Peptide therapeutics for treating Alzheimer's disease and related conditions

Abstract

Peptide compounds derived from cathepsin G (CG), neutrophil elastase (NE), and 37 kDa cationic antimicrobial protein (CAP37), and methods of their use in inhibiting and / or reversing the polymerization of amyloid beta peptide into oligomers and / or fibrils and / or disaggregating amyloid beta plaques, and methods of using said peptide compounds in the treatment of, for example, Alzheimer's disease, cerebral amyloid angiopathy, dementia, and / or neuroinflammation, and / or symptoms thereof.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Patent Application No. 17 / 115, 321, filed on 8 December 2020, which is expressly incorporated herein by reference in whole.

[0002] government support This invention was made with government support under authorization numbers R21EY026229 and P20GM103640-06 granted by the National Institutes of Health. The government has certain rights in this invention. [Background technology]

[0003] Alzheimer's disease (AD) is the leading cause of dementia among people aged 65 and older and the sixth leading cause of death in the United States. Currently, approximately 5 million people are affected, and that number is projected to double in the next 20 years. Currently, AD cannot be cured, prevented, or slowed, highlighting the critical need for the development of effective treatment approaches.

[0004] It has been previously shown that the 37kDa cationic antimicrobial protein (CAP37) is significantly upregulated in neuronal and hippocampal vascular structures in patients with AD compared to age-matched controls. It has also been shown that the CAP37 protein binds to and cleaves amyloid beta (amyloid β, or Aβ) (the major component of amyloid plaques found in the brains of patients with AD), and that the CAP37 protein interacts with pro-inflammatory receptors of advanced glycation end products (RAGE). Aβ accumulates in the brain early in the progression of AD and is a key, prominent feature of the AD pathogenesis. As its concentration increases, Aβ polymerizes, initially forming soluble oligomers and fibrils, which then precipitate to form insoluble amyloid plaques. Aβ accumulation in the brain may result from (1) increased production from the cleavage of amyloid precursor protein (APP), (2) increased influx from the blood, and / or (3) decreased clearance.

[0005] RAGE belongs to the immunoglobulin superfamily of transmembrane receptors and mediates pro-inflammatory signals. Aβ (in monomer, oligomer, and fibril forms) is a known ligand for RAGE. Other known ligands for RAGE include advanced glycation end products (AGEs), S100 / cargranulin, and the high-mobility group box 1 (HMGB1) protein. RAGE is expressed at low levels in adult tissues and is upregulated by a positive feedback loop as RAGE ligand concentrations increase. RAGE expression is upregulated in the brains of patients with AD. In AD development, RAGE is thought to play a role in mediating the toxic effects of Aβ oligomers and fibrils in neurons and microglial cells. In neurons, RAGE can mediate the induction of oxidative stress and tau hyperphosphorylation. RAGE also mediates the activation of microglial cells, perpetuating the neuroinflammation underlying chronic conditions associated with AD. In the blood-brain barrier, RAGE activation is thought to be involved in barrier dysfunction and other pathological features associated with AD. Finally, RAGE mediates the influx of Aβ from the circulation into the brain via transcellular transport mechanisms.

[0006] Previous findings have established the following: (1) CAP37 expression is increased in neurons of patients with Alzheimer's disease (AD) to a greater extent than in patients of the same age who have not been diagnosed with AD. This established a positive correlation between the amount of CAP37 in the brain and the disease. This was not the case for two closely related proteins, neutrophil elastase (NE) and cathepsin G (CG). (2) The regulation of CAP37, NE, and CG expression generally correlates with the regulation of proteins that act as ligands for pro-inflammatory receptors of advanced glycation end products (RAGE); (3) CAP37 binds to RAGE, as does Aβ peptide, a known ligand for RAGE, and NE and CG also bind to RAGE; (4) CAP37-RAGE and NE-RAGE interactions are inhibited by certain RAGE inhibitors; (5) CAP37, NE, and CG bind to Aβ; and (6) CAP37, NE, and CG exert protease activity against Aβ, cleaving it at several identified cleavage sites.

[0007] Thus, both Aβ and RAGE have been identified as important pharmacological targets in the development of AD, and this disclosure concerns the identification of drugs that can target Aβ and RAGE and therefore be used to treat patients with AD and / or neuroinflammation. [Brief explanation of the drawing]

[0008] [Figure 1] This figure shows the binding of amyloid β1-42 (Aβ1-42) to the full-length CAP37, NE, and CG proteins. Binding is stronger with respect to CAP37 and CG and does not change significantly with the addition of antiproteases (phenylmethylsulfonyl fluoride (PMSF) or chymostatin). Binding of Aβ1-42 to NE is almost undetectable, but increases significantly in the presence of PMSF, which inhibits cleavage of Aβ1-42 by NE. [Figure 2] This figure shows the binding levels of Aβ1-42 to peptides 20-44 (20-44 CC, 95-122, and 120-146 QR) derived from wild-type CAP37 and peptides derived from mutant CAP37 (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. [Figure 3] Figure 2 shows the level of Aβ1-42 binding to the "5RMP" modified CAP37-derived peptide (two AEEA mini-PEG molecules and four consecutive arginine molecules linked to the N-terminus of the peptide, and one arginine molecule linked to the C-terminus of the peptide). [Figure 4] This graph shows the level of fibrillation (aggregation) of Aβ1-42 in the presence of the full-length CAP37, NE, or CG at the indicated concentrations. The time to complete fibrillation of Aβ1-42 (without protein addition) is indicated by the vertical dotted line. [Figure 5] This graph shows the level of Aβ1-42 fibrillation (aggregation) in the presence of full-length CAP37, NE, or CG at various protein concentrations. NE has the highest inhibitory effect on Aβ1-42 fibrillation, followed by CG, and then CAP37. [Figure 6] This figure shows that the protease inhibitor PMSF impairs the inhibitory effect of CAP37 on Aβ1-42 fibrosis. The IC50 of CAP37 is increased 3.5 times in the presence of PMSF. This indicates that the inhibitory effect of CAP37 on fibrosis is at least partially due to the cleavage of Aβ1-42. [Figure 7] This figure shows that the protease inhibitor PMSF impairs the inhibitory effect of NE on Aβ1-42 fibrosis. The IC50 of NE increases 36-fold in the presence of PMSF. This indicates that the inhibitory effect of NE on fibrosis is at least partially due to the cleavage of Aβ1-42. [Figure 8]This figure shows the direct correlation between NE-mediated cleavage of Aβ1-42 and inhibition of fibrosis. This correlation supports the idea that NE-mediated inhibition of fibrosis is primarily mediated by Aβ cleavage. This experiment was conducted in the absence of PMSF. [Figure 9] This figure shows the direct correlation between NE-mediated cleavage of Aβ1-42 and inhibition of fibrosis. This experiment was conducted in the presence of PMSF. [Figure 10] This figure shows that the protease inhibitor chymostatin impairs the inhibitory effect of CG on Aβ1-42 fibrosis. The IC50 of CG is increased 17-fold in the presence of chymostatin. This indicates that the inhibitory effect of CG on fibrosis is at least partially due to the cleavage of Aβ1-42. [Figure 11] This figure shows the direct correlation between CG-mediated cleavage of Aβ1-42 and inhibition of fibrosis. This correlation supports the idea that CG-mediated inhibition of fibrosis is primarily mediated by Aβ cleavage. This experiment was conducted in the absence of chymostatin. [Figure 12] This figure shows the direct correlation between CG-mediated cleavage of Aβ1-42 and inhibition of fibrosis. This experiment was performed in the presence of chymostatin. [Figure 13] This figure shows that peptides 120-146 WR and 120-146 WR 5RMP derived from CAP37 inhibit fibrosis. Peptide 95-122 5RMP does not inhibit fibrosis. [Figure 14] This figure shows that the Aβ1-42 oligomer is more toxic to neurons than the fibril. The monomeric form of Aβ1-42 has the lowest toxicity. [Figure 15] This figure shows that when incubated under conditions favorable to fibril formation, NE and CG inhibit the toxicity of Aβ1-42, but CAP37 does not. [Figure 16] This figure shows the effects of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of NE. NE significantly inhibited the toxicity of Aβ1-42 oligomers in a dose-dependent manner. [Figure 17]This figure shows the effects of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CG. CG significantly inhibited the toxicity of Aβ1-42 oligomers in a dose-dependent manner. [Figure 18] This figure shows the effects of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CAP37 protein. Full-length CAP37 does not inhibit the toxicity of Aβ1-42 oligomers. [Figure 19] This figure shows the effects of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CAP37-derived peptide 120-146 WR. Modified CAP37 peptides significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 20] This figure shows the effects of Aβ1-42 oligomers on neuronal cells in the presence of various concentrations of CAP37-derived peptides 120-146 WR-5RMP. The 5RMP-modified CAP37 peptide variant significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 21] This figure shows the binding levels of Aβ1-42 to wild-type CAP37-derived peptides 95-122 and CG-derived peptides CG119-144, as well as mutant CG-derived peptides (CG119-144 ΔR129V130, CG119-144 D137R, and CG119-144 E141PR). The highest levels of binding to Aβ1-42 are shown for peptides CG119-144, CG119-144 D137R, and CG119-144 E141PR. [Figure 22] This figure shows the level of Aβ1-42 fibrillation, recorded as relative fluorescence units (Rfu), in the presence of the indicated concentrations of wild-type and mutant CG peptides. The mutants CG119-144 D137R and CG119-144 E141PR exhibit inhibitory effects on Aβ1-42 fibrillation. Wild-type CG119-144 and mutant CG119-144 ΔR129V130 do not exhibit inhibitory effects. [Figure 23] Peptide CG119-144 D137R inhibits Aβ1-42 fibrillation in a dose-dependent manner. [Figure 24] This figure shows the effects of Aβ1-42 oligomers on neurons in the presence of various concentrations of the CG-derived mutant peptide CG119-144 D137R. This mutant CG peptide significantly inhibited Aβ1-42 oligomer toxicity in a dose-dependent manner. [Figure 25] This figure shows the effect of pre-incubation of CAP37-derived and CG-derived peptides with Aβ1-42 oligomers on neurotoxicity. The oligomers were initially formed, and then incubated for 3 days in or without the presence of CG mutant CG119-144 D137R or CAP37 mutant 120-146 WR-5RMP before treatment of nerve cells. Both peptides significantly reversed the neurotoxicity of the Aβ1-42 oligomers. [Figure 26] This figure shows the effects of CG peptide CG119-144D137R and CAP37 peptide CAP37120-146WR on the formation of Aβ1-42 oligomers. In the absence of the peptides, the Aβ1-42 monomer spontaneously forms oligomers. When combined with the Aβ1-42 monomer, CG peptide CG119-144D137R and CAP37 peptide CAP37120-146WR inhibit the formation of Aβ1-42 oligomers in a dose-dependent manner. [Modes for carrying out the invention]

[0009] Detailed explanation This disclosure relates, in at least certain embodiments, to peptide compounds containing peptide sequences obtained from or derived from three neutrophil granule proteins, cathepsin G (CG), neutrophil elastase (NE), and a 37 kDa cationic antimicrobial protein (CAP37), methods of using them to inhibit amyloid beta (Aβ) fibrillation and oligomerization, and methods of using them as therapeutic agents to treat, alleviate, and / or inhibit Aβ-related diseases and conditions, including, for example, Alzheimer's disease (AD), cerebral amyloid angiopathy, dementia, and / or neuroinflammation, and / or their symptoms. CAP37, NE, and CG are Aβ 1-42 It was discovered that the 42-amino acid form of Aβ can be cleaved at different sites and at different rate structures. It was also observed that CAP37 is upregulated in the brains of patients with AD, while NE and CG are not. CAP37 has been found to bind to RAGE, as well as to the Aβ peptide, a known ligand for RAGE. NE and CG also bind to RAGE. The CAP37-RAGE interaction and the NE-RAGE interaction are inhibited by certain RAGE inhibitors. 1-42 It is hypothesized that the binding and cleavage of Aβ would inhibit its ability to form toxic oligomers and profibrils. This disclosure describes CAP37, NE, and CG, as well as peptide derivatives, that can inhibit Aβ aggregation (profibril formation). The oligomeric and polymerized forms of Aβ are considered to be more toxic than the monomeric form. Therefore, inhibiting Aβ oligomerization and aggregation can reduce the toxicity of Aβ and slow the progression of AD.

[0010] While not intending to be bound by theory, the peptide compounds of this disclosure are thought to be able to bind to RAGE at the blood-brain barrier, thereby inhibiting the binding of systemically produced Aβ and thus inhibiting its transport to the brain. Subsequently, the peptides, not Aβ, would be transported to the brain across the blood-brain barrier via RAGE-mediated transcellular transport. Once in the brain, the peptides inhibit the oligomerization and / or polymerization of Aβ, causing deaggregation of aggregated Aβ (depolymerizing Aβ oligomers and fibrils), thereby reducing and reversing both the neurotoxic effects of Aβ and the deposition of Aβ in amyloid plaques. Furthermore, the peptides bind to RAGE and TLR4 on neuronal and microglial cells, which are mediated by aggregated Aβ and other agonists, antagonizing their pro-inflammatory effects and thereby reducing neuroinflammation.

[0011] Before further detailed descriptions of various embodiments of the compositions and methods of use of the Disclosure, it should be understood that the Disclosure is not limited to its application to the details of methods and compositions as shown in the following description. The descriptions provided herein are intended for illustrative purposes only and are not intended to be construed as limiting. Other embodiments of the Disclosure are possible, or can be carried out or performed in various ways. As such, the language used herein is intended to give the broadest possible range and meaning, and the embodiments are intended to be illustrative and not exhaustive. It should also be understood that the expressions and terminology used herein are for illustrative purposes only and should not be considered as limiting unless otherwise indicated. Furthermore, in the detailed description below, numerous specific details are given to provide a more complete understanding of the Disclosure. However, it will be apparent to those skilled in the art that various embodiments of the Disclosure can be carried out without these specific details. In other examples, features well known to those skilled in the art are not described in detail to avoid unnecessarily complicating the description. All substitutes, replacements, modifications, and equivalents obvious to those skilled in the art are intended to be included within the scope of the Disclosure as defined herein. Accordingly, the following examples, including specific embodiments, serve to illustrate the practice of the Disclosure, and it is understood that the details shown are presented as examples and for illustrative purposes only of specific embodiments, and are intended to provide a useful and easily understandable explanation of the procedures and the principles and conceptual aspects of the inventive concept. Accordingly, although the compositions and methods of the Disclosure are described in relation to specific embodiments, it will be apparent to those skilled in the art that modifications can be applied to the compositions and / or methods described herein, and to the steps or sets of steps of such methods, without departing from the concept, spirit, and scope of the inventive concept disclosed herein.

[0012] All patents, published patent applications, and non-patent publications referenced herein represent the level of skill of those skilled in the art to which this disclosure relates. Each patent, published patent application, and non-patent publication referenced anywhere in this application is expressed herein by reference to the same extent as if each individual patent, published patent application, or non-patent publication were specifically and individually indicated and incorporated by reference. In particular, U.S. Patent Application No. 17 / 115,321 filed on 8 December 2020, International Application No. PCT / US2019 / 036281 brochure filed on 10 June 2019, U.S. Provisional Patent Application No. 62 / 682,455 filed on 8 June 2018, International Publication Nos. 2014 / 089088 and 2014 / 172747 brochures, and U.S. Patents Nos. 9,096,679; 9,603,896; 9,624,283; and 9,862,748 are expressly incorporated herein by reference as a whole.

[0013] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings generally understood by those skilled in the art. Furthermore, unless otherwise required by context, singular terms shall include plural forms, and plural terms shall include singular forms.

[0014] When used in accordance with the methods and compositions of this disclosure, the following terms shall be understood to have the following meanings unless otherwise indicated:

[0015] The words “a” or “an” when used with the term “comprising” in the claims and / or specification may mean “one,” but it also coincides with the meanings of “one or more,” “at least one,” and “one or more.” The use of the term “or” in the claims is used to mean “and / or,” even if this disclosure supports a definition that refers only to substitutes and “and / or,” unless it is expressly indicated to refer only to substitutes, or unless the substitutes are mutually exclusive. The use of the term “at least one” is understood to include one, as well as any quantity more than one, including but not limited to 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 100, or any integer contained therein. The term "at least one" can expand to a maximum of 100 or 1000 or more, depending on the term it is attached to; in addition, the quantity of 100 / 1000 should not be considered restrictive, as higher limits may also yield satisfactory results. Furthermore, the use of the term "at least one of X, Y, and Z" is understood to include X alone, Y alone, and Z alone, as well as any combination of X, Y, and Z.

[0016] As used in this specification and claims, the words “comprising” (and any form of “comprising,” such as “comprise” and “comprises”), “having” (and any form of “having,” such as “have” and “has”), “including” (and any form of “including,” such as “includes” and “include”), or “containing” (and any form of “containing,” such as “contains” and “contain”) are comprehensive or open-ended and do not exclude additional, unenumerated elements or method steps.

[0017] As used herein, the term “or any combination thereof” refers to all permutations and combinations of the enumerated items preceding the term. For example, “A, B, C, or any combination thereof” is intended to include at least one of the following: A, B, C, AB, AC, BC, or ABC, and also, where the order is important in a particular context, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing this example, combinations containing repetitions of one or more items or terms are clearly included, such as BB, AAA, AAB, BBC, AAABCCCC, CBBAAA, CABABB, etc. Typically, those skilled in the art will understand that there is no limit to the number of items or terms in any combination unless otherwise evident from the context.

[0018] Throughout this application, the terms “about” or “approximately” are used to indicate that a value includes inherent variations in errors of a composition, a method used to administer the composition, or variations present among the subjects of study. As used herein, the modifying phrases “about” or “approximately” are intended to include not only exact values, quantities, degrees, orientations, or other recognized characteristics or values, but also some slight variations due to, for example, measurement errors, manufacturing tolerances, observer errors, and combinations thereof. As used herein when referring to measurable values ​​such as quantities or temporal durations, the terms “about” or “approximately” are intended to include variations of, for example, ±20%, ±10%, ±5%, ±1%, or ±0.1% from a specified value, as is reasonable for carrying out the disclosed method and as is understood by those skilled in the art. As used herein, the term “substantially” means that the event or situation described thereafter is occurring entirely, or that the event or situation described thereafter is occurring to a large extent or degree. For example, the term “substantially” means that the event or situation described thereafter occurs at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0019] Where used herein, any reference to “one embodiment” or “an embodiment” means that certain elements, features, compositions, structures, or properties described in relation to an embodiment are included in at least one embodiment. The phrase “in one embodiment” appearing in various places herein does not necessarily refer to the same embodiment.

[0020] As used herein, the terms “AEEA,” “AEEEA,” “AEEP,” and “AEEEP” refer to short polyethylene glycol molecules ("miniPEG") that, in certain embodiments, constitute a part of the compounds disclosed herein and have the following specific meanings:

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

[0022] The term AEEEA represents the -NH(CH2CH2O)3CH2CO- or NH2(CH2CH2O)3CH2CO-, which is the N or C-terminal group derived from {2-[2-(2-amino-ethoxy)-ethoxy]-ethoxy}acetic acid (also known as 11-amino-3,6,9-trioxaundecanoic acid).

[0023] The term AEEP refers to the N or C-terminal group derived from [3-(2-amino-ethoxy)-ethoxy]-propanoic acid (also known as 9-amino-4,7-dioxanonic acid), which is -NH(CH2CH2O)2CH2CH2CO- or NH2(CH2CH2O)2CH2CH2CO-.

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

[0025] The following is a list of other abbreviations used herein, as well as the complete words or phrases they represent: AGEs: Advanced Glycation End Products Aβ: Aminoid 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: Bicinconinic acid BSA: Bovine serum albumin DAB: Diaminobenzidine tetrahydrochloride BPI: Enhances bactericidal permeability DAG: Diacylglycerol EDTA: Ethylenediaminetetraacetic acid EGF: Epidermal growth factor EGFR: Epidermal Growth Factor Receptor ERK: Extracellular Signal Regulatory Kinase FBS: Fetal Bovine Serum 5RMP:5 Arginine-Mini PEG 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 equilibrium 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-inducing protein-10 KC: Keratinocyte-derived chemokine KSFM: Serum-free keratinocyte 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: Holball 12-Millistate 13-Acetate PMSF: Phenylenmethylsulfonyl Fluoride RAGE: Advanced Glycation End Product Receptor RIPA: Radioactive Immunoprecipitation 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, 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 CAP37 Protein Amino Acids 120-146 CG 20-47 : Amino acids 20-47 of cathepsin G protein NE 20-44 : Amino acids 20-44 of neutrophil elastase CAP37 20-44 :CAP37 Protein Amino Acids 20-44

[0026] The following standard amino acid abbreviations are used herein: Alanine:ala:A, cysteine:cys:C, aspartic acid:asp:D, glutamic acid: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:pro:P, arginine:arg:R, serine:ser:S, threonine:thr:T, valine:val:V, tryptophan:trp:W, and tyrosine:tyr:T.

[0027] The term “variant” is intended to refer to a protein, peptide, nucleic acid, or organism having at least one amino acid or nucleotide that differs from the wild-type version of the protein, peptide, nucleic acid, or organism, and includes, but is not limited to, point substitutions, multiple consecutive or discontinuous substitutions, insertions, chimeric or fusion proteins, and the nucleic acids that encode them. Examples of conserved amino acid substitutions include, but are not limited to, substitutions occurring within the same group, such as within the group of 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 asparagine), hydrophobic amino acids (e.g., leucine, isoleucine, and valine), aromatic amino acids (e.g., phenylalanine, tryptophan, tyrosine), and small amino acids (e.g., glycine, alanine, serine, threonine, methionine). Other examples of possible substitutions are listed below.

[0028] As used herein, the terms “peptide,” “peptide analog,” “peptide derivative,” or “peptide compound” may refer to a molecule containing only amino acids, or a molecule containing amino acids and one or more non-amino acid structures (e.g., PEG units). The terms “peptide,” “peptide analog,” “peptide derivative,” or “peptide compound” may refer to a variant ("mutant") of a “wild-type” peptide, or a molecule containing amino acids and one or more non-amino acid structures (e.g., PEG units).

[0029] The term "amyloid beta" (Aβ), unless otherwise specified, refers to an Aβ peptide or other form containing either 1 to 40 or 1 to 42 amino acids.

[0030] The term "pharmaceutically acceptable" refers to compounds and compositions that are suitable for administration to humans and / or animals, without excessive adverse side effects such as toxicity, irritation, and / or allergic reactions, and that meet a reasonable benefit-to-risk ratio.

[0031] "Bioactive" means the ability of an active agent to modify the physiological system of an organism, regardless of how the active agent produces its physiological effects.

[0032] As used herein, “pure” or “substantially pure” means that the species of interest is the dominant species present (i.e., on a molar basis, it is more abundant in the composition than any other species of interest), and in particular, that the substantially purified fraction is composed of at least about 50 percent (on a molar basis) of the species of interest of all the polymer species present. Generally, a substantially pure composition consists of more than about 80%, more particularly, more than about 85%, more than about 90%, more than about 95%, or more than about 99% of all the polymer species present in the composition. The terms “pure” or “substantially pure” also refer to a preparation in which the target species (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 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 are understood to refer to warm-blooded animals, particularly mammals. Non-restrictive 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, cattle, sheep, zoo animals, camels, llamas; non-human primates, including Old World and New World monkeys and non-human primates (e.g., crab-eating macaques, chimpanzees, rhesus macaques, orangutans, and baboons); and humans.

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

[0035] "Treatment" refers to a therapeutic procedure. "Prevention" refers to preventive or protective measures. The term "to treat" refers to administering a composition to a patient for therapeutic purposes.

[0036] The terms “therapeutic composition” and “pharmaceutical composition” refer to active agent-containing compositions that can be administered to a subject by any method known in the art or by another method contemplated herein, wherein the administration of the composition produces a therapeutic effect as described elsewhere herein. In addition, the compositions of this disclosure may be designed using formulation techniques known in the art to provide delayed, controlled, extended, and / or sustained release.

[0037] When used in the context of the inventive concept, the term "effective dose" refers to the amount of active agent sufficient to produce a detectable therapeutic effect, without excessive adverse side effects (e.g., toxicity, irritation, and allergic responses), commensurate with a reasonable benefit-to-risk ratio. The effective dose for a patient depends on the patient's type, size and health, the nature and severity of the condition to be treated, the method of administration, the duration of treatment, the nature of any concomitant therapies, and the specific formulation used. Therefore, it is not possible to determine the exact effective dose in advance. However, the effective dose for a given situation can be determined by those skilled in the art using routine experiments based on the information provided herein.

[0038] The term “remission” means a detectable or measurable improvement in a condition, disease, or its symptoms. A detectable or measurable improvement may include a subjective or objective reduction, decrease, inhibition, suppression, limitation, or control of the incidence, frequency, severity, progression, or duration of the condition or disease, or an improvement in the symptoms, underlying causes, or consequences of the disease, or an improvement in the disease itself. A successful treatment outcome may result in a “therapeutic effect” or “benefit” of remitting, reducing, decreasing, inhibiting, suppressing, limiting, controlling, or preventing the incidence, frequency, severity, progression, or duration of the disease or condition, or the prognosis of the disease or condition in the subject.

[0039] A reduction or decrease in worsening, such as stabilizing a condition or disease, is also a successful treatment outcome. Therefore, therapeutic benefit does not require the complete disappearance or improvement of any one, most, or all adverse symptoms, complications, prognosis, or underlying cause of the disease or condition, or any one, most, or all adverse symptoms, complications, prognosis, or underlying cause associated with the disease or condition. Thus, a satisfactory endpoint may be achieved if there is a partial reduction, decrease, inhibition, suppression, limitation, control, or prevention of the incidence, frequency, severity, progression, or duration of the condition or disease, or a gradual improvement (e.g., stabilization) over a short or long duration (hours, days, weeks, months, etc.) of the condition or disease. The effectiveness of methods or uses, such as treatments, that result in potential therapeutic benefits or improvements in a condition or disease can be confirmed by various methods and test assays.

[0040] The terms “peptide” or “peptide sequence” are used herein to refer to a series of amino acid residues linked together. In natural ("wild-type") peptides, amino acids are linked by peptide bonds between their alpha-amino groups and the carbonyl groups of adjacent amino acids to form an amino acid sequence. In certain embodiments, a peptide may have a length of 5 to 15, 25, 40, 60, 75 or more amino acids, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27 , 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or in the range of 60-75-100 or more amino acids. The terms “polypeptide” or “protein” are used herein to refer to a series of amino acid residues that are longer than a single peptide and are typically linked from one to the other by a peptide bond between the alpha-amino group and carbonyl group of adjacent amino acids. “Fusion protein” or “fusion polypeptide” refers to (and may be used interchangeably) a protein or polypeptide that is constructed by recombinant or synthetic methods to combine peptides in a continuous stereochemistry. In certain embodiments, the CAP37, NE, and CG peptide derivatives described elsewhere in this Specification may be shortened by, for example, one, two, three, four, or five amino acids at one or both ends, as long as the resulting variant peptide retains the biological activity described herein relating to the inhibition of oligomerization and / or polymerization of Aβ and / or deaggregation of Aβ. In other embodiments, the variant peptide may include the insertion of one, two, or three amino acids.

[0041] As used herein, the term “wild-type” refers to an amino acid sequence or peptide that exists naturally or under natural conditions, as opposed to a “non-wild-type” amino acid sequence or peptide that does not exist naturally or under natural conditions. A non-wild-type amino acid sequence or peptide may differ from the wild-type amino acid sequence or peptide, for example, by substitution, deletion, or insertion of one or more natural (alpha) amino acids at one or more amino acid positions of the wild-type amino acid sequence or peptide. A non-natural amino acid sequence or peptide may also differ from the wild-type amino acid sequence or peptide by only one or more substitutions with respect to its corresponding D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. For example, if a wild-type peptide contains serine, the non-wild-type may instead contain D-serine, β-serine, homoserine, β-homoserine, or a peptoid monomer analog of serine. A non-wild-type may differ from the wild-type by only one amino acid position, a subset of amino acid positions, or all amino acid positions. For example, peptides containing only D-amino acids are known as retro-inverso peptides. Non-wild-type amino acids or peptides may also be those in which two or more amino acid monomers are linked by non-peptide bonds, such as peptoid bonds. In certain embodiments, non-wild-type non-peptide bonds may be of the following types: reverse peptide bond (-NH-CO-), -CH2-NH-, -CH2S-, -CH2CH2-, -CH=CH-, -COCH2-, -CH(OH)CH2-, and -CH2SO-. Exemplary non-peptide bonds that may be used are described, for example, in U.S. Patents 4,897,445 and 10,064,926. The peptide compounds of this disclosure may also include wild-type amino acid sequences or peptides conjugated with other natural peptides or amino acid sequences, which, when conjugated together, include synthetic or non-natural amino acid sequences or peptides.The peptide compounds of this disclosure may also include wild-type amino acid sequences or peptides conjugated with non-natural amino acid sequences or peptides, such as one or more D-amino acids, β-amino acids, homoamino acids, β-homoamino acids, or peptoid monomer analogs. The peptide compounds of this disclosure may include wild-type amino acid sequences or peptides conjugated with non-amino acid molecules, such as PEG molecules, to enhance solubility, permeability, or resistance to enzymatic degradation. In another embodiment, peptides may be “stapled” to a specific helical configuration. As used herein, the term “staple” generally refers to a covalent bond between the side chains or functional groups of two amino acids in a peptide. Such “stapling” in a peptide is also considered non-wild-type, distinct from the wild-type peptide. Stapled peptides are discussed in further detail below.

[0042] The terms “synthetic amino acids” and “non-natural amino acids” may be used in place of the term “non-wild type” and also refer to organic compounds that have a structure similar to natural amino acids in order to mimic the structure and reactivity of natural amino acids. Synthetic amino acids, as defined herein, generally increase or enhance the properties of a peptide (e.g., selectivity, stability) when they are either substituted for a natural amino acid or incorporated into a peptide.

[0043] Therefore, a non-wild-type homolog of a wild-type amino acid sequence or peptide refers to a non-wild-type amino acid sequence or peptide that has at least one difference from the wild-type amino acid sequence or peptide in at least one manner as described above.

[0044] When used in reference to a single amino acid molecule, the term "non-wild type" can also refer to a single amino acid or amino acid analog, such as a peptoid monomer, that does not exist naturally or under natural conditions, in contrast to wild-type amino acids.

[0045] The non-wild-type amino acid sequences and peptides of this disclosure may include, but are not limited to, common natural amino acids, alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, and tyrosine, as well as less common naturally occurring amino acids, modified amino acids, or synthetic compounds, including: alpha-asparagine, 2-aminobutanoic acid, 2-aminobutyric acid, 4-aminobutyric acid, 2-aminocaproic acid (2-aminodecanoic acid), 6-aminocaproic acid, alpha-glutamine, 2-aminoheptanoic acid, 6-aminohexanoic acid, alpha-aminoisobutyric acid (2-aminoalanine), 3-aminoisobutyric acid, beta-alanine, allo-hydroxylysine, allo-isoleucine, 4-amino-7-methylheptanoic acid, 4-amino-5-phenylpentanoic acid, 2-aminopimelic acid, gamma-amino-beta-hydroxybenzenepentanoic acid, 2-aminosuberic acid, 2-carboxyazetidine, beta-alanine, beta-aspartic acid, biphenylalanine, 3,6-diaminohexanoic acid, butanoic acid, cyclobutylalanine, cyclohexylalanine, cyclohexylglycine, N5-aminocarbonylornithine, cyclopentylalanine, cyclopropylalanine, 3-sulfoalanine, 2,4-diaminobutanoic acid, diaminopropionic acid, 2,4-diaminobutyric acid, diphenylalanine, NN-dimethylglycine, diaminopimelic acid, 2,3-Diaminopropanoic acid, S-ethylthiocysteine, N-ethylasparagine, N-ethylglycine, 4-azaphenylalanine, 4-fluorophenylalanine, gamma-glutamic acid, gamma-carboxyglutamic acid, hydroxyacetic acid, pyroglutamic acid, homoarginine, homocysteineic acid, homocysteine, homohistidine, 2-hydroxyisovaleric acid, homophenylalanine, homoleucine, homoproline, homoserine, homoserine, 2-hydroxypentanoic acid, 5-hydroxylysine, 4-hydroxyproline, 2-cabroxyoctahydroindole, 3-carboxyisoquinoline, isovaline, 2-hydroxypropanoic acid (lactic acid), Mercaptoacetic acid, mercaptobutanoic acid, sarcosine, 4-methyl-3-hydroxyproline, mercaptopropanoic acid, norleucine, nipecoic acid, nortyrosine, norvaline, omega-amino acids, ornithine, penicillamine (3-mercaptovaline), 2-phenylglycine, 2-carboxypiperidine, sarcosine (N-methylglycine), 2-amino-3-(4-sulfophenyl)propionic acid, 1-amino-1-carboxycyclopentane, 3-thienylalanine, epsilon-N-trimethyllysine, 3-thiazolylalanine, thiazolidinedione-4-carboxylic acid, alpha-amino-2,4-dioxopyrimidinepropanoic acid, and 2-naphthylalanine.

[0046] The peptides and nucleic acids encoded herein include peptide and nucleic acid variants containing natural amino acid or base substitutions (conservative or non-conservative). For example, peptide variants include, but are not limited to, those having additional amino acid residue substitutions (conservative or non-conservative) that are not exactly the same as the sequences disclosed herein, but which do not substantially impair the activity or properties of the variants described herein, in addition to the substitutions explicitly described for the various sequences enumerated herein. Examples of such conserved amino acid substitutions include: ala to gly, ser, or thr; arg to gln, his, or lys; asn to asp, gln, his, lys, ser, or thr; asp to asn or glu; cys to ser; gln to arg, asn, glu, his, lys, or met; glu to asp, gln, or lys; gly to pro or ala; his to arg, asn, gln, or tyr; ile to leu, met, or va Possible but not limited to: to l; from leu to ile, met, phe, or val; from lys to arg, asn, gln, or glu; from met to gln, ile, leu, or val; from phe to leu, met, trp, or tyr; from ser to ala, asn, met, or thr; from thr to ala, asn, ser, or met; from trp to phe or tyr; from tyr to his, phe, or trp; and from val to ile, leu, or met.

[0047] Those skilled in the art will readily know how to construct, identify, select, or test for binding activity to the same receptor targeted by non-variant peptides. Specific examples of conserved amino acid substitutions include, but are not limited to, gly:ala substitution; val:ile:leu substitution; asn:glu:his substitution; asp:glu substitution; ser:thr:met substitution; lys:arg:his substitution; and phe:tyr:trp substitution. Other types of substitutions, alterations, additions, deletions, and derivatives that result in functional variant peptides are also encompassed by this disclosure, and those skilled in the art will readily know how to construct, identify, or select such variants or derivatives, and how to test for receptor binding activity of those variants. Examples of conservative amino acid substitutions include, but are not limited to, substitutions occurring within the same group, such as within basic amino acids (arginine, lysine, histidine, etc.), acidic amino acids (glutamic acid and aspartic acid, etc.), polar amino acids (glutamine and asparagine, etc.), hydrophobic amino acids (leucine, isoleucine, and valine, etc.), aromatic amino acids (phenylalanine, tryptophan, tyrosine, etc.), and small amino acids (glycine, alanine, serine, threonine, methionine, etc.). Other examples of possible substitutions are listed below.

[0048] The term “mutant” or “variant” is intended to refer to a protein, peptide, nucleic acid, or organism that has at least one amino acid or nucleotide that differs from the wild-type version of the protein, peptide, nucleic acid, or organism, including, but not limited to, point substitutions, multiple consecutive or non-consecutive substitutions, insertions, chimeric or fusion proteins, and the nucleic acids that encode them, or other non-wild-type features as described above.

[0049] As used herein, the terms “peptide,” “peptide analog,” “peptide derivative,” or “peptide compound” may refer to a molecule containing only amino acids, or a molecule containing amino acids and one or more non-amino acid structures (e.g., PEG units). The terms “peptide,” “peptide analog,” “peptide derivative,” or “peptide compound” may refer to a variant ("mutant") of a “wild-type” peptide, or a molecule containing amino acids and one or more non-amino acid structures (e.g., PEG units).

[0050] The terms “polypeptide” or “protein” are used herein to refer to a series of amino acid residues that are longer than a single peptide and are typically linked from one to the other by a peptide bond between an alpha-amino group and the carbonyl group of an adjacent amino acid, or by other linkages as described herein. The peptide compounds of the present disclosure may be peptide conjugates, which are compounds comprising the peptide of the present disclosure that are conjugated (e.g., covalently linked directly or indirectly via a linker sequence) with another molecule, such as (non-limitingly) a protein or other polymer molecule, such as a carrier molecule such as a serum albumin molecule or an antibody, or (non-limitingly) a drug, or another therapeutic compound such as an imaging or diagnostic moiety, even when the peptide is conjugated with the molecule, and which retain its activity (e.g., binding, targeting, imaging, or inhibitory) even when conjugated with the molecule. The peptides of the present disclosure may be produced using any nucleotide sequence encoding a desired amino acid sequence. Any of the peptides or active variants described herein may be used to construct the peptide conjugates of this disclosure.

[0051] As used herein, the terms “homologous” or “% identity” mean a nucleic acid (or a fragment thereof) or peptide having a degree of homology to the corresponding natural reference nucleic acid or peptide, which may be more than 60%, or more than 65%, or more than 70%, or more than 75%, or more than 80%, or more than 85%, or more than 90%, or more than 91%, or more than 92%, or more than 93%, or more than 94%, or more than 95%, or more than 96%, or more than 97%, or more than 98%, or more than 99%, or any other specific percentage described herein. For example, with respect to peptides, the percentage of homology or identity described herein is typically calculated (as shown by Dayhoff, in Atlas of Protein Sequence and Structure, Vol. 5, p. 124, National Biochemical Research Foundation, Washington, DC (1972)) as the percentage of amino acid residues found in the shorter of the two aligned sequences with respect to identical amino acid residues in the sequences to be compared, provided that four gaps per 100 amino acids are introduced to aid in the alignment. In one embodiment, such percentage homology is calculated as the percentage of components found in the shorter of the two sequences that may also be found in the longer of the two sequences (with the introduction of gaps), where a component is defined as a sequence of four consecutive amino acids. Any protein product that can be isolated thanks to cross-reactivity with antibodies against native protein products is also included as substantially homologous. Sequence identity or homology can be determined by comparing sequences when they are aligned to maximize overlap and identity while minimizing sequence gaps. In particular, sequence identity can be determined using one of several mathematical algorithms.An unrestricted example of a mathematical algorithm used for comparing two arrays is the algorithm in Karlin & Altschul, Proc. Natl. Acad. Sci. USA 1990, 87, 2264-2268, which is modified as shown in Karlin & Altschul, Proc. Natl. Acad. Sci. USA 1993, 90, 5873-5877.

[0052] In at least one embodiment, "% identity" represents the number of amino acids or nucleotides that are identical at corresponding positions in two sequences of nucleic acids encoding a peptide or a similar peptide. For example, two amino acid sequences, each having 15 residues, have at least 60% identity if at least 9 of the amino acids at corresponding positions are the same, at least 66% identity if at least 10 of the amino acids at corresponding positions are the same, at least 73% identity if at least 11 of the amino acids at corresponding positions are the same, at least 80% identity if at least 12 of the amino acids at corresponding positions are the same, at least 86% identity if at least 13 of the amino acids at corresponding positions are the same, and at least 93% identity if at least 14 of the amino acids at corresponding positions are the same. In another example, two amino acid sequences, each having 19 residues, are at least 73% identical if at least 14 of the amino acids at their corresponding positions are the same, at least 78% identical if at least 15 of the amino acids at their corresponding positions are the same, at least 84% identical if at least 16 of the amino acids at their corresponding positions are the same, at least 89% identical if at least 17 of the amino acids at their corresponding positions are the same, and at least 94% identical if at least 18 of the amino acids at their corresponding positions are the same.

[0053] Similarly, two amino acid sequences, each having 20 residues, will have at least 95% identity if 19 of the amino acids at their corresponding positions are the same, or at least 90% identity if at least 18 of the amino acids at their corresponding positions are the same, or at least 85% identity if at least 17 of the amino acids at their corresponding positions are the same, or at least 80% identity if at least 16 of the amino acids at their corresponding positions are the same. In other non-limiting examples, two amino acid sequences, each having 100 residues, will have 95% identity if 95 of the amino acids at their corresponding positions are the same. Two amino acid sequences, each having 100 residues, will have at least 90% identity if at least 90 of the amino acids at their corresponding positions are the same. Furthermore, where a sequence is described herein as having "at least X% identity" with respect to a reference sequence, this is intended to include all percentages higher than X%, e.g., (X+1)%, (X+2)%, (X+3)%, (X+4)%, etc., up to 100%, unless otherwise indicated.

[0054] In at least certain embodiments, the Disclosure relates to peptides comprising at least 25 amino acids, wherein the peptide comprises at least 1 to at least 25 D-amino acids. In certain embodiments, the peptides of the Disclosure have sequences selected from SEQ ID NOs: 1 to 6, wherein the sequences may contain at least 1 or more D-amino acids, or may consist entirely of D-amino acids. Peptides comprising 1 or more D-amino acids have certain properties compared to peptides composed solely of L-amino acids, including, but not limited to, (1) the ability to withstand all natural peptidases that can act on L-amino acid-containing peptides; (2) the ability to withstand gastrointestinal exposure to natural peptidases and limited acid hydrolysis; and (3) the possibility of oral, nasal, or alveolar administration.

[0055] As discussed above, in certain embodiments, the peptides of this disclosure comprise one or more β-amino acids in which the primary amino group of an amino acid is bonded to the β-carbon of the amino acid, rather than to an α-amino acid as in natural amino acids. Such β-amino acids are commercially available. In alternative embodiments, the amino acids may be homoamino acids, or β-homoamino acid versions of the α-amino acids described herein.

[0056] As discussed above, in certain embodiments, as used herein, the term “peptide compound” is also intended to include peptoid versions of the peptides described herein. Peptoids include peptoid monomer analogs of α-amino acids described herein. All or part of the amino acids of the peptides of this disclosure may be peptoid monomer equivalents of α-amino acids described elsewhere herein. Peptoids include a class of peptide mimetic compounds composed of poly-N-substituted glycine monomers, in which the side chains are bonded to the nitrogen atom of the peptide backbone rather than to the α-carbon (where they are located within amino acids). Peptoids lack amide hydrogens, which are responsible for many of the secondary structural elements in peptides and proteins. Like D-amino acid peptides and β-amino acid peptides, peptoids are resistant to proteolysis and are therefore advantageous for therapeutic applications where proteolysis is the primary concern.

