Peptide compositions and methods for treating tauopathies

Abstract

The present invention relates to the treatment of tauopathies, such as Alzheimer's disease, using systemic administration of SCO-Spondin-derived peptides to patients. The peptides have the amino acid sequence X1-WS-A1-WS-A2-CS-A3-A4-CG-X2, where A1, A2, A3, and A4 each consist of an amino acid sequence of 1 to 5 amino acids, and X1 and X2 each consist of an amino acid sequence of 1 to 6 amino acids, or X1 and X2 are absent. The N-terminal amino acid can be acetylated, the C-terminal amino acid can be amidated, or both the N-terminal and C-terminal amino acids can be acetylated and amidated.

Description

[Technical Field]

[0001] The present invention relates to peptides, peptide compositions, and methods for the treatment of tauopathies, such as Alzheimer's disease (AD). [Background technology]

[0002] Tauopathy is a class of neurodegenerative disorders associated with the pathological aggregation of tau protein. Tau is a microtubule-associated protein that is abundant in the central nervous system and is mainly expressed in axons. Tau protein may play a role in stabilizing the microtubule network in neurons. Excessive or abnormal phosphorylation of tau can cause tau aggregation, which is believed to be the main cause of disease progression.

[0003] Tau phosphorylation is a normal metabolic process important in regulating tau binding to microtubules and occurs constantly in the brain. In both aging and neurodegenerative diseases, tau can become hyperphosphorylated and aggregate as detectable fibrillar deposits in tissues.

[0004] Tauopathies include Alzheimer's disease (AD), progressive supranuclear palsy (PSP), tau-positive frontotemporal dementias such as Pick's disease, dementia with Lewy bodies, corticobasal degeneration, Niemann-Pick disease type C, chronic traumatic encephalopathy including dementia pugilistica, and postencephalitic parkinsonism. In some tauopathies, such as AD, aggregation of amyloid beta peptides is observed in addition to tau aggregation.

[0005] Treatment options for tauopathies are very limited. Acetylcholinesterase inhibitors (AChEIs) (donepezil, galantamine, and rivastigmine) are the mainstay of symptomatic treatment for AD and increase the availability of acetylcholine by inhibiting its breakdown in synapses. However, some patients fail to respond to or benefit from treatment over time. Furthermore, some patients may experience side effects. As a result, acetylcholinesterase inhibitors are contraindicated for some patients. Therefore, there is a significant medical need for innovative treatment options for tauopathies.

[0006] SCO-Spondin-derived peptides have been described for their neuroregenerative properties, particularly for their ability to improve cell survival and neurite outgrowth in vitro. The use of SCO-spondin-derived peptides for the treatment of spinal cord injury has been investigated in animal models. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO99 / 03890 [Patent Document 2] US6,995,140 [Patent Document 3] WO2018146283 [Patent Document 4] WO2017 / 051135 [Non-patent literature]

[0008] [Non-Patent Document 1] Carsten Henneges et al., Journal of Alzheimer's Disease 52 (2016), 1065–1080 [Non-patent document 2] Bart Sheehan, Therapeutic Advances in Neurological Disorders 2012, 5(6), 349–358 [Non-patent document 3] www.rxlist.com / aricept-drug.htm#indications [Non-patent document 4] Biophysical Journal, Volume 95, November 2008, pp. 4879-4889 Summary of the Invention [Problem to be solved by the invention]

[0009] It is an object of the present invention to provide compositions and methods useful for the treatment of tauopathies and / or disorders in which tau protein plays a role in the pathology.

[0010] Another object of the present invention is to provide compositions and methods useful for the treatment of tauopathies and / or disorders in which amyloid beta protein and tau protein are involved in the pathology.

[0011] Another object is to provide compositions and methods useful for treating Alzheimer's disease (AD), progressive supranuclear palsy (PSP), tau-positive frontotemporal dementia such as Pick's disease, dementia with Lewy bodies, corticobasal degeneration, Niemann-Pick disease type C, chronic traumatic encephalopathy including dementia pugilistica, and postencephalitic parkinsonism.

[0012] Another object of the present invention is to provide compositions and methods for reducing or disrupting tau protein aggregation, reducing tau protein, and / or reducing excessive and / or abnormal phosphorylation of tau protein in a subject. Yet another object is to provide such compositions and methods that may be beneficial for subjects suffering from diseases or disorders characterized by or at least partially caused by tau protein, e.g., elevated or pathological tau protein levels, and / or aggregation of tau protein, and / or excessive and / or abnormal phosphorylation of tau protein.

[0013] Another object of the present invention is to provide such compositions and methods which can further interfere with the formation of amyloid-beta plaques, prevent the aggregation of amyloid-beta, and / or depolymerize already formed amyloid-beta deposits or amyloid-beta aggregates.

[0014] Another object of the present invention is to provide compositions and methods that are compatible with the systemic administration of active ingredients for the treatment of tauopathies.

[0015] Another object of the present invention is to provide compositions and methods useful in the treatment of tauopathies in patients in whom acetylcholinesterase inhibitors are contraindicated.

[0016] Another object of the present invention is to provide compositions and methods useful in the treatment of tauopathies in patients in combination with other agents indicated for the treatment of tauopathies. [Means for solving the problem]

[0017] The present inventors have made the unexpected discovery that peptides derived from SCO-Spondin may be useful for treating tauopathies. In particular, the present inventors have made the unexpected discovery that peptides derived from SCO-Spondin regulate tau, particularly its phosphorylation, and reduce tau protein accumulation or tau protein aggregation. In addition, the present inventors have demonstrated that this beneficial effect can be obtained through systemic administration of the peptides. Furthermore, the present inventors have discovered that compositions or methods according to the present invention may be advantageous for patients in whom treatment with acetylcholinesterase inhibitors is contraindicated, or even in cases of resistance to acetylcholinesterase inhibitors.

