New peptide pp1 of FKBP52 and derivatives and use of the therapeutic peptides that inhibit pathological TAU aggregation for therapeutic purposes

Abstract

The invention is directed to peptide fragments of FKBP52 that inhibit Tau protein aggregation and ameliorate tauopathies like Alzheimer's Disease (AD). It also involves modifications to these peptides to improve their pharmacokinetic and pharmacodynamic properties.

Description

[0001] NEW PEPTIDE PPI OF FKBP52 AND DERIVATIVES AND USE OF THE THERAPEUTIC PEPTIDES THAT INHIBIT PATHOLOGICAL TAU AGGREGATION FOR THERAPEUTIC PURPOSES

[0002] REFERENCE TO A SEQUENCE LISTING

[0003] In accordance with 37 CFR §1.833-1835 and 37 CFR§ 1.77(b)(5) , the specification makes reference to a Sequence Listing submitted electronically as a .xml file named "E_sequence listing PI025882.WO.xml". The .xml file was generated on April 10, 2025 and is 5,630 bytes in size. The entire contents of the Sequence Listing are hereby incorporated by reference.

[0004] FIELD OF THE INVENTION

[0005] The invention relates to the fields of peptide chemistry and medicine including to peptide- based neuroprotection and treatment of tauopathies using specific peptides derived from peptide PPI of FK506 binding protein 52 identified herein as FKBP52.

[0006] BACKGROUND OF THE INVENTION

[0007] A large number of neurodegenerative diseases, including Alzheimer Disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), and frontotemporal lobar degeneration due to the mutation of Tau gene (FTLD-Tau) are characterized by intra-neuronal aggregates of Tau, which are hallmarks of those disorders of the human brain now called tauopathies. In FTLD-Tau, different Tau mutations have been described (i.e.,Tau- P301S, Tau-P301L) and about half of the known mutations show effects at the level of the Tau protein, reducing its normal function and increasing its propensity to assemble into abnormal filaments; Goedert M, Tau gene mutations and their effects. Mov DISORD, 2005, 20 Suppl 12, S45- 52. Tau proteins are widely expressed in the central nervous system, predominantly in neurons where they play a key role in regulating microtubule dynamics, axonal transport and neurite outgrowth; Weingarten MD, Lockwood AH, Hwo SY, Kirschner MW (1975) A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A 72, 1858-1862. Avila J, Lucas JJ, Perez M, Hernandez F, Role of tau protein in both physiological and pathological conditions. PHYSIOL REV, 2004, 84, 361-384. Alternative splicing of Tau results in six isoforms present in the adult brain that differ in their sizes and in their effects on microtubular dynamics; Goedert M, Jakes R (1990) Expression of separate isoforms of human tau protein: correlation with the tau pattern in brain and effects on tubulin polymerization. EMBO J 9, 4225-4230. Tau is a natively unfolded protein; its aggregation is a multistep process converting a soluble and monomeric Tau form into an insoluble, hyperphosphorylated and filamentous form;. Barghom S, Mandelkow E (2002) Toward a unified scheme for the aggregation of tau into Alzheimer paired helical filaments . BIOCHEMISTRY 41, 14885-14896.

[0008] During this process, transient small oligomeric species do form filaments that specifically present a twist appearance in AD brains and are the major source of higher Tau aggregates called NeuroFibrillary Tangles (NFTs). Growing evidence suggest that NFTs do not appear to be the main toxic entities leading to disease but it is rather the intermediate entity, such as oligomeric soluble forms of Tau, that could be responsible for toxicity and disease; Santacruz K, Lewis J, Spires T, Paulson J, Kotilinek L, Ingelsson M, Guimaraes A, DeTure M, Ramsden M, McGowan E, Forster C, Yue M, Ome J, Janus C, Mariash A, Kuskowski M, Hyman B, Hutton M, Ashe KH, Tau suppression in a neurodegenerative mouse model improves memory function. SCIENCE 2005, 309, 476-481. Huang Y, Wu Z, Zhou B, ) Behind the curtain of tauopathy: a show of multiple players orchestrating tau toxicity. CELL MOL LIFE SCI 2016, 73, 1-21.

[0009] The inventors have previously discovered that the protein named FKBP52 interacts physically and functionally with Tau showing an antagonist effect of FKBP52 on Tau’s tubulin assembly; Chambraud B, Sardin E, Giustiniani J, Dounane O, Schumacher M, Goedert M, Baulieu EE, A role for FKBP52 in Tau protein function. PROC NATL ACAD SCI U S A, 2010 107, 2658- 2663. The inventors considered that FKBP52 might represent a promising target, leading to innovative therapeutic attempts centered on Tau. FKBPs (FK506 Binding Proteins) are a family of ubiquitously expressed protein folding chaperones with a peptidyl prolyl cis / trans isomerase (PPiase) activity. The FKBPs differ by the structure of domains and show different subcellular localizations, strongly suggesting specific functions for each FKBP; Gothel SF, Marahiel MA (1999) Peptidyl -prolyl cis-trans isomerases, a superfamily of ubiquitous folding catalysts. Cell Mol Life Sci 55, 423-436. These proteins are particularly abundant in the nervous system and involved in neurodegenerative disorders; Chattopadhaya S, Harikishore A, Yoon HS (2011) Role of FK506 binding proteins in neurodegenerative disorders. CurrMed Chem 18, 5380-5397. FKBP52 (FKBP of MW 52 kDa) was first cloned in our laboratory and its 3D structure revealed a modular organization with four functional domains; Chattopadhaya S, Harikishore A, Yoon HS (2011) Role of FK506 binding proteins in neurodegenerative disorders. Curr Med Chem 18, 5380-5397.

[0010] Two consecutive FKBP domains have been determined (FK1 AA 31-139, FK2 AA 149- 267). While the second FK2 domain shares 34% of identity with FK1, only FK1 presents peptidyl- prolyl isomerase (PPiase) activity; Chambraud B, Rouviere-Fourmy N, Radanyi C, Hsiao K, Peattie DA, Eivingston DJ, Baulieu EE (1993) Overexpression of p59-HBI (FKBP59), full length and domains, and characterization of PPiase activity. Biochem Biophys Res Commun 196, 160- 166. The C-Terminal part of FKBP52 contains additional functional domains such as a tetratricopeptide repeat domain that displays a co-chaperon activity and which serves as binding site for molecular chaperone HSP90; Bose S, Weikl T, Bugl H, Buchner J (1996) Chaperone function of Hsp90-associated proteins. Science 274, 1715-1717. Radanyi C, Chambraud B, Baulieu EE (1994) The ability of the immunophilin FKBP59-HBI to interact with the 90-kDa heat shock protein is encoded by its tetratricopeptide repeat domain. Proc Natl Acad Sci U S A 91, 11197-11201.

[0011] Finally an a-helix in its extreme C-terminus includes a putative calmodulin binding site; Massol N, Lebeau MC, Renoir JM, Faber LE, Baulieu EE (1992) Rabbit FKBP59-heat shock protein binding immunophillin (HBI) is a calmodulin binding protein. Biochem Biophys Res Commun 187, 1330-1335. FKBP52 protein expression is strongly decreased in the frontal cortex of AD and FTLD-Tau brains and this decrease is highly correlated with the accumulation and aggregation of pathological Tau; Giustiniani J, Sineus M, Sardin E, Dounane O, Panchai M, Sazdovitch V, Duyckaerts C, Chambraud B, Baulieu EE (2012) Decrease of the immunophilin FKBP52 accumulation in human brains of Alzheimer's disease and FTDP-17. J Alzheimers Dis 29, 471-483. The inventors previously reported that FKBP52 is able to induce Tau oligomerization depending of the nature of Tau suggesting an involvement of FKBP52 in Tau aggregation; Giustiniani J, Guillemeau K, Dounane O, Sardin E, Huvent I, Schmitt A, Hamdane M, Buee L, Landrieu I, Lippens G, Baulieu EE, Chambraud B (2015) The FK506-binding protein FKBP52 in vitro induces aggregation of truncated Tau forms with prion-like behavior. FASEB J. Giustiniani J, Chambraud B, Sardin E, Dounane O, Guillemeau K, Nakatani H, Paquet D, Kamah A, Landrieu I, Lippens G, Baulieu EE, Tawk M (2014) Immunophilin FKBP52 induces Tau-P301L filamentous assembly in vitro and modulates its activity in a model of tauopathy. Proc Natl Acad Sci U S A 111, 4584-4589. Recently, the inventors have increased their knowledge of Tau-FKBP52 interaction using different biochemical and biophysical approaches including Nuclear Magnetic Resonance (NMR) studies; Kamah A, Cantrelle FX, Huvent I, Giustiniani J, Guillemeau K, Byrne C, Jacquot Y, Landrieu I, Baulieu EE, Smet C, Chambraud B, Lippens G (2016) Isomerization and Oligomerization of Truncated and Mutated Tau Forms by FKBP52 are Independent Processes. J Mol Biol 428, 1080-1090. Indeed, FK1 and FK2 domains of FKBP52 are able to interact with one hexapeptide of Tau (AA 306-311) called PHF6 known to be highly involved in Tau aggregation; Kamah, A, et al. supra, von Bergen M, Friedhoff P, Biemat J, Heberle J, Mandelkow EM, Mandelkow E (2000) Assembly of tau protein into Alzheimer paired helical filaments depends on a local sequence motif ((306)VQIVYK(311)) forming beta structure. Proc Natl Acad Sci U S A 97, 5129-5134.

[0012] This hexapeptide, which self-aggregates, is largely used to study amyloid formation; Schirmer C, Lepvrier E, Duchesne L, Decaux O, Thomas D, Delamarche C, Gamier C (2016) Hsp90 directly interacts, in vitro, with amyloid structures and modulates their assembly and disassembly. Biochim Biophys Acta 1860, 2598-2609.

[0013] The peptides according to the invention are characterized by their properties of active peptides corresponding to their ability to form a complex with Tau.

[0014] SUMMARY OF THE INVENTION

[0015] The invention is broadly directed to PPI of FKBP52 (Gene ID: 2288) that inhibit Tau protein aggregation and in order to ameliorate tauopathies like Alzheimer’s Disease (AD). It also involves modifications to these peptides to improve their pharmacokinetic and pharmacodynamic properties. The invention is also broadly directed to prevention or treatment of tauopathies and other diseases, disorders or conditions affected by aggregation of Tau protein. Non-limiting aspects of the invention include the following embodiments. The active peptide according to the invention concerns molecules having the capacity to be bound to Tau protein motifs. Such active peptides can be PPI or a fragment of PPI (SEQ ID NO: 1) that contains the 15 amino acids of N- terminal part of PPI linked to a peptidic sequence excluding PPI sequences. One aspect of the invention is directed to a method for preventing, reducing the severity of, or treating a tauopathy comprising administering to a subject in need thereof a peptide fragment of FKBP52 or a modified form thereof. The tauopathy may be Alzheimer’ s Disease (AD), familial FTLD-Tau or progressive supranuclear palsy (PSP) or the other tauopathy or tau-related disorders described herein.

[0016] In one embodiment, the peptide fragment used in this method comprises, consists essentially of, or consists of PPI (SEQ ID NO: 1). In some embodiments, the peptide fragment comprises, consists essentially of, consists of a modified form of PPI (SEQ ID NO: 1) having backbone protection or a modified N or C terminus or consists of a modified form of PPI (SEQ ID NO: 1) that further comprises a cell penetrating peptide linked to PPI such as YARAAARQARA (SEQ ID NO: 2). As used herein the term “consists essentially of’ refers to a peptide that prevents or reduces the severity of a tauopathy or its symptoms or that decreases the incidence or degree of aggregation of Tau protein or of NFTs.

[0017] Another aspect of the invention is directed to a peptide or peptide product such as a modified PPI peptide comprising a fragment of FKBP52 that inhibits Tau protein aggregation. In some preferred embodiments, the peptide or modified peptide is PPI. The modified peptide many have a modified backbone or a modified C or N terminus. In one embodiment, the modified peptide can be CPP1 that has an N terminal cysteine residue and do not contain le peptide P35 (containing 35 amino acids) or the peptide P50 (containing 50 amino acids). In some embodiments, the modified peptide may further comprise a cell penetrating peptide, such as the cell penetrating peptide YARAAARQARA (SEQ ID NO: 2).

[0018] Another aspect of the invention is the delivering of peptides of the present invention using one or more modalities. The present invention provides vectors that package nucleic acid of the invention encoding peptides. Vectors of the present invention may be used to deliver the nucleic acid or modified nucleic acid encoding a peptide of interest to a cell such as neurons, a local tissue site such as brain or a subject. These vectors may be of any kind, including DNA vectors, RNA vectors, plasmids, viral vectors and particles. Viruses, which are useful as vectors include, but are not limited to lentiviral vectors, adenoviral vectors, adeno-associated viral (AAV) vectors.

[0019] Vectors can comprise native or non-native promoters operably linked to the nucleic acid coding for peptides of the invention. The promoters selected may be strong, weak, constitutive, inducible, tissue specific, development stage- specific, and / or organism specific.

[0020] In some embodiments, the optimal promoter may be selected based on its ability to achieve minimal expression of the peptide of interest of the invention.

[0021] In some embodiments, lentiviral vehicles / particles may be used as delivery modalities. Lentiviruses are subgroup of the Retroviridae family of viruses, named because reverse transcription of viral RNA genomes to DNA is required before integration into the host genome. As such, the most important features of lentiviral vehicles / particles are the integration of their genetic material into the genome of a target / host cell. Some examples of lentivirus include the Human Immunodeficiency Viruses: HIV-1 and HIV-2, the Simian Immunodeficiency Virus (SIV), feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV).

[0022] Typically, lentiviral particles making up the gene delivery vehicle are replication defective on their own (also referred to as “self-inactivating”). Lentiviruses are able to infect both dividing and non-dividing cells by its entry through the intact host nuclear envelope (Naldini L et al., Curr. Opin. Biotechnol, 1998, 9: 457-463).

[0023] Lentivirus vectors used may be selected from, but are not limited to pLenti, pLenti6, pULTRA, pInducer20, pl 97. Delivery of any nucleic acid coding for peptides of the present invention may be achieved using recombinant adeno-associated viral (AAV) vectors. Such vectors or viral particles may be designed to utilize any of the known serotype capsids or combinations of serotype capsids. Capsids may include but not limited to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, and AAV12.

[0024] AAV vectors include not only single stranded vectors but self-complementary AAV vectors (scAAVs). scAAV vectors contain DNA which anneals together to form double stranded vector genome. By skipping second strand synthesis, scAAVs allow for rapid expression in the cell.

[0025] In some embodiments, nucleic acid coding for the peptide of interest may be administered in one or more AAV particles.

[0026] BRIEF DESCRIPTION OF THE FIGURES

[0027] Figures 1A-1E: FKBP52 inhibits PHF6 aggregation in vitro. FIGURE 1A: The three dimensional structure of FKBP52 (AA 1-459) showing the main structural domains. Functional domains are colored: FK1 (red), FK2 (green) and TPR (TetratricoPeptideRepeats, purple) domains. The cartoon is generated with Pymol v0.99 and DOG 2.0. Figure IB: ThT fluorescence analysis of PHF6 (200pM, blue curve) aggregation with or without FKBP52 (2.5pM, grey curve) over time (60min). Figure 1C: Electron microscopy images obtained for the corresponding samples (Blue for PHF6 and grey for PHF6 / FKBP52). Figure ID: ThT fluorescence analysis of PHF6 (200pM, blue curve) aggregation stopped by the adding of FKBP52 (2.5pM, grey curve) during elongation phase assembly (see black arrow) over time (60min). Figure IE: Electron microscopy images of the corresponding samples (Blue / top panel for PHF6 and grey / bottom panel for PHF6 / FKBP52). Figures 2A-2F: FK2 domain of FKBP52 inhibits PHF6 aggregation in vitro. Figures 2A- 2C: ThT fluorescence analysis of PHF6 (200|iM, blue / top curve) aggregation incubated with FKBP52 (5|iM, grey / bottom curve), FK1 (5pM, red / mid curve), FK2 (2.5pM, greenbottom curve) or FK1+2 (2.5|iM, yellow curve) over time (60min). Figure 2D: Superposition of the three dimensional structure of FK1 (red) and FK2 (green). The cartoon is generated with Pymol v0.99. Figure 2E: Electron microscopy images obtained for the corresponding samples. Scale bar: 0.5pm. Figure 2F: ThT fluorescence analysis of PHF6 (200pM, blue / top curve) aggregation stopped by the adding of FK2 (2.5pM, green / lowern curve) during elongation phase assembly (see black arrow) over time (60min).

[0028] Figures 3A-3C: FK2 domain inhibits Tau-P301L aggregation in vitro. Figure 3A, panel 1: ThT fluorescence analysis of Tau-P301L (5pM, blue / top curve) aggregation (co-incubation with Tau seeds) with or without FK2 (50pM, green / mid curve) over time (24 hours). Incubation of Tau - P301L without Tau seeds does not show aggregation (grey / bottom curve). Figure 3A, panel 2: Histograms represent the fluorescence intensity measured at 24h. Statistical analysis was performed using Student’s t-test; n = 3, ***p < 0.001; ± SEM. Figure 3B, top panel: Spin-down assays of Tau-P301L. The soluble fraction of Tau-P301L is analyzed by western blot. Figure 3B, bottom panel: Quantification of Tau signals is represented by histograms; the incubation of Tau- P301L alone shows 100% of soluble proteins after centrifugation (grey / 3rd histogram). Statistical analysis was performed using Student’s t-test; n = 4, ***p < 0.001; ± SEM. Figure 3C, comparative panels 1-3: Electron microscopy images obtained for the corresponding samples shows the presence of Tau fibers (see magnification) excepting in Tau-P301L incubated alone. Scale bar =0.5pm. Figures 4A-4B: Synthetic peptides obtained from FK2 domain inhibit Tau-P301L aggregation in vitro. Figure 4A: Three dimensional structure of FK2 (AA 146-526) and PPI (AA 147-211) is generated with Pymol v0.99. Figure 4B, top panel: ThT fluorescence analysis of Tau- P301L (5pM, blue / 2ndfrom top curve) aggregation (co-incubation with Tau seeds) with or without PPI (50pM, green / 3rdfrom top curve) or PPI scramble (50 pM, yellow / top curve) over time (24 hours). Incubation of Tau-P301L without Tau seeds does not show aggregation (grey / bottom curve). Figure 4B, lower panel: Histograms represent the fluorescence intensity measured at 24h. Statistical analysis was performed using Student’s t-test; n = 3,*p<0.05, ***p < 0.001; ± SEM.

[0029] Figures 5A-5B: Lentiviral (Figure 5A) or AAV (Figure 5B) plasmid coding for the peptide PPI and respectively named P197-PP1 and AAV-hSyn-PPl. MND is a promotor. An IRES (internal ribosome entry site) is used to co-express different under the same promotor.

[0030] Materials and methods

[0031] Protein purification. Recombinant wild-type Tau and Tau with P301L mutation proteins was expressed in Escherichia coli (E. coli, BL21) and purified as described; Goedert M, Jakes R (1990) Expression of separate isoforms of human tau protein: correlation with the tau pattern in brain and effects on tubulin polymerization. EMBO J 9, 4225 -4230. Combs B, Tieman CT, Hamel C, Kanaan NM (2017) Production of recombinant tau oligomers in vitro. Methods Cell Biol 141, 45-64.

[0032] Recombinant FKBP52 or FK1 (EGVDISPKQD

[0033] EGVLKVIKREGTGTEMPMIGDRVFVHYTGWLLDGTKFDSSLDRKDKFSFDLGKGEVIK AWDIAIATMKVGEVCHITCKPEYAYGSAGSPPKIPPNATLVFEVELF)(SEQ ID NO: 4); FK2 (DLTEEEDGGIIRRIQTRGEGYAKPNEGAIVEVALEGYYKDKLFDQ

[0034] RELRFEIGEGENLDLPYGLERAIQRMEKGEHSIVYLKPSYAFGSVGKEKFQIPPNAELKY ELHLKSFEKAKE)(SEQ ID NO: 5) or FK1-FK2 domains are expressed in E. coli as fusion proteins tagged with a GST and then purified through affinity, size exclusion and anion exchange column steps in a final buffer MES 50pM PH 6.4. Quantification of proteins was carried out using BCA (Thermofisher Company) quantification assay.

[0035] Peptides synthesis. PPI

[0036] (DGGIIRRIQTRGEGYAKPNEGAIVEVALEGYYKDKLFDQRELRFEIGEGENLDLPYGLER AIQRM)(SEQ ID NO: 1) and PPI Sc (scramble:

[0037] LKYTEIRGQDLIIVARRKENPGYDARELGEAPGQRGEMAFREIVGELIYGQFEDRLKYLE GGDNI)(SEQ ID NO: 3 were synthesized by Proteogenix (Schiltigheim, FRANCE) with a purity >95% as determined by mass spectrometry.

[0038] PHF6 peptide synthesis and peptide assembly assays. Lyophilized N-acetyl peptide amide Ac-VQIVYK-NH2 is prepared as described previously by Schirmer, et al., supra. Briefly 2mg of peptide was resuspended in 400pl of water and then centrifuged at 100,000g for lOmn at room temperature. Peptide concentration in supernatant is obtained by measuring the absorbance at 280nm using a molar absorption coefficient of 1450 M-l.cm-1. Assembly assays were performed in polymerization buffer: lOmM MOPS, 150mM NaCl Ph7.4 (Pl-Buffer) at 25°C as described

[0021] . Pre-dilution of 200pM of PHF6 in water are maintained 2 hours at room temperature and then incubated at 25 °C in spectrophotometer (Spetramax Molecular Devices) before adding a mix containing 2.5pM of protein or polypeptides (FKBP52, FK1, FK2, FK1-FK2, PPI) in Pl-Buffer containing lOpM Thioflavine-T (ThT) (Abeam). Measure of fluorescence was monitored every

[0039] 30s during 1 hour at 480nm (excitation at 450nm). Proteins or peptides were added at different time points of PHF6 polymerization: at the beginning of PHF6 polymerization (co-polymerization assay) or during PHF6 elongation (elongation assay).

[0040] Effect of FK2, PPI on Tau-P301L aggregation. Tau-P301L protein is used in all aggregation assays. Tau seeds are prepared during a first polymerization: 15pM of monomeric Tau-P301L was incubated at 37°C in polymerization buffer (Pl-buffer): NaH2PO4 25mM, Na2HPO4 25mM, NaCl 25mM, EDTA 5mM PH 6,6 with heparin 7.5pM, Thioflavine-T 50pM, and Dithiothreitol (DTT) 0.3mM in a final volume of lOOpl. Fluorescence was monitored every 30 minutes during 25 hours at 480nm (excitation at 450nm) using Perkin-Elmer Endvision spectrophotometer. After polymerization, samples (lOOpl) were centrifuged at 20,000 g for 1 hour at 4°C. The pellets were resuspended in lOOpl of Pl -buffer and sonicated (Branson sonicator) at 40%, 3 times of 1 second, in order to obtain small pre-formed Tau fibrils used as seeds for the second polymerization assay. A volume of 18% of Tau seeds was pre-incubated in the same buffer of the first polymerization, with or without polypeptides, during Ih at 37°C. Subsequently, 5pM of monomeric Tau-P301L was added and fluorescence was monitored in the same conditions.

[0041] Spin-down assays. After Tau-P301L aggregation for 24h at 37°C, the samples were collected and centrifuged at 28,000 rpm during 30 min. The supernatant corresponding to the soluble fractions was analyzed by SDS-PAGE. Western-blot was carried out with Tau5 antibody (1:1000; Abeam).

[0042] Transmission Electron Microscopy (TEM). A volume of 10 pl of the sample was placed on a Formvar 400 mesh hexagonal grid (Pelannes Instruments, FCF400-Ni-50) for 30 s. The sample on the grid was stained with uranyl acetate 2% for 1 min. At each step, excess stain was removed by blotting the edge of the grid with filter paper. Grids were observed with a Jeol 1011 transmission electron microscope. Acquisitions were performed with a an Erlangshen charge coupled device camera (Gatan, Pleasanton, CA, USA) by use of Digital Micrograph software.

[0043] Lentiviral cell transduction. In case of the selected peptides are unmodified in their amino acid sequences, we have also the possibility to express them by cellular internalization of a lentivirus vector engineered to produce specific cDNA encoding proteins or peptides of interest and simultaneously a non-chimeric ZsGreen fluorescent protein. The simultaneous detection of ZsGreen proteins will help us to detect transduced neurons that express peptides of interest. Those vectors have been already designed and are currently available. The cDNA encoding human FKBP52 or PPI was inserted into the restriction site of the 197 plasmid (pRRLsin-MND-MCS- Ires2-ZsGreenWPRE, Vectorology platform Vect’UB, U1035 Inserm, Bordeaux) to obtain the concomitant expression of FKBP52 or PPI and ZsGreen proteins. Lentiviruses containing the cDNA coding for FKBP52 or derived-peptides were generated at the vectorology platform Vect’UB (U1035 Inserm, Bordeaux).

[0044] Conjugation of peptides to Cell penetrating peptides: Conjugation of CPP1 with cell penetrating peptides was carried out in a mixture of acetonitrile / 10 mM ammonium acetate. To CPP1 (2 mg, 1 equivalent) under an atmosphere of Argon was added degassed acetonitrile (300 pL), was added 10 mM ammonium acetate (700 pL). To this solution was slowly added C(Npys)YARA (1 equivalent) in degassed 10 mM ammonium acetate (1 mL). The solution immediately turned bright yellow. The solution was mixed for 5 minutes and purified by RP-HPLC on a Cl 8 column with an acetonitrile gradient. The eluted purified peptide was freeze-dried and identified by MALDI-TOF mass spectrometry.

[0045] Amino acids (single and 3-letter codes) and protecting groups for Boc Solid Phase Peptide Synthesis:

[0046] A Ala Alanine Boc-Alanine-OH C Cys Cysteine (for CPPs) Boc-Cys(Npys)-OH

[0047] D Asp Aspartic acid Boc-Asp(Bzl)-OH

[0048] E Glu Glutamic acid Boc-Glu(Bzl)-OH

[0049] F Phe Phenyalanine Boc-Phe-OH

[0050] G Gly Glycine Boc-Gly-OH,

[0051] I He Isoleucine Boc-Ile-OH Boc-Lys(2-Cl-Z)-OH

[0052] L Leu Leucine Boc-Leu-OH

[0053] M Met Methionine Boc-Met-OH

[0054] Asn Asparagine Boc-Asn(Xan)-OH

[0055] P Pro Proline Boc-Pro-OH

[0056] Q Gin Glutamine Boc-Gln-OH

[0057] R Arg Arginine Boc-Arg(Tos)-OH

[0058] V Vai Valine Boc-Val-OH

[0059] Y Tyr Tyrosine Boc-Tyr(2-Br-Z)-OH

[0060] Amino acid (single and 3-letter codes) and protecting groups for Fmoc Solid Phase Peptide Synthesis:

[0061] A Ala Alanine Fmoc-Ala-OH

[0062] C Cys Cysteine Fmoc-Cys(Trt)-OH

[0063] D Asp Aspartic acid Fmoc-Asp(tBu)-OH

[0064] E Glu Glutamic acid Fmoc-Glu(tBu)-OH

[0065] F Phe Phenyalanine Fmoc-Phe-OH

[0066] G Gly Glycine Fmoc-Gly-OH,

[0067] I He Isoleucine Fmoc-Ile-OH

[0068] K Lys Lysine Fmoc-Lys(Boc)-OH

[0069] L Leu Leucine Fmoc-Leu-OH

[0070] M Met Methionine Fmoc-Met-OH

[0071] N Asn Asparagine Fmoc-Asn(Trt)-OH P Pro Proline Fmoc-Pro-OH

[0072] Q Gin Glutamine Fmoc-Gln(Trt)-OH

[0073] R Arg Arginine Fmoc-Arg(Pbf)-OH

[0074] V Vai Valine Fmoc-Val-OH

[0075] Y Tyr Tyrosine Fmoc-Tyr(tBu)-OH

[0076] Abbreviations:

[0077] 2-C1-Z 2-chlorobenzyloxycarbonyl

[0078] 2-Br-Z 2-bromobenzyloxycarbonyl

[0079] Ac Acetyl

[0080] AciO Acetic anhydride

[0081] Boc tert-butyloxycarbonyl

[0082] Bzl Benzyl

[0083] DCC Dicyclohexylcarbodiimide

[0084] DCM Dichloromethane

[0085] DIC Diisopropylcarbodiimide

[0086] DIPEA N,N-Diisoproylethylamine

[0087] DMF Dimethylformamide

[0088] DMS Dimethylsulfide

[0089] Fmoc Fluorenylmethoxycarbonyl

[0090] HB TU TV, TV, TV', TV'-Tetramethyl- O- ( 1 / / -benzotriazol- 1 -yl)uronium hexafluoropho sphate

[0091] MAEDI-TOF Matrix Assisted Easer Desorption Ionisation Time Of Flight mass spectrometry MeOH Methanol

[0092] Mob 4-Methoxybenzyl

[0093] Npys 3-nitro-2-pyridylthio

[0094] Oxyma Ethyl cyano(hydroxyimino)acetate

[0095] Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl RP-HPLC Reversed phase high pressure liquid chromatography tBu tert-butyl

[0096] TIS Triisopropylsilane

[0097] TFA Trifluoroacetic acid

[0098] .TFA Trifluoroacetate salt

[0099] Tos Tosyl

[0100] Trt Trityl

[0101] Xan Xanthyl

[0102] EXAMPLE 1: Results and Discussion of Figures 1 and 2.

[0103] FK2 domain of FKBP52 is able to mimic the effects of the whole protein on PHF6 aggregation in vitro.

[0104] Considering our previous results of FKBP52 / Tau interaction by NMR studies

[0019] , we wondered if FKBP52 and its FK1 / FK2 domains (Fig. 1 A) are able to modulate the aggregation of Tau “PHF6” sequence in vitro. A typical sigmoid curve is obtained during the fibril formation of PHF6 including three phases: a lag-time before growth, called nucleation phase, a fibrillar growth phase named elongation reaching a near complete aggregation to give a plateau phase (Fig. IB, blue curve; Schirmer, et al., supra, Wang CK, Northfield SE, Huang YH, Ramos MC, Craik DJ (2016) Inhibition of tau aggregation using a naturally- occurring cyclic peptide scaffold. Eur J Med Chem 109, 342-349). Using Thioflavine T fluorescence assays, we have shown that FKBP52 is able to fully inhibit PHF6 assembly (Fig. 1 B, grey curve). At the end of the incubation, the corresponding samples were analyzed by electron microscopy confirming the absence of PHF6 filaments when incubated with FKBP52 (Fig. 1C). Also, the adding of FKBP52 is able to stop PHF6 assembly during elongation phase showing the capacity of FKBP52 to inhibit PHF6 aggregation at different step of PHF6 aggregation (Fig. 1C).

[0105] We have next evaluated the effects of purified FK1 and FK2 domains individually or linked together (FK1+2) on PHF6 aggregation. Interestingly, while incubation of FK1 had no apparent effects on PHF6 aggregation (Fig. 2A, Red curve), incubation of FK2 mimicked the inhibition effects of the whole FKBP52 protein (Fig. 2B, Green curve). The addition of FK1+2 showed moderate effects (Fig. 2C, Yellow curve) thus suggesting a possible competition of both domains for PHF6 interaction. Moreover, we cannot exclude the possibility that FK1 might modulate PHF6 or FK2 structures decreasing the ability of FK2 to have its inhibitory aggregation effects. Despite the fact that both FK1 and FK2 domains can interact with PHF6 motifs of Tau and show 34 % of identity (Fig. 2D, [ Wu, et al. supra', Kamah, et al., supra ), our observations highlight the different behavior and contribution of these domains in the modulation of PHF6 aggregation. As expected, electron microscopy analysis of the corresponding samples showed an absence of PHF6 fibers when incubated with FKBP52 or FK2 (Fig. 2E). The presence of PHF6 aggregates in FK1 or FK1+2 conditions did not show particular shapes when compared to the incubation of PHF6 alone (Fig. 2E). Interestingly, ThT fluorescence intensity showed that FKBP52 and FK2 added during PHF6 elongation phase assembly stopped PHF6 aggregation. Altogether, our results highlight the potential of FKBP52 and FK2 to inhibit initiation and elongation steps of PHF6 aggregation (Fig. 2F, black arrow).

[0106] EXAMPLE 2: Results and Discussion of figure 3.

[0107] Both FKBP52 and its FK2 domain inhibit Tau-P301L aggregation in vitro induced by preformed Tau fibrils. Given our previous results, we wondered if FK2 domain is also able to inhibit the aggregation of the whole Tau protein. In order to answer this question, we monitored in vitro the aggregation of the mutated Tau-P301L protein which quickly aggregates in the presence of pre-formed Tau fibrils (seeds) and in a reproducible manner. Using Thioflavine T fluorescence assays, we have shown that FK2 is also able to inhibit Tau-P301L assembly but in a lesser extent than PHF6 (Fig. 3A). Indeed, while FK2 inhibits PHF6 aggregation in a 1:80 stoichiometric ratio, we observed an inhibition of Tau-P301L aggregation in a 10:1 stoichiometric ratio. In these conditions, we observed a decrease of 40.1% (+ / - 8.7%) of Tau-P301L aggregates in the presence of FK2 in comparison to Tau-P301L only incubated with seeds (Fig. 3 A, green and blue curves / histograms; t-test,*** p<0.001). In order to evaluate the solubility of Tau in these conditions, the samples are then centrifuged at the end of the kinetic and the soluble fraction analyzed by western blot (Fig. 3B). While 42.8 % (+ / - 4 %, sem) of Tau-P301L is soluble after aggregation (Blue histogram) in comparison to the control (Grey histogram, 100%), we observed an increase of Tau solubility of 65.3 % (+ / - 4.1 %, sem) when FK2 is added (Green histogram; t- test, *** p<0.001). Electron microscopy analysis of the corresponding samples did not show apparent differences in the shape of Tau aggregates (Fig. 3C). Altogether, our observations suggest that FK2 is able to partially inhibit Tau-P301L aggregation without affecting Tau aggregate structures.

[0108] EXAMPLE 3: Results and Discussion of figure 4.

[0109] Synthetic peptides obtained from FK2 domain inhibit Tau-P301L aggregation in vitro.

[0110] Given the ability of FK2 to slow down Tau-P301L aggregation in vitro, we wondered if smaller amino acid sequences obtained from this domain might reproduce the same effects. FK2 is a structured domain of FKBP52 which is composed of 110 amino acids (Figure 4, AA 146-256). Different truncated parts of FK2 domain were then generated and tested on their solubility and efficacy to inhibit Tau aggregation in vitro. As shown Fig. 4, we have identified a peptide of 65 AA called PPI that is able to prevent Tau-P301L assembly (Fig. 4A and B). Using Thioflavine T fluorescence assays, we observed that Tau-P301L aggregate formation is decreased when incubated in the presence of PPI (61.5%, + / - 3.2 %, sem) in comparison to PPI scramble (PPlsc, 134% (+ / - 28%, sem)) used as control (Fig. 4 B, green and yellow curves / histograms; t-test, * p<0.05).

