rats, monkeys, and baboons; and humans.

[0034] As used herein, the term "active agent" refers to a biological agent as described herein. It refers to a compound or composition having a biological activity.

[0035] "Treatment" refers to therapeutic treatment. "Prevention" refers to prophylactic or preventative measures. The term "treating" refers to the administration of a composition to a patient for therapeutic purposes.

[0036] The terms "therapeutic composition" and "pharmaceutical composition" refer to a composition that is prepared by any method known in the art. or any other method contemplated herein, Administration of the composition induces a therapeutic effect as described elsewhere herein. In addition, the compositions of the present disclosure may be formulated to provide delayed, controlled release, or other therapeutic effects using formulation techniques well known in the art. The compositions may be designed to provide extended, prolonged, and / or sustained release.

[0037] The term "effective amount," when used in the context of an inventive concept, is a reasonable amount commensurate with a reasonable benefit / risk ratio. without excessive adverse side effects (e.g., toxicity, irritation, and allergic responses). An effective amount for a patient refers to the amount of active agent that is sufficient to produce a therapeutic effect. the type of patient, the size and health of the patient, the nature and severity of the condition to be treated, The method of administration, duration of treatment, nature of concomitant therapy (if any), specific drug product used, etc. Thus, it is not possible to specify an exact effective amount in advance. However, an effective amount for a given situation can be determined using routine practice based on the information provided herein. This can be determined experimentally by one of ordinary skill in the art.

[0038] The term "ameliorate" refers to the reduction or amelioration of a detectable or measurable decrease in a subject's condition, disease, or symptoms thereof. A detectable or measurable improvement means an improvement that is more pronounced than the onset of a condition or disease. A subjective or objective decrease, reduction, or inhibition of the rate, frequency, severity, progression, or duration of , inhibit, limit, or control, or the symptoms or underlying causes or consequences of disease Successful treatment results in the improvement or reversal of a disease or condition. The prognosis, incidence, frequency, severity, progression, or duration of a disease or condition in a subject To ameliorate, reduce, decrease, inhibit, suppress, limit, control, or A “therapeutic effect” or “benefit” may be achieved by preventing or treating a disease.

[0039] A reduction or decrease in deterioration, such as stabilization of a condition or disease, is also a successful treatment outcome. Thus, the therapeutic benefit is a benefit to a disease or condition or to a disease or condition associated with the disease or condition. Any one, most, or all of the adverse symptoms, complications, prognosis, or underlying It does not require complete disappearance or reversal of a cause. Points are calculated based on the incidence, frequency, severity, progression, or duration of a condition or disease. partially reduce, decrease, inhibit, suppress, limit, control, or prevent the Inhibition or improvement (e.g., of symptoms) over short or long durations (hours, days, weeks, months, etc.) This may be achieved if there is gradual improvement (e.g., stabilization) of the underlying condition or disease. The efficacy of a method or use, such as a treatment, to provide a therapeutic benefit or improvement may be evaluated in various ways and This can be confirmed by test assays.

[0040] The term "peptide" or "peptide sequence" typically refers to a contiguous peptide that forms a sequence of amino acids. On the other hand, peptide bonds between the alpha-amino and carbonyl groups of the selected amino acids It is used herein to refer to a series of amino acid residues connected from one to another. In this embodiment, the peptide is from 5 to 15, up to 25, up to 40 amino acids in length. up to 60, up to 75 or more amino acids, e.g., 5, 6, 7 , 8 pieces, 9 pieces, 10 pieces, 11 pieces, 12 pieces, 13 pieces, 14 pieces, 15 pieces, 16 pieces, 17 pieces, 1 8 pieces, 19 pieces, 20 pieces, 21 pieces, 22 pieces, 23 pieces, 23 pieces, 24 pieces, 25 pieces, 26 pieces, 2 7 pieces, 28 pieces, 29 pieces, 30 pieces, 31 pieces, 32 pieces, 33 pieces, 34 pieces, 35 pieces, 36 pieces, 3 7 pieces, 38 pieces, 39 pieces, 40 pieces, 41 pieces, 42 pieces, 43 pieces, 44 pieces, 45 pieces, 46 pieces, 4 7 pieces, 48 ​​pieces, 49 pieces, 50 pieces, 51 pieces, 52 pieces, 53 pieces, 54 pieces, 55 pieces, 56 pieces, 5 7, 58, 59, or 60-75 or more amino acids. The term "polypeptide" or "protein" refers to any molecule that is longer than a single peptide in length. A peptide bond, typically between the alpha-amino and carbonyl groups of adjacent amino acids is used herein to refer to a series of amino acid residues connected one to another by A "fusion protein" or "fusion polypeptide" is a peptide that is fused together in a contiguous configuration. Proteins or polypeptides that have been produced by recombinant or synthetic methods to be combined in In certain embodiments, the present invention refers to a polypeptide (and may be used interchangeably). The CAP37, NE, and CG peptide derivatives described elsewhere in the specification were The variant peptides inhibit Aβ oligomerization and / or polymerization and / or As long as the compound retains the biological activity described herein related to Aβ disaggregation, e.g., Or it may be truncated by 1, 2, 3, 4, or 5 amino acids on either end. In one embodiment, the variant peptide has one, two, or three amino acid insertions. may include.

[0041] The peptides of the present disclosure, and the nucleic acids encoding them, may contain any of the naturally occurring amino acids or bases. Included are peptide and nucleic acid variants that contain substitutions (conservative or non-conservative). For example, peptide variants include those that are not exactly the same as the sequences disclosed herein, but In addition to the substitutions explicitly stated for the various sequences listed herein, Amino acid residues (conservative or otherwise) that do not substantially impair the activity or properties of the described variants. Conservative or non-conservative (Ala- ... Examples of such conservative amino acid substitutions include ala to gly, ser, or thr. ;arg to gln, his, or lys;asn to asp, gln, his, l to ys, ser, or thr; asp to asn or glu; cys to ser ;gln to arg, asn, glu, his, lys, or met;glu to a sp, gln, or lys; gly to pro or ala; his to arg, to asn, gln, or tyr; ile to leu, met, or val; leu from ile, met, phe, or val; from lys to arg, asn, gln, or or glu; met to gln, ile, leu, or val; phe to leu , met, trp, or tyr; ser to ala, asn, met, or thr to;thr to ala, asn, ser, or met;trp to phe or ty r; tyr to his, phe, or trp; and val to ile, leu, or or met, but are not limited to these.

[0042] Those skilled in the art will appreciate that such variants can be generated, identified, selected, or non-variant. The peptides are then tested for their binding activity to the same receptors targeted by the peptides. Specific examples of conservative amino acid substitutions include gly:ala substitutions; al:ile:leu replacement;asn:glu:his replacement;asp:glu replacement;ser :thr:met substitution; lys:arg:his substitution; and phe:tyr:trp substitution Other types of recombinant peptides that result in functional variant peptides include, but are not limited to, recombinant peptides. Substitutions, modifications, additions, deletions, and derivatives are also encompassed by the disclosure and will be apparent to those of skill in the art. Methods for making, identifying, or selecting such variants or derivatives, and methods for producing, identifying, or selecting such variants or derivatives One would readily know how to test the agonist for receptor binding activity.

[0043] In certain embodiments, the present disclosure provides a method for the preparation of a peptide having one or more of the following amino acid sequences: The present invention relates to peptide compounds which are fabricated in whole or in part by: CG 119-144 :GTLCTVAGWGRVSMRRGTDTLREVQL (sequence number No. 1), NE 119-145 :GVQCLAMGWGLLGRNRGIASVLQELNV(Array Number 2), CAP37 120-46 :GTRCQVAGWGSQRSGGRLSRFPRFVNV( SEQ ID NO: 3), CG 20-47 :IQSPAGQSRCGGFLVREDFVLTAAHCWG(sequence number No. 4), NE 20-44 : LRGGHFCGATLIAPNFVMSAAHCVA (SEQ ID NO: 5) , and CAP37 20-44 :NQGRHFCGGALIHARFVMTAASCFQ(sequence number No. 6).

[0044] More particularly, the disclosure includes peptide variants that include specific substitutions such as: However, it is not limited to these.

[0045] In certain non-limiting embodiments, CG containing the following substitutions: 119-144 (Sequence number The variants of No. 1) may be used to form the peptide compounds used herein. It can be synthesized by using one or more of the four basic arginine residues (positions 11, 15, 16, and 22). The acidic residue aspartate at position 19 can be changed to lysine or histidine. The glutamic acid can be modified to glutamic acid and / or the glutathione at position 23 can be modified to glutamic acid. Amino acids at positions 2, 5, 18, and 20 can be changed to aspartic acids. One or more of the threonine residues in the ribozyme are hydrophilic residues (asparagine, glutamine, selenite, etc.). to another small residue (serine, alanine, glycine, valine, proline) Two other hydrophilic residues, serine at position 13 and One or more of the glutamines at position 5 may also be replaced by another hydrophilic residue (asparagine, glutamine, serine, threonine, or tyrosine). One or more of the other residues in this sequence is another hydrophobic residue (alanine, Lysine, cysteine, valine, proline, leucine, isoleucine, methionine, phenyl It can be substituted with alpha-aminobutyric acid (e.g., tryptophan, alpha-aminobutyric acid, etc.).

[0046] In other non-limiting embodiments, NE includes the following substitutions: 119-145 (SEQ ID NO:2) Variants of the above can be used to form the peptide compounds used herein. The aromatic tryptophan residue at position 9 can be substituted with another aromatic residue (tyrosine or phenyl). The acidic glutamic acid residue at position 24 can be changed to aspartic acid. Two basic arginines at positions 14 and 16 can be changed to paragic acid. One or more of the residues can be changed to another basic amino acid (lysine or histidine). The polar residues glutamine (positions 3 and 23), asparagine (position 15), and One or more of the serines (position 20) may be substituted with any other polar residue (asparagine, glutamine, etc. , serine, threonine, or tyrosine). The amino acid sequence was also changed to another small residue (threonine, alanine, glycine, valine, proline). One or more of the other residues in this sequence (which are hydrophobic) may be Hydrophobic residues (alanine, glycine, cysteine, valine, proline, leucine, isopropyl It can be substituted with any of the following amino acids: isine, methionine, phenylalanine, tryptophan.

[0047] In another non-limiting embodiment, CAP37 containing the following substitutions: 120-146 (Sequence number The variants of No. 3) may be used to form the peptide compounds used herein. This peptide contains small hydrophobic glycines at positions 1, 8, 10, 15, and 16. One or more of the residues may be substituted with another hydrophobic amino acid (alanine, cysteine, valine, proline, etc. leucine, isoleucine, methionine, phenylalanine, or tryptophan) , or another small amino acid (alanine, serine, threonine, valine, or proline) Similarly, positively charged arginines at positions 3, 13, 17, 20, and 23 can be One, two, three, four or all of the lysine residues may be replaced by another basic amino acid (lysine or Histidine). Other examples of this sequence include the hydrophilic amino acid, Threonine at position 2, glutamine at positions 5 and 12, and amino acids 14 and 19. One approach is to change one or more of the serines at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, Threonine can be changed to asparagine, glutamine, serine, or tyrosine. glutamine can be changed to asparagine, serine, threonine, or tyrosine and serine can be converted to asparagine, glutamine, threonine, or tyrosine. One or more of the other residues in this sequence (which are hydrophobic) can be changed to another Hydrophobic residues (alanine, glycine, cysteine, valine, proline, leucine, isopropyl It can be substituted with any of the following amino acids: isine, methionine, phenylalanine, tryptophan.

[0048] In other non-limiting embodiments, CG 20-47 (SEQ ID NO:4) Alternative variants form the peptide compounds used herein. In this sequence, two negatively charged residues, glutamic acid and and aspartic acid (positions 17 and 18, respectively) are replaced by two glutamic acids or two or aspartic acid and The two nucleotides at positions 9 and 16 of this peptide can be converted to glutamic acid. One or more of the positively charged arginine residues are conservatively changed to lysine or histidine The histidine residue at position 25 can be changed to arginine or lysine. One or more of the polar glutamine residues at positions 2 and 7 can be replaced by asparagine , serine, threonine, or tyrosine; and One or more of the polar serine residues in or tyrosine (which is hydrophobic). or more than one may be another hydrophobic residue (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan) The aromatic residues phenylalanine 13 and 19, and tryptophan 2 One or more of 7 may be replaced by another aromatic residue (phenylalanine, tryptophan, or thiamine). It can be changed to Rosin.