[0018] Systemic administration is unexpectedly effective for the peptides of the present invention in this particular context. Systemic administration offers significant advantages over direct administration at sites where tau is not modulated or directly into the CSF (intrathecal or intracerebral injection) because it is safer and more convenient for the patient being treated. An important or advantageous aspect of this mode of administration is that it allows for repeated administration over time by medical professionals to a given patient. The peptides can be easily administered via injection or infusion, including via perfusion. The inventors have also found that the bioavailability of these peptides after systemic administration or their biological effect at the lesion level does not require the administration of excessively large amounts of peptide. Unexpectedly, the peptides (native or unmodified) exert their biological effects after systemic administration. Also unexpectedly, biological effects are observed in the CNS despite the peptides' very short half-lives (approximately 12 minutes in monkeys and approximately 19 minutes in humans). Also unexpectedly, despite this very short half-life, efficacy of the peptides was observed when the peptides were administered daily or every other day.

[0019] This specification describes a composition comprising a peptide derived from SCO-Spondin for treating tauopathy.This use can reduce the total level of tau protein, the level of phosphorylated tau protein, tau aggregation, or a combination thereof in cells.This use can reduce toxic tau oligomers, and treat, slow down, or prevent tauopathy and its progression.

[0020] In this specification, unless contraindicated, all disclosed features apply to the various subjects: "Compositions for Use", "Methods of Use", and "Use of said peptides for the manufacture of a medicament".

[0021] In one aspect, the present invention relates to a composition comprising at least one SCO-Spondin-derived peptide for use in treating a tauopathy in a subject, and a method comprising administering at least one SCO-Spondin-derived peptide. The use and method may comprise administering an effective or sufficient amount of at least one SCO-Spondin-derived peptide to the subject. In one embodiment, systemic administration of at least one SCO-Spondin-derived peptide is performed.

[0022] In another aspect, the present invention relates to a composition comprising at least one SCO-Spondin-derived peptide for use in reducing or disrupting tau aggregation in a subject and / or treating a tauopathy in said subject, and a method comprising administering at least one SCO-Spondin-derived peptide. The uses and methods may comprise administering to the subject an effective or sufficient amount of at least one SCO-Spondin-derived peptide. In one embodiment, systemic administration of at least one SCO-Spondin-derived peptide is performed.

[0023] In another aspect, the present invention relates to a composition comprising at least one SCO-Spondin-derived peptide for use in reducing tau protein and / or hyperphosphorylation of tau protein in a subject and / or treating a tauopathy in said subject, and a method comprising administering at least one SCO-Spondin-derived peptide. The use and method may comprise administering to the subject an effective or sufficient amount of at least one SCO-Spondin-derived peptide. In one embodiment, systemic administration of at least one SCO-Spondin-derived peptide is performed.

[0024] In another embodiment, the present invention relates to a composition comprising at least one SCO-Spondin-derived peptide for use in treating a tauopathy through systemic administration to a subject. In another embodiment, the present invention relates to a method for treating a tauopathy comprising administering to a subject at least one SCO-Spondin-derived peptide via a systemic route.

[0025] In another aspect, the present invention relates to compositions comprising at least one SCO-Spondin-derived peptide for use in treating a subject for whom an acetylcholinesterase inhibitor is contraindicated, or is not or no longer tolerated, or is not sufficiently active or is no longer active, and methods comprising administering at least one SCO-Spondin-derived peptide.

[0026] In another aspect, the present invention relates to use of at least one SCO-Spondin-derived peptide for the manufacture of a pharmaceutical composition for treating a tauopathy. In one embodiment, the composition is for systemic administration. In one embodiment, the composition includes a pharmaceutical excipient or vehicle for systemic administration.

[0027] In some embodiments of these different aspects, the tauopathy is selected from the group consisting of Alzheimer's disease (AD), progressive supranuclear palsy (PSP), tau-positive frontotemporal dementia such as Pick's disease, dementia with Lewy bodies, corticobasal degeneration, Niemann-Pick disease type C, chronic traumatic encephalopathy including dementia pugilistica, and postencephalitic parkinsonism.

[0028] In one embodiment of these different aspects, the tauopathy is AD. The AD can be one in which treatment with an acetylcholinesterase inhibitor (e.g., donepezil) is contraindicated, treatment with an acetylcholinesterase inhibitor is not sufficiently active or is no longer active, the patient has developed resistance to the acetylcholinesterase inhibitor, or the patient has developed intolerance to the acetylcholinesterase inhibitor.

[0029] In one embodiment, the use may further interfere with the formation of amyloid-beta plaques, prevent the aggregation of amyloid-beta, and / or depolymerize already formed amyloid-beta deposits or amyloid-beta aggregates.

[0030] In one embodiment, the subject is also treated with a sufficient amount of an acetylcholinesterase inhibitor.

[0031] In another aspect, the present invention relates to a combination of at least one SCO-Spondin-derived peptide and an acetylcholinesterase inhibitor, preferably DPZ, for use in a method for treating tauopathy, wherein the peptide is administered to a subject via a systemic route. The administration of both active ingredients can be, in particular, separate, simultaneous, or sequential.

[0032] In another aspect, the present invention relates to a pharmaceutical composition comprising at least one SCO-Spondin-derived peptide, an acetylcholinesterase inhibitor, preferably DPZ, and a pharmaceutically acceptable vehicle, carrier, or excipient. Preferably, the composition is suitable for systemic administration. In one embodiment, the composition is for use in treating tauopathy, and the composition is administered to a subject via a systemic route. DETAILED DESCRIPTION OF THE INVENTION

[0033] Tauopathy The compositions and methods detailed herein can be used to treat tauopathies.

[0034] In some embodiments, the compositions and methods detailed herein modulate tau protein to treat tauopathy. Tau is a protein that binds to and stabilizes microtubules. Tau is also called microtubule-associated protein tau (MAPT). Tau protein can also interact with tubulin to stabilize microtubules and promote the assembly of tubulin into microtubules. There are six tau isoforms. Tau protein is abundant in neurons of the central nervous system and is expressed at very low levels in astrocytes and oligodendrocytes of the central nervous system (CNS). Tau protein may play a role in stabilizing the microtubule network within neurons.

[0035] Altered phosphorylation of tau, including excessive phosphorylation (hyperphosphorylation) and aberrant phosphorylation, can disrupt the organization, accumulation, and / or aggregation of tau protein on microtubules. In some embodiments, tau aggregates do not function properly. For example, tau aggregates do not properly stabilize microtubules. Examples of tau aggregates include PHF-tau (paired helical filaments), NFTs (neurofibrillary tangles), and gliofibrillary tangles. Tau aggregates can also be described as monomers or high molecular weight multimers and oligomers. Tau aggregates can be insoluble. Tau aggregates can be present in the brain. Tau protein can accumulate in the form of inclusions within swollen neurons. Aggregation of tau into oligomeric species can lead to various pathologies called tauopathies and may be a major cause of disease progression.

[0036] Tauopathies are a class of neurodegenerative diseases associated with altered phosphorylation of tau and / or pathological aggregation of tau.

[0037] The compositions and methods detailed herein may inhibit or reduce tau protein levels, total tau protein levels in cells, phosphorylated tau protein levels, inhibit, reduce, or disrupt tau protein aggregation, or a combination thereof. The levels may be reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 98%. Tau aggregation may be reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 98%.

[0038] The compositions and methods detailed herein may inhibit or reduce levels of tau protein, may inhibit or reduce total intracellular tau protein levels, may inhibit or reduce levels of phosphorylated tau protein, may inhibit, reduce or disrupt tau protein aggregation, or a combination thereof, to treat a tauopathy.

[0039] "Systemic administration" refers to any mode or route of administration in which a majority or sufficient amount of the administered peptide or peptide compound reaches the blood circulation after such administration. Intrathecal administration and all means of systemic administration that do not target the peptide to the blood circulation are excluded. The systemic administration routes of the present invention qualify as "systemic administration routes that target the blood."

[0040] The administration or use of a peptide ("peptide" or "peptides" or "peptide(s)") is a general expression, and the present invention encompasses the administration or use of one single peptide or two or more single peptides, i.e., the administration or use of at least two peptides according to the present disclosure. Thus, in the present disclosure, the singular or plural is not limited unless otherwise indicated and may encompass one single peptide or at least two peptides each time. The same applies to the similar expression "peptide compound," which may be used interchangeably with "peptide."

[0041] "Treatment" refers to the delivery of an amount of a peptide compound according to the present invention to a subject. As used herein, these terms refer to a treatment aimed at slowing (reducing) an undesirable physiological condition, disorder, or disease, or achieving a beneficial or desired clinical outcome. For purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; attenuation of the severity of the condition, disorder, or disease; stabilization (i.e., not worsening) of the condition, disorder, or disease state; delay in onset or slowing of progression of the condition, disorder, or disease; improvement of the condition, disorder, or pathology; and detectable or undetectable remission (partial or complete remission) or improvement or amelioration of the condition, disorder, or disease. Treatment also includes extending lifespan compared to the expected lifespan if not receiving treatment. The term "treatment" includes preventing, suppressing, arresting, ameliorating, or completely eliminating a disease. Prevention of a disease can involve administering a composition of the present invention to a subject prior to the onset of the disease. Suppressing the disease can involve administering a composition of the invention to a subject after induction of the disease but before its clinical manifestation. Arresting or ameliorating the disease can involve administering a composition of the invention to a subject after clinical manifestation of the disease. In one embodiment, the beneficial or desired clinical outcome includes a beneficial effect on memory loss and / or cognitive impairment, e.g., a reduction, delay, or partial or complete reversal.

[0042] The term "inhibit" means that activity is reduced or prevented in the presence of an inhibitor as opposed to in its absence. The term "inhibition" refers to eliminating or minimizing the expression or activity of a biomolecule or polypeptide, reducing or downregulating a process, or removing a stimulus for a process. Inhibition can be direct or indirect. Inhibition can be specific, i.e., an inhibitor inhibits one biomolecule or polypeptide and not others.

[0043] Similar terms, such as "effective amount," "sufficient amount," and "therapeutically effective amount," are used interchangeably herein unless otherwise defined, and refer to an amount of one or more peptides of the present invention effective for a period of time necessary to achieve a desired therapeutic result. An effective dosage can be determined by one skilled in the art and may vary according to factors such as the individual's condition, age, sex, and weight, as well as the ability of the agent to elicit a desired response in the individual. As used herein, the term also refers to an amount effective to produce a desired in vivo effect in a subject. A therapeutically effective amount may be administered in one or more administrations (e.g., the composition may be administered at any stage of disease progression, pre- or post-symptomatically, as a prophylactic treatment, or therapeutically), application, or dosage, and is not intended to be limited to a particular formulation, combination, or systemic administration route. It is within the scope of the present disclosure that the peptide may be administered at various times during the course of treatment of a subject. The administration time and dosage used will depend on several factors, such as the goal of treatment (e.g., treatment vs. prevention), the condition of the subject, and can be readily determined by one skilled in the art. A therapeutically effective amount is also an amount in which any toxic or detrimental effects of a substance are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, a prophylactic dose is used in subjects before or at an early stage of disease, so the prophylactically effective amount may be less than the therapeutically effective amount. When considering the amount of a peptide individually and / or in combination with another active ingredient, an "effective amount," "sufficient amount," may also consider combinations of different peptides, for example, because the dose of one or two agents in the combination may be reduced as a result of combined or synergistic effects.

[0044] As used herein, the terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variations thereof are intended to be open-ended transitional phrases, expressions, or words that do not exclude the possibility of additional actions or products, e.g., peptides, compounds, or agents. The singular forms "a," "and," and "the" include plural references unless the context clearly indicates otherwise. The present disclosure also contemplates other embodiments that "comprise," "consist of," and "consist essentially of" the embodiments or elements presented herein, whether or not expressly indicated.

[0045] "Patient or subject" means an animal, particularly a mammal, including a human. In one embodiment, the subject is a human. In other embodiments, the subject is a large or livestock animal, a companion animal (e.g., a cat, a dog), or a sport animal (e.g., a horse).

[0046] "Acetylcholinesterase inhibitors" that may be used in combination with the peptides of the present invention include donepezil, galantamine, and rivastigmine, or any other acetylcholinesterase inhibitor approved by a regulatory agency. In one embodiment, such inhibitors, particularly donepezil, are used in combination with NX210, NX218, or both NX210 and NX218 (SCO-Spondin-derived peptides).

[0047] As used herein, "concurrently" or "concurrently" refers to the administration of two agents in parallel, while the term "in combination" refers to the administration of two agents "sequentially" within a time frame in which they are both available to act therapeutically within the same time frame if they are not administered simultaneously. Thus, in "sequentially" or "sequential" administration, one agent may be administered within 5 minutes, 10 minutes, or even hours after the other agent is administered, provided that the circulating half-life of the first-administered agent allows both agents to be present in parallel in therapeutically effective amounts. The time delay between the administration of the components varies depending on the exact nature of the components, the interaction between them, and their respective half-lives. "Separately" or "separately" refers to a significant interval, i.e., hours, days, weeks, or months, between the administration of one agent and the administration of the other agent, which may include cases in which the first-administered agent is no longer present in the circulation in therapeutically effective amounts by the time the second agent is administered.

[0048] SCO-Spondin-derived peptides "SCO-Spondin" is a glycoprotein specific to the central nervous system and is present in all vertebrates, from the prenotochord to humans. It is known as an extracellular matrix molecule secreted by the subcommissural organ, a specific organ located in the roof of the third ventricle. It is a large molecule. It consists of more than 4,500 amino acids and has a multimodular structure containing various conserved protein patterns, including 26 TR or TSR patterns. It is known that certain peptides derived from SCO-Spondin that begin with the TSR pattern have biological activity in nerve or neuronal cells (described in particular in WO99 / 03890).

[0049] "TSR or TR patterns" are protein domains consisting of approximately 55-60 residues based on the alignment of the conserved amino acids cysteine, tryptophan, and arginine. These patterns were first isolated in TSP-1 (thrombospondin 1), a molecule involved in coagulation. These patterns were subsequently described in many other molecules, such as SCO-spondin. Indeed, this thrombospondin type 1 unit (TSR) contains approximately 55-60 amino acids (AA), and some of these amino acids, such as cysteine ​​(C), tryptophan (W), serine (S), glycine (G), arginine (R), and proline (P), are highly conserved in all proteins studied and previously mentioned.

[0050] SCO-Spondin peptides or peptide compounds are used in the practice of the present invention (for various purposes of the invention, e.g., peptides or compositions for use, methods of use, methods of treatment, use of peptides to manufacture medicaments, etc.).

[0051] In particular, the present invention provides a compound having the sequence X1-WS-A1-WS-A2-CS-A3-A4-CG-X2 (SEQ ID NO: 1) Using the peptide During the ceremony, A1, A2, A3, and A4 each consist of an amino acid sequence consisting of 1 to 5 amino acids, The two cysteines may or may not form a disulfide bridge, X1 and X2 each consist of an amino acid sequence consisting of 1 to 6 amino acids, or X1 and X2 are absent; The N-terminal amino acid can be acetylated (e.g., to have H3CCOHN-), the C-terminal amino acid can be amidated (e.g., to have -CONH2), or both the N-terminal amino acid can be acetylated and the C-terminal amino acid can be amidated.

[0052] In one embodiment, in the formula of SEQ ID NO: 1, X1 or X2, or both X1 and X2 are absent. In one embodiment, when X1 and / or X2 are absent, the N-terminal W is acetylated and / or the C-terminal G is amidated. Preferably, both X1 and X2 are absent, the N-terminal W is acetylated, and the C-terminal G is amidated.

[0053] In particular, the present invention provides a compound having the sequence WS-A1-WS-A2-CS-A3-A4-CG (SEQ ID NO: 2) A peptide of the formula: A1, A2, A3, and A4 each consist of an amino acid sequence consisting of 1 to 5 amino acids, The two cysteines may or may not form a disulfide bridge.

[0054] In one embodiment of the formulas of SEQ ID NO: 1 and 2, the peptide is a linear peptide or the cysteines present in the peptide formulas of SEQ ID NO: 1 and 2 do not form disulfide bridges (reduced form).

[0055] In another embodiment, the two cysteines present in the peptide formulas of SEQ ID NOs: 1 and 2 form a disulfide bridge (oxidized form).

[0056] Preferably, in the formulae of SEQ ID NOs: 1 and 2, A1, A2, A3 and / or preferably A4 preferably consist of one or two amino acids, more preferably consist of one amino acid.

[0057] Preferably, A1 is selected from G, V, S, P and A, more preferably G and S.

[0058] Preferably, A2 is selected from G, V, S, P and A, more preferably G and S.

[0059] Preferably, A3 is selected from R, A, and V, more preferably R, V.

[0060] Preferably, A4 is selected from S, T, P and A, more preferably S, T.

[0061] Preferably, A1 and A2 are independently selected from G and S.

[0062] Preferably, A3-A4 is selected from RS or VS or VT or RT.

[0063] Preferably, X1, X2, A1, A2, A3, and A4 do not contain cysteine.

[0064] When X1 is an amino acid sequence consisting of 1 to 6 amino acids, the amino acids may be any amino acids, and are preferably selected from V, L, A, P, and any combination thereof.

[0065] When X2 is an amino acid sequence consisting of 1 to 6 amino acids, the amino acids may be any amino acids, and are preferably selected from L, G, I, F, and any combination thereof.

[0066] In one embodiment, the peptide of SEQ ID NO: 1 or 2 is one in which A1 and A2 are independently selected from G and S, and A3-A4 are selected from RS, VS, VT, or RT. In a specific manner, the peptide is further acetylated and / or amidated. In one embodiment, the peptide is a linear peptide or the cysteines do not form disulfide bridges. In another embodiment, the peptide has two cysteines forming disulfide bridges (C-terminal cyclization). In another embodiment, the peptide used in the present invention or administered to a patient via a systemic route includes both oxidized and linear peptide forms.

[0067] For purposes of the present invention, the term "amino acid" refers to both naturally occurring and unnatural amino acids, and amino acid changes, including natural to unnatural changes, can be routinely made by those skilled in the art while maintaining the function or effectiveness of the original peptide. "Natural amino acids" refer to the L-form amino acids found in natural proteins, i.e., alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. "Unnatural amino acids" refer to the aforementioned amino acids in their D-form, as well as certain amino acids in their homoform, e.g., arginine, lysine, phenylalanine, and serine, and leucine or valine in their nor-form. This definition also includes other amino acids, such as alpha-aminobutyric acid, agmatine, alpha-aminoisobutyric acid, sarcosine, statin, ornithine, and desaminotyrosine. The nomenclature used to describe peptide sequences is the International Nomenclature, which uses a one-letter code with the amino terminus shown on the left and the carboxy terminus shown on the right. A dash "-" indicates a conventional peptide bond joining the amino acids in the sequence.

[0068] In one embodiment, a peptide according to the present invention, for example any one of the peptides of the sequences of SEQ ID NOs: 1 to 63, comprises an N-terminal acetylation, a C-terminal amidation, or both an N-terminal acetylation and a C-terminal amidation.

[0069] In a different embodiment, the present invention relates to the use of a polypeptide consisting essentially of or consisting of the following amino acid sequence (Table 1):

[0070] [Table 1]

[0071] In one embodiment, the peptides of the sequences SEQ ID NOs: 3-34 disclosed in Table 1 are linear peptides, or the cysteines do not form disulfide bridges (reduced peptides). In another embodiment, the peptides of the sequences SEQ ID NOs: 3-34 disclosed in the aforementioned tables have two cysteines oxidized to form disulfide bridges (oxidized peptides). In another embodiment, the peptides used in the present invention or administered to patients via a systemic route comprise both oxidized and linear peptide forms of the same peptide sequence. In yet another embodiment, the peptides used in the present invention or administered to patients via a systemic route comprise a mixture of at least two of these different peptides selected from the sequences SEQ ID NOs: 3-34, where the mixture can be a mixture of at least two linear peptides, or a mixture of at least two oxidized peptides, or a mixture of at least one linear peptide and at least one oxidized peptide, for example, having the same amino acid sequence.

[0072] In a preferred embodiment, the peptide consists of the amino acid sequence WSGWSSCSRSCG (SEQ ID NO: 3). In one embodiment, the peptide is a linear peptide or the cysteines do not form disulfide bridges (reduced form, designated NX210). In another embodiment, the peptide has two cysteines oxidized to form disulfide bridges (oxidized form), designated NX218. In another embodiment, the peptide used in the present invention or administered to a patient via a systemic route comprises both the oxidized and reduced forms.

[0073] In one embodiment of the peptide of SEQ ID NO: 1, X1 is a hydrogen atom or P or AP or LAP or VLAP, and / or X2 is a hydrogen atom or L or LG or LGL or LGLI or LGLIF.

[0074] In a different embodiment, the present invention therefore relates to the use of a polypeptide consisting of or consisting essentially of the following amino acid sequence (Table 2):

[0075] [Table 2]

[0076] In one embodiment, the peptides of SEQ ID NOS: 35-63 disclosed in Table 2 or SEQ ID NOS: 3-63 disclosed in Tables 1 and 2 are linear peptides or have no cysteines forming disulfide bridges (reduced peptides). In another embodiment, the peptides have two cysteines oxidized to form disulfide bridges (oxidized peptides). In another embodiment, the peptides used in the present invention or administered to patients via a systemic route comprise both oxidized and linear peptide forms of the same peptide sequence. In yet another embodiment, the peptides used in the present invention or administered to patients via a systemic route comprise a mixture of at least two of these different peptides selected from the sequences of SEQ ID NOS: 35-63 or 3-63, where the mixture can be a mixture of at least two linear peptides, or a mixture of at least two oxidized peptides, or a mixture of at least one linear peptide and at least one oxidized peptide, for example, having the same amino acid sequence.

[0077] Each of the peptides of the sequences of SEQ ID NOs: 3 to 63 may be acetylated, amidated, or acetylated and amidated.

[0078] In the present invention, the peptides used in the present invention or administered to patients via a systemic route are defined by their amino acid sequences. The peptide used can be one peptide disclosed herein or a mixture of at least two peptides disclosed herein. The mixture also includes a mixture of linear and oxidized peptides of the same or different amino acid sequences. When 100% pure peptides are used in accordance with the present invention, it is possible for the peptide to have a purity of more than 80%, preferably more than 85%, more preferably more than 90%, and even more preferably 95, 96, 97, 98, or 99% or more, which is encompassed by the present invention. Conventional purification methods, such as by chromatography, can be used to purify the desired peptide compound.

[0079] In one embodiment, the peptides used in the present invention or administered to patients via the systemic route contain both oxidized peptide (Op) and linear peptide (Lp) forms, e.g., in similar or different amounts, e.g., (by number): Op: 10, 20, 25, 30, 40, 50, 60, 70, 80, or 90%, with the remainder up to 100% being Lp. The combined oxidized and linear peptides may be of the same sequence or different sequences. For example, the oxidized and linear forms of the peptide of sequence SEQ ID NO: 3 may be combined, e.g., in the ratios disclosed above (NX210 and NX218). The same applies to any one of the peptides of sequences SEQ ID NOs: 4-34 and 35-63.

[0080] The pharmaceutical compositions used in the present invention comprise as active ingredients the peptides or peptide mixtures described above, e.g., peptides of different amino acid composition or peptides of the same amino acid composition, in oxidized and linear forms, and one or more pharmaceutically acceptable vehicles, carriers, or excipients.

[0081] The peptide compounds according to the invention can be used in pharmaceutical compositions or in the manufacture of medicaments, in which the active ingredient can be incorporated into the composition in various forms, i.e. in the form of a solution, usually an aqueous solution, or in lyophilized form, or in the form of an emulsion, or in any other pharmaceutically and physiologically acceptable form suitable for systemic administration routes.

[0082] According to an important feature of the present invention, the peptide compound, or a composition containing it, is administered via a systemic route. The following injection or administration routes may be mentioned in particular: intravenous, intraperitoneal, intranasal, subcutaneous, intramuscular, sublingual, oral, and combinations thereof.

[0083] For the intravenous route, injection, ie administration using an injection device (eg using a syringe, pump) or gravity perfusion may be selected.

[0084] In the subcutaneous route, the composition is administered as a bolus into the subcutaneous tissue. Injection sites can be the lateral area of ​​the upper arm, the abdomen from the costal margin to the iliac crest, the anterior thigh, the back, or the buttocks.

[0085] For the intranasal or nasal route, nasal insufflation (where the composition is blown into the nose, especially using a gas, powder or vapor), nasal inhalation or nasal drops may be chosen.

[0086] Compositions containing one or more of the peptides disclosed herein are sterile. These compositions are suitable for administration to deliver the peptide into the blood circulation. Delivery into the blood circulation is delivery of a sufficient amount of peptide into the blood circulation, which is correlated with a beneficial effect in the CNS. In other words, delivery into the blood circulation is delivery of a sufficient amount of peptide into the blood circulation and delivery of a sufficient "pharmaceutical effect" and / or "improvement of cognitive function" in the CNS, as defined herein below. As disclosed herein, a "pharmaceutical effect" may include inhibiting or reducing the level of tau protein, inhibiting or reducing the level of total tau protein in cells, inhibiting or reducing the level of phosphorylated tau protein, inhibiting, reducing, or disrupting the aggregation of tau protein, or a combination thereof. In some embodiments, the active ingredient in the pharmaceutical composition consists of (1) a linear peptide disclosed herein, (2) an oxidized peptide disclosed herein, (3) NX210, (4) NX218, or (5) a mixture of linear and oxidized peptides, in similar or dissimilar amounts, as disclosed above, e.g., particularly NX210 and NX218.

[0087] The active ingredient can be administered to animals and humans in unit dosage forms as a mixture with conventional pharmaceutical supports, carriers, excipients, or vehicles. Suitable unit dosage forms include oral route forms, such as tablets, gel capsules, powders, granules, and oral suspensions or solutions; sublingual and buccal dosage forms, aerosols; implants; subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subcutaneous, transdermal, and intranasal dosage forms; and rectal dosage forms.

[0088] Preferably, the pharmaceutical composition contains a pharmaceutically acceptable carrier, excipient, or vehicle for a liquid formulation that can be injected to deliver the active ingredient into the bloodstream. These can be, in particular, ready-to-use solutions, such as isotonic solutions, sterile solutions, saline (monosodium phosphate or disodium phosphate, sodium chloride, potassium chloride, calcium chloride, or magnesium chloride, and the like, or mixtures of such salts), or dry compositions, in particular lyophilized compositions, which, when added with sterile water or saline, allow the constitution of an injectable solution.

[0089] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations containing sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be as fluid as possible for easy syringability. The form must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[0090] Sterile injectable solutions are prepared by incorporating the active polypeptide in the required amount in the appropriate solvent, followed by filtered sterilization.

[0091] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation, and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, although drug release capsules and the like are also available.

[0092] For appropriate administration in aqueous solution, for example, the solution should be appropriately buffered if necessary, and the liquid diluent should first be made isotonic with sufficient saline or glucose.These particular aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration.In this regard, available sterile aqueous media will be clear to those skilled in the art in light of this disclosure.In addition to the compounds of the present invention formulated for administration by injection, such as intravenous or intramuscular injection, other pharmaceutically acceptable forms include, for example, tablets or other solids for oral administration; liposome preparations; sustained-release capsules; and any other form currently used to deliver active ingredients into the bloodstream.

[0093] One dose, expressed as the mass of peptide per kg of patient body weight, can be in the range of about 1 μg / kg to about 1 g / kg, particularly about 10 μg / kg to about 100 mg / kg, for example about 50 μg / kg to about 50 mg / kg.

[0094] The dosing regimen may include a single administration or multiple administrations. According to one embodiment, multiple administrations may include administering one dose per treatment day, e.g., every day or every two or three days, over the treatment period. According to another embodiment, multiple administrations may include administering at least two doses per treatment day, e.g., two, three, or more doses per day, over the treatment period. In these embodiments, the treatment period may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or more days (e.g., up to six months). The treatment is designed so that the patient maintains a "benefit" from the treatment over a "period." The "benefit" may include the "pharmaceutical effect" or "improvement of cognitive function" described above. This "duration" may depend on the medication regimen and the patient himself, for example, the severity of the disease and the patient's responsiveness to the regimen dose. "Improvement of cognitive function" includes "partial cognitive recovery" and "complete cognitive recovery." Cognitive recovery is "partial" when a subject shows partial recovery of memory alterations, and there is a significant improvement in memory compared to the subject's initial situation before treatment, but this remains significantly lower than that of healthy subjects. "Complete recovery" means that the subject has recovered memory, or that the memory level is not significantly different from that of healthy subjects.

[0095] Improvement in cognitive function can be measured in subjects using the MMSE (Mini-Mental State Examination; see Carsten Henneges et al., Journal of Alzheimer's Disease 52 (2016), 1065-1080) and / or the ADAS-cog (Alzheimer's Disease Assessment Scale; see Bart Sheehan, Therapeutic Advances in Neurological Disorders 2012, 5(6), 349-358).

[0096] The MMSE is a brief, 30-item test with total scores ranging from normal (30) to severe impairment (0). Questions are grouped into seven categories, with each subscore representing a specific aspect of cognitive function: orientation to time (score range 0-5), orientation to place (score range 0-5), memory (score range 0-3), attention and concentration (counting backwards or spelling, score range 0-5), recall (score range 0-3), language (score range 0-8), and drawing (score range 0-1). MMSE criteria (the spectrum of AD) that may be used herein are mild AD (MMSE 21-26), mild-moderate AD (MMSE 15-20), and moderate / severe (MS / S) AD (MMSE <15).

[0097] The ADAS-Cog is widely used to measure cognitive performance in clinical trials and assesses multiple cognitive domains, including memory, orientation, visuospatial ability, language, and performance. It measures the following cognitive domains: word recall task (score range 0–10), object naming and pointing (score range 0–5), following verbal commands (score range 0–5), constructional behavior (score range 0–5), conceptual behavior (score range 0–5), orientation (score range 0–8), word recognition task (score range 0–12), oral language ability (score range 0–5), oral language comprehension (score range 0–5), word finding difficulty (score range 0–5), and recall of task instructions (score range 0–5).

[0098] In one embodiment, an MMSE criterion or ADAS-cog is measured in a subject before starting treatment with a composition disclosed herein, and then the same subject subsequently has this criterion measured one or more times (at least twice) either during the protocol or after treatment has stopped, or during the protocol, after stopping the protocol, and after a lag period without treatment.

[0099] Advantageously, baseline measurements are taken to determine whether to continue ongoing treatment or to stop treatment if the baseline indicates sufficient improvement in cognitive function, whether partial or complete, or to resume treatment if the improvement disappears after a lag period after treatment has stopped.

[0100] In one embodiment, this "pharmaceutical effect" or "improvement in cognitive function" continues for a certain period of time, which can be monitored or predicted (e.g., based on experience or practice). In some patients, it may be advantageous to administer such treatment sequentially, e.g., administer a first treatment, a treatment-free period (lag period), and a second or subsequent treatment, and the sequential treatments and lag periods can increase over the patient's lifespan or according to the patient's needs therefor. The dosing regimen may thus include an initial or first-applied dosing regimen, a treatment-free period (recovery period), a second dosing regimen, a second recovery period, optionally followed by a third dosing regimen, and a third recovery period, etc. The sequential dosing regimen is as disclosed above.

[0101] The "period" or lag period (which may be referred to as a recovery period) during which a patient maintains the benefit of treatment according to the dosing regimen of the present invention can be monitored by the patient themselves, for example, using the MMSE and ADAS-cog methods described above, or based on experience with similar cases. This lag or recovery period (during which the patient experiences improved cognitive function or another benefit, e.g., inhibition or reduction of tau protein levels, total tau protein levels in cells, phosphorylated tau protein levels, or inhibition or reduction or disruption of tau protein aggregation, or a combination thereof) may last at least 14 days, or 1, 2, 3, 4, 5, 6, 7, or more months.

[0102] In one embodiment, the dose is administered by perfusion, which can last for a few minutes, tens of minutes, several hours, and up to 24 hours per day.

[0103] The uses according to the present invention and the treatment methods according to the present invention may be characterized by allowing the delivery of an amount of a peptide compound according to the present invention in a subject and obtaining a favorable effect on tauopathy and / or obtaining one or more of the reduction or disruption of tau protein aggregation, reduction of tau protein levels and / or reduction of hyperphosphorylation and reduction of abnormal phosphorylation of tau protein.

[0104] In one embodiment, the use of treatment or method of treatment has the effect of aiding or leading to recovery of function, i.e., the patient recovers all or part of the function lost as a result of tauopathy and / or one or more of tau protein aggregation, increased tau protein levels, hyperphosphorylation, and abnormal phosphorylation of tau protein.

[0105] In one embodiment, the use or method of treatment has the effect of arresting or inhibiting tauopathy and / or loss of function resulting from one or more of tau protein aggregation, increased tau protein levels, hyperphosphorylation of tau protein, and abnormal phosphorylation.

[0106] In one embodiment, the present invention relates to a combination of at least one SCO-Spondin-derived peptide of the present invention and an acetylcholinesterase inhibitor, preferably donepezil (DPZ), for use in a method for treating tauopathy as disclosed herein, wherein the peptide is administered to a subject via a systemic route. The administration of both active ingredients can be separate, simultaneous, or sequential. The combination of active ingredients can have a synergistic effect on the efficacy or benefit of treatment.

[0107] In one embodiment, DPZ is administered to a subject according to standard of care, specifically the following (www.rxlist.com / aricept-drug.htm#indications): - Mild to Mild-Moderate Alzheimer's Disease Dosing: The recommended starting dose of ARICEPT® is 5 mg once daily in the evening, immediately before bedtime. The recommended maximum dose of ARICEPT® in patients with mild to mild-moderate Alzheimer's disease is 10 mg per day. The 10 mg dose should not be administered until the patient has been receiving a daily dose of 5 mg for 4 to 6 weeks. - Dosing in Mild-Moderate to Severe Alzheimer's Disease: The recommended starting dose of ARICEPT® is 5 mg once daily in the evening, immediately before bedtime. The recommended maximum dose of ARICEPT® in patients with mild-moderate to severe Alzheimer's disease is 23 mg per day. The 10 mg dose should not be administered until the patient has been receiving a 5 mg daily dose for 4 to 6 weeks. The 23 mg per day dose should not be administered until the patient has been receiving a 10 mg daily dose for at least 3 months.

[0108] For example, in the case of a synergistic effect between DPZ and a peptide of the present invention, such as NX210, NX218, or NX210+NX218, the dose of the acetylcholinesterase inhibitor, such as DPZ, and / or the dose of the peptide may be reduced compared to the dose for a single molecule. The dose of DPZ may therefore be reduced compared to standard therapy when DPZ and a peptide of the present invention are administered to the same subject (simultaneously or sequentially). The dose (partial dose) of DPZ may be, for example, 3 mg / day or less, and in particular 2.5, 2, 1.5, or 1 mg / day or less. The dosing or administration regimen may be 0.5-3, 0.5-2.5, 0.5-2, 0.5-1.5, or 0.5-1 mg / day. In these intervals, the minimum value of 0.5 may be replaced by 0.6, 0.7, 0.8, 0.9, or 1.

[0109] In one embodiment, the SCO-Spondin-derived peptide of the present invention and an acetylcholinesterase inhibitor, preferably donepezil (DPZ), are used separately in the same patient. In particular, the SCO-Spondin-derived peptide of the present invention is used as a second-line treatment for tauopathy in a patient who has been initially treated with an acetylcholinesterase inhibitor, preferably DPZ, as a first-line treatment. This first-line treatment may be standard treatment. See above. The present invention therefore relates to a combination of at least one SCO-Spondin-derived peptide of the present invention and an acetylcholinesterase inhibitor, preferably DPZ, for use in a method for treating a tauopathy as disclosed herein, wherein the peptide is administered via a systemic route to a subject or patient previously treated with the inhibitor.

[0110] Alternatively, the second-line treatment is a combination of the SCO-Spondin-derived peptides of the present invention with an acetylcholinesterase inhibitor, preferably donepezil (DPZ). "Combination" can include simultaneous or sequential administration of the agents. In one embodiment, partial doses of the SCO-Spondin-derived peptides of the present invention and / or an acetylcholinesterase inhibitor, preferably donepezil (DPZ), are used.

[0111] The method may be further described as follows: - selecting a patient who has received therapeutic treatment with said acetylcholinesterase inhibitor, preferably wherein the patient has been treated according to a treatment protocol (preferably a standard or approved treatment protocol) using said inhibitor, - optionally determining that the patient is in a phase in which the patient no longer responds to the treatment and / or shows intolerance to said treatment, - subjecting said patient to a therapeutic treatment or treatment protocol of the invention using a composition according to the invention or a combination as defined immediately above, which treatment may comprise at least one dosing regimen as disclosed above, which dosing regimen may be followed by a recovery period and optionally a second dosing regimen, a second recovery period, etc., as disclosed above.

[0112] In one embodiment, the method comprises: - administering therapeutic treatment to a patient in need thereof with said acetylcholinesterase inhibitor, preferably wherein the patient is treated according to a therapeutic protocol using said inhibitor; - optionally determining that the patient is in a phase in which the patient no longer responds to the treatment and / or shows intolerance to said treatment, - subjecting said patient to a therapeutic treatment or treatment protocol of the invention using a composition according to the invention or a combination as defined immediately above, which treatment may comprise at least one dosing regimen as disclosed above, which dosing regimen may be followed by a recovery period and optionally a second dosing regimen, a second recovery period, etc., as disclosed above. Includes:

[0113] For reasons of completeness, various aspects of the invention are set out in the following numbered paragraphs, and the features described also apply to other aspects of the invention, e.g., "for use" and "use for producing a medicament."

[0114] Item 1. A method for treating a tauopathy in a subject, comprising administering to the subject a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient via a systemic route.

[0115] Item 2. A method for reducing or disrupting tau aggregation in a subject, comprising administering to the subject via a systemic route a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient.

[0116] Item 3. A method for reducing tau protein in a subject, comprising administering a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient to the subject via a systemic route.

[0117] Item 4. The method of any one of items 1 to 3, wherein the method comprises at least one treatment period and at least one recovery period, wherein the treatment period comprises administering at least one dosing regimen to the subject, and the recovery period is a period of time during which no treatment is administered.

[0118] Item 5. The method according to any one of Items 1 to 4, wherein the level of phosphorylated tau protein is reduced.

[0119] Item 6. The method according to any one of Items 1 to 5, wherein the total tau protein level is reduced.

[0120] Item 7. The method of item 5 or 6, wherein the level is reduced by at least 10%.

[0121] Item 8. The method of item 7, wherein the level is reduced by at least 50%.

[0122] Item 9. The method of item 8, wherein the level is reduced by at least 80%.

[0123] Item 10. The method according to any one of Items 1 to 9, wherein tau aggregation is reduced.

[0124] Item 11. The method of Item 10, wherein tau aggregation is reduced by at least 10%.

[0125] Item 12. The method of Item 11, wherein tau aggregation is reduced by at least 50%.

[0126] Item 13. The method of Item 12, wherein tau aggregation is reduced by at least 80%.

[0127] Item 14. The method according to any one of Items 1 to 13, which results in improