Peptides for the treatment or prevention of 4R tauopathies

Abstract

One aspect of the present invention relates to a pharmaceutical composition or food composition for preventing, treating, or ameliorating a 4R tauopathy, comprising as an active ingredient one or more selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to said amino acid sequence, and peptide fragments thereof; or a medicinal use of said peptide for preventing, treating, or ameliorating a 4R tauopathy.

Description

[Technical Field]

[0001] The present invention relates to peptides that are effective in treating or preventing 4R tauopathies and compositions containing the same, and more particularly to peptides selected from telomerase that are effective in treating or preventing tauopathies, or compositions containing the same. [Background technology]

[0002] Neurons are a variety of cell types that are produced by differentiation of neural stem cells. Neural stem cells differentiate into various cells, such as neurons, glial cells, astrocytes, and oligodendrocytes, which make up the nervous system.

[0003] Neurological disorders are those that cause abnormalities in neurological function and include stroke, brain tumors, epilepsy, migraines, sleep disorders, Alzheimer's disease, and tauopathy. Tauopathies include progressive supranuclear palsy, corticobasal degeneration, Pick's disease, frontotemporal lobar degeneration (FTLD-tau), argyrophilic grain disease, subacute sclerosing panencephalitis, Christianson syndrome, post-encephalitic parkinsonism, Guadeloupean parkinsonism, spinocerebellar ataxia type 11, chronic traumatic encephalopathy (includes traumatic brain injury and dementia pugilistica), and globular glial tauopathy. These include primary age-related tauopathy (ARTAG), aging-related tau astrogliopathy includes globular glial tauopathy (ARTAG), and primary age-related tauopathy (PART, which includes tangle-predominant dementia and clinically asymptomatic cases).

[0004] Tauopathy is a general term for neurodegenerative diseases caused by the aggregation of tau protein in neurofibrillary tangles (NFTs) in the human brain. Tauopathy begins with the accumulation of excessive phosphates on tau protein, which leads to hyperphosphorylation and aggregation into neurofibrillary tangles (NFTs). These tau aggregates resemble compressed bundles of filaments, resembling twisted threads within brain cells. These abnormal tau aggregates, collectively known as tauopathies, are a key feature of neurodegenerative diseases (Brandt R, Hundelt M, Shahani N (2005) Tau alteration and neuronal degeneration in tauopathies: mechanisms and models. Biochimica.et biophysica.acta.1739:331-354).

[0005] In healthy neurons, tau stabilizes microtubules by promoting axonal growth and neuronal polarization. Pathological hyperphosphorylation of tau results in tau dissociation from microtubules and formation of insoluble aggregates (Gendron TF, Petrucelli L (2009) The role of tau in neurodegeneration. Mol. Neurodegener. 4:13).

[0006] Over the years, a structural scaffold for tau aggregation has been proposed, with evidence showing that 10 soluble monomers form insoluble filaments that associate into higher-order structures called neurofibrillary tangles (NFTs).

[0007] Tauopathies present with distinct neurodegenerative disorders depending on the predominant tau protein isoform involved in the disease, i.e., 4R tau or 3R tau (Front. Neurol., 05 November 2020 | https: / / doi.org / 10.3389 / fneur.2020.599384; Neuron 90, 941-947, June 1, 2016).

[0008] According to this, tauopathies can be classified into primary tauopathies, in which tau deposition is the primary etiology, and secondary tauopathies, in which additional etiologies (such as amyloid, trauma, and autoimmunity) are involved in addition to tau deposition. Primary tauopathies are divided into 4R tauopathies and 3R tauopathies, depending on whether 4R tau or 3R tau is predominant. Alzheimer's disease, known as the most common tauopathy, is a secondary tauopathy that manifests when amyloid deposition is involved in tau deposition, and in this respect is distinguished from the primary tauopathies. In particular, Alzheimer's disease is a secondary tauopathy in which amyloid beta is considered the primary etiological factor, which has been consistently proven by genetic and biochemical evidence (Rosler TW et al., Four-repeat tauopathies. Prog Neurobiol. 2019, 180:101644 https: / / pubmed.ncbi.nlm.nih.gov / 31238088 / ).

[0009] Among the primary tauopathies, 4R tauopathies include progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain disease, globular glial tauopathy, and aging-related tau astrogliopathy including globular glial tauopathy (ARTAG) (Olfati N et al., Clinical Spectrum of Tauopathies. Front Neurol. 2022, 13:944806). In particular, progressive supranuclear palsy is classified as a typical disease manifested by 4R tauopathy.

[0010] The number of diseases associated with 4R tauopathy is increasing with the aging population, but no drugs have been found to be effective in treating or preventing these diseases to date. Therefore, there is a need for drugs that can be effectively used to treat, improve, or prevent 4R tauopathy.

[0011] Meanwhile, it has been reported that PEP1 (SEQ ID NO: 1), a 16-amino acid peptide selected from human telomerase reverse transcriptase (hTERT), has anti-cancer, anti-inflammatory, and antioxidant effects, and is also effective in treating Alzheimer's disease (WO2013 / 167574). Furthermore, PEP1 has completed toxicity testing in numerous clinical trials targeting multiple diseases, confirming its stability against side effects.

[0012] LMTM (WO2021-001306) is known as a tau aggregation inhibitor, and is currently undergoing phase 3 clinical trials for tauopathies. However, the specific efficacy of LMTM, particularly for 4R tauopathies, has not been mentioned or tested in the prior art. In addition, LMTM has a tau aggregation inhibitory interval (EC 50 =2.2±0.2μM) and cytotoxicity (GI 50 = 6.4 ± 0.3 μM) have been reported to exhibit adverse effects (Yun Kyung Kim et al. Levosimendan inhibits disulfide tau oligomerization ameliorating tau pathology in TauP301L-BiFC mice, Experimental & Molecular Medicine volume 55, pages 612-627 (2023)).

[0013] Furthermore, LMTM is associated with the most common gastrointestinal and urinary side effects, which are reported to be the most common reason for discontinuing high-dose LMTM (Gauthier S et al., Efficacy and safety of tau-aggregation inhibitor therapy in patients with mild or moderate Alzheimer's disease: a randomized, controlled, double-blind, parallel-arm, Phase 3 trial. Lancet. 2016, 388(10062):2873-2884). [Prior art documents] [Patent documents]

[0014] [Patent Document 1] WO2013 / 167574 [Patent Document 2] WO2021 / 001306 [Non-patent literature]

[0015] [Non-licensed document 1] Brandt R,Hundelt M,Shahani N(2005)Tau alteration and neuronal degeneration in tauopathies:mechanisms and models.Biochimica.et biophysica.acta.1739:331-354 [Non-licensed document 2] Gendron TF,Petrucelli L(2009)The role of tau in neurodegeneration.Mol.Neurodegener.4:13 [Non-licensed document 3] Front.Neurol.,05 November 2020 https: / / doi.org / 10.3389 / fneur.2020.599384

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

[0016] One aspect of the present invention is to provide peptides that are effective in treating or preventing 4R tauopathies.

[0017] Another aspect of the present invention is to provide a pharmaceutical composition for treating or preventing a 4R tauopathy, comprising the peptide.

[0018] Another aspect of the present invention is to provide a food composition for improving or preventing 4R tauopathy, comprising the peptide.

[0019] Another aspect of the present invention is to provide a pharmaceutical use of said peptide for the treatment or prevention of 4R tauopathies.

[0020] Another aspect of the present invention is to provide a method for treating or preventing a 4R tauopathy, comprising the step of administering the peptide to a subject in need thereof. [Means for solving the problem]

[0021] One aspect of the present invention provides a pharmaceutical composition for preventing or treating 4R tauopathy, comprising, as an active ingredient, one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity with the amino acid sequence, and peptides that are fragments thereof.

[0022] Another aspect of the present invention provides a food composition for preventing or ameliorating 4R tauopathy, comprising as an active ingredient one or more selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity with said amino acid sequence, and peptides that are fragments thereof.

[0023] Another aspect of the present invention provides one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof, for the prevention or treatment of 4R tauopathy.

[0024] Another aspect of the present invention provides a pharmaceutical use of one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof, for the prevention or treatment of 4R tauopathy.

[0025] Another aspect of the present invention provides a method for preventing or treating a 4R tauopathy, comprising the step of administering to a subject in need thereof a pharmaceutically effective amount of one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof. [Effects of the Invention]

[0026] One or more peptides selected from the group consisting of peptides containing the amino acid sequence of SEQ ID NO: 1, peptides sharing 80% or more sequence identity with said amino acid sequence, and peptide fragments thereof, showed significant improvements in motor performance and cognitive performance compared to control and reference drug-treated groups in a 4R-Tau-BiFC mouse model carrying the P301L mutation, known to induce 4R tauopathy. Furthermore, brain tissue imaging and brain lysate analysis of the 4R-Tau-BiFC mouse model confirmed superior suppression of 4R tau formation and hyperphosphorylation of 4R tau, which are the cause of 4R tauopathy, compared to control and reference drug-treated groups. Therefore, because one or more peptides selected from the group consisting of peptides containing the amino acid sequence of SEQ ID NO: 1, peptides sharing 80% or more sequence identity with said amino acid sequence, and peptide fragments thereof, were confirmed to effectively suppress 4R tauopathy, they are expected to be useful for treating, ameliorating, or preventing 4R tauopathy. [Brief explanation of the drawings]

[0027] [Figure 1] FIG. 1 is a diagram illustrating the drug dose and administration schedule in the animal-based behavioral evaluation of PEP1 according to Experimental Example 2. [Figure 2] Figure 2 shows the results of the rota-rod test to assess the motor performance improvement in the 4R TauP301L-BiFC mouse model. Analysis was performed by two-way ANOVA with Tukey's multiple comparisons (*p<0.05; **p<0.01; ns: not significant compared to vehicle). [Figure 3]Figure 3 shows the results of evaluating the effect of novel object recognition on cognitive performance in the 4R TauP301L-BiFC mouse model. Black bars indicate the results for the learned object, and hatched bars indicate the results for the novel object. Analysis was performed by two-way ANOVA with Sidak's multiple comparisons test (*p<0.05; ***p<0.0001). [Figure 4] FIG. 4 is a diagram illustrating the drug dose and administration schedule in the animal-based evaluation of the inhibitory effect of PEP1 on 4R-tauopathy according to Experimental Example 3. [Figure 5] FIG. 5 shows photographs of (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining results in the sensorimotor cortex region, as well as graphs showing the results of quantitative analysis of the respective fluorescence levels. [Figure 6] FIG. 6 shows photographs of (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining results in the motor cortex region, as well as graphs showing the results of quantitative analysis of the respective fluorescence levels. [Figure 7] FIG. 7 shows photographs of (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining results in the hippocampal region, as well as graphs showing the results of quantitative analysis of the respective fluorescence levels. [Figure 8] Figure 8 shows photographs of (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining in the substantia nigra, as well as graphs showing the quantitative analysis of the fluorescence levels of each (Figures 5 to 8 were analyzed by one-way ANOVA with Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001; ns: not significant compared to vehicle). [Figure 9a]Brain lysates were obtained from TauP301L-BiFC mice treated for 5 months, from 7 to 12 months of age. The soluble and non-soluble fractions (Fig. 9a, 9b, 9c) were immunoblotted with total tau (Tau5) and phosphorylated tau (pS199, pS396) antibodies. The results are shown in Fig. 9c. Photographs of immunoblots for the sensorimotor cortex and motor cortex, respectively; Fig. 9b, 9c ... [Figure 9b] Brain lysates were obtained from TauP301L-BiFC mice treated for 5 months, from 7 to 12 months of age. The soluble and non-soluble fractions (Fig. 9a, 9b, 9c) were immunoblotted with total tau (Tau5) and phosphorylated tau (pS199, pS396) antibodies. The results are shown in Fig. 9c. Photographs of immunoblots for the sensorimotor cortex and motor cortex, respectively; Fig. 9b, 9c ... [Figure 9c]Brain lysates were obtained from TauP301L-BiFC mice treated for 5 months, from 7 to 12 months of age. The soluble and non-soluble fractions (Fig. 9a, 9b, 9c) were immunoblotted with total tau (Tau5) and phosphorylated tau (pS199, pS396) antibodies. The results are shown in Fig. 9c. Photographs of immunoblots for the sensorimotor cortex and motor cortex, respectively; Fig. 9b, 9c ... [Figure 10] Brain lysates were obtained from TauP301L-BiFC mice treated for 5 months, from 7 to 12 months of age. The soluble and non-soluble fractions (Fig. 9a, 9b, 9c) were immunoblotted with total tau (Tau5) and phosphorylated tau (pS199, pS396) antibodies. The results are shown in Fig. 9c. Photographs of immunoblots for the sensorimotor cortex and motor cortex, respectively; Fig. 9b, 9c ... DETAILED DESCRIPTION OF THE INVENTION

[0028] The present invention will now be described in more detail to facilitate understanding of the present invention.

[0029] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those of ordinary skill in the art related to the present invention. Although preferred methods and samples are described herein, similar or equivalent values ​​are also included within the scope of the present invention. Furthermore, numerical values ​​described herein are considered to include the meaning of "about" even if not explicitly stated. The contents of all publications referenced herein are incorporated herein by reference in their entirety.

[0030] As used herein, the term "4R tauopathy" refers to a neurodegenerative disease caused by the predominance of 4R tau aggregation among tau protein isoforms, including, but not limited to, progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain disease, globular glial tauopathy, and ARTAG (aging-related tau astrogliopathy includes globular glial tauopathy).

[0031] The present inventors have found that PEP1 (hereinafter also referred to as "GV1001"), a peptide selected from telomerase, is effective in suppressing 4R tauopathy. More specifically, behavioral tests and brain tissue analysis using a 4R-Tau-BiFC mouse model (i.e., an animal model of 4R tauopathy) carrying the P301L mutation, which is known to induce 4R tauopathy (Acta Neuropathol (2017) 133:665-704), demonstrated that PEP1 suppressed the formation and hyperphosphorylation of 4R tau, which are the cause of 4R tauopathy (Experimental Example 3), and actually improved motor and cognitive abilities in the animal model of 4R tauopathy (Experimental Example 2). Furthermore, compared with LMTM, a tauopathy drug known for its cytotoxicity and gastrointestinal and urinary side effects, PEP1 was confirmed to inhibit the formation and hyperphosphorylation of 4R tau, which are the cause of 4R tauopathy, while improving cognitive and motor abilities in 4R tauopathy, even at significantly lower doses. GV1001 is known as a highly stable peptide with no side effects (Hyun-Hee Park et al., Neurobiology of Aging (2014) 35(6):1255-74; Hyun-Hee Park et al., Neurotoxicology (2016) 55:131-141). The efficacy of GV1001 against 4R tauopathy is significant in that it can be safely and effectively used to treat or prevent tauopathies, especially as the first to demonstrate its efficacy in 4R tauopathy, compared with LMTM, a conventional, simple tauopathy drug.

[0032] Specifically, in Experiment 2, a 4R TauP301L-BiFC mouse model was administered the drug for 5 months from 7 to 12 months of age. The results of the rotarod test (Figure 2) showed that the PEP1 1 and 2 mg / kg groups showed significantly improved motor performance compared to the vehicle (control group) and the reference compound LMTM. Furthermore, the results of the novel object recognition test (Figure 3) showed that the PEP1-treated group showed significantly improved cognitive performance for novel objects compared to the vehicle and the reference compound LMTM.

[0033] In Experiment 3, we performed (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining on brain tissue from a 4R TauP301L-BiFC mouse model treated with the drug for 5 months from 7 to 12 months of age. Quantitative analysis of Tau-BiFC fluorescence levels in the somatosensory cortex, motor cortex, hippocampus (CA1), and substantia nigra revealed that 4R tauopathy is predominantly caused by the formation of tau oligomers (Figures 5-8). The PEP1-treated group showed a significant decrease in fluorescence intensity compared to the vehicle and the reference compound LMTM-treated group, demonstrating its inhibitory effect on tau oligomer formation. Furthermore, immunofluorescence staining using the tau hyperphosphorylation antibody AT8 (phospho-Tau S202 / T205) showed a significant decrease in AT8 fluorescence intensity in the PEP1-treated group compared with the vehicle and the reference compound LMTM-treated group, demonstrating its inhibitory effect on 4R tau hyperphosphorylation. Furthermore, after obtaining brain lysates from a TauP301L-BiFC mouse model treated with the drug for 5 months from 7 to 12 months of age, we immunoblotted i) soluble fraction samples and ii) insoluble fraction samples with total tau (Tau5) and phosphorylated tau (pS199, pS396) antibodies (Figures 9 and 10). As a result, in the soluble fraction samples, the PEP1-treated group showed a significant decrease in total tau and phosphorylated tau compared to the vehicle and reference compound LMTM-treated groups, and in the insoluble fraction samples, the PEP1-treated group showed a significant inhibition of the formation of insoluble tau aggregates compared to the vehicle and reference compound LMTM-treated groups.

[0034] In one aspect, the present invention provides a pharmaceutical composition for preventing or treating a 4R tauopathy, comprising, as an active ingredient, one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof.

[0035] In another aspect of the present invention, there is provided a food composition for preventing or ameliorating 4R tauopathy, comprising as an active ingredient one or more selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity with said amino acid sequence, and peptides that are fragments thereof.

[0036] In one embodiment, the fragment is a peptide, that is, a fragment consisting of three or more amino acids.

[0037] The 4R tauopathy is selected from the group consisting of, but is not limited to, progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain disease, globular glial tauopathy, and aging-related tau astrogliopathy includes globular glial tauopathy (ARTAG), and includes any neurodegenerative disease caused by a predominance of 4R tau aggregation.

[0038] The peptides disclosed herein can include peptides with 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more sequence identity. The peptides disclosed herein can also include peptides comprising SEQ ID NO: 1 or fragments thereof, and peptides with one or more amino acid changes, two or more amino acids, three or more amino acids, four or more amino acids, five or more amino acids, six or more amino acids, or seven or more amino acids.

[0039] In one embodiment, the amino acid changes may be those that alter the physicochemical properties of the peptide, such as improving the thermal stability of the peptide, altering its substrate specificity, or changing its pH optimum.

[0040] As used herein, the term "amino acid" includes not only the 22 standard amino acids naturally incorporated into peptides, but also D-isomers and modified amino acids. Thus, in one embodiment, the peptide may include D-amino acids.

[0041] In one embodiment, the peptide may include non-standard amino acids that have undergone post-translational modifications. Examples of post-translational modifications include phosphorylation, glycosylation, acylation (e.g., acetylation, myristoylation, and palmitoylation), alkylation, carboxylation, hydroxylation, glycation, biotinylation, ubiquitinylation, chemical changes (e.g., beta-elimination deimidation, deamidation), and structural changes (e.g., disulfide bridge formation). The peptide may also include amino acid changes, such as changes in the amino group, carboxy group, or side chain, that occur due to chemical reactions that occur during conjugation with a crosslinker to form a peptide conjugate.

[0042] The peptide may be a wild-type peptide identified and isolated from a natural source. Alternatively, the peptide may be an artificial variant, comprising an amino acid sequence in which one or more amino acids have been substituted, deleted, and / or inserted compared to the peptide of SEQ ID NO: 1. Amino acid changes in wild-type polypeptides, as well as artificial variants, include conservative amino acid substitutions that do not significantly affect protein folding and / or activity. Examples of conservative substitutions are within the following groups: basic amino acids (arginine, lysine, and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine, valine, and methionine), aromatic amino acids (phenylalanine, tryptophan, and tyrosine), and small amino acids (glycine, alanine, serine, and threonine). Generally, amino acid substitutions that do not alter specific activity are known in the art. The most commonly occurring substitutions are Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu, and Asp / Gly, and their opposites. Other examples of conservative substitutions include those in Table 1 below.

[0043] [Table 1]

[0044] Substantial variations in the biological properties of peptides are achieved by selecting substitutions that differ significantly in (a) their effect on maintaining the structure of the polypeptide backbone in the region of substitution, e.g., sheet or helix conformation, (b) their effect on maintaining the charge or hydrophobicity of the molecule at the target site, or (c) their effect on maintaining the bulk of the side chain. Natural residues can be divided into the following groups based on common side chain properties:

[0045] (1) Hydrophobic: norleucine, met, ala, val, leu, ile; (2) Neutral hydrophilic: cys, ser, thr; (3) Acidic: asp, glu; (4) basic: asn, gln, his, lys, arg; (5) residues that influence chain orientation: gly, pro; and (6) Aromatics: trp, tyr, phe.

[0046] Non-conservative substitutions are made by exchanging a member of one of these classes for another. Cysteine ​​residues not involved in maintaining the proper conformation of the peptide are usually substituted with serine to improve the oxidative stability of the molecule and prevent aberrant cross-linking. Conversely, cysteine ​​bond(s) can be added to the peptide to improve its stability.

[0047] Another type of amino acid variant of a peptide is one in which the glycosylation pattern of the antibody is altered by deleting one or more carbohydrate residues found in the peptide and / or adding one or more glycosylation sites that are not present in the peptide.

[0048] Glycosylation of peptides typically refers to either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate residue to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate residue to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of either the sugar N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.

[0049] Glycosylation sites can be added to the peptide by altering the amino acid sequence so that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). Such alterations can also be made by adding or substituting one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).

[0050] In addition, the active ingredient, a peptide containing the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence homology with the amino acid sequence, and peptides that are fragments thereof, have the advantage of low toxicity within cells and high stability in the body.

[0051] Peptides having the amino acid sequence of SEQ ID NO: 1 are shown in Table 2 below. The "names" in Table 2 below are given to distinguish the peptides. In Table 2 below, the peptide of SEQ ID NO: 2 indicates the full-length peptide of human telomerase. In one embodiment, a peptide comprising the amino acid sequence of SEQ ID NO: 1 includes the peptide of SEQ ID NO: 2. In one embodiment, a peptide comprising the amino acid sequence of SEQ ID NO: 1 includes a peptide composed of the amino acid sequence of SEQ ID NO: 1. Peptides comprising the amino acid sequence of SEQ ID NO: 1, peptides having 80% or more sequence homology to the amino acid sequence, and peptides that are fragments thereof include "synthetic peptides" synthesized by selecting peptides at the corresponding positions from peptides contained in telomerase. SEQ ID NO: 2 indicates the amino acid sequence of full-length telomerase.

[0052] [Table 2]

[0053] In one embodiment, the pharmaceutical composition contains a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, or a peptide fragment thereof at a concentration of 0.01 mg / mL to 100 mg / mL, preferably 0.1 mg / mL to 10 mg / mL. However, if there are differences in efficacy depending on the dose for each treatment subject, the concentration can be adjusted appropriately. Containing a peptide at or below this range can favorably exhibit the intended effects of the present invention while ensuring both the stability and safety of the composition. Furthermore, containing a peptide at this range is also appropriate from the perspective of cost-effectiveness.

[0054] The pharmaceutical composition can be applied to all animals, including humans, dogs, chickens, pigs, cows, sheep, guinea pigs, or monkeys.

[0055] The pharmaceutical composition can be administered orally, rectally, transdermally, intravenously, intramuscularly, intraperitoneally, intramedullary, intrathecally, intradermally or subcutaneously, preferably intraperitoneally, intradermally, intravenously or subcutaneously, more preferably subcutaneously.

[0056] Formulations for oral administration may be, but are not limited to, tablets, pills, soft or hard capsules, granules, powders, liquids, or emulsions, while formulations for parenteral administration may be, but are not limited to, injections, drops, lotions, ointments, gels, creams, suspensions, emulsions, suppositories, patches, or aerosols.

[0057] The pharmaceutical composition may contain, as needed, additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, flavors, or sweeteners. The pharmaceutical composition may be prepared by conventional pharmaceutical methods known in the art.

[0058] The pharmaceutical composition may be administered alone or in combination with one or more additional therapeutic agents. In one embodiment, the one or more additional therapeutic agents are active ingredients conventionally known as therapeutic agents for neurodegenerative diseases. The term "co-administration" refers to administering a pharmaceutical composition according to one aspect of the present invention and one or more additional therapeutic agents to an individual or patient to be treated simultaneously or at different times and in any order with a time interval. Thus, each component may be administered separately or at a time sufficiently close to achieve the desired therapeutic effect.

[0059] The dosage of the pharmaceutical composition may vary depending on the age, sex, and weight of the patient, the condition and its severity, the route of administration, and the prescriber's discretion. Determining the dosage based on these factors is within the skill of a person skilled in the art. The daily dosage is, for example, 0.0001 mg / kg / day to 100 mg / kg / day, preferably 0.001 mg / kg / day to 10 mg / kg / day, and more preferably 0.001 mg / kg / day to 1 mg / kg / day. However, if the effect differs depending on the dosage, the dosage can be adjusted appropriately.

[0060] Determining the dosage and administration interval of the active ingredient of the pharmaceutical composition is within the level of a person skilled in the art. For example, the unit administration dose for humans is about 0.1 to 100 mg per kg, more specifically about 0.1 to 50 mg, 0.1 mg to 30 mg, or 0.5 mg to 10 mg, based on an adult weighing 60 kg, and the administration interval is, but is not limited to, once to four times a day or once to four times a week.

[0061] In another aspect, the present invention provides one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof, for use in the prevention or treatment of a 4R tauopathy.

[0062] In yet another aspect, the present invention provides one or more pharmaceutical uses selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof, for the prevention or treatment of 4R tauopathy.

[0063] In yet another aspect, the present invention provides a method for treating or preventing a 4R tauopathy, comprising the step of administering to a subject in need of prevention or treatment of a 4R tauopathy a pharmaceutically effective amount of one or more peptides selected from the group consisting of a peptide comprising the amino acid sequence of SEQ ID NO: 1, a peptide having 80% or more sequence identity to the amino acid sequence, and peptides that are fragments thereof.

[0064] The details of the peptide for use in the prevention or treatment of 4R tauopathy, the pharmaceutical use for the prevention or treatment of 4R tauopathy, and the method for treating or preventing 4R tauopathy according to the above-mentioned embodiment are the same as those of the pharmaceutical composition according to one embodiment of the present invention.

[0065] Furthermore, the peptide for use in the prevention or treatment of 4R tauopathy according to the above-mentioned embodiment, the pharmaceutical use for the prevention or treatment of 4R tauopathy, and the administration method used in the method for treating or preventing 4R tauopathy are the same as those described above in terms of the dosage, administration interval, administration frequency, etc. of the pharmaceutical composition, as they are amounts effective for the treatment or prevention of an individual or patient.

[0066] The configuration and effects of the present invention will be explained in more detail below with reference to examples and experimental examples. However, the following examples and experimental examples are provided to facilitate understanding of the present invention and are not intended to limit the scope of the present invention.

[0067] Experimental Example 1: Synthesis of peptide PEP-1 A peptide consisting of 16 amino acids selected from human telomerase and having the following structural formula 1 and SEQ ID NO: 1 (PEP-1) was synthesized.

[0068] [Chemical formula 1] [ka]

[0069] The peptide PEP1 of SEQ ID NO: 1 was prepared by a conventional solid-phase peptide synthesis method. Specifically, the peptide was synthesized by coupling amino acids one by one from the C-terminus using Fmoc solid-phase peptide synthesis (SPPS) using an ASP48S (Peptron, Inc., Daejeon, Republic of Korea). The peptide was used with the first amino acid at the C-terminus attached to the resin, as follows: NH2-Lys(Boc)-2-chloro-Trityl Resin NH2-Ala-2-chloro-Trityl Resin NH2-Arg(Pbf)-2-chloro-Trityl Resin

[0070] All amino acid starting materials used in peptide synthesis were N-terminally protected with Fmoc, and the residues were protected with Trt, Boc, t-butylester (t-Bu), 2,2,4,6,7-pentamethyl dihydrobenzofuran-5-sulfonyl (Pbf), etc., which can be removed by acid. For example,

[0071] Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Pro-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH , Fmoc-Lys(Boc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Met-OH, Fmoc-Asn(Trt)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ahx-OH, Trt-Mercaptoacetic acid.

[0072] The coupling reagent used was HBTU [2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetamethylaminium hexafluorophosphate] / HOBt [N-Hydroxybenzotriazole] / NMM [4-Methylmorpholine]. Fmoc was removed using piperidine in 20% DMF. The synthesized peptide was separated from the resin and the protecting groups of the residues were removed using a cleavage cocktail [TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H2O = 92.5 / 2.5 / 2.5 / 2.5].

[0073] Each peptide was synthesized by repeatedly reacting the starting amino acid with a protecting group bound to the solid support, washing with solvent, and deprotecting it. The synthesized peptide was cleaved from the resin, purified by HPLC, confirmed by MS, and lyophilized.

[0074] The specific synthesis process of PEP1 is as follows.

[0075] 1) Coupling An amino acid (8 equivalents) protected with NH2-Lys(Boc)-2-chloro-Trityl Resin and coupling reagents HBTU (8 equivalents), HOBt (8 equivalents), and NMM (16 equivalents) were dissolved in DMF and added, and the mixture was reacted at room temperature for 2 hours. The mixture was then washed with DMF, MeOH, and DMF, in that order.

[0076] 2) Fmoc deprotection 20% piperidine in DMF was added, and the mixture was reacted twice for 5 minutes at room temperature, and then washed with DMF, MeOH, and DMF in that order.

[0077] 3) The peptide backbone was created by repeating the reactions 1) and 2) above.

[0078] 4) Cleavage: After synthesis, a cleavage cocktail was added to the peptide resin to separate the peptide from the resin.

[0079] 5) Cooling diethyl ether was added to the resulting mixture, and the resulting peptide was precipitated by centrifugation.

[0080] 6) After purification by Prep-HPLC, the molecular weight was confirmed by LC / MS, and the product was frozen and produced as a powder.

[0081] Experimental Example 2: Animal-based behavioral evaluation of GV1001 Experimental Example 2-1: Experimental Overview The experimental animals and the drugs administered thereto are as shown in Table 3 below and FIG.

[0082] [Table 3]

[0083] Experimental Example 2-2: Experimental Method 1) Rotarod test method (1) After three days of adaptation training, behavioral experiments were conducted on the fourth day. (2) One hour before the experiment, the experimental mice were allowed to adapt to the space where the behavioral experiment would be conducted. (3) During the adaptation training period, mice were forced to walk on a treadmill at speeds of 5, 15, and 20 / 25 rpm for 360 seconds. If the mice lost balance and fell to the floor, they were forced to walk again. (4) After the adaptation training, a behavioral experiment was conducted by gradually increasing the treadmill speed from 5 rpm to 35 rpm for 480 seconds. When the treadmill was running, the time (seconds) until the experimental mice lost their balance and fell to the floor was measured.

[0084] 2) Novel object recognition test (1) On the first day, the subjects were trained to adapt to a new open space. On the second day, two similar objects were added to the open space, and the subjects were given time to adapt. On the third day, an experiment was conducted to evaluate the subjects' cognitive ability regarding the newly replaced objects. (2) One hour before the experiment, the experimental mice were allowed to adapt to the space where the behavioral experiment would be conducted. (3) On day 1, the experimental mice were exposed to an open space measuring 40 × 40 cm, and then behavioral evaluation was performed for 10 minutes. (4) On the second day, two similar objects were placed in the same open space and the rats were allowed to learn the objects for 10 minutes. (5) On the third day, one of the two learned objects was replaced with a new object in the same open space, and cognitive ability regarding the replaced object was assessed.

[0085] Experimental Example 2-3: Results of behavioral experiment 1) Evaluation experiment of motor ability improvement using the rotarod test The results of evaluating the effect of improving motor ability in the 4R TauP301L-BiFC mouse model using the rotarod test are shown in Table 4 and Figure 2 below. The experiment was conducted using TauP301L-BiFC mouse models administered with the drug for 5 months from 7 to 12 months of age. Analysis was performed using two-way ANOVA with Tukey's multiple comparison test ( *p<0.05; ** p<0.01; ns not significant compared to vehicle).

[0086] [Table 4]

[0087] According to Table 4 and FIG. 2, the results of the motor evaluation in the 4R TauP301L-BiFC mouse model are as follows:

[0088] Exercise capacity was assessed in transgenic mouse models aged 11.6 to 12.1 months, 59 days after administration. Exercise capacity testing using a rotarod treadmill showed that the drug group exhibited relatively higher exercise capacity than the vehicle group.

[0089] -In the group administered the reference compound LMTM (15 mg / kg), a tendency for improvement in motor ability was observed, but the difference was not significant.

[0090] -GV1001 at 1 and 2 mg / kg showed significant improvements in motor performance compared to vehicle. In the GV1001 at 0.1 mg / kg group, improvements in motor performance were observed, but the difference was not statistically significant compared to vehicle.

[0091] 2) Experiment to evaluate cognitive ability improvement through novel object recognition test Figure 3 shows the results of evaluating the effect of improving cognitive ability in the 4R TauP301L-BiFC mouse model using a novel object recognition test. The experiment was conducted using TauP301L-BiFC mouse models administered the drug for 5 months from 7 to 12 months of age. The black bars indicate the results for the learned object, and the shaded bars indicate the results for the novel object. Analysis was performed using two-way ANOVA with Sidak's multiple comparisons test ( * p<0.05; *** p<0.0001).

[0092] According to Figure 3, the comparative results of cognitive ability in the 4R TauP301L-BiFC mouse model are as follows:

[0093] Cognitive ability was evaluated in transgenic mouse models aged 12.1 to 12.4 months, 62 days after treatment. Cognitive ability was assessed based on behavior after replacing a learned object with a new object. Behavioral testing of the treated group showed that, compared to the vehicle group, the exploration time for the new object was longer than the exploration time for the learned object.

[0094] -The reference compound LMTM (15 mg / kg) significantly improved cognitive performance for novel objects.

[0095] -GV1001 at 1 mg / kg significantly improved cognitive performance. At 0.1 mg / kg, there was no significant difference compared to vehicle. At 2 mg / kg, there was a trend toward improvement in cognitive performance, but the difference was not statistically significant.

[0096] Experimental Example 3: Evaluation of the inhibitory effect of GV1001 on animal-based 4R tauopathy Experimental Example 3-1: Experimental Overview The experimental animals and the drugs administered thereto are as shown in Table 5 below and FIG.

[0097] [Table 5]

[0098] The experimental reagents used are as follows: 1) Brain tissue analysis

[0099] [Table 6]

[0100] 2) Analysis of brain lysates (immunoblots)

[0101] [Table 7]

[0102] Experimental Example 3-2: Experimental Method A. Experimental protocol for analyzing TauP301L-BiFC mouse-based brain tissue images 1) Experimental method for brain tissue sampling (1) On day 1, the TauP301L-BiFC mouse model, which had completed the 63-dose administration schedule, was anesthetized and prepared for cardiac laparotomy. (2) Saline was injected into the heart of the TauP301L-BiFC mouse model, and blood was removed from the body using a perfusion pump. (3) The brains of the mice were removed and weighed. (4) The brains used in tissue staining experiments were immersed in 4% PFA solution and fixed for 24 hours. (5) On the second day, the fixed brain tissue was immersed in a 20% sucrose solution and dehydrated for 24 hours. (6) On the third day, the brain tissue was immersed in 30% sucrose solution and dehydrated for 48 hours. (7) On day 5, the dehydrated brain tissue was removed and excess water was removed. The brain tissue was then placed in a prepared mold with OCT compound and frozen as quickly as possible on dry ice while protecting the tissue. The frozen brain tissue mold was stored in a deep freezer at -80°C. The tissue was then cut into 30 μm-thick sections using a cryostat and stored in 0.05% sodium azide solution.

[0103] 2) Experimental method for AT8 immunofluorescence staining of brain tissue (1) On day 1, 30 μm-thick cryosectioned brain tissue samples were selected for each brain region and immersed in PBS solution and washed three times for 10 minutes. (3) The tissue was fixed with 3.7% formaldehyde for 5 minutes. (4) Washed with PBS solution three times for 10 minutes. (5) Brain tissue samples were permeabilized with 0.3% PBS-T solution. (6) The sections were immersed in a 5% BSA in PBS solution and blocked for 1 hour. (7) The brain tissue was placed in a solution of 3% BSA and 0.1% Tween-20 in PBS in which the primary antibody (AT8) was diluted at a ratio of 1:200, and binding was carried out overnight. (8) On the second day, the brain tissue was immersed in PBS solution and washed three times for 10 minutes. (9) The brain tissue was placed in a solution of 3% BSA and 0.1% Tween-20 in PBS in which the secondary antibody was diluted at a ratio of 1:500, and binding was allowed to occur at room temperature for 1 hour. (10) The brain tissue was immersed in PBS solution and washed once for 10 minutes. (11) The brain tissue was immersed in a 1:2000 diluted Hoechst solution (stock concentration 1 mg / mL) in PBS to stain the nuclei. (12) The brain tissue was immersed in PBS solution and washed three times for 10 min. (13) Fluorescence imaging of brain tissue was performed using a slide scanner.

[0104] 3) Experimental method for brain tissue lipid staining for Tau-BiFC imaging (1) On day 1, Sudan Black B staining was performed on 30 μm-thick cryosectioned brain tissue samples. (2) A 0.05% Sudan Black B solution in 70% EtOH was stirred overnight (O / N), and then the particles were filtered through a 0.22 μm filter. (3) The selected brain tissue samples were immersed in distilled water (DW) and washed three times for 5 minutes. (4) The brain tissue was immersed in the prepared 0.05% Sudan Black B solution and stained for 10 minutes. (5) The brain tissue stained with Sudan black B was immersed in 0.1% PBS-T solution and washed at 30-second intervals to adjust the degree of staining. (6) Washed with DW three times for 5 minutes. (7) The brain tissue was immersed in a Hoechst solution in distilled water at a concentration of 0.2 μg / mL, and nuclei were stained for 20 minutes. (8) Washed with DW three times for 5 minutes. (9) Tau-BiFC imaging was performed using a slide scanner.

[0105] 4) How to interpret fluorescent images of brain tissue (1) To obtain fluorescent images of the stained brain tissue, brain tissue slides were scanned using a Zeiss Axio Scan (Zeiss, Oberkochen, Germany). -BiFC fluorescence: λex = 460-490nm, λem = 500-550nm -AT8 fluorescence: λex = 620-640nm, λem = 650-700nm -Hoechst fluorescence: λex = 330-375nm, λem = 430-470nm (3) Fluorescence values ​​were calculated for each brain tissue region obtained by imaging using Image J software (NIH).

[0106] B. TauP301L-BiFC Mouse-Based Brain Lysate Analysis Experimental Protocol 1) Experimental method for brain soluble fraction sampling (1) After administration, the TauP301L-BiFC mouse model was anesthetized and prepared for cardiac laparotomy. (2) Saline was injected into the heart of the TauP301L-BiFC mouse model, and blood was removed from the body using a perfusion pump. (3) After removing the blood, the brain was removed and weighed. (4) The excised brain was homogenized by adding 1 mL of RIPA buffer (with protease / phosphatase inhibitor cocktail). (5) The homogenized brain lysate was transferred to an EP-tube and then incubated on an orbital shaker at 4°C for 2 hours. (6) After centrifugation at 13,000 rpm for 20 minutes at 4°C, the supernatant (soluble fraction) was collected into a new EP tube and stored at -80°C. (7) To prepare immunoblot samples, the concentration of the supernatant (soluble fraction) samples was quantified using the Bradford protein assay. (8) RIPA buffer and 4X SDS Laemmli sample buffer (w / 2.5% β-mercaptoethanol for reducing conditions) were added to each sample to a final concentration of 2 mg / mL. (9) The samples were boiled at 97°C for 5 minutes.

[0107] 2) Experimental method for sampling insoluble brain fraction (1) The pellet remaining after the supernatant (soluble fraction) was transferred was dissolved in 1 mL of 1 M sucrose and DNase I (conc. 1 mg / mL) in RIPA buffer. (2) After centrifugation at 13,000 rpm for 20 minutes at 4°C, the supernatant was removed. (3) The pellet was resuspended in 2% SDS solution (1 ml per gram of tissue) and then incubated at room temperature for 1 hour. (4) After centrifugation at 13,000 rpm for 1 minute at room temperature, the supernatant (insoluble fraction) was collected and transferred to a new EP tube, and an equal volume of 2x SDS Laemmli sample buffer (w / 2.5% β-mercaptoethanol for reducing conditions) was added. (5) The sample was boiled at 97°C for 5 minutes.

[0108] 3) Tau immunoblotting experimental method (1) Day 1: 20 μg of brain lysate sample (soluble / soluble fractions) was loaded onto a 10% SDS-PAGE gel. (2) Electrophoresis was carried out at 80 V for 30 minutes, then changed to 90 V and electrophoresis was carried out for 1 hour. -SDS-PAGE running buffer: 1X Tris / Glycine / SDS buffer in triple distilled water (3) A PVDF membrane cut to the size of a transfer tray was immersed in 100% methanol for 1 minute, autoclaved triple-distilled water for 3 minutes, and cold 1X transfer buffer for 10 minutes. Transfer buffer: 1X Tris / Glycine buffer with 20% methanol in triple distilled water (1X Tris / Glycine buffer with 20% methanol in 3'DW) (4) Wet transfer was performed at 100 V for 1 hour and 20 minutes. (5) The membrane was immersed in a 5% BSA in TBS-T (w / 0.1% Tween-20) solution and blocked at room temperature for 1 hour. (6) The membrane was immersed in a 2.5% BSA in TBS-T (w / 0.1% Tween-20) solution containing the primary antibody, and allowed to bind overnight (O / N) on an orbital shaker at 4°C. -Anti-Tau(Tau5)antibody(ab80579, 1:5000) -Anti-Tau(phospho S199)antibody(ab109390, 1:5000) -Anti-Tau(phospho S396)antibody(ab81268, 1:5000) (7) Day 2: The membrane was immersed in TBS-T (w / 0.1% Tween-20) solution and washed three times for 10 minutes. (8) The membrane was immersed in a 2.5% BSA in TBS-T (w / 0.1% Tween-20) solution containing a secondary antibody and allowed to bind at room temperature for 1 hour. -Goat Anti-Mouse IgG H&L(HRP)secondary antibody(ab6789, 1:10000) -Goat Anti-Rabbit IgG H&L(HRP)secondary antibody(ab6721, 1:10000) (9) The membrane was immersed in TBS-T (w / 0.1% Tween-20) solution and washed three times for 10 minutes. (10) The bands on the membrane were detected by ChemiDoc using ECL solution, an HRP substrate. (11) The intensity of the detected bands was quantified using Image J software (NIH).

[0109] Experimental Example 3-3: Evaluation of the animal-based 4R-tauopathy inhibitory effect of GV1001 A. Evaluation of tauopathy improvement effects using Tau-BiFC imaging and AT8 immunofluorescence staining (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining were performed using brain tissue from a TauP301L-BiFC mouse model administered the drug for 5 months from 7 to 12 months of age.

[0110] Photographs of (A) Tau-BiFC imaging and (B) AT8 immunofluorescence staining results for the sensorimotor cortex, motor cortex, hippocampus, and substantia nigra are shown in Figures 5 to 8, along with graphs showing the quantitative analysis results of the respective fluorescence levels: Figure 5 (sensorimotor cortex), Figure 6 (motor cortex), Figure 7 (hippocampus), and Figure 8 (substantia nigra). The results are also shown in Table 8 below. Analysis was performed using one-way ANOVA with Dunnett's multiple comparison test ( * p<0.05, ** p<0.01, *** p<0.001, ns not significant compared to vehicle).

[0111] [Table 8]

[0112] Interpretation of brain tissue imaging results in the 4R TauP301L-BiFC mouse model (Tau-BiFC) Brain tissue was collected from a TauP301L-BiFC mouse model that had been administered 63 times over a 5-month period, from -7 to 12 months of age. Autofluorescence in the brain tissue was removed with Sudan black B stain, and Tau-BiFC fluorescence imaging was performed. In a 12-month-old TauP301L-BiFC mouse model, Tau-BiFC fluorescence levels were quantitatively analyzed in the somatosensory cortex, motor cortex, and hippocampus (CA1), where 4R-tauopathy due to the formation of tau oligomers, as expressed by Tau-BiFC fluorescence, is predominant. The GV1001 drug group showed a significant inhibitory effect on tau oligomer formation compared to the vehicle group. Furthermore, the evaluation of motor performance in Experimental Example 2 showed that the GV1001 group showed significantly improved motor performance compared to the vehicle group. Therefore, we further examined the Tau-BiFC fluorescence level in the substantia nigra, a region located in the basal ganglia that plays an important role in regulating physical activity. As a result, the Tau-BiFC fluorescence level was significantly reduced in the 0.1 mpk (mg / kg) and 2 mpk GV1001 drug groups compared to the vehicle group. This confirmed the inhibitory effect of tau oligomer formation.

[0113] -Tau-BiFC fluorescence levels were significantly reduced in the motor cortex, hippocampus, and substantia nigra in the group treated with the reference compound LMTM (15 mpk) compared to the vehicle group, confirming its inhibitory effect on tau oligomer formation.

[0114] Specifically, in the somatosensory cortex, the 0.1 mpk, 1 mpk, and 2 mpk GV1001 drug groups were found to significantly reduce Tau-BiFC fluorescence levels by 0.71-fold, 0.68-fold, and 0.63-fold, respectively, compared to the vehicle group.

[0115] In the motor cortex, the 1 mpk and 2 mpk GV1001 drug groups were found to significantly reduce Tau-BiFC fluorescence levels by 0.56-fold and 0.73-fold, respectively, compared with the vehicle group.

[0116] In the hippocampus (CA1) region, the 1 mpk and 2 mpk GV1001 drug groups were found to significantly reduce Tau-BiFC fluorescence levels by 0.77-fold and 0.54-fold, respectively, compared with the vehicle group.

[0117] In the substantia nigra region, the 0.1 mpk and 2 mpk GV1001 drug groups were found to significantly reduce Tau-BiFC fluorescence levels by 0.56-fold and 0.68-fold, respectively, compared to the vehicle group.

[0118] Interpretation of brain tissue imaging results (AT8 immunofluorescence staining) in the 4R TauP301L-BiFC mouse model Brain tissue was collected from TauP301L-BiFC mice, which were administered 63 doses over a 5-month period from 7 to 12 months of age. Immunofluorescence staining was performed using the tau hyperphosphorylation antibody AT8 (phospho-Tau S202 / T205), followed by quantitative analysis of AT8 fluorescence levels in the sensory motor cortex, motor cortex, hippocampus (CA1), and substantia nigra using brain tissue fluorescence imaging. The GV1001 drug group demonstrated significantly greater suppression of tau hyperphosphorylation than the vehicle group. In the group treated with the reference compound LMTM (15 mpk), AT8 fluorescence levels were significantly reduced in the sensorimotor cortex, motor cortex, and hippocampus compared to the vehicle group, confirming the inhibitory effect of tau hyperphosphorylation.

[0119] Specifically, in the somatosensory cortex, the 0.1 mpk, 1 mpk, and 2 mpk GV1001 drug groups were found to significantly reduce AT8 fluorescence levels by 0.75-fold, 0.63-fold, and 0.61-fold, respectively, compared to the vehicle group.

[0120] In the motor cortex region, the 0.1 mpk, 1 mpk, and 2 mpk GV1001 drug groups were found to significantly reduce AT8 fluorescence levels by 0.74-fold, 0.74-fold, and 0.50-fold, respectively, compared to the vehicle group.

[0121] In the hippocampus (CA1) region, it was confirmed that the 1 mpk and 2 mpk GV1001 drug groups significantly reduced AT8 fluorescence levels by 0.64-fold and 0.66-fold, respectively, compared to the vehicle group.

[0122] In the substantia nigra region, the 2mpk GV1001 drug group was found to significantly reduce AT8 fluorescence levels by 0.42-fold compared to the vehicle group.

[0123] B. Analysis of brain lysates from TauP301L-BiFC mice Brain lysates were collected from TauP301L-BiFC mice that had been administered the drug for 5 months, from 7 to 12 months of age. 1) Soluble fraction samples and 2) Insoluble fraction samples were immunoblotted using antibodies against total tau (Tau5) and phosphorylated tau (pS199, pS396). The results are shown in Figure 9 (Figure 9a: sensorimotor cortex, motor cortex; Figure 9b: hippocampus, substantia nigra) and Figure 10. The results of quantitative analysis of the levels of total tau and phosphorylated tau are also shown in the graphs. The results are shown in Table 9 below. Analysis was performed by one-way ANOVA with Dunnett's multiple comparison test ( * p<0.05, ** p<0.01, *** p<0.001, ns not significant compared to vehicle).

[0124] [Table 9]

[0125] Tau immunoblot analysis using brain lysates from the 4R TauP301L-BiFC mouse model Brain lysates from TauP301L-BiFC mice were administered 63 doses over a 5-month period, from 7 to 12 months of age. Soluble and insoluble fractions were collected. Tau immunoblotting was performed using antibodies against total tau (Tau5) and phosphorylated tau (pS199, pS396). The band intensities of total and phosphorylated tau were quantitatively analyzed. Tau immunoblotting of brain lysates from the -TauP301L-BiFC mouse model revealed bands of 85 kDa and 76 kDa in human Tau-BiFC (hTau-VN173, hTau-VC155), and bands of 50-70 kDa in mouse Tau (mTau). Tau immunoblotting using the soluble fraction showed a significant decrease in total and phosphorylated tau levels in the GV1001 drug group compared to the vehicle group, confirming the inhibitory effect on the formation of soluble tau oligomers and phosphorylated tau. Furthermore, tau immunoblotting using the insoluble fraction also showed a significant decrease in total tau levels in the GV1001 drug group compared to the vehicle group, confirming that GV1001 drug inhibits the formation of insoluble tau aggregates.

[0126] -The reference compound LMTM (15mpk) significantly reduced total and phosphorylated tau levels in mice compared to the vehicle group.

[0127] More specifically, the soluble fraction was as follows:

[0128] For total tau, the 2 mpk GV1001 drug group significantly reduced total hTau by 0.79-fold compared to the vehicle group, and the 0.1 mpk, 1 mpk, and 2 mpk GV1001 drug groups significantly reduced total mTau by 0.77-fold, 0.69-fold, and 0.71-fold, respectively.

[0129] For -phosphorylated tau (pS199), hTau pS199 was significantly reduced by 0.74-fold and 0.57-fold in the 0.1mpk and 2mpk GV1001 drug groups compared to the vehicle group, while mTau pS199 was significantly reduced by 0.58-fold, 0.56-fold, and 0.46-fold in the 0.1mpk, 1mpk, and 2mpk GV1001 drug groups, respectively, compared to the vehicle group.

[0130] For -phosphorylated tau (pS396), hTau pS396 was significantly reduced by 0.67-fold, 0.68-fold, and 0.51-fold in the 0.1mpk, 1mpk, and 2mpk GV1001 drug groups compared to the vehicle group, respectively, and mTau pS396 was significantly reduced by 0.62-fold, 0.62-fold, and 0.50-fold in the 0.1mpk, 1mpk, and 2mpk GV1001 drug groups, respectively, compared to the vehicle group.

[0131] The insoluble fraction was as follows:

[0132] In the 0.1mpk, 1mpk, and 2mpk GV1001 drug groups, total tau levels were significantly reduced by 0.19-fold, 0.34-fold, and 0.29-fold, respectively, compared with the vehicle group.

[0133] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Terms preceded by a noun without a number are not intended to denote a limitation of quantity, but rather to denote the presence of one or more of the referenced noun item.

[0134] Reciting a range of values ​​is merely an easy way of referring to each separate value falling within the range individually, and unless expressly stated otherwise, each value is incorporated herein as if it were individually set forth in the specification. All range endpoints are included within the range and are independently combinable.

[0135] All methods recited herein can be performed in any suitable order unless otherwise noted or clearly contradicted by context. The use of any and all embodiments or exemplary language (e.g., "such as"), unless included in the claims, is merely to better describe the invention and does not limit the scope of the invention. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs.

[0136] Preferred embodiments of the present invention include the best mode known to the inventors for carrying out the invention. Variations of the preferred embodiments will become apparent to those skilled in the art upon reading the foregoing description. The inventors expect that such variations will be readily apparent to those skilled in the art, and also expect that the invention may be practiced otherwise than as described herein. Accordingly, this invention includes equivalents of the subject matter recited in the appended claims and all modifications thereof as permitted by patent law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless expressly stated to the contrary herein or clearly contradicted by context. While the present invention has been particularly shown and described with reference to exemplary embodiments, those skilled in the art will recognize that various changes in form and detail can be made therein without departing from the spirit and scope of the invention as defined by the following claims.

Claims

1. A pharmaceutical composition for preventing or treating 4R tauopathy, comprising a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient, The pharmaceutical composition, wherein the 4R tauopathy is progressive supranuclear palsy.

2. A food composition for preventing or ameliorating 4R tauopathy, comprising a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient, The food composition, wherein the 4R tauopathy is progressive supranuclear palsy.

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