[0111] DETAILED DESCRIPTION OF THE INVENTION

[0112] Tauopathies. This term refers to a group of neurodegenerative diseases characterized by the abnormal accumulation add aggregation of tau proteins in the brain or nervous system leading to neuronal dysfunction and / or degeneration. A large number of neurodegenerative diseases, including Alzheimer Disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), and frontotemporal lobar degeneration due to the mutation of Tau gene (FTLD-Tau) are characterized by intra-neuronal aggregates of Tau, which are hallmarks of those disorders of the human brain now called tauopathies. Features of tauopathies include misfolding and aggregation of tau proteins which normally stabilize microtubules, formation of neurofibrillary tangles or NFTs composed of abnormal tau proteins and neuronal toxicity and degeneration due to disruption of normal tau function. The methods disclosed herein may treat one or more of the above-mentioned features. Other tauopathies are argyrophilic grain disease (AGD), globular glial tauopathy (GGT), and Primary age-related tauopathy (PART). Besides those tauopathies described above, the invention contemplates treatment of other diseases, disorders, or conditions associated with dysregulation or aggregation of tau protein function or tauopathies associated with various isoforms of tau protein including 3R, 4R or mixed 3R / 4R tau isoforms. Treatment of conditions such as normal brain aging or tauopathy resulting from exposure to biological, chemical or pharmaceutical agents, including alcohol, cannabis, tobacco, cocaine, or other illegal or recreational drugs, which are characterized by at least a degree of tauopathy are also contemplated. In some embodiments, the method disclosed herein will reduce the severity of a tauopathy or its symptoms or decrease the incidence or degree of aggregation of Tau protein or of NFTs, for example, byl, 2, 5, 10, 15, 20% or more.

[0113] According to a particular aspect, the present invention further relates to a peptide fragment of FKBP52 or a modified form thereof, in particular PPI of FK506-binding protein (FKBP52) or a derived active peptide of PPI containing at least 15 Nter sequence of PPI, for preventing, reducing the severity of, or treating a tauopathy in a subject in need thereof. The present invention further provides the use of a peptide fragment of FKBP52 or a modified form thereof, in particular PPI of FK506-binding protein (FKBP52) or a derived active peptide of PPI containing at least 15 Nter sequence of PPI, for preventing, reducing the severity of, or treating a tauopathy in a subject in need thereof.

[0114] It is further provided the use of a peptide fragment of FKBP52 or a modified form thereof, in particular PPI of FK506-binding protein (FKBP52) or a derived active peptide of PPI containing at least 15 Nter sequence of PPI, for the manufacture of a medicament for preventing, reducing the severity of, or treating a tauopathy in a subject in need thereof.

[0115] The peptide fragment and the modified form thereof may be as described throughout the specification.

[0116] The tauopathy and the subject in need thereof may be as described throughout the specification. According to another aspect, the present invention provides an in vitro method for screening active peptides that can prevent, reduce the severity of, or treat a tauopathy, said method comprising the steps of:

[0117] (i) contacting a candidate peptide with a Tau protein, in particular a Tau-P301L protein and / or with a TauA291-397 protein,

[0118] (ii) measuring the level of aggregation of the Tau protein, in particular the Tau-P301L protein and / or the TauA291-397 protein in the presence of said candidate peptide, and

[0119] (iii) comparing the level of aggregation of the Tau protein, in particular the Tau-P301L protein and / or the TauA291-397 protein measured in step (ii) with a control level.

[0120] (iv) selecting the candidate peptide as an active peptide that can prevent, reduce the severity of, or treat a tauopathy depending on the comparison obtained of step (iii).

[0121] In the context of the present invention, the terms “screening”, “identifying”, “selecting” and “testing” candidate peptides can be used indifferently throughout the specification with relation to the method.

[0122] Screening methods may utilize high-throughput techniques, binding assays, functional assays, computational modeling, or other analytical methodologies to identify candidate peptides with optimal performance or functionality for further development.

[0123] According to a particular embodiment, the method for screening is a high-throughput method for screening.

[0124] The level of aggregation of a Tau protein, may include the incidence or degree of aggregation of Tau protein. In a particular embodiment, the candidate peptide is a peptide fragment of FKBP52 or a modified form thereof.

[0125] According to a particular embodiment, the control level is the level of aggregation of the Tau protein, in particular the Tau-P301L protein and / or the TauA291-397 protein, measured in the absence of a candidate peptide or in the presence of a scrambled peptide of the candidate peptide.

[0126] A “scrambled peptide” of a reference peptide as used herein is a peptide having the same amino acid composition as the reference peptide but in a different order.

[0127] In this case, the candidate peptide will be selected in step (iv) as an active peptide that can prevent, reduce the severity of, or treat a tauopathy when the level of aggregation measured in step (ii) is significantly lower than the control level.

[0128] According to a particular embodiment, the control level is the level of aggregation of the Tau protein, in particular the Tau-P301L protein and / or the TauA291-397 protein, measured in the presence of a treatment that is known and proven to be efficient against the aggregation of a Tau protein, in particular the Tau-P301L protein and / or the TauA291-397 protein.

[0129] In this case, the candidate peptide will be selected in step (iv) as an active peptide that can prevent, reduce the severity of, or treat a tauopathy when the level of aggregation measured in step (ii) is at least as low as the control level. In particular, the candidate peptide will be selected in step (iv) as an active peptide that can prevent, reduce the severity of, or treat a tauopathy when the level of aggregation measured in step (ii) is significantly lower than the control level.

[0130] According to a particular embodiment, the control level is the level of aggregation of a Tau protein measured in a cell from a healthy individual. By “healthy” individual, it is understood an individual that is not suffering from a tauopathy, or any illness that may affect the aggregation of the Tau protein in the individual.

[0131] In this case, the candidate peptide will be selected in step (iv) as an active peptide that can prevent, reduce the severity of, or treat a tauopathy when the level of aggregation measured in step (ii) is at least as low as the control level.

[0132] According to a particular embodiment, the control level is the level of aggregation of a Tau protein measured in a cell from an individual suffering from a tauopathy or any illness that may affect the aggregation of the Tau protein in the individual.

[0133] In this case, the candidate peptide will be selected in step (iv) as an active peptide that can prevent, reduce the severity of, or treat a tauopathy when the level of aggregation measured in step (ii) is significantly lower than the control level.

[0134] Methods for measuring the aggregation level of a Tau protein, in particular a Tau-P301L protein and / or a TauA291-397 protein are well known in the art. Such methods include, but are not limited to ThT (Thioflavine T) fluorescence analysis. A level of aggregation is “significantly lower than” a control level if established by a statistical test, in particular by a T-test (e.g. p<0.05).

[0135] A level of aggregation is “at least as low as” a control level if the aggregation level is not significantly different or is significantly lower than the control level as established by a statistical test, in particular by a T-test (e.g. p<0.05). The TauA291-397 protein as used herein may be as described in Lo vestram el al. (Assembly of recombinant tau into filaments identical to those of Alzheimer’s disease and chronic traumatic encephalopathy; ELIFE 2022; l l:e76494. DOI: https: / / doi.org / 10.7554 / eLife.76494).

[0136] Symptoms of tauopathies. The methods as disclosed herein may prevent, treat, ameliorate or reduce the severity of one or more symptoms of a tauopathy. Such symptoms include cognitive / behavioral disorders, such as problems with cognition, vision, language, executive function, and social cognition; changes in personality, social behavior and decision-making abilities; or dementia and memory impairment. Other symptoms include movement disorders such as Parkinsonism, apraxia, alien limb phenomena, and gait disturbances and freezing. Language disorders are also symptoms of some tauopathies and include non-fluent agrammatic primary or progressive aphasia, apraxia of speech. Other symptoms include psychiatric or amnesic symptoms, late-onset cerebellar ataxia and primary lateral sclerosis. Methods for objectively characterizing and quantifying these symptoms are known.. For example, in tauopathies, regional topography of Tau pathology correlates well with clinical symptoms. In vivo molecular imaging using radiotracers provide useful techniques to diagnose the intensity of Tau accumulation and the progression of the tauopathy in the brain of suffering patients. The invention may provide significant relief for one or more symptoms, for example, a reduction in intensity, frequency or progression of such symptoms by at least 1, 2, 5, 10, 15, 20% or more as determined by objective or subjective medical diagnostic procedures known in the art.

[0137] FKBP52 (FK506 Binding Proteins of -52 kDa) is a peptidyl-prolyl cis / trans isomerase implicated in the folding and function of its target protein. Besides its enzymatic activity, FKBP52 also displays a chaperone activity. While FKBP52 has mainly been described as a chaperone of steroid receptors, it is also involved in many other biological processes. FKBP52 is largely distributed and particularly abundant in the nervous system. The inventors have previously shown that FKBP52 prevents in vitro microtubule formation through its interaction with Tubulin and Tau and that FKBP52 is able to interact with the “PHF6” sequences of Tau by NMR experiments. Moreover, they have shown that FKBP52 levels are strongly decreased in the frontal cortex of AD and familial FTLD-Tau brains and that this decrease is highly correlated with the accumulation and aggregation of pathological Tau. Both mechanisms and timeline of this FKBP52 decrease in AD and other tauopathic brains are still unknown.

[0138] Peptides and Peptide-related products. These terms refer to peptides, peptide fragments such as subsections of a longer, full-length peptide, modified peptides such as peptides substituted with one or more D-amino acid residues or D-amino acid peptides, peptides incorporating nonnatural residues such as beta-amino acids, methylation of backbone residues of a peptide, modification of the N-terminus or C-terminus of a peptide, including acetylation of the N-terminus or amidation of the C-terminus, peptide stapling to form or retain secondary, tertiary or quaternary structures, conjugation to targeting moieties such as transferin or insulin receptors to enhance Blood Brain Barrier (BBB) penetration, conjugation of C or N terminus to a fatty acid, or conjugation or linkage of a fluorescent dye to a peptide, or addition of an affinity tag to the N or C terminus. The peptides disclosed herein such as PPI may be administered as peptides or as any of the peptide products disclosed above, for example, PPI may be conjugated to a CPP or have their N or C terminals modified. The term “modified form” describes a peptide that has been chemically modified or that is structurally distinct from its natural form, for example, a modified form of the PPI peptide excludes the natural unmodified amino acid sequence of these peptide fragments, but would include PPI peptides that have modified N or C termini, have modified peptide backbones (e.g., via methylation), or that are conjugated to other peptides or proteins such as PP1-CPP.

[0139] Peptide fragments or other constructs based on PPI. In some embodiments the peptide or peptide product of the invention will consist or comprise a fragment containing the 15 amino peptide sequence DGGIIRRIQTRGEGY of Nter of PPI or modified fragment of PPI peptides, of sufficient length to protect or reduce the severity of a tauopathy, said peptide fragment of PPI containing at least the five N-terminal residues of PPI between 5, 10, 15, 20, 25 and 30residues in length.

[0140] In another embodiment, the PPI may consist of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or 30 additional residues of the FKBP52 upstream from the N-terminal or downstream from the C terminal of PPI. Peptide products with such additional residues typically will be adequate to protect or reduce the severity of a tauopathy.

[0141] In another embodiment, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or 30 additional peptide residues, which may or may not correspond to the adjacent residues in FKBP52, may be linked to either the N or C terminal of PPI for example, protein tags, CPP sequences, etc. Peptide products with such additional residues typically will be adequate to protect or reduce the severity of a tauopathy.

[0142] In another embodiment, 1, 2, 3, 4, 5 up to 20 residues of PPI may be deleted or substituted with conservative or non-conservative amino acid residues. In another embodiment, 1, 2, 3, 4, 5 up to 20 residues may be inserted into the internal sequences of PPI. Peptide products with such additional modification typically will be adequate to protect or reduce the severity of a tauopathy.

[0143] Some non-limiting examples of the peptides or peptide products described herein are shown below. PPI

[0144] (DGGIIRRIQTRGEGYAKPNEGAIVEVALEGYYKDKLFDQRELRFEIGEGENLDLPYGLER AIQRM) (SEQ ID NO: 1)

[0145] YARA-PP1 (YARAAARQARA-PP1) (SEQ ID NO: 2)

[0146] Cell penetrating peptides or CPPs are short peptides that assist intake or uptake of a peptide product (or nucleic acid) to which they are associated or covalently bound. CPPs may be cationic (e.g. penetratin, oligoarginine), amphipatic (e.g., transportin, TP10, Pep-1), hydrophobic (e.g., azurin, crotamine, maurocalcine, lycosin-1), or chimeric (transportan). CPPs include the YARA peptides disclosed herein as well as other known CPPs, such as those described by, and incorporated by reference to Reissmann, S. et al., New generation of cell-penetrating peptides: functionality and potential clinical application,. J. PEPTIDE SCI. 2021, 27(5), J. Xie, et al, Cellpenetrating peptides in diagnosis and treatment of human diseases: from preclinical research to clinical application, FRONT. PHARMCOL., 2020, 11 ; or H. Derakhshankhah, et al., Cell penetrating peptides: a concise review with emphasis on biomedical applications. BIOMEDICINE & PHARMACOTHERAPY, 2018, 108, 1090.

[0147] Nucleic acids encoding the peptides or peptide products described herein include vectors carrying a coding sequence for the peptides or peptide products disclosed herein, such as those encoding PPI peptides. In some embodiments, a nucleic acid or modified nucleic acid encoding a peptide of interest, such as PPI may be administered instead of a peptide. Typically, the nucleic acid will encode the peptide of interest once it is transported into the cell. Vectors that can transform a target cell such as a neuron with a sequence encoding a peptide as disclosed herein such as PPI include plasmid-based expression vectors, viral expression vectors such as adeno associated virus, herpes simplex virus, or lentiviruses. Such vectors may be delivered to target cells in combination with CPPs, lipid nanoparticles, or synthetic cationic polymers. Modes of administration. The peptides and peptide-related products described herein may be administered to a subject in need of prophylaxis against or treatment for a tauopathy may be administered by any route that contacts the peptide or peptide-related product with nervous system tissue such as neurons. These include systemic administration, direct intranasal administration for example to target a nose-to-brain pathway bypassing the BBB which may optionally be in combination with chitosan or another absorption enhancer, administration in combination with a cell penetrating peptide or other transporter molecules that facilitate neuronal uptake, administration in an encapsulated form, such as in a liposome or nanoparticle, administration in a form that provides receptor-mediated transcytosis across the BBB, administering targeting axonal import (TAxl) using peptides that deliver payloads to specific neuronal tissues. These are other modes of administration are described by and incorporated by reference to Salameh, T.S. et al, Delivery of therapeutic peptides and proteins to the CNS, ADV. PHARMACOL, 2014, 71, 277; as described by and incorporated by reference to Lalatsa, et al., Strategies to deliver peptide drugs to the brain, MOL. PHARMACEUTICS, 2014, 11(4), 1081; or as described by M. H. Baig, et al., Peptide based therapeutics and their use for the treatment of neurodegenerative and other diseases, BIOMED. & PHARMACOTHERAPY, 2018, 103, 574. The peptides, peptide products, and nucleic acids disclosed herein that encode peptides such as PPI may be administered in vivo, ex vivo, or in vitro.

[0148] Functional effects of administering PPI or PPI modified active peptides derived from FKBP52 include prevention of FKBP52 decrease, increases in free FKBP52, inhibition of interaction between FKBP52 and Tau proteins, blockade of Tau aggregation.

[0149] Peptides of interest are able to interact with Tau proteins. These peptides likely inhibit Tau aggregation by interacting with the polymerizing side of the Tau aggregates. Terminology. The description and specific examples, while indicating embodiments of the technology, are intended for purposes of illustration only and are not intended to limit the scope of the technology. Moreover, recitation of multiple embodiments having stated features is not intended to exclude other embodiments having additional features, or other embodiments incorporating different combinations of the stated features. Specific examples are provided for illustrative purposes of how to make and use the compositions and methods of this technology and, unless explicitly stated otherwise, are not intended to be a representation that given embodiments of this technology have, or have not, been made or tested.

[0150] As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0151] It will be further understood that the terms “comprises” and / or “comprising,” when used in this specification, specify the presence of stated features, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof.

[0152] As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items and may be abbreviated as “ / ”.

[0153] The terms “we” and “us” refer to the inventors or to the contributions of one or more inventors.

[0154] All publications and patent applications mentioned in this specification are herein incorporated by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference, especially referenced is disclosure appearing in the same sentence, paragraph, page or section of the specification in which the incorporation by reference appears. The citation of references herein does not constitute an admission that those references are prior art or have any relevance to the patentability of the technology disclosed herein. Any discussion of the content of references cited is intended merely to provide a general summary of assertions made by the authors of the references and does not constitute an admission as to the accuracy of the content of such references.

[0155] The present invention comprises the following claims / embodiments:

[0156] 1. Embodiment 1 : An embodiment of the present invention is directed to a method for preventing, reducing the severity of, or treating a tauopathy comprising administering to a subject in need thereof PPI of FK506-binding protein (FKBP52) or a derived active peptide of PPI containing at least 15 Nter sequence of PPI .

[0157] 2. Embodiment 2: concerns a method of embodiment 1, wherein the tauopathy is Alzheimer’s Disease (AD), familial FTED-Tau or progressive supranuclear palsy (PSP)

[0158] 3. Embodiment 3: concerns a method of embodiments 1 or 2, wherein the peptide fragment consists of PPI (SEQ ID NO: 1).

[0159] 4. Embodiment 4: concerns a method of embodiments 1, 2 or 3, wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) having backbone protection or a modified N or C terminus.

[0160] 5. Embodiment 5: concerns a method of embodiments 1, 2, 3 or 4, wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) that further comprises a cell penetrating peptide linked to PPI. Embodiment 6: concerns a method of embodiments 1, 2, 3, 4 or 5, wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) that further comprises cell penetrating peptide YARAAARQARA (SEQ ID NO: 2). Embodiment 7: concerns a peptide or modified peptide comprising a fragment of FKBP52 that inhibits Tau protein aggregation. Embodiment 8: concerns a peptide or modified peptide of Embodiment 7 that is PPI. Embodiment 9: concerns a modified peptide of Embodiments 7 or 8 that has a modified backbone or a modified C terminus. Embodiment 10 concerns a modified peptide of Embodiments 7, 8, or 9that is PPI that has an N terminal cysteine residue. Embodiment 11 concerns a modified peptide of Embodiments7, 8, 9 or 10 that further comprises a cell penetrating peptide. Embodiment 12 concerns a modified peptide of Embodiments 7, 8, 9, 10 or 11 that further comprises cell penetrating peptide YARAAARQARA (SEQ ID NO: 2). Embodiment 13 concerns a peptide or modified peptide according to at least one of the Embodiments 7 to 11 wherein the active amino-acid sequence comprises at least one TRG amino acid sequence and the said sequence is less than 30 amino acids. Embodiment 14 concerns a pharmaceutical composition comprising in a peptide according to at least one active peptide or modified peptide of any one of Embodiments! to 7 associated with a non replicative adenoviral vector or with a non- replicative retroviral vector penetrating in eukaryotic cells and expressing the said peptide. The above written description of the invention provides a manner and process of making and using it such that any person skilled in this art is enabled to make and use the same, this enablement being provided in particular for the subject matter of the appended claims, which make up a part of the original description. The above description is presented to enable a person skilled in the art to make and use the invention and is provided in the context of a particular application and its requirements. Various modifications to the preferred embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments and applications without departing from the spirit and scope of the invention. Thus, this invention is not intended to be limited to the embodiments shown but is to be accorded the widest scope consistent with the principles and features disclosed herein. Having generally described this invention, a further understanding can be obtained by reference to certain specific examples or data or technical information, which are provided herein for purposes of illustration only, and are not intended to be limiting unless otherwise specified.

Claims

CLAIMS1. A method for preventing, reducing the severity of, or treating a tauopathy comprising administering to a subject in need thereof PPI of FK506-binding protein (FKBP52) or a derived active peptide of PPI containing at least 15 Nter amino acids sequence of PPI .

2. The method of claim 1, wherein the tauopathy is Alzheimer’s Disease (AD), familial FTLD-Tau, progressive supranuclear palsy (PSP) or other tauopathies.

3. The method of any one of claims 1-2, wherein the peptide fragment consists of PPI (SEQ ID NO: 1).

4. The method of any one of claims 1-3 , wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) having backbone protection or a modified N or C terminus.

5. The method of any one of claims 1-4 , wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) that further comprises a cell penetrating peptide linked to PPI.

6. The method of any one of claims 1-5 , wherein the peptide fragment consists of a modified form of PPI (SEQ ID NO: 1) that further comprises cell penetrating peptide YARAAARQARA (SEQ ID NO: 2).

7. A peptide or modified peptide comprising a fragment of FKBP52 that inhibits Tau protein aggregation.

8. The peptide or modified peptide of claim 7 that is PPI.

9. The modified peptide of claims 7 or 8 that has a modified backbone or a modified C or N terminus.

10. The modified peptide of any one of claims 7-9 is CPP1 that has an N terminal cysteine residue.

11. The modified peptide of any one of claims 7-10 that further comprises a cell penetrating peptide.

12. The modified peptide of any one of claims 7-11 that further comprises cell penetrating peptide YARAAARQARA (SEQ ID NO: 2).

13. A peptide or modified peptide according to any one of the claim 7 to 12 wherein the active amino-acid sequence comprises at least one TRG amino acid sequence and the sequence of the modified is less than 30 amino acids.

14. A pharmaceutical composition comprising in the active according to at least one active peptide or modified peptide of any one of claims 1 to 13 associated with a non replicative adenoviral vector or with a non-replicative retroviral vector penetrating in eukaryotic cells and expressing the said peptide.

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