[0057] Therefore, wherever 20 natural amino acids are identified herein, each is intended to be read not only as a natural L-amino acid, but also, alternatively, as its D-amino acid, as a β-amino acid, as a homoamino acid, as a β-homoamino acid, or as a peptoid monomer analog.

[0058] In other non-limiting embodiments, the amino acid sequences of the peptide compounds currently disclosed may be “stapled” to a specific helical configuration, meaning that certain non-adjacent residues, regions, parts, or domains of the peptide are connected to one another. Stapled peptides generally exhibit increased α-helix stability compared to the corresponding non-stapled peptides. In other embodiments, stapled peptides exhibit increased thermal stability compared to the corresponding non-stapled peptides. In yet another embodiment, stapled peptides exhibit increased biological activity compared to the corresponding non-stapled polypeptides. In yet another embodiment, stapled peptides exhibit increased resistance to proteolysis compared to the corresponding non-stapled peptides. In yet another embodiment, stapled peptides exhibit increased ability to penetrate living cells compared to the corresponding non-stapled peptides.

[0059] As used herein, the term “stapled peptide” refers to a peptide having a selected number of standard (wild-type or natural) or non-standard (non-wild-type or non-natural) amino acids, and further having at least two moieties capable of undergoing a reaction that promotes carbon-carbon bond formation, for example, modulating peptide stability, wherein the at least two moieties are in contact with a reagent that generates at least one crosslinker between them. As used herein, the term “stapling” refers to introducing at least two moieties capable of undergoing a reaction that promotes carbon-carbon bond formation into a peptide, which can be in contact with a reagent that generates at least one crosslinker between the at least two moieties. Stapling imparts constraints to a secondary structure, such as an α-helix structure. The length and geometry of the crosslinker can be optimized to improve the yield of the desired secondary structure content. The given constraints can, for example, prevent the secondary structure from unfolding and / or enhance the shape of the secondary structure. A secondary structure that is prevented from unfolding is, for example, more stable. Further examples of peptide stapling are shown in U.S. Patents No. 8,586,707; No. 10,011,631; No. 10,308,684; and No. 10,682,388, each of which is expressly incorporated herein by reference as a whole.

[0060] In certain embodiments, the i-position (where i is an integer) and i+4-position (or i+7 and / or i+11-position, or other positions) of the α-helix can be stapled using various covalent bonding methods. Specifically, amino acids at one or more positions selected from the group consisting of i, i+3, i+4, i+7, i+8, i+10, and i+11 can be stapled. Stapled peptides have increased cell permeability compared to non-stapled versions. In some cases, two or more amino acid positions selected from the group consisting of i, i+3, i+4, i+7, i+8, i+10, and i+11 can be stapled. Typically, two amino acids can be linked to each other by disulfide bonds, carbon-carbon double bonds, or amide bonds. Examples of methods for linking two amino acids together include the introduction of a disulfide between the two amino acid positions, the introduction of a carbon-carbon double bond by metathesis, the introduction of an amide bond, the introduction of a short linker by Michael's reaction, and other similar methods.

[0061] The term "peptide stapling" can encompass the linking of two double-bonded side chains, two triple-bonded side chains, or side chains containing one double bond and one triple bond, which may be present within a polypeptide chain, using several reaction conditions and / or catalysts that facilitate such a reaction, providing a single "stapled" polypeptide. In certain embodiments, the introduction of staples requires a modification of standard peptide synthesis, in which α-methyl, α-alkenyl amino acids are introduced at two positions along the peptide chain, separated by either three or six (i+4 or i+7) intervening residues. These spacings place the stapling amino acids on the same plane of the α-helix, spanning one (i+4) or two (i+7) helix turns. Additionally, the term "peptide stitching" refers to multiple, serial "stapling" events in a single peptide chain that provide a "stitched" (multi-stapled) polypeptide. International Publication No. 2008 / 121767, which is explicitly incorporated herein by reference in its entirety, provides specific examples of stitched peptide technology.

[0062] In a particular embodiment, the stapling process may be carried out using Aileron Therapeutics' "stapled peptide" technology, in which two key residues within the peptide are modified by the attachment of linkers through their amino acid side chains. Once synthesized, the linkers are catalyzed, thereby creating crosslinks that physically bind the peptide to its native α-helix shape.

[0063] In certain embodiments, if the peptide has a helical secondary structure, the staple is a macrocyclic ring, which is either an exogenous (not part of the helical peptide structure) core or an endogenous (non-stapled) helical peptide structure. The macrocyclic ring consists of a pyrazoline ring and incorporates at least two amino acids of the peptide. The size of the macrocyclic ring is determined by the number of helical peptide amino acids in the ring (y) and the number of methyl groups (m and n) in the portion connecting the pyrazoline group to the peptide. A stapled peptide has at least one peptide. In various embodiments, a stapled peptide has one, two, or three staples.

[0064] The peptide compounds of this disclosure may further contain a labeling substance conjugated to the peptide. Various methods for labeling peptides are known in the art and may be used in accordance with this disclosure. Examples of labeling for peptides and polypeptides include, but are not limited to, radioisotopes or radionuclides, fluorescent labeling, chemiluminescence labeling, and other similar methods. In some embodiments, the labeling substance may be attached to the peptide by spacers or linkers of varying lengths to reduce potential steric hindrance. In addition, the terms “labeling,” “labeling substance,” “detection marker,” “detection portion,” and “reporter molecule” are used interchangeably herein. These conjugates are useful in a variety of diagnostic methods. [Examples]

[0065] Examples are provided below. However, it should be understood that this disclosure is not limited in its application to any specific experiments, results, and experimental procedures disclosed below or elsewhere in this specification. Rather, each example is provided as one of many possible embodiments and is intended to be illustrative and not exhaustive.

[0066] In certain embodiments, the present disclosure is directed to peptide compounds that are wholly or partially made up of one or more of the following amino acid sequences: CG 119-144 :GTLCTVAGWGRVSMRRGTDTLREVQL (SEQ ID NO: 1), NE 119-145 :GVQCLAMGWGLLGRNRGIASVLQELNV (SEQ ID NO: 2), CAP37 120-46 :GTRCQVAGWGSQRSGGRLSRFPRFVNV (SEQ ID NO: 3), CG 20-47 :IQSPAGQSRCGGFLVREDFVLTAAHCWG (SEQ ID NO: 4), NE 20-44 :LRGGHFCGATLIAPNFVMSAAHCVA (SEQ ID NO: 5), and CAP37 20-44 :NQGRHFCGGALIHARFVMTAASCFQ (SEQ ID NO: 6).

[0067] More particularly, the present disclosure includes, but is not limited to, peptide variants of SEQ ID NOs: 1-6 that contain certain substitutions as described below.

[0068] <00007​The variant of (SEQ ID NO: 1) can be used to form the peptide compounds used herein: one or more of the four basic arginine residues (positions 11, 15, 16, and 22) can be changed to lysine or histidine. Arginine at position 11 may be deleted. Valine at position 12 may be deleted. Both arginine at position 11 and valine at position 12 may be deleted. Glycine at position 17 may be deleted. Threonine at position 18 may be deleted. Both glycine at position 17 and threonine at position 18 may be deleted. The acidic residue aspartic acid at position 19 can be changed to glutamic acid, arginine, lysine, or histidine. Glutamic acid at position 23 can be changed to aspartic acid or a proline-arginine ("PR") pair. One or more threonine residues at positions 2, 5, 18, and 20 can be replaced with another hydrophilic residue (asparagine, glutamine, serine, tyrosine) or another small residue (serine, alanine, glycine, valine, proline). Similarly, one or more of the two other hydrophilic residues, serine at position 13 and glutamine at position 25, can be replaced with another hydrophilic residue (asparagine, glutamine, serine, threonine, or tyrosine). One or more of the other (hydrophobic) residues in this sequence can be replaced with another hydrophobic residue (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan). Other possible substitutions or changes are described in more detail below (see Sequence ID 7). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids can be covalently linked, i.e., "stapled".

[0069] In other non-limiting embodiments, NE includes one or more of the following substitutions. 119-145The variant of (SEQ ID NO: 2) can be used to form the peptide compounds used herein: the aromatic tryptophan residue at position 9 can be replaced with another aromatic residue (tyrosine or phenylalanine). The acidic residue glutamic acid at position 24 can be replaced with aspartic acid. One or more of the two basic arginine residues at positions 14 and 16 can be replaced with another basic amino acid (lysine or histidine). One or more of the polar residues glutamine (positions 3 and 23), asparagine (position 15), and serine (position 20) can be replaced with any other polar residue (asparagine, glutamine, serine, threonine, or tyrosine). The serine at position 20 can also be replaced with another small residue (threonine, alanine, glycine, valine, proline). One or more of the other residues in this (hydrophobic) sequence may be substituted with other hydrophobic residues (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan). Other possible substitutions or alterations are described in more detail below (see Sequence ID No. 8). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids may be covalently linked, i.e., “stapled.”

[0070] In other non-limiting embodiments, CAP37 includes one or more of the following substitutions. 120-146The variant of (SEQ ID NO: 3) can be used to form the peptide compounds used herein: one or more of the small hydrophobic glycine residues at positions 1, 8, 10, 15, and 16 of this peptide can be replaced with another hydrophobic amino acid (alanine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, or tryptophan) or another small amino acid (alanine, serine, threonine, valine, or proline). Similarly, one, two, three, four, or all of the positively charged arginine residues at positions 3, 13, 17, 20, and 23 can be replaced with another basic amino acid (lysine or histidine). Other examples of this sequence include changing one or more hydrophilic amino acids, threonine at position 2, glutamine at positions 5 and 12, and serine at positions 14 and 19, to different hydrophilic residues. For example, threonine can be changed to asparagine, glutamine, serine, or tyrosine; glutamine can be changed to asparagine, serine, threonine, or tyrosine; and serine can be changed to asparagine, glutamine, threonine, or tyrosine. One or more of the other (hydrophobic) residues in this sequence can be substituted with other hydrophobic residues (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan). Other possible substitutions or changes are described in more detail below (see Sequence ID No. 9). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids can be covalently linked, i.e., "stapled".

[0071] In other non-limiting embodiments, CGs include one or more of the following substitutions. 20-47Alternative variants of (SEQ ID NO: 4) can be used to form the peptide compounds used herein: In this sequence, two charged residues, glutamic acid and aspartic acid (positions 17 and 18, respectively), can be changed to two glutamic acid residues or two aspartic acid residues, or reversed to aspartic acid and glutamic acid, respectively. One or more of the two positively charged arginine residues at positions 9 and 16 of this peptide can be conservatively changed to lysine or histidine, and the histidine residue at position 25 can be changed to arginine or lysine. One or more polar glutamine residues at positions 2 and 7 can be changed to asparagine, serine, threonine, or tyrosine; and one or more polar serine residues at positions 3 and 8 can be changed to asparagine, glutamine, threonine, or tyrosine. One or more of the other residues in this (hydrophobic) sequence may be substituted with another hydrophobic residue (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan). One or more of the aromatic residues phenylalanine 13 and 19, and tryptophan 27, may be changed with another aromatic residue (phenylalanine, tryptophan, or tyrosine). Other possible substitutions or changes are described in more detail below (see Sequence ID No. 10). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids may be covalently linked, i.e., “stapled.”

[0072] In other non-limiting embodiments, NE includes one or more of the following substitutions. 20-44Alternative variants of (SEQ ID NO: 5) can be used to form the peptide compounds used herein: the positively charged arginine residue at position 2 of this peptide can be conservatively replaced with lysine or histidine, and one or more of the two histidine residues at positions 5 and 22 can be replaced with arginine or lysine. One or more of the two polar residues asparagine 15 and serine 19 can be replaced with any other polar residue (asparagine, glutamine, serine, threonine, or tyrosine). One or more of the other residues in this sequence (which are hydrophobic) can be replaced with another hydrophobic residue (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, or tryptophan). One or more of the phenylalanine residues at positions 6 and 16 can be replaced with another aromatic residue, either tryptophan or tyrosine. Other possible substitutions or alterations are described in more detail below (see SEQ ID NO: 11). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids can be covalently linked, i.e., "stapled".

[0073] In other non-limiting embodiments, CAP37 includes one or more of the following substitutions. 20-44Alternative variants of (SEQ ID NO: 6) can be used to form the peptide compounds used herein: one or more of the two positively charged arginine residues at positions 4 and 15 of this peptide can be conservatively replaced with lysine or histidine, and one or more of the two histidine residues at positions 5 and 13 can be replaced with arginine or lysine. One or more of the three aromatic phenylalanine residues at positions 6, 16 and 24 can be replaced with other aromatic residues such as tryptophan or tyrosine. One or more of the polar residues asparagine 1, glutamine 2 and 25, threonine 19, and serine 22 can be replaced with any other polar residue (asparagine, glutamine, serine, threonine, or tyrosine). One or more of the other residues in this (hydrophobic) sequence may be substituted with other hydrophobic residues (alanine, glycine, cysteine, valine, proline, leucine, isoleucine, methionine, phenylalanine, tryptophan). Other possible substitutions or alterations are described in more detail below (see Sequence ID No. 12). Each of the amino acids identified above is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog. In certain embodiments, the side chains of two adjacent or non-adjacent amino acids may be covalently linked, i.e., “stapled.”

[0074] Furthermore, peptide compounds are CG as shown in the following sequence (the residues in the bold form represent the amino acids of the wild-type sequence). 119-144 Peptides, NE 119-145 CAP37 120-146 CG 20-47 , NE 20-44 , or CAP37 20-44 It may contain peptides having an amino acid sequence derived from [the source].

[0075] In at least one embodiment, the peptide compound is a peptide CG having the following amino acid sequence (SEQ ID NO: 7) 119-144Includes 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 array, X1 is G, X2 is selected from A, F, G, I, L, M, P, S, T (bold), W, and V. X3 is selected from A, F, G, H, I, K, L (bold), M, N, P, Q, R, V, and W. X4 is selected from A, C (bold), S, T, and V. X5 is selected from A, F, G, I, L, M, P, S, T (bold), W, and V. X6 is selected from A, F, G, I, L, M, P, W, and V (bold type). X7 is selected from A (bold), F, G, I, L, M, P, W, and V. X8 is G, X9 is selected from A, F, G, I, L, M, P, W (bold), and V. X 10 G is, X 11 is selected from A, C, F, G, H, I, K, L, M, N, P, Q, R (bold), S, T, V, W, and Y, or does not exist. X 12 It is selected from A, F, G, I, L, M, P, W, and V (bold), or does not exist. X 13 The letters are selected from A, C, F, G, I, L, M, N, P, Q, S (bold), T, V, W, and Y. X 14The letters are selected from A, F, G, I, L, M (bold), P, W, and V. X 15 The font is selected from H, K, and R (bold type). X 16 The letters are selected from C, H, K, N, Q, R (bold), S, T, and Y. X 17 is either G or does not exist X 18 It is selected from G, C, N, Q, S, T (bold), and Y, or does not exist. X 19 The font is selected from D (bold), E, ​​H, K, and R. X 20 The letters are selected from A, C, F, G, I, L, M, N, P, Q, S, T (bold), V, W, and Y. X 21 The letters are selected from A, C, F, G, I, L (bold), M, N, P, Q, S, T, V, W, and Y. X 22 The letters are selected from A, F, G, H, I, K, L, M, P, R (bold), W, and V. X 23 The font is selected from L, C, D, E (bold), F, G, H, I, K, A, M, N, P, Q, R, PR, S, T, V, W, and Y. X 24 The font is selected from A, F, G, I, L, M, P, W, and V (bold). X 25 The letters are selected from C, N, Q (bold), S, T, and Y, and X 26 The font is selected from A, F, G, I, L (bold), M, P, W, and V.

[0076] Each of the amino acids identified above or below for Sequence ID No. 7 is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog.

[0077] In an alternative embodiment, X of Sequence ID No. 7 17 and / or X 18 and / or X 23 The position may change due to deletions, insertions, or substitutions as shown in Table 1 below.

[0078] [Table 1]

[0079] Position X in Sequence ID 7 shown in Table 1 17 / X 18 / X 23 In an alternative embodiment of optional deletion, insertion, and / or replacement at position X 11 Or X 12 Either of the above, or X 11 and X 12 Amino acids can be deleted in both of these.

[0080] In certain embodiments of the peptide, X 23 If it is PR, X 19 However, examples include those selected from (a) R, H, and K, or (b) R, H, K, D, and E.

[0081] Other specific embodiments of the peptide include X 11 Or X 12 is missing, or X 11 and X 12 If both are missing, X 19 However, examples include those selected from (a) R, H, and K, or (b) R, H, K, D, and E.

[0082] Other specific embodiments of the peptide include X 23 is R, H, or K, and X 11 Or X 12 is missing, or X 11 and X 12 If both are missing, X 19Those selected from (a) R, H, and K, or (b) R, H, K, D, and E, are included.

[0083] In other specific embodiments of the peptide, X 23 When X is PR, X 19 is selected from (a) R, H, and K, or (b) R, H, K, D, and E, and X 11 or X 12 is deleted, or X 11 and X 12 are both deleted.

[0084] In other specific embodiments of the peptide, X 23 is selected from L, C, D, E, F, G, H, I, K, A, M, N, P, Q, R, PR, S, T, V, W, and Y, and when X 23 is PR, X 19 is selected from (a) R, H, and K, or (b) R, H, K, D, and E.

[0085] In other specific embodiments of the peptide, X 23 is selected from L, C, D, E, F, G, H, I, K, A, M, N, P, Q, R, PR, S, T, V, W, and Y, and X 11 or X 12 is deleted, or when X 11 and X 12 are both deleted, X 19 is selected from (a) R, H, and K, or (b) R, H, K, D, and E.

[0086] In at least one embodiment, the peptide compound comprises a derivative of peptide NE 119-145 having the following amino acid sequence (SEQ ID NO: 8): X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X16 X 17 X 18 X 19 X 20 X 21 X 22 X 23 X 24 X 25 X 26 X 27 In the array, X1 is in G (bold type), X2 is selected from A, F, G, I, L, M, P, S, T, W, and V (bold type). X3 is selected from A, F, G, H, I, K, L, M, N, P, Q (bold), R, V, and W. X4 is selected from A, C (bold), S, T, and V. X5 is selected from A, F, G, I, L (bold), M, P, S, T, W, and V. X6 is selected from A (bold), F, G, I, L, M, P, W, and V. X7 is selected from A, F, G, I, L, M (bold), P, W, and V. X8 is in G (bold font), X9 is selected from A, F, G, I, L, M, P, W (bold), and V. X 10 This is G (bold type), X 11 The letters are selected from A, F, G, I, L (bold), M, P, W, and V. X 12 The letters are selected from A, C, F, G, I, L (bold), M, N, P, Q, S, T, V, W, and Y. X 13 The letters are selected from A, F, G (bold), I, L, M, P, W, and V. X 14 The font is selected from H, K, and R (bold type). X 15 The letters are selected from C, H, K, N (bold), Q, R, S, T, and Y. X 16The letters are selected from A, F, G, H, I, K, L, M, P, R (bold), W, and V. X 17 This is G (bold type), X 18 The letters are selected from D, A, E, F, G, I (bold), L, M, P, W, and V. X 19 The font is selected from A (bold), C, F, G, I, L, M, N, P, Q, S, T, V, W, and Y. X 20 The letters are selected from A, C, F, G, I, L, M, N, P, Q, S (bold), T, V, W, and Y. X 21 The fonts are selected from A, F, G, H, I, K, L, M, P, R, W, and V (bold). X 22 The letters are selected from A, C, F, G, I, L (bold), M, N, P, Q, S, T, V, W, and Y. X 23 The letters are selected from C, N, Q (bold), S, T, and Y, and X 24 The letters are selected from A, C, D, E (bold), F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y. X 25 The letters are selected from A, F, G, I, L (bold), M, P, W, and V. X 26 It is selected from C, N (bold), Q, S, T, and Y, and X 27 The font is selected from A, F, G, I, L, M, P, W, and V (bold).

[0087] Each of the amino acids identified above or below for Sequence ID No. 8 is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog.

[0088] In an alternative embodiment, X of Sequence ID No. 816 and / or X 17 The position may change due to deletions, insertions, or substitutions as shown in Table 2 below.

[0089] [Table 2]

[0090] In at least one embodiment, the peptide compound has the following amino acid sequence (SEQ ID 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 array, X1 is in G (bold type), X2 is selected from A, F, G, I, L, M, P, S, T (bold), W, and V. X3 is selected from A, F, G, H, I, K, L, M, N, P, Q, R (bold), V, and W. X4 is selected from A, C (bold), S, T, and V. X5 is selected from A, F, G, I, L (bold), M, N, P, Q (bold), S, T, W, and V. X6 is selected from A, F, G, I, L, M, P, W, and V (bold type). X7 is selected from A (bold), F, G, I, L, M, P, W, and V. X8 is in G (bold font), X9 is selected from A, F, G, I, L, M, P, W (bold), and V, X 10 is G (bold), X 11 is selected from A, C, F, G, I, L, M, N, P, Q, S (bold), T, V, W, and Y, X 12 is selected from W, A, C, F, G, I, L, M, N, P, Q (bold), S, T, V, and Y, X 13 is selected from C, H, K, N, Q, R (bold), S, T, and Y, X 14 is selected from C, H, K, N, Q, R, S (bold), T, and Y, X 15 is G (bold), X 16 is selected from A, F, G (bold), I, L, M, P, S, T, W, and V, X 17 is selected from D, A, E, F, G, I, K, L, M, P, R (bold), S, T, V, W, and Y, X 18 is selected from A, C, F, G, I, L (bold), M, N, P, Q, S, T, V, W, and Y, X 19 is selected from A, C, F, G, I, L, M, N, P, Q, S (bold), T, V, W, and Y, X 20 is selected from A, F, G, H, I, K, L, M, P, R (bold), W, and V, X 21 is selected from A, F (bold), G, I, L, M, P, V, W, and Y, X 22 is selected from A, C, D, E, F, G, H, I, K, L, M, N, P (bold), Q, R, S, T, V, W, and Y, X 23 is selected from A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R (bold), S, T, V, W, and Y, X 24 is selected from A, F (bold), G, I, L, M, P, W, and V, X 25 is selected from A, F, G, I, L, M, P, W, and V (bold), X 26 is selected from C, N (bold), Q, S, T, and Y, and X 27 is selected from A, F, G, I, L, M, P, W, and V (bold).

[0091] Each of the amino acids identified above or below for SEQ ID NO:9 is intended to be read not only as an L-α-amino acid but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptidomonomer analog.

[0092] In an alternative embodiment, the X of SEQ ID NO:9 15 and / or X 16 position may be changed by deletions, insertions, or substitutions as shown in Table 3 below.

[0093]

Table 3

[0094] In at least one embodiment, the peptide compound comprises a derivative of peptide CG 20-47 having the following amino acid sequence (SEQ ID NO:10): 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 25X 26 X 27 X 28 In the array, X1 is selected from F, Y, W, A, V, I (bold), L, and M. X2 is selected from N, S, T, and Q (bold type). X3 is selected from N, S (bold), T, and Q. X4 is selected from P (bold), F, Y, W, A, V, I, L, M, and G. X5 is selected from F, Y, W, A (bold), V, I, L, and M. X6 is selected from G (bold), A, and V. X7 is selected from N, S, T, and Q (bold type). X8 is selected from N, S (bold), T, and Q. X9 is selected from R (bold), H, and K. X 10 It is selected from C (bold), S, and T. X 11 It is selected from G (bold), A, and V, X 12 It is selected from G (bold), A, and V, X 13 The font is selected from F (bold), Y, W, A, V, I, L, and M. X 14 The font is selected from F, Y, W, A, V, I, L (bold), and M. X 15 The letters are selected from F, Y, W, A, V (bold), I, L, and M. X 16 It is selected from R (bold), H, and K. X 17 It is selected from E (bold) and D, X 18 It is selected from E and D (bold type), X 19 The font is selected from F (bold), Y, W, A, V, I, L, and M. X 20 The letters are selected from F, Y, W, A, V (bold), I, L, and M. X 21 The font is selected from F, Y, W, A, V, I, L (bold), and M. X 22 The font is selected from N, S, T (bold), and Q. X 23 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 24 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 25 It is selected from R, H (bold), and K. X 26 It is selected from C (bold), S, and T. X 27 These are selected from F, Y, W (bold), A, V, I, L, and M, and X 28 The letter is selected from G (bold), A, and V.

[0095] Each of the amino acids identified above or below for Sequence ID No. 10 is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog.

[0096] In at least one embodiment, the peptide compound has the following amino acid sequence (SEQ ID 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 X23 X 24 X 25 In the array, X1 is selected from F, Y, W, A, V, I, L (bold), and M, X2 is selected from R (bold), H, and K, X3 is selected from G (bold), A, and V, X4 is selected from G (bold), A, and V, X5 is selected from R, H (bold), and K, X6 is selected from F (bold), Y, W, A, V, I, L, and M, X7 is selected from C (bold), S, and T, X8 is selected from G (bold), A, and V, X9 is selected from F, Y, W, A (bold), V, I, L, and M, X 10 is selected from N, S, T (bold), and Q, X 11 is selected from F, Y, W, A, V, I, L (bold), and M, X 12 is selected from F, Y, W, A, V, I (bold), L, and M, X 13 is selected from F, Y, W, A (bold), V, I, L, and M, X 14 is selected from P (bold), F, Y, W, A, V, I, L, M, and G, X 15 is selected from N (bold), S, T, and Q, X 16 is selected from F (bold), Y, W, A, V, I, L, and M, X 17 is selected from F, Y, W, A, V (bold), I, L, and M, X 18 is selected from F, Y, W, A, V, I, L, and M (bold), X 19The font is selected from N, S (bold), T, and Q. X 20 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 21 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 22 It is selected from R, H (bold), and K. X 23 It is selected from C (bold), S, and T. X 24 These are selected from F, Y, W, A, V (bold), I, L, and M, and X 25 The letter is selected from F, Y, W, A (bold), V, I, L, and M.

[0097] Each of the amino acids identified above or below for Sequence ID No. 11 is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog.

[0098] In at least one embodiment, the peptide compound has the following amino acid sequence (SEQ ID 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 array, X1 is selected from N (bold), S, T, and Q. X2 is selected from N, S, T, and Q (bold type). X3 is selected from G (bold), A, and V. X4 is selected from R (bold), H, and K. X5 is selected from R, H (bold), and K. X6 is selected from F (bold), Y, W, A, V, I, L, and M. X7 is selected from C (bold), S, and T. X8 is selected from G (bold), A, and V. X9 is selected from G (bold), A, and V. X 10 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 11 The font is selected from F, Y, W, A, V, I, L (bold), and M. X 12 The letters are selected from F, Y, W, A, V, I (bold), L, and M. X 13 It is selected from R, H (bold), and K. X 14 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 15 It is selected from R (bold), H, and K. X 16 The font is selected from F (bold), Y, W, A, V, I, L, and M. X 17 The letters are selected from F, Y, W, A, V (bold), I, L, and M. X 18 The font is selected from F, Y, W, A, V, I, L, and M (bold). X 19 The font is selected from N, S, T (bold), and Q. X 20 The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 21The letters are selected from F, Y, W, A (bold), V, I, L, and M. X 22 The font is selected from N, S (bold), T, and Q. X 23 It is selected from C (bold), S, and T. X 24 The font is selected from F (bold), Y, W, A, V, I, L, and M, and X 25 The font is selected from N, S, T, and Q (bold).

[0099] Each of the amino acids identified above or below for Sequence ID No. 12 is intended to be read not only as an L-α-amino acid, but also, alternatively, as its D-amino acid, β-amino acid, homoamino acid, β-homoamino acid, or peptoid monomer analog.

[0100] Peptide compounds may be administered alone or in combination with other drug therapies, and may be administered via various routes of administration. Generally speaking, the methods of administration may be carried out using any medically acceptable mode of administration (meaning any mode that produces an effective level of the active compound without causing clinically unacceptable adverse effects). Administration may be, for example, oral, intraperitoneal, (non-limited) intracavitary, transdermal, topical, skin, nasal, inhalation, mucosal, intradermal, intraperitoneal, or parenteral routes. The term “parenteral” includes subcutaneous, intravenous, intramuscular, or infusion. In certain embodiments, it may be appropriate to administer the compound by continuous infusion, for example, every few days, or once a week, every few weeks, or once a month or every other month. Intravenous or intramuscular routes may be used, in particular, in emergencies. Oral or intranasal administration may be used for prophylactic treatment for the convenience of the patient and the medication plan. Similarly, sustained-release devices may be useful in certain embodiments, for example, for prophylactic or postoperative treatment.

[0101] In certain embodiments, the peptide compound is administered intranasally or intravenously.

[0102] Currently, specific pharmaceutical formulations of the disclosed peptide compositions include, but are not limited to, sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Non-limited examples of non-aqueous solvents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil (non-limited), and injectable organic esters such as ethyl oleate (non-limited). Aqueous carriers include water, alcoholic / aqueous solutions, emulsions, or suspensions, such as saline and buffering media. Parenteral vehicles include, for example, sodium chloride solution, glucose-added Ringer's solution, glucose and sodium chloride, lactated Ringer's solution, or fixative oils. Intravenous vehicles include fluid nutritional fluids, electrolyte fluids (such as those based on glucose-added Ringer's solution), and others of the same kind. Preservatives and other additives may also be present, for example (but not limited to), antimicrobial agents, antioxidants, chelating compounds, inert gases, and others of the same kind. Pharmaceutical compositions may conveniently be provided in unit dosage forms and may be prepared by any method well known in the field of pharmacy.

[0103] Compositions suitable for oral administration may (not limited to) include individual units such as capsules, pills, tablets, lozenges, melts, powders, suspensions, solutions, elixirs, or emulsions, each containing a predetermined amount of the peptide composition. Other compositions include (not limited to) suspensions in aqueous or non-aqueous liquids such as syrups, elixirs, or emulsions. In yet another embodiment, a particular vehicle is a biocompatible microparticle or implantable tablet suitable for implantation in a mammalian recipient.

[0104] Other embodiments of the peptide composition include pharmaceutically acceptable compositions comprising one or more therapeutically effective amounts of the compounds described herein, formulated with one or more pharmaceutically acceptable carriers (additives) and / or diluents. As described in detail herein, pharmaceutically acceptable compositions may be specifically formulated for administration in solid or liquid form, including, but not limited to, those adapted for: oral administration, e.g., aqueous or non-aqueous solutions or suspensions, tablets, e.g., targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., sterile solutions or suspensions, or sustained-release formulations; topical application, e.g., creams, ointments, or controlled-release patches or sprays applied to the skin; vaginal or rectal administration, e.g., creams or foams; sublingual administration; ocular administration; transdermal administration; or nasal administration.

[0105] Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, the following: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose, and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; water free of pyrogens; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyacid anhydrides; and other non-toxic, suitable substances used in pharmaceutical formulations.

[0106] In solid dosage forms (capsules, tablets, pills, powders, granules, and other similar forms) of peptide compositions for oral administration, the compound may be mixed with one or more pharmaceutically acceptable carriers, the carriers of which include, but are not limited to, sodium citrate or calcium hydrogen phosphate, and / or any of the following: fillers or bulking agents such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia gum; humectants such as glycerol; disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; dissolution retarders such as paraffin; absorption enhancers such as quaternary ammonium compounds; and wetting agents such as cetyl alcohol, glycerol monostearate, and nonionic surfactants. Agents; absorbents such as kaolin and bentonite clay; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; and colorants. In the case of capsules, tablets, and pills, the pharmaceutical composition may also contain buffers. Similar types of solid compositions may also be used as fillers in soft gelatin capsules and hard gelatin capsules, with excipients such as lactose or milk sugar, plus high molecular weight polyethylene glycol and other similar substances.

[0107] Liquid dosage forms for oral or nasal administration of peptide compounds include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may contain (but not limited to) inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, as well as mixtures thereof. In addition to inert diluents, the oral composition may also contain (non-limited) auxiliary agents such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, colorants, fragrances, and preservatives. In addition to the active compound, the suspension agent may contain, for example, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum hydroxide (aluminum metahydroxide), bentonite, agar, and tragacanth, as well as mixtures thereof.

[0108] Generally, the therapeutically effective dose of the peptides disclosed herein varies depending on the age, condition, and sex of the subject, as well as the nature and severity of the disease in the subject (all of which can be determined by those skilled in the art). The dosage may be adjusted by the individual physician or veterinarian, especially if any complications occur. The therapeutically effective dose is typically, but not limited to, amounts ranging from 0.1 μg / kg to about 2000 mg / kg, or from 1.0 μg / kg to about 1000 mg / kg, or from about 0.1 mg / kg to about 500 mg / kg, or from about 1.0 mg / kg to about 100 mg / kg, administered once or multiple times daily for more than one day. If necessary, the effective daily dose of the active compound may be administered as two, three, four, five, six, or more sub-doses, for example, separately at appropriate intervals throughout the day, optionally in unit dosage forms. In some embodiments, the composition is administered for longer than 7 days, longer than 10 days, longer than 14 days, or longer than 20 days. In other embodiments, the peptide is administered over several weeks or months. In other embodiments, the peptide is delivered every other day, for example, the active ingredient may be delivered every 2 days, or every 3 days, or every 4 days, or every 5 days, or every 6 days, or weekly, or monthly.

[0109] In certain non-limiting embodiments, an effective dose or therapeutic dose of the peptide compound of the Disclosure contains enough active agent to deliver about 0.001 μg / kg to about 100 mg / kg (weight of active agent / body weight of subject). For example, a composition delivers about 0.01 μg / kg to about 50 mg / kg, more particularly about 0.1 μg / kg to about 10 mg / kg, and more particularly about 1 μg / kg to about 1 mg / kg. Practice of the methods of the Disclosure may involve administering an effective dose of the peptide compound in any suitable systemic and / or topical formulation to a subject in an amount effective to deliver a therapeutic dose of the active agent. In certain embodiments, the effective dose may be in the range of about 1 μg / kg to about 1 mg / kg of the active agent.

[0110] The practice of the methods of this disclosure may involve administering a therapeutically effective dose of the peptide compound in any suitable systemic and / or topical formulation in an amount effective to deliver the doses listed above. The dose may be administered as a single dose, multiple times (for example, 1 to 5 times per day, or once or twice per week), or continuously by intravenous infusion, depending on the desired therapeutic effect, for example. In one non-limiting example of the therapeutic methods of this disclosure, the variant peptide compound is delivered once daily by IV infusion in the range of approximately 0.01 mg / kg body weight to approximately 10 mg / kg body weight.

[0111] Oral formulations can be formulated such that the peptide compound passes through the digestive system before being released, for example, so that it cannot be released until it reaches the small intestine or colon.

[0112] The solid unit dosage form may be a conventional gelatin capsule containing, for example, a surfactant, a lubricant, and an inert filler such as lactose, sucrose, and corn starch, or the dosage form may be a sustained-release preparation. The pharmaceutical composition may contain a solid carrier such as gelatin or an adjuvant. Tablets, capsules, and powders may contain about 0.05% to about 95% of the active substance compound by dry weight. When administered in liquid form, a liquid carrier such as water, petroleum, peanut oil, mineral oil, soybean oil, or animal or plant-derived oils such as sesame oil, or synthetic oils may be added. The pharmaceutical composition in liquid form may further contain physiological saline, dextrose or other sugar solutions, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains about 0.005% to about 95% by weight of the active substance. For example, a dose of approximately 10 mg to 1000 mg may be administered orally once or twice a day.

[0113] In another embodiment, the peptide compounds of this disclosure can be tableted on conventional tablet bases such as lactose, sucrose, and corn starch, in combination with a binder such as acacia gum, corn starch, or gelatin, a disintegrant such as potato starch or alginic acid, and a lubricant such as stearic acid or magnesium stearate. Liquid preparations are prepared by dissolving the peptide compounds in an aqueous or non-aqueous pharmaceutically acceptable solvent, which may also contain suspending agents, sweeteners, flavoring agents, and preservatives, as is known in the art.

[0114] For parenteral administration, for example, peptide compounds may be dissolved in a physiologically acceptable pharmaceutical carrier and administered as either a solution or a suspension. Examples of suitable pharmaceutical carriers include water, saline solution, dextrose solution, fructose solution, ethanol, or oils of animal, plant, or synthetic origin. Pharmaceutical carriers may also contain preservatives and buffers, as is known in the art.

[0115] When an effective amount of a peptide compound is administered intravenously, cutaneously, or subcutaneously, the compound is, in particular, in the form of a parenterally acceptable aqueous solution or suspension that does not contain pyrogens. The preparation of such parenterally acceptable solutions, taking into account pH, isotonicity, stability, etc., is well within the skill of those skilled in the art. Specific pharmaceutical compositions for intravenous, cutaneous, or subcutaneous injection may contain, in addition to the peptide compound, an isotonic vehicle such as sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, lactated Ringer's injection, or other vehicles known in the art. The pharmaceutical compositions of this disclosure may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those skilled in the art.

[0116] As stated above, the specific amount and mode of administration can be determined by those skilled in the art. Those skilled in the art who prepare formulations can, for example, refer to Remington: The Science and Practice of Pharmacy, 22nd Using the formulation technologies known in the art as described in the ed., the appropriate form and mode of administration can be easily selected depending on the specific properties of the selected peptide compound, the condition to be treated, the stage of the condition, and other relevant circumstances.

[0117] Additional pharmaceutical methods may be used to control the duration of action of peptide compounds. Increased half-life and / or sustained-release preparations can be achieved through the use of proteins or polymers that conjugate, complex with, and / or absorb the peptide compound, as previously discussed herein. Controlled delivery and / or increased half-life can be achieved by selecting appropriate polymers (but not limited to, polysaccharides, polyesters, polyamino acids, homopolymers, polyvinylpyrrolidone, ethylene vinyl acetate, methylcellulose or carboxymethylcellulose, and acrylamides such as N-(2-hydroxypropyl)methacrylamide) for controlling release, as well as appropriate concentrations of the polymers and methods of incorporation.

[0118] Another possible method useful in controlling the duration of action of peptide compounds by sustained-release preparations and half-lives is the incorporation of peptide compounds or their functional derivatives into particles of polymer materials such as polyesters, polyamides, polyamino acids, hydrogels, poly(lactic acid), ethylene vinyl acetate copolymers, and, for example, copolymer micelles of polyethylene glycol (PEG) and poly(l-aspartamide).

[0119] It is also possible to capture peptide compounds within microcapsules (e.g., hydroxymethylcellulose or gelatin microcapsules, and poly(methyl methacrylate) microcapsules, respectively) prepared by coacervation technology or interfacial polymerization, within colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or within macroemulsions. Such techniques are well known to those skilled in the art.

[0120] When peptide compounds are intended for use as injection materials, they can be formulated into conventional injection carriers. Suitable carriers include biocompatible and pharmaceutically acceptable phosphate-buffered saline, particularly isotonic.

[0121] For the reconstruction of lyophilized products according to this disclosure, sterile diluents may be used that may contain materials as generally recognized to approximate physiological conditions and / or as required by government regulations. In this regard, sterile diluents may contain buffers to obtain a physiologically acceptable pH, such as sodium chloride, saline, phosphate-buffered saline, and / or other physiologically acceptable and / or safe for use. Generally, materials for intravenous injection in humans should comply with regulations established by the U.S. Food and Drug Administration, which are available to those skilled in the art. Pharmaceutical compositions may also be in the form of aqueous solutions containing many of the same substances as described above for the reconstruction of lyophilized products.

[0122] Peptide compounds can also be administered as pharmaceutically acceptable acids or base addition salts formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with inorganic bases such as sodium hydroxide, ammonium hydroxide, and potassium hydroxide, as well as organic bases such as mono-, di-, tri-alkyl and allylamines, and substituted ethanolamines.

[0123] In a particular embodiment, the disclosure includes a peptide compound composition comprising a peptide compound, wherein at least one peptide compound is directly or indirectly coupled to a carrier molecule, the at least one peptide of the peptide compound having a length of 20 to 50 amino acids.

[0124] In at least certain embodiments, the Disclosure relates to the use of full-length CAP37, NE, and CG proteins and / or peptide fragments thereof, and / or derivatives of such peptides, alone, in combination, or as peptide components of peptide compounds containing a non-peptide portion (e.g., a PEG molecule), for the inhibition of the pro-inflammatory and neurotoxic effects of Aβ in Alzheimer's disease, cerebral amyloid angiopathy, and other neuroinflammatory conditions associated with Aβ. In non-limiting examples, peptide compounds of the Disclosure include peptides having the amino acid sequences disclosed above, SEQ ID NOs: 1-12.

[0125] Examples of CG-based, NE-based, and CAP37-based variant peptides that can be used in various embodiments of the peptide compounds of this disclosure are, but are not limited to, those shown in Table 4.

[0126] [Table 4-1]

[0127] [Table 4-2]

[0128] [Table 4-3]

[0129] As further details below, the peptides of this disclosure may be modified by the addition of R, K, and / or H moieties (e.g., 1 to 10 amino acids linked to the N-terminal and / or C-terminal portions of the peptide) and “mini-PEG” moieties (AEEA, AEEEA, AEEP, and AEEEP) linked to the N-terminal and / or C-terminal portions of tagged or untagged peptide sequences. Examples of such mini-PEG moieties are shown, for example, in International Publication No. 2014 / 089088 and Brochure No. 2014 / 172747, and in U.S. Patent No. 9,624,283 and Specification No. 9,862,748.

[0130] In a particular embodiment, the peptide compound of this disclosure is represented by formula (I): SX R -Pep-Y R (I) In the formula, Pep is an amino acid sequence disclosed or described herein (e.g., SEQ ID NOs: 1-46); X R and Y R These are, independently, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 R, K, or H residues, for example, X R +Y R = up to 10; and S is a solubilized portion made up of any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (i.e., 1 to 10) short PEG subunits, such as AEEA, AEEP, AEEEA, and AEEEP as defined elsewhere herein.

[0131] Peptide (XR -Pep-Y R -O(CH2) of the PEG subunit (S) attached to ) q The CO- portion ("head group") is the (CH2) of the head group. q The part may be further modified to include several alternative embodiments. These alternative embodiments, as well as those described above, may be characterized by formula (II): -O(CR1R2) q CO- (II) In formula II, R1 and R2 are side-chain groups on C (i.e., "R1-C-R2"), q=0 to 4, and R1 and R2 are selected from H, CH3, and CH2CH3. When q=1, one or both of R1 and R2 may be selected from H, CH3, and CH2CH3. 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, any of the three R1 groups and any of the three R2 groups may be selected from H, CH3, and CH2CH3. When q=4, any of the four R1 groups and any of the four R2 groups may be selected from H, CH3, and CH2CH3. Thus, in these non-limiting embodiments, the head group may include one or more substitutions for H on the carbon of the carbon backbone when C=1 to 4. Examples of embodiments of the head group having formula II include, but are not limited to, the following: 1. -OCH2CH2CO- (for example, as in AEEP) 2. -OCH2CO- (for example, 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 R1 and R2=CH3 on the second carbon, and q=2) 7.-OCH2CH2CH2CH(CH3)CO-(R1=H and R2=CH3 on one carbon, and R1 and R2=H on all other carbons, and q=4) 8.-OCH2CH2CH2C(CH3)2CO- (R1 and R2 on one carbon are CH3, and R1 and R2 on all other carbons are H) 9.-OCH2CH2CH2CH(CH2CH3)CO-(R1=H and R2=CH2CH3 on one carbon, and R1 and R2=H on all other carbons) 10.-OCH(CH3)CH2CH2CH(CH2CH3)CO-(R1=H and R2=CH3 on one carbon, R1=H and R2=CH2CH3 on one carbon, and R1 and R2=H on all other carbons)

[0132] All stereoisomers of peptide compounds formed based on the solubilization units described for Formula II are intended to be included in this disclosure.

[0133] [Examples] method Binding assay The binding of Aβ to CAP37, CAP37-derived peptide, NE, CG, and BSA (used as a negative control) was quantified using enzyme-linked immunosorbent assay (ELISA). CAP37, NE, and CG were purified from human neutrophils and purchased from Athens Research and Technology. BSA was from Sigma Aldrich. CAP37-derived peptide was synthesized by CS Bio Co with a purity of ≥95%. Nunc Maxisorp plates were coated with 100 μl of neutrophil protein, peptide, or BSA diluted at 5 μg / ml in PBS containing or without 1 mM PMSF (for experiments involving CAP37, NE, and BSA) purchased from Sigma Aldrich, or ±125 μM chymostatin (for experiments involving CG and BSA) purchased from Santa Cruz. Plate coating was performed at room temperature for 2 hours with shaking, followed by overnight at 4°C. The plate was washed three times in PBST (PBS + 0.05% tween), and then blocked at room temperature for 1 hour with shaking in 3% BSA in PBST. The plate was washed three times in PBST and 150 nM Aβ 1-42The amyloid precursor protein (purchased from Bachem and reconstituted in 1 mM NaOH) was added to 0.1% BSA in PBST and incubated at 37°C for 70 minutes. The plate was then washed four times with PBST, and monoclonal mouse anti-amyloid precursor protein / amyloid beta antibody purchased from Cell Signaling Technology was added to 1% BSA in PBST at a dilution of 1:500 and incubated at room temperature for 1 hour with shaking. The plate was then washed four times with PBST, and donkey anti-mouse HRP conjugate secondary antibody from Jackson ImmunoResearch Laboratories was added to PBST at a dilution of 1:10,000 and incubated at room temperature for 1 hour with shaking, and then washed four times with PBST. A substrate buffer (49 mM citrate, 102 mM sodium phosphate, pH adjusted to 5.0 with HCl, with freshly added 7.5 mM Kodak substrate o-phenylenediamine and 0.024% final H2O2) was added to the plate and incubated in the dark for 30 minutes. The reaction was stopped by adding sulfuric acid (5N) to each well, and the absorbance at 492 nm was read using a Biotek Synergy II plate reader.

[0134] Amyloid-beta (Aβ) fibrosis assay I purchased the Amyloid Beta (1-42) Thioflavin T Aggregation Kit from Anaspec and used it according to the manufacturer's instructions for use, with some modifications. 1-42 The formation of peptide fibrils was monitored. Thioflavone T dye was diluted to 250 μM in fibrillation buffer A and kept in the dark until use. Aβ 1-42The peptide was suspended in Buffer A at 250 μg / ml (or 55 μM), incubated on ice for 5 minutes, mixed by inverting the tube multiple times, sonicated in a water bath sonicator at 4°C for 5 minutes, mixed again, and centrifuged at 10,000 xg for 5 minutes. The supernatant was transferred to a new tube and stored at 4°C until use. The aggregation inhibitor, CAP37, CAP37-derived peptide, NE, CG, and BSA (used as a negative control) were diluted in water at 20 × the desired final concentration. Under low light, 10 μl of thioflavin T dye was added to a Corning low-protein-binding black plate, and 85 μl of Aβ prepared in Buffer A was added. 1-42 5 μl of water or an agglutination inhibitor to be tested was also added. Neutrophil proteins were tested in the absence and presence of the protease inhibitor. PMSF was dissolved in 100% ethanol and used at a final concentration of 1 mM in the reactions for CAP37 and NE. Chymostatin was dissolved in DMSO and used at a final concentration of 125 μM in the reaction for CG. To ensure mixing, the reactants were pipetted up and down twice in the well using a multichannel pipette, and fluorescence (λ) was detected. Ex =440nm, λ Em The wavelength (485nm) was read every 5 minutes for 3 hours using a BioTek Synergy 2 plate reader with Gen 5 software, at a sensitivity set to 65.

[0135] Enzyme activity assay Aβ treated with hexafluoroisopropanol 1-42 The enzyme was purchased from Bachem and reconstituted in anhydrous DMSO at 5 mM. The reaction was carried out using an increasing concentration of the enzyme (CAP37, NE, or CG) and 10 μM Aβ in PBS. 1-42 The reaction was set with the substrate and incubated at 37°C for 1 hour. The reaction was stopped by adding acetonitrile and trifluoroacetic acid (TFA) to final concentrations of 7.5% and 0.5%, respectively. An internal standard (Bachem Aβ) dissolved in 10% acetic acid was used. 40-1The solution was added to achieve a final concentration of 8 μg / ml. A 10 μl aliquot of this mixture was desalted using an Agilent Technologies C18 OMIX tip. First, the C18 resin was rinsed twice with 10 μl of 50% methanol, then twice with 10 μl of equilibrium buffer (5% acetonitrile, 0.5% TFA). The sample was then applied to the resin by pipetting through it 10 times, washed twice with equilibrium buffer, and eluted with 75% acetonitrile, 0.1% TFA.

[0136] Desalted samples were analyzed by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MARLDI-TOF) using a Bruker Ultraflex II instrument and Flex Control Software. The sample was spotted onto a steel plate with 1 μl of α-cyano-4-hydroxycinnamic acid matrix in 30% acetonitrile 0.1% TFA. Each sample was spotted in a triple configuration. The plates were dried in a vacuum desiccator. Each chromatograph was the average of 5 × 200 shots at 28% laser power in linear positive mode. The chromatographs were analyzed using Flex analysis software. The chromatographs were for Aβ 40-1 The peak around 4,330 MHz, corresponding to the internal standard, and Aβ 1-42 It had a peak around 4,515 m / z corresponding to the substrate. Aβ divided by the internal standard peak intensity. 1-42 The ratio of peak intensities was calculated for each spot, and the amount of uncleaved Aβ in each enzyme reaction was determined by averaging the results across the three consecutive spots.

[0137] Cell culture and neurotoxicity assays The mouse neuroblastoma cell line Neuro-2a from ATCC was used in these experiments. The cells were grown and seeded in 96-well plates at a rate of 20,000 cells per well in complete medium (Gibco MEMα, nucleoside, and phenol red-free, supplemented with 10% heat-inactivated fetal bovine serum from Gibco and penicillin / streptomycin from Lonza). After 24 hours post-seeding, the cells were washed with warmed MEMα and then 90 μl of MEMα + N2 supplement + 20 μl of test sample (Aβ prepared as described below). 1-42 The cells were incubated in ) and then removed by vigorously pipetting up and down. 10 μl of cells were added to 10 μl of BioRad trypan blue dye (0.4% solution), mixed, and counted with a BioRad TC20 cell counter. The percentage of live and dead cells was recorded for each well. For water and Triton X100 controls, 20 μl of MEMα+N2 supplement was added instead of the test sample, and immediately before counting the cells, 20 μl of medium was removed and replaced with 20 μl of water or Triton X100 at a final concentration of 0.1%.

[0138] Preparation of test samples for Aβ, monomers, protofibrils, and oligomers. Bachem monomer Aβ 1-42 (Hexafluoroisopropanol-treated Aβ 1-42 ) was reconstituted in 5 mM anhydrous DMSO. Untreated Aβ from CS Bio 1-42 It was also used in these experiments. First, it was treated with 1 mM hexafluoroisopropanol from Sigma Aldrich, incubated at room temperature for 30 minutes, and dried overnight in a fume hood. Then, it was vacuum-dried at room temperature for 1 hour. Aβ 1-42 The dried film was stored at -20°C until use to retain it in monomer form. Aβ from CS Bio. 1-42 Furthermore, it was reconstituted in 5 mM anhydrous DMSO before use.

[0139] Aβ 1-42 The toxicity of the monomer was determined by a newly prepared Aβ 1-42 The test was performed by diluting (5 mM in DMSO) to a final concentration of 18 μM in MEMα+N2 supplement. Using this mixture, the complete growth medium was changed in cells 24 hours after seeding, and Aβ 1-42 The toxicity of the monomer was measured after a 24-hour incubation.

[0140] To prepare test samples of profibrils and oligomers, monomer Aβ 1-42 The material was placed under conditions favorable to the formation of either profibrils or oligomers. To form profibrils, 100 μM Aβ was added. 1-42 The proteins were added to Anaspec Fibrosis Buffer A ± Neutrophil Protein and incubated at 37°C for 24 hours, after which they were added to cells at a final concentration of 18 μM. Neutrophil proteins were used at the Aβ / protein ratio that yielded the previously determined 50% inhibition of fibrillation (IC50) for each protein in the fibrillation assay. To form oligomers, MEMα+N2 supplements were used instead of Fibrosis Buffer A ± increasing concentrations of neutrophil protein or CAP37-derived peptides. Each mixture was then added to freshly prepared Aβ at a final concentration of 100 μM in DMSO. 1-42 The sample was added to a tube containing the substance and incubated at room temperature for 15 minutes. These test samples were then tested on cells at a final concentration of 18 μM.

[0141] Amyloid beta (Aβ) 1-42 Quantification of oligomer formation Oligomers were formed in the presence and absence of increasing concentrations of the peptide inhibitor. MEMα was first mixed with increasing concentrations of the peptide inhibitor, and then with freshly prepared Aβ. 1-42 The monomer was added to a final concentration of 100 μM. These preparations were incubated at room temperature for 15 minutes, and Aβ 1-42The oligomers are formed, then transferred to ice, and 2 × 10⁻¹⁶ units are used to ensure that the Aβ concentration falls within the linear detection range of the Biosensis (Thebarton, Australia) oligomer amyloid-β (o-Aβ) ELISA kit. 5 The solution was diluted to twice its original concentration. ELISA quantification was performed in two separate lines, as recommended by the manufacturer, and the absorbance was read at 450 nm.

[0142] result Figure 1 shows Aβ 1-42 This shows the binding of Aβ to the full-length CAP37, NE, and CG proteins. The binding is stronger with respect to CAP37 and CG and is not significantly altered by the addition of antiproteases (phenylmethylsulfonyl fluoride (PMSF) or chymostatin). 1-42 The binding of to NE is almost undetectable, but the binding of Aβ by NE is 1-42 In the presence of PMSF, which inhibits cleavage, the increase is significant. These results indicate stronger Aβ to CAP37 and CG. 1-42 Figure 2 shows the binding of Aβ to peptides 20-44 (20-44 CC, 95-122, and 120-146 QR) from wild-type CAP37 and peptides from mutant CAP37 (20-44 CS, 20-44 SC, 120-146 WR, 120-146 QH, and 120-146 WH). 1-42 This indicates the level of binding. Aβ 1-42 It did not bind to peptides 95-122. Figure 3 shows the Aβ binding to the "5RMP" modified CAP37-derived peptide from Figure 2 (two AEEA miniPEG molecules and four consecutive arginine residues linked to the N-terminus of the peptide, and one arginine linked to the C-terminus of the peptide). 1-42 This shows the level of binding. These results are Aβ 1-42 This shows that, regardless of the presence or absence of "5RMP" modification, peptides 20-44 and 120-146 from the wild-type and mutant CAP37 bind, but do not bind to peptide 95-122, regardless of the presence or absence of "5RMP" modification.

[0143] Figures 4-7, 10, and 13 show the relative fluorescence units (Rfu) recorded for Aβ in the absence and presence of three neutrophil granule proteins and specific wild-type and mutant CAP37 peptides at elevated concentrations. 1-42 This shows fibrosis. Figure 4 shows Aβ in the presence of full-length CAP37, NE, or CG at the indicated concentrations. 1-42 This indicates the level of fibrillation (aggregation) of Aβ (when no protein is added). 1-42 The time to complete fibrillation is indicated by the vertical dotted line. Fibrillation is inhibited either partially or completely in the presence of these proteins. Figure 5 shows the full length of Aβ in the presence of CAP37, NE, or CG at various concentrations of the proteins. 1-42 This indicates the level of fibrosis (aggregation). NE is Aβ 1-42 The inhibitory effect on fibrosis is highest with CG, followed by CAP37. Figure 5 shows the dose response for each full-length protein. The percentage of fibrosis shown in Figure 5 corresponds to Aβ as shown by the dotted line in Figure 4. 1-42 These were calculated at the time point corresponding to complete fibrosis of the control. These dose-response experiments showed that NE was Aβ 1-42 Highest risk of fibrosis (1 / 4,167 NE / Aβ) 1-42 The study shows that CG (with an IC50 of 2.4 nM corresponding to the molar ratio of 1 / 2,632) and CAP37 (with an IC50 of 876 nM; with a molar ratio of 1 / 11) have inhibitory effects.

[0144] Figure 6 shows that the protease inhibitor PMSF inhibits CAP37 Aβ 1-42 This indicates that the inhibitory effect on fibrosis is impaired. The IC50 of CAP37 is increased 3.5 times in the presence of PMSF. This suggests that the inhibitory effect of CAP37 on fibrosis is impaired, at least partially, by Aβ 1-42 This indicates that it is due to cleavage of NE Aβ. Figure 7 shows that the protease inhibitor PMSF has an effect on NE Aβ 1-42This indicates that the inhibitory effect on fibrosis is impaired. The IC50 of NE is increased 36-fold in the presence of PMSF. This suggests that the inhibitory effect of NE on fibrosis is impaired, at least partially by Aβ 1-42 This indicates that it is due to cleavage of the three neutrophil granule proteins, Aβ 1-42 It has been previously found that it can be cut, and therefore, in Figures 6, 7, and 10, Aβ 1-42 To determine whether cleavage of Aβ is necessary to inhibit fibrosis, CAP37, NE, and CG were tested in the absence and presence of sufficient protease inhibitors (PMSF for CAP37 and NE, and chymostatin for CG). In all three experiments, Aβ 1-42 Inhibition of the enzymatic cleavage of Aβ also impaired the inhibitory effect of these proteins on fibrillation. This is because the inhibitory effect on fibrillation was at least partially impaired. 1-42 This indicates that the cleavage is due to Aβ. Figure 13 shows the dose-response observed with increasing concentrations of CAP37-derived peptides 95-122 5RMP, 120-146 WR 5RMP, and 120-146 WR. Peptides 95-122 5RMP are Aβ 1-42 They did not interact with Aβ1-42 (Figure 3) and did not inhibit fibrosis. Both peptides 120-146 WR 5RMP and 120-146 WR interacted with Aβ1-42 (Figures 2 and 3) and were able to inhibit fibrosis. The inhibitory effect of these CAP37-derived peptides is due to the peptides interacting with Aβ 1-42 Despite lacking enzymatic activity against Aβ, it was similar to that of full-length CAP37 (IC50 of approximately 1000 nM; molar ratio of approximately 1 / 10). While I don't intend to be bound by theory, this suggests that Aβ 1-42 The cleavage of Aβ 1-42 This indicates that Aβ is not the only mechanism for inhibiting fibrosis. 1-42 Direct binding to it also appears to play a role in inhibiting fibrosis.

[0145] Figures 8, 9, 11, and 12 show the percentage of fibrosis and the corresponding Aβ on the right y-axis in experiments performed with increased concentrations of full-length neutrophil granule protein in the absence and presence of protease inhibitors. 1-42 This shows the percentage cleavage of Aβ by NE. Figure 8 shows Aβ by NE. 1-42 This shows a direct correlation between cleavage and inhibition of fibrosis. This correlation supports the idea that NE inhibition of fibrosis is primarily mediated by Aβ cleavage. This experiment was conducted in the absence of PMSF. Figure 9 shows Aβ cleavage by NE. 1-42 This shows a direct correlation between cleavage and inhibition of fibrosis. This experiment was performed in the presence of PMSF. Figure 11 shows Aβ induced by CG. 1-42 This shows a direct correlation between cleavage and inhibition of fibrosis. This correlation supports the idea that the inhibition of CG-mediated fibrosis is primarily mediated by Aβ cleavage. This experiment was conducted in the absence of chymostatin. Figure 12 shows Aβ cleavage by CG. 1-42 This shows a direct correlation between cleavage and inhibition of fibrosis. This experiment was performed in the presence of chymostatin. These results establish a direct correlation between cleavage and inhibition of fibrosis, and Aβ 1-42 This supports the idea that cleavage of this protein is the main mechanism explaining the inhibition of full-length protein-mediated fibrillation.

[0146] Figure 14 shows various higher-order structures of Aβ in neurons. 1-42 It exhibits toxicity. Aβ 1-42 Conditions favorable to the higher-order structure of the monomer, oligomer, or protofibril were used. Figure 14 shows Aβ 1-42 The oligomer has been shown to be more toxic to neurons than the fibril. 1-42 The monomeric form is the least toxic. These results confirm findings previously published by other groups, which indicated that the oligomer is the most toxic, followed by the protofiber, and then the monomer.

[0147] Figure 15 shows that when incubated under conditions favorable to fibril formation, NE and CG are Aβ 1-42Figure 15 shows that it inhibits the toxicity of [protein Aβ], but not CAP37. Figure 15 shows the Aβ formed in the absence or in the presence of neutrophil granule protein, used at concentrations (IC50) previously determined for each protein in Figure 5 that inhibit approximately 50% of fibrosis. 1-42 This demonstrates the toxicity of the fibril. This experiment shows significant inhibition of fibril toxicity by NE and CG. CAP37 exhibits Aβ under these conditions. 1-42 It appears to have no effect against toxicity. In experiments with fibrils, all three neutrophil granule proteins were treated with Aβ for 24 hours before being added to cells. 1-42 Aβ is pre-incubated during this 24-hour fibrosis step. 1-42 Percentage cleavage is likely to be higher than 50%. For NE and CG, cleavage is important for inhibiting fibrillary toxicity. In contrast, Aβ by CAP37 1-42 The cleavage of Aβ is involved in inhibiting fibrosis, as determined in Figure 6, but when pre-incubated under fibrotic conditions, 1-42 It is not possible to inhibit the toxicity of Aβ. This is because NE and CG do not inhibit the toxicity of Aβ. 1-42 This can be explained by the fact that it has more cleavage sites than CAP37 and produces smaller products. Therefore, the larger products produced by CAP37 cleavage may still be able to form toxic aggregates under fibrous conditions.

[0148] Figures 16-20 show the increased concentrations of Aβ formed in the absence or presence of three neutrophil granule proteins and two CAP37-derived peptides found to inhibit fibrosis in Figure 13. 1-42 This shows the toxicity of the oligomer. Figure 16 shows Aβ in the presence of various concentrations of NE. 1-42 This shows the effect of oligomers on neuronal cells. NE reacts in a dose-dependent manner with Aβ 1-42 The oligomer toxicity was significantly inhibited. NE was used at 0.5, 1, 2, and 4 times the IC50 determined in Figure 5. The results show significant inhibition of oligomer toxicity by NE, starting from 2 × IC50.

[0149] Figure 17 shows Aβ in the presence of various concentrations of CG. 1-42 This shows the effect of oligomers on neuronal cells. CG is dose-dependent, and Aβ 1-42 The oligomer toxicity was significantly inhibited. The same conditions as in Figure 16 were used for CG in Figure 17, showing stronger inhibition of oligomer toxicity by CG, starting from 0.5 × IC50.

[0150] Figure 18 shows Aβ in the presence of various concentrations of CAP37 protein. 1-42 This study demonstrates the effects of oligomers on neuronal cells. Under the conditions used, full-length CAP37 showed effects on Aβ 1-42 It does not inhibit oligomer toxicity. Notably, in these experiments on oligomers, all three neutrophil granule proteins showed Aβ only for 15 minutes before being added to cells. 1-42 It is pre-incubated. Aβ during the 15-minute oligomerization step. 1-42 The percentage cleavage is unknown. As discussed above, the larger products resulting from CAP37 cleavage may still be capable of forming toxic aggregates under oligomerization conditions.

[0151] Figure 19 shows the effects of Aβ in the presence of CAP37-derived peptides 120-146 WR at various concentrations. 1-42 This shows the effect of the oligomer on neuronal cells. The modified CAP37 peptide acts in a dose-dependent manner on Aβ 1-42 It significantly inhibited oligomer toxicity. Figure 20 shows the effects of Aβ in the presence of various concentrations of CAP37-derived peptides 120-146 WR-5RMP. 1-42 This shows the effect of the oligomer on neuronal cells. The 5RMP-modified CAP37 peptide variant acts in a dose-dependent manner on Aβ 1-42 It significantly inhibited oligomer toxicity. In Figures 19 and 20, both show Aβ 1-42Interacting with (Figures 2 and 3) and inhibiting fibrosis (Figure 13), peptides 120-146 WR and 120-146 WR 5RMP were also able to significantly reduce the toxicity of the oligomer. The peptides affect Aβ 1-42 It does not have enzymatic activity against Aβ, and therefore, 1-42 The cleavage of Aβ 1-42 This suggests that it is not the only mechanism for oligomerization and inhibition of toxicity. 1-42 Direct binding to it may inhibit oligomerization.

[0152] Figure 21 shows Aβ 1-42 This shows the binding levels of wild-type and mutant CG-derived peptides. 119-144 Each of the two load residues within is used in peptide CG. 119-144 D137R and CG 119-144 The replacement of the positive charge R in E141PR with Aβ 1-42 The binding with peptide CG was increased. 119-144 The deletion of the positive charge R129 in ΔR129V130 is equivalent to Aβ 1-42 The connection with it was removed.

[0153] Figures 22 and 23 show the effects of Aβ in the presence and absence of increasing concentrations of CG-derived peptides. 1-42 This shows fibrosis. Figure 23 shows the mutant CG. 119-144 Aβ by D137R 1-42 It shows dose-dependent inhibition of fibrosis. CG 119-144 The IC50 of the D137R is 1 / 3 CG 119-144 D137R / Aβ 1-42 This corresponds to a molar ratio of 3.1 μM.

[0154] Figure 24 shows peptide CG at various concentrations. 119-144 Aβ in the presence of D137R 1-42 This shows the effect of oligomers on nerve cells. Aβ 1-42 In addition to interacting with (Figure 21) and inhibiting fibrosis (Figures 22 and 23), this CG-derived mutant was also able to significantly reduce the toxicity of the oligomer.

[0155] MEMα+N2 supplement contains 100μM Aβ 1-42 Oligomers were formed by incubation at 4°C for 3 days. Subsequently, the peptide was added with a vehicle (water) or at a final concentration of 2×IC50 and incubated with the oligomers at 4°C for 3 days. Finally, these oligomers were added to neurons at a final concentration of 18 μM, and percentage cell death was measured at 24 hours. Figure 25 shows the Aβ of CG and CAP37 peptides. 1-42 It shows an effect on oligomers. Aβ 1-42 These peptides interact with (Figures 2 and 20) and inhibit fibrosis (Figures 13, 19, 22, and 23), and also affect Aβ 1-42 We were able to significantly reduce the toxicity of the oligomers.

[0156] Figure 26 shows CG peptide CG. 119-144 D137R and CAP37 peptide CAP37 120-146 WR's Aβ 1-42 It shows the effect on oligomer formation. In the absence of the peptide, Aβ 1-42 Monomers spontaneously form oligomers. Aβ 1-42 When combined with monomers, CG peptide CG 119-144 D137R and CAP37 peptide CAP37 120-146 WR is Aβ 1-42 The formation of oligomers was inhibited in a dose-dependent manner.

[0157] Uptake of peptides into the brain Peptide CG 119-144 Experiments were conducted using D137R (SEQ ID NO: 15) to demonstrate the uptake of peptides into the brain. This peptide was first treated with hydrazinonicotinamide (HYNIC) (technetium-99m( 99m The peptide was coupled with an efficient bifunctional chelator used to label biomolecules in Tc). Subsequently, at the College of Pharmacy Research Imaging Facility, the peptide was processed. 99mThe peptides were labeled with Tc and used in mice on the same day. Intravenous injection into the tail vein and intranasal administration of the labeled peptides were performed in four mice (two males and two females) for each administration route. The inventors administered 40 mg of the labeled peptide to each animal, and four hours after administration, they measured the percentage of total injected radioactivity per gram of tissue in the blood and brain. Subsequently, the inventors calculated the ratio of peptide concentration in blood to brain. This ratio was three times higher when using intranasal administration (brain / blood ratio 1 / 10) than when using intravenous administration (brain / blood ratio 1 / 30). These results demonstrate the peptide's ability to penetrate the brain. The estimated total peptide concentration in the brain was 5.5 nM four hours after intranasal delivery and 2.2 nM four hours after intravenous delivery.

[0158] While this disclosure is described in a manner that allows for a more complete understanding and recognition of its aspects in relation to certain particular embodiments, it is not intended to be limited to these particular embodiments. Rather, all substitutes, modifications, and equivalents are intended to be within the scope of this disclosure. Accordingly, it is understood that the above examples, including certain embodiments, serve to illustrate the practice of this disclosure, and that the details shown are presented as examples and for illustrative purposes only of certain embodiments, and are intended to provide what is considered to be the most useful and easily understandable explanation of the procedures, as well as the principles and conceptual aspects of the methods and compositions of this disclosure. Modifications can be made in the formulations of the various compositions described herein, in the methods described herein, or in the steps or sets of steps of the methods described herein, without departing from the spirit and scope of this disclosure.

Claims

1. A peptide compound comprising a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:43, SEQ ID NO:47, SEQ ID NO:48, and SEQ ID NO:

49.

2. The peptide compound of claim 1, wherein one or more of the amino acid positions comprises a wild type difference selected from the group consisting of D-amino acids, β-amino acids, homoamino acids, β-homoamino acids, peptoid monomers, and peptide staples.

3. The peptide compound of claim 1 or 2, wherein the peptide compound is effective in inhibiting and / or reversing the polymerization of amyloid beta (Aβ) peptides into Aβ oligomers and / or Aβ fibrils, and / or causing disaggregation of Aβ plaques.

4. The Aβ peptide is Aβ １－４０ and Aβ １－４２ The peptide compound according to claim 3, which is at least one of:

5. The peptide compound of claim 1, wherein the peptide compound is conjugated to, complexed with, and / or absorbed into a polymer to increase half-life and / or control release.

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

7. A peptide compound according to any one of claims 1 to 6, wherein the peptide consists of up to 50 amino acids.

8. A pharmaceutical composition comprising a peptide compound described in any one of claims 1 to 7.

9. The pharmaceutical composition of claim 8, wherein the peptide compound is administered in an amount effective to inhibit and / or reverse the formation of Aβ oligomers and / or Aβ fibrils from Aβ peptide.

10. 10. The pharmaceutical composition of claim 9, wherein inhibiting and / or reversing polymerization of Aβ peptide comprises causing disaggregation of at least one of Aβ oligomers, Aβ fibrils, or Aβ plaques.

11. The pharmaceutical composition described in claim 8, wherein the pharmaceutical composition is for use in treating a disease or condition associated with amyloid beta (Aβ) peptide, and the disease or condition is Alzheimer's disease, cerebral amyloid angiopathy, dementia, and / or neuroinflammation, and / or symptoms thereof.

12. An ex vivo method for inhibiting the formation of amyloid beta (Aβ) oligomers and / or Aβ fibrils, comprising the step of exposing Aβ peptides and / or Aβ oligomers to a peptide compound described in any one of claims 1 to 7.

13. An ex vivo method for reversing polymerization of Aβ peptides by causing disaggregation of at least one of Aβ oligomers, Aβ fibrils, and / or Aβ plaques, comprising the step of exposing Aβ oligomers, Aβ fibrils, and / or Aβ plaques to a peptide compound described in any one of claims 1 to 7.

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