[0049] In other non-limiting embodiments, NE includes the following substitutions: 20-44 (SEQ ID NO:5) Substitution variants can be used to form the peptide compounds used herein. The positively charged arginine residue at position 2 of this peptide can be converted to a lysine or histidine. Conservative modifications may be made to one or more of the two histidine residues at positions 5 and 22. The two polar residues, asparagine, can be changed to arginine or lysine. One or more of serine 15 and serine 19 may be substituted with any other polar residue (asparagine, glutamate, can be changed to amino acids 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, One or more of the other residues in this sequence may be another hydrophobic residue (alanine, glycine, etc. leucine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine The phenyl groups at positions 6 and 16 can be substituted with One or more of the amino acids are substituted with another aromatic residue, either tryptophan or tyrosine. can be changed.

[0050] In another non-limiting embodiment, CAP37 containing the following substitutions: 20-44 (SEQ ID NO:6 Alternative variants of the above may be used to form the peptide compounds used herein. One of the two positively charged arginine residues at positions 4 and 15 of this peptide or more can be conservatively modified to lysine or histidine at positions 5 and 13 One or more of the two histidine residues in One or more of the three aromatic phenylalanine residues at positions 6, 16, and 24 can be The aromatic residues can be changed to tryptophan or tyrosine. Residues asparagine 1, glutamines 2 and 25, threonine 19, and serine 22 One or more of the amino acids may be any other polar residue (asparagine, glutamine, serine, threonine, The other amino acids in this sequence can be changed to amino acids (hydrophobic amino acids, tyrosine, or tyrosine). One or more of the residues may be another hydrophobic residue (alanine, glycine, cysteine, valine, Proline, leucine, isoleucine, methionine, phenylalanine, tryptophan) can be replaced with

[0051] Additionally, the peptide compounds have the following sequences (residues in bold are amino acids of the wild type sequence): CG as shown in 119-144 Peptide, NE 119-145 , C AP37 120-146 , C.G. 20-47 , N.E. 20-44 , or CAP37 20-4 The present invention may include a peptide having an amino acid sequence derived from SEQ ID NO:4.

[0052] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 7), peptide CG 119-144 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 X 26 In the sequence, [ka]

[0053] In an alternative embodiment, X of SEQ ID NO:7 17 and / or X 18 The rankings are shown in Table 1 below. The sequence may be altered by deletion, insertion, or substitution as shown in .

[0054] [Table 1]

[0055] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 8), peptide NE 119-145 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19X 20 X 21 X 22 X 23 X 24 X 25 X 26 X 27 In the sequence, [ka]

[0056] In an alternative embodiment, X of SEQ ID NO:8 16 and / or X 17 The rankings are shown in Table 2 below. The sequence may be altered by deletion, insertion, or substitution as shown in .

[0057] [Table 2]

[0058] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 9), peptide CAP37 120-146 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 X 26 X 27 In the sequence, [ka]

[0059] In an alternative embodiment, X of SEQ ID NO:9 15 and / or X 16 The rankings are shown in Table 3 below. The sequence may be altered by deletion, insertion, or substitution as shown in .

[0060] [Table 3]

[0061] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 10) peptide CG 20-47 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 X 26 X 27 X 28 In the sequence, [ka]

[0062] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 11) peptide NE 20-44 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 In the sequence, [ka]

[0063] In at least one embodiment, the peptide compound has the following amino acid sequence: No. 12), peptide CAP37 20-44 Includes derivatives of: X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 In the sequence, [ka]

[0064] In certain non-limiting embodiments, an effective amount or therapeutic dose of a peptide compound of the present disclosure The dosage is about 0.001 μg / kg to about 100 mg / kg (weight of active agent / body weight of subject). For example, the composition may contain about 0.01 μg / kg to about 0.1 μg / kg of active agent. About 50 mg / kg, more particularly about 0.1 μg / kg to about 10 mg / kg, more particularly about 1 The practice of the disclosed methods delivers therapeutic dosages of active agent. The peptide in any suitable systemic and / or topical formulation in an amount effective to deliver In certain embodiments, the method may include administering to the subject an effective amount of a tide compound. Effective dosages can range from about 1 μg / kg to about 1 mg / kg of active agent.

[0065] The practice of the methods of the present disclosure may involve administering to a subject, any suitable subject, an amount effective to deliver the dosages recited above. administering to the subject a therapeutically effective amount of the peptide compound in an appropriate systemic and / or topical formulation; Dosages may include, but are not limited to, depending on the desired therapeutic effect. Depending on the dosage, the drug may be administered once or multiple times (for example, but not limited to, 1 to 5 times per day or once or twice a week) or continuously via intravenous infusion In one non-limiting example of the therapeutic method of the present disclosure, the variant peptide The compound is administered intravenously once daily at a dose ranging from about 0.01 mg / kg body weight to about 10 mg / kg body weight. The input is supplied at the input.

[0066] The administration of peptide compounds for use in pharmaceutical compositions or for practicing the methods of the present disclosure. Administration may be by any means, including, but not limited to, oral, inhalation, rectal, topical, nasal, or oral route. It can be administered in a variety of conventional ways, such as by injection into the skin, subcutaneously, intraperitoneally, vaginally, or intravenously. Oral formulations allow the peptide compound to pass through parts of the digestive system before being released. For example, the compound may be formulated so that it is not released until it reaches the small intestine or colon. It can be done.

[0067] When an effective amount of the peptide compound is administered orally, it may be administered in the form of a capsule, pill, tablet, or the like. Lozenges, melts, powders, suspensions, solutions, elixirs, or emulsions The solid unit dosage form may be in the form of any solid or liquid preparation. agents, and inert fillers such as lactose, sucrose, and corn starch The dosage form may be a capsule of the usual gelatin type containing the agent, or the dosage form may be a sustained release preparation. The pharmaceutical composition may contain a solid carrier such as gelatin or an adjuvant. The tablets, capsules, and powders contain from about 0.05% to about 95% of the active compound by dry weight. When administered in liquid form, the liquid may contain water, petroleum, peanut oil, mineral oil, soybean oil, etc. or oils of animal or vegetable origin, such as sesame oil, or synthetic oils, are added. Liquid pharmaceutical compositions may be prepared by dissolving in saline, dextrose or other sugar solutions. Liquid, or ethylene glycol, propylene glycol, or polyethylene glycol When administered in liquid form, the pharmaceutical composition may further comprise a glycol, such as glycerol. The active ingredient is about 0.005% to about 95% by weight. For example, once a day or Two doses of about 10 mg to about 1000 mg may be administered orally.

[0068] In another embodiment, the peptide compounds of the present disclosure are selected from the group consisting of gum acacia, gum corn denaturant, and the like. A binder such as bread or gelatin, a disintegrating agent such as potato starch or alginic acid and in combination with a lubricant such as stearic acid or magnesium stearate. Tableting with traditional tablet bases such as lactose, sucrose, and corn starch The liquid preparations may contain suspending agents, sweeteners, flavoring agents, etc., as known in the art. and a preservative. The mixture is prepared by dissolving the mixture.

[0069] For parenteral administration, for example, the peptide compounds may be administered in a physiologically acceptable pharmaceutical carrier. It can be dissolved and administered either as a solution or as a suspension. Examples of such solutions include water, saline, dextrose solution, fructose solution, ethanol, or or oils of animal, vegetable, or synthetic origin. Pharmaceutical carriers also include those known in the art. Preservatives and buffers may be included as necessary.

[0070] When an effective amount of the peptide compound is administered by intravenous, cutaneous, or subcutaneous injection, The composition is preferably a pyrogen-free, parenterally acceptable aqueous solution or suspension. Such parenterally acceptable solutions are selected taking into consideration pH, isotonicity, stability, etc. The preparation of solutions is well within the skill of one in the art. Certain pharmaceutical compositions of the present invention contain, in addition to the peptide compounds, sodium chloride injection, Ringer's injection Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer's It may contain an isotonic vehicle, such as an injection solution, or other vehicle as known in the art. The pharmaceutical compositions of the present disclosure may also contain stabilizers, preservatives, buffers, antioxidants, or other additives of ordinary skill in the art. It may also contain other additives known in the art.

[0071] As stated, the particular amounts and modes of administration can be determined by one of skill in the art. Those skilled in the art can prepare the above-mentioned compositions by referring to, for example, Remington: The Science and Practice of Pharmacy, 22 nd ed The selected peptide is prepared using formulation techniques known in the art, as described in . The specific characteristics of the compound, the condition to be treated, the stage of the condition, and other relevant circumstances. Depending on the type and dosage, one can easily select an appropriate form and mode of administration.

[0072] Additional pharmaceutical methods can be used to control the duration of action of the peptide compounds. Increased half-life and / or sustained release preparations may be used as previously discussed herein. , conjugate, complex with, and / or absorb peptide compounds. This can be achieved through the use of proteins or polymers that can be used to control and / or reduce delivery. The increase in phase can be achieved by the addition of suitable polymers (e.g., but not limited to, Polysaccharides, polyesters, polyamino acids, homopolymers, polyvinylpyrrolidone, ethylene acetate Vinyl acetate, methylcellulose or carboxymethylcellulose, and N-(2-hydroxyethyl) (e.g., propyl)acrylamide) and the appropriate concentration of polymer This can be achieved by selecting the gene expression vector, as well as the method of integration.

[0073] Controlling the duration of action of peptide compounds through sustained release preparations and half-lives Another possible method that is useful for this purpose is the use of polyesters, polyamides, polyamino acids, hydrogels, etc. , poly(lactic acid), ethylene vinyl acetate copolymers, e.g., polyethylene glycol (P to particles of polymeric materials such as copolymer micelles of EG) and poly(l-aspartamide). or a functional derivative thereof.

[0074] Microcavities prepared, for example, by coacervation techniques or by interfacial polymerization. capsules (e.g., hydroxymethylcellulose or gelatin microcapsules, respectively) colloidal drug delivery system (PCD, PEG, and poly(methyl methacrylate) microcapsules) (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in macroemulsions. Such techniques are well known to those skilled in the art.

[0075] If the peptide compound is to be used as an injectable material, it should be administered in a manner similar to that of conventional The composition may be formulated into an injectable carrier. Suitable carriers include, in particular, isotonic, biocompatible, pharmaceutical Examples of suitable buffered saline include physiologically acceptable phosphate buffered saline.

[0076] For reconstitution of the lyophilized product according to the present disclosure, approximating physiological conditions is generally Sterile dilutions that may contain materials as recognized and / or required by government regulations. In this regard, sterile diluents include sodium chloride, saline, phosphate buffered saline, etc. saline solution, and / or any solution that is physiologically acceptable and / or safe for use. It may contain buffering agents, such as certain other substances, to obtain a physiologically acceptable pH. Generally, materials for intravenous injection in humans are available to those skilled in the art. Pharmaceutical compositions should also comply with regulations established by the Department of Pharmacy. It may be in the form of an aqueous solution containing many of the same materials as described above for composition.

[0077] Peptide compounds are also hydrochloric, hydrobromic, perchloric, nitric, thiocyanic, sulfuric, and Inorganic acids such as acetic acid, propionic acid, glycolic acid, lactic acid, and pyridine. with organic acids such as benzoic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid. or by the reaction of sodium hydroxide, ammonium hydroxide, potassium hydroxide, etc. Chemical bases, as well as mono-, di-, tri-alkyl and aryl amines, and substituted ethanoates. Pharmaceutically acceptable acids or bases formed by reaction with organic bases such as diamines It may also be administered as an addition salt.

[0078] In certain embodiments, the present disclosure provides a method for the preparation of a medicament for the treatment of cancer, comprising: coupling directly or indirectly to a carrier molecule; At least one peptide compound that has been subjected to a peptide synthesis test, A peptide compound composition comprising a peptide compound, the peptide being 20 to 50 amino acids long. Includes things.

[0079] In at least certain embodiments, the present disclosure provides a method for treating Alzheimer's disease, cerebral amyloidosis, Proinflammatory and neurotoxic properties of Aβ in vasculopathy and other Aβ-associated neuroinflammatory conditions For inhibition of the inflammatory effect, either alone, in combination, or in combination with a non-peptide moiety (e.g., PEG As peptide components of a peptide compound containing full-length CAP37, NE, and CG protein and / or peptide fragments thereof, and / or such peptides In a non-limiting example, the peptide compounds of the present disclosure include derivatives of the above. and peptides having the amino acid sequences disclosed in SEQ ID NOs: 1 to 12.

[0080] The amino acid sequences described elsewhere herein may be modified by the addition of R, K, and / or H moieties. Addition (e.g., 1 to 10 amino acids linked to the N- and / or C-terminal portions of the peptide) ) and linked to the N- and / or C-terminal portions of the tagged or untagged peptide sequence The "mini-PEG" (AEEA, AEEEA, AEEP, and AEEEP) parts Examples of such mini-PEG moieties can be found, for example, in WO 2014 / 08 No. 9088 and No. 2014 / 172747, as well as U.S. As shown in Patent Nos. 9624283 and 9862748.

[0081] CG-based, NE-based, and other peptides that can be used in various embodiments of the peptide compounds of the present disclosure. Examples of CAP37-based variant peptides include those shown in Table 4. These include, but are not limited to:

[0082] [Table 4-1]

[0083] [Table 4-2]

[0084] In certain embodiments, the peptide compounds of the present disclosure are represented by formula (I): : SX R -Pep-Y R (I) where Pep is an amino acid sequence disclosed or described herein; R and Y R are, independently, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten R, K, or H residues, e.g., X R +Y R = Maximum 10 and S is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 PEG-100 (i.e., 1-10) short PEG subunits, e.g., Any of the following, as defined: AEEA, AEEP, AEEEA, and AEEEP The solubilizing portion is made up of a combination of the above.

[0085] Peptide (X R -Pep-Y R ) of the -O(C H2) q The CO- moiety ("head group") is the (CH2) q Some alternative implementations of parts These alternative embodiments, as well as those described above, may be further modified to include the following forms: The compound may be characterized by formula (II): -O(CR1R2) q CO- (II) where R1 and R2 in Formula II are side chain groups on C (i.e., "R1-C-R2" ), q=0-4, and R1 and R2 are selected from H, CH3, and CH2CH3. When q=1, R1 and / or R2 can be H, CH3, and CH2C. H3. When q=2, one of the two R1 groups and one of the two R2 groups may be selected from H, CH3, and CH2CH3. When q=3, the three R1 groups Any and any of the three R2 groups are selected from H, CH3, and CH2CH3. When q=4, any of the four R1 groups and any of the four R2 groups can be H, CH3, and CH2CH3. Accordingly, these non-limiting embodiments In the formula, when C=1-4, the head group is one or more H atoms on the carbon of the carbon backbone. It may include the substitution:

[0086] Examples of embodiments of head groups having formula II include, but are not limited to, the following: stomach: 1. -OCH2CH2CO- (e.g., as in AEEP) 2. -OCH2CO- (e.g., as in AEEA) 3. -OCH2CH2CH2CO-(q=3) 4. -OCH2CH2CH2CH2CO- (q=4, i.e., pentane head group) 5. -OC(CH3)2CO- (R1 and R2 = CH3, and q = 1) 6. -OCH2C(CH3)2CO- (R1 and R2=H on the first carbon and R2=H on the second carbon) (R1 and R2=CH3 on the second carbon, and q=2) 7. -OCH2CH2CH2CH(CH3)CO- (R1=H on one carbon and R2=CH3, and R1 and R2=H on every other carbon, and q=4) 8. -OCH2CH2CH2C(CH3)2CO- (R1 and R2 on one carbon) =CH3, and R1 and R2 =H on all other carbons) 9. -OCH2CH2CH2CH(CH2CH3)CO- (R1=H on one carbon) and R2=CH2CH3, and R1 and R2=H on all other carbons) 10. -OCH(CH3) CH2CH2CH(CH2CH3)CO- (on one carbon) R1=H and R2=CH3 on one carbon, R1=H and R2=CH2CH3 on the other (R1 and R2 = H on all other carbons)

[0087] All of the peptide compounds formed based on the solubilizing unit described in formula II are intended to be included in this disclosure. EXAMPLES

[0088] method Binding assays Using enzyme-linked immunosorbent assay (ELISA), we investigated the CAP37 and CAP37-induced Aβ expression. Binding to the peptide, NE, CG, and BSA (used as a negative control) was quantified. CAP37, NE, and CG were purified from human neutrophils and collected at the Athens Research Center. BSA was purchased from Sigma Aldrich and Technology. The CAP37-derived peptides were purchased from CS Bio Co. with purity of ≥95%. 1 mM PMSF (CAP37, purchased from Sigma Aldrich) NE, and BSA) or Santa ±125 μM chymostatin (for experiments with CG and BSA) was purchased from Cruz. 100 μl neutrophil protein, peptide, or Maxisorp plates (Nunc) were coated with 100% ribosomal IgG or BSA. The plating was carried out at room temperature with shaking for 2 hours and then at 4°C overnight. Wash three times in BST (PBS + 0.05% Tween) and then in 3% BST in PBST. The plate was blocked with PBST for 1 hour at room temperature with shaking. , 150nM Aβ 1-42 (purchased from Bachem and reconstituted in 1 mM NaOH) The resulting solution was added to 0.1% BSA in PBST and incubated at 37°C for 70 minutes. Then, wash the plate 4 times with PBST and immerse in Cell Signaling Technology. Monoclonal mouse anti-amyloid precursor protein / amyloid purchased from logy Beta antibody was added at a 1:500 dilution in 1% BSA in PBST and incubated at room temperature with shaking. Plates were washed 4 times in PBST and incubated with Jackson Donkey anti-mouse HRP conjugate from ImmunoResearch Laboratories Add conjugated secondary antibodies at 1:10,000 in PBST and incubate at room temperature with shaking. Incubated for 1 hour and then washed 4 times in PBST. M citric acid, 102 mM sodium phosphate, pH adjusted to 5.0 with HCl, Kodak Freshly added 7.5 mM substrate o-phenylenediamine and 0.024% final H2O2 Sulfuric acid (5N) was added to each well and incubated for 30 minutes in the dark. The reaction was stopped by adding 100 mM NaCl to the wells, and the absorbance at 492 nm was measured using a Biotek Syringe. The plates were read on a nergy II plate reader.

[0089] Amyloid beta (Aβ) fibrillization assay Amyloid beta (1-42) Thioflavin T Aggregation Kit was purchased from Anaspec. , and then used to elucidate Aβ 1-42 The formation of peptide fibrils was monitored by adding Thioflavin T dye to fibrillation buffer A at 2 The Aβ was diluted to 50 μM and kept in the dark until use. 1-42 Peptides were dissolved in Buffer A. Suspend the samples at 250 μg / ml (or 55 μM) in 100% PBS, incubate on ice for 5 minutes, and then Mix by inverting the tube twice and place in a water bath sonicator at 4°C for >5 min. Sonicate, mix again and centrifuge at 10,000xg for 5 min. Transfer the supernatant to a new tube. The mixture was transferred to a microtube and stored at 4°C until use. Tide, NE, CG, and BSA (used as a negative control) were diluted in water at 20x the desired final concentration. In low light, 10 μl Thioflavin T dye was added to a Corning low-temperature Add 85 μl Aβ prepared in Buffer A to the protein-binding black plate. 1- 42 1 μl of water or the aggregation inhibitor to be tested was also added. PMSF was dissolved in 100% ethanol and tested in the absence and presence of protease inhibitors. It was dissolved and used at 1 mM final concentration in reactions involving CAP37 and NE. Chin was dissolved in DMSO and used at 125 μM final concentration in reactions with CG. To ensure mixing, the reactions were pipetted into the wells using a multichannel pipette. Pipette up and down twice to measure fluorescence (λ Ex = 440 nm, λ Em =48 5 nm) on a BioTek Synergy 2 plate reader. Using the software, readings were taken every 5 minutes for 3 hours with the sensitivity set at 65.

[0090] Enzyme activity assay Aβ treated with hexafluoroisopropanol 1-42 Purchased from Bachem The reaction was performed using 100 mM glycerol (GlcNAc) and 100 mM glycerol (GlcNAc) in 100 mM MgCl2. G) Increasing concentrations of Aβ, and 10 μM Aβ in PBS 1-42 Set up with substrate and incubate at 37 °C for 1 h. The mixture was incubated for 1 h in acetonitrile and trifluoroacetic acid (TFA), respectively. The reaction was stopped by adding 100 mM NaCl to final concentrations of 7.5% and 0.5%. Internal standard (Aβ from Bachem) dissolved in 10% acetic acid 40-1 ) to a final concentration of 8 μg / A 10 μl aliquot of this mixture was added to an Agilent Te The desalted solution was then desalted using a C18 OMIX tip from chnologies. First, the C18 resin was 2x with 100 μl of 50% methanol, then 10 μl of equilibration buffer (5% acetonite The resin was rinsed twice with 0.5% TFA and then the sample was pipetted 10 times through the resin. The resin was then washed twice with equilibration buffer and incubated for 75 Eluted with 0.1% acetonitrile and 0.1% TFA.

[0091] Desalted samples were analyzed using a Bruker Ultraflex II instrument and Flex Co. Matrix-assisted laser desorption / ionization (MALD) time-of-flight analysis was performed using the ntrol software. The mixture was analyzed by mass spectrometry (MARLDI-TOF). The samples were then washed with 1 μl of α-cyano-4-hydroxycinnamic acid matrix in 10% TFA. Each sample was spotted in triplicate on a steel plate. The plate was vacuum-sealed. Each chromatograph was run in linear positive mode. The average value of 5 × 200 shots at 28% laser power was obtained. The chromatograph was analyzed using the ex analysis software. 40-1 Corresponding to internal standards Peaks around 4,330 m / z and Aβ 1-42 4,515m corresponding to the substrate The Aβ peaks were divided by the internal standard peak intensity. 1-42 Peak Intensity The ratio of cleavage in each enzyme reaction was calculated for each spot and averaged over triplicate spots. The amount of unblocked Aβ was determined.

[0092] Cell culture and neurotoxicity assays The mouse neuroblastoma cell line Neuro-2a (ATCC) was used in these experiments. Cells were grown in complete medium (10% heat-inactivated fetal bovine serum and Lo Gibco MEMα, Nucleotide, and Penicillin / Streptomycin supplemented with NZA 96-well plate with 20,000 cells per well in 100% ethanol (leucoside, no phenol red) After 24 hours from seeding, the cells were washed with warmed MEMα and then Then, 90 μl MEMα + N2 supplement + 20 μl test sample (prepared as follows Aβ 1-42 The cells were incubated at 37°C for an additional 24 hours. Then, remove the sample by pipetting up and down vigorously. 10 μl of cells were incubated with 10 μl of trypan blue dye (0.4% solution) from BioRad. The cells were added to the 500 ml tube, mixed and counted in a BioRad TC20 cell counter. The percentage of dead cells was recorded for each well. In the control case, 20 μl of MEMα+N2 supplement was added instead of the test sample. Just before counting the cells, remove 20 μl of medium and add 20 μl water or 0.1% final It was replaced with Triton X100 at 100% concentration.

[0093] Preparation of Aβ, monomers, fibrils, and oligomers test samples Monomeric Aβ from Bachem 1-42 (Hexafluoroisopropanol treated Aβ 1-42 ) was reconstituted at 5 mM in anhydrous DMSO. Untreated Aβ from CS Bio 1-42 was also used in these experiments. It was first purchased from Sigma Aldrich Treat with 1 mM hexafluoroisopropanol and incubate for 30 min at room temperature. It was then left open in a fume hood overnight to dry. The mixture was dried in vacuum for 1 hour. 1-42 of the dried film, keeping it in monomeric form. The Aβ antibody was purchased from CS Bio and stored at -20°C until use. 1-42 Also, before use , reconstituted at 5 mM in anhydrous DMSO.

[0094] Aβ 1-42 Toxicity of monomers was evaluated using freshly prepared Aβ 1-42 (5 mM in DMSO) The antibodies were tested by dilution in MEMα+N2 supplement to a final concentration of 18 μM. The complete growth medium was replaced in cells 24 hours after seeding with a mixture of 1- 42 The toxicity of the monomers was measured after 24 hours of incubation.

[0095] To prepare fibrillar and oligomeric test samples, monomeric Aβ 1-42 The original line The mixture was subjected to conditions that favored the formation of either fibrils or oligomers. To do this, 100 μM Aβ 1-42 Anaspec Fibrillation Buffer A ± Neutrophil Protein The protein was added to the protein and incubated at 37°C for 24 hours, after which it was added to the cells at a final concentration of 18 μM. Neutrophil proteins were added according to the previously determined values ​​for each protein in the fibrosis assay. The Aβ / protein ratio that resulted in 50% inhibition of fibrillation (IC50) was used. To form fibrillation buffer A ± increasing concentrations of neutrophil protein or CA Instead of the P37-derived peptide, MEMα+N2 supplement was used. , freshly prepared Aβ at a final concentration of 100 μM in DMSO. 1-42 Chew containing The test samples were then added to the 18 μL plate and incubated at room temperature for 15 minutes. M final concentrations were tested on the cells.

[0096] result Figure 1 shows Aβ 1-42 We demonstrated binding of CAP37 to full-length CAP37, NE, and CG proteins. The binding is stronger for CAP37 and CG, and is more consistent with the antiprotease (phenyl fluoride). is not significantly altered by the addition of PMSF or chymostatin Aβ 1-42 Binding of Aβ to NE was almost undetectable, whereas NE-mediated Aβ binding was 1-4 The expression of CAP37 was significantly increased in the presence of PMSF, which inhibits the cleavage of CAP37. and stronger Aβ to CG 1-42 The results show that NE binds to β-lactams, as well as weaker binding to NE. FIG. 2 shows the wild-type CAP37-derived peptide 20-44 (20-44 CC, 95-12 2, and 120-146 QR) and mutant CAP37-derived peptide (20-44 CS, 20-44 SC, 120-146 WR, 120-146 QH, and 120 -146 WH) to Aβ 1-42 The levels of Aβ binding are shown. 1-42 Peptide 9 5-122. Two AEEA mini-PEG molecules linked to the N-terminus of the peptide and four consecutive arginine residues, as well as one arginine linked to the C-terminus of the peptide) -42 These results show that Aβ 1-42 However, the presence or absence of "5RMP" modification Regardless, wild-type and mutant CAP37-derived peptides 20-44 and 120-1 46, but not peptide 95-122, with or without the "5RMP" modification. This indicates that there is no

[0097] Figures 4-7, 10, and 13 show increasing concentrations of three neutrophil granule proteins and Relative fluorescence intensity in the absence and presence of certain wild-type and mutant CAP37-derived peptides. Aβ recorded as Rfu 1-42 Figure 4 shows the complete fibrosis of the indicated concentrations. Aβ in the presence of full-length CAP37, NE, or CG 1-42 Level of fibrosis (aggregation) Aβ (when no protein was added) 1-42 The time to complete fibrillation of the Fibrillization was either partial or complete in the presence of these proteins. Figure 5 shows the effect of various concentrations of full-length protein on either the Aβ in the presence of CAP37, NE, or CG 1-42 The level of fibrosis (aggregation) of NE is an Aβ 1-42 The highest effect on fibrosis was observed in CG, followed by CAP37. FIG. 5 shows the dose response performed for each full-length protein. The percent fibrillation measured was the percentage of Aβ 1-42 versus These dose-response experiments demonstrated that NE Aβ 1-42 The highest incidence of fibrosis (1 / 4,167 NE / Aβ 1-42 Molar ratio for ) followed by CG (IC50 of 3.8 nM; 1 / 2,632 molar ratio) and CAP37 (IC50 of 876 nM; 1 / 11 molar ratio). This indicates that it has a certain inhibitory effect.

[0098] Figure 6 shows that the protease inhibitor PMSF inhibits Aβ binding to CAP37. 1-42 Inhibition of fibrosis The IC50 of CAP37 was increased 3.5-fold in the presence of PMSF, impairing the anti-inflammatory effect of PMSF. This suggests that the inhibitory effect of CAP37 on fibrosis is, at least in part, due to A β 1-42 Figure 7 shows that the protease inhibitor PMSF inhibits N E Aβ 1-42 The IC50 of NE was 1.001 mg / kg / day, which is 1.001 mg / kg / day, and 1.001 mg / kg / day. In the presence of SF, the fibrosis-inhibiting effect of NE was increased by 36-fold. At least in part, Aβ 1-42 This indicates that the cleavage of three neutrophil granule proteins The protein is Aβ 1-42 It has previously been found that the cleavage of and 10, Aβ 1-42 To determine whether cleavage of β-lactamase is required for inhibition of fibrillation, To achieve this, CAP37, NE, and CG were incubated with sufficient protease inhibitors (CAP37 and Tests were performed in the absence and presence of PMSF for NE and chymostatin for CG. In all three experiments, Aβ 1-42 Inhibition of the enzymatic cleavage of these proteins also This impairs the inhibitory effect of fibrosis on the protein. At least in part, Aβ 1-42 Figure 13 shows that the increased concentration CAP37-derived peptides 95-122 5RMP, 120-146 WR 5RMP Dose responses performed with peptides 95-122 5R and 120-146 WR are shown. MP is Aβ 1-42 (Fig. 3) and did not inhibit fibrosis. Both 0-146 WR 5RMP and 120-146 WR correlated with Aβ1-42. These CAP37-derived peptides interacted with each other (Figs. 2 and 3) and inhibited fibrosis. The inhibitory effect of the peptide is that the peptide inhibits Aβ 1-42 Although it has no enzymatic activity against , similar to that of full-length CAP37 (IC50 of approximately 1000 nM; approximately 1 / 10 the molar ratio). Without wishing to be bound by theory, this is 1-42 The cleavage of Aβ 1-42 This suggests that Aβ is not the only mechanism for the inhibition of fibrillation. 1-4 Direct binding to 2 also appears to play a role in inhibiting fibrosis.

[0099] Figures 8, 9, 11, and 12 show the increased concentrations of β-lactamase in the absence and presence of protease inhibitors. Percent fibrosis and right-handedness in experiments performed with full-length neutrophil granule protein Corresponding Aβ on the y-axis 1-42 Figure 8 shows the percent cleavage of Aβ by NE. 1- 42 This correlation shows a direct correlation between cleavage of NE and inhibition of fibrosis. This experiment supports the idea that the effect of PMSF on the inflammatory response is primarily mediated by cleavage of Aβ. Figure 9 shows the effect of NE on Aβ 1-42 Direct interaction between cleavage of β-lactamase and inhibition of fibrosis This experiment was performed in the presence of PMSF. 1- 42This correlation shows a direct correlation between the cleavage of CG and the inhibition of fibrosis. This supports the idea that the effect is primarily mediated by cleavage of Aβ. FIG. 12 shows the effect of CG on Aβ 1-42 between cleavage and inhibition of fibrillation This experiment was performed in the presence of chymostatin. Establish a direct correlation between Aβ inhibition and inhibition of fibrillation 1-42 Cleavage of the full-length protein These results support the idea that this is the primary mechanism explaining the inhibition of fibrosis mediated by IL-1.

[0100] Figure 14 shows the various higher-order structures of Aβ in neurons. 1-42 It shows the toxicity of Aβ. 1- 42 Conditions were used that favored monomeric, oligomeric, or fibril conformations of . 4 is Aβ 1-42 Oligomers have been shown to be more toxic to neurons than fibrils. Aβ 1-42 The monomeric form of β-lactam β-lactam 1 was the least toxic. These results are consistent with those of other groups. The study confirmed previously published results, with oligomers showing the highest toxicity, followed by This indicates that it is a fibril and then a monomer.

[0101] Figure 15 shows that NE and C, when incubated under conditions that favor fibril formation, G is Aβ 1-42 These results show that the cytotoxicity of CAP37 is inhibited by CAP41, but not by CAP37. 5 inhibited approximately 50% of fibrillation, as previously determined for each protein in FIG. Formation in the absence or presence of neutrophil granule proteins used at concentrations that are deleterious to neutrophil granules (IC50). Aβ 1-42This experiment shows the fibril toxicity caused by NE and CG. CAP37 inhibits Aβ expression under these conditions. 1-42 Toxicity of In the fibril experiments, all three neutrophil granules Granule proteins were incubated with Aβ for 24 h before addition to cells. 1-42 Preincubated with During this 24-h fibrillation step, Aβ 1-42 The percent cleavage of is higher than 50% In the case of NE and CG, cleavage is likely important for inhibition of fibril toxicity. In contrast, CAP37 inhibits Aβ 1-42 The cut is made by the line, as determined in Fig. 6. When preincubated under fibrillogenic conditions, Aβ 1- 42 This is because NE and CG are unable to inhibit the toxicity of Aβ. 1-42 Above, This is explained by the fact that it has more cleavage sites than CAP37, resulting in smaller products. Thus, the larger product generated by CAP37 cleavage is still As such, it is possible that they can form toxic aggregates under fibrotic conditions.

[0102] 16-20 show increasing concentrations of three neutrophil granule proteins, and In the absence or presence of two CAP37-derived peptides that were found to inhibit fibrosis Formed by Aβ 1-42 Figure 16 shows the toxicity of oligomers in the presence of various concentrations of NE. Aβ in the presence of 1-42 The effect of oligomers on neuronal cells is shown. NE was dose-dependent. In a cellular manner, Aβ 1-42 The toxicity of oligomers was significantly inhibited. NE was determined in Figure 5. IC50 concentrations of 0.5, 1, 2, and 4 were used. Results start from 2×IC50. These results show that NE significantly inhibits oligomer toxicity.

[0103] FIG. 17 shows the Aβ 1-42 Oligomeric neuronal cells The effect of CG on Aβ in a dose-dependent manner was 1-42 Significantly inhibits oligomer toxicity The same conditions as in FIG. 16 were used in FIG. 17 for the CG, with 0.5×IC5 Starting from 0, it shows a stronger inhibition of oligomer toxicity by CG.

[0104] FIG. 18 shows the Aβ expression profile in the presence of various concentrations of CAP37 protein. 1-42 Oligomer Effects on neuronal cells. Under the conditions used, full-length CAP37 inhibited Aβ1 -42 Notably, these studies of oligomers did not inhibit oligomer toxicity. In the experiment, all three neutrophil granule proteins were incubated for 15 min prior to addition to the cells. Aβ 1-42 Preincubated with A during the 15 min oligomerization step. β 1-42 The percent cleavage of ribozymes is unknown. Therefore, inhibition of oligomerization and toxicity The significance of the cleavage for toxicity is unknown. As discussed above, CAP37 cleavage may be involved in The larger products generated by this reaction still form toxic aggregates under oligomerization conditions. It may also be possible to do so.

[0105] FIG. 19 shows the A in the presence of various concentrations of the CAP37-derived peptide 120-146 WR. β 1-42 The effect of oligomers on neuronal cells. Modified CAP37 peptide in a dose-dependent manner, 1-42 Figure 20 shows that the various Aβ in the presence of various concentrations of the CAP37-derived peptide 120-146 WR-5RMP 1- 42 The effect of oligomers on neuronal cells. The dovariant inhibited Aβ in a dose-dependent manner. 1-42 The toxicity of oligomers was significantly inhibited. 19 and 20, both of which are Aβ 1-42 (Figures 2 and 3) (Figure 13), peptides 120-146 WR and 120-146 WR 5 RMP was also able to significantly reduce the toxicity of oligomers. -42 It has no enzymatic activity against Aβ 1-42 The cleavage of Aβ 1-42 O This suggests that this is not the only mechanism for inhibition of ligomerization and toxicity. Aβ 1-42 Direct binding to may inhibit oligomerization.

[0106] The present disclosure may be more fully understood and appreciated in connection with certain specific embodiments. Although described herein, it is not intended that the disclosure be limited to these particular embodiments. On the contrary, all alternatives, modifications, and equivalents are intended to be included within the scope of the present disclosure. Accordingly, the above examples, including specific embodiments, are intended to be included in the scope of the present disclosure. The details shown serve to illustrate the practice and are provided by way of example and of specific embodiments only. The present disclosure is based on a set of principles and methods for the preparation and use of the disclosed compositions. To provide what is believed to be the most useful and easily understood explanation of the concepts and conceptual aspects of the It is understood that the formulation of the various compositions described herein is provided herein. In the method or steps of the method described herein No variations or modifications may be made in the steps or sequence of steps without departing from the spirit and scope of the disclosure. Changes can be made.

Claims

1. A composition for use in inhibiting and / or reversing polymerization of amyloid beta (Aβ) peptide in a subject in need of such treatment, the composition comprising a peptide compound comprising a peptide having the amino acid sequence of SEQ ID NO:

29.

2. 2. The composition of claim 1, wherein inhibiting and / or reversing polymerization of Aβ peptide comprises inhibiting the formation of at least one of Aβ oligomers and Aβ fibrils.

3. 2. The composition of claim 1, wherein inhibiting and / or reversing polymerization of Aβ peptide comprises causing disaggregation of at least one of Aβ oligomers, Aβ fibrils, and Aβ plaques.

4. The composition of claim 1 , wherein the peptide compound is conjugated to, incorporated into, complexed with, or entrapped within a polymer.

5. The composition of claim 1 , wherein the peptide consists of up to 50 amino acids.

6. The composition of claim 1 , wherein the peptide compound comprises a polyethylene glycol moiety conjugated to a peptide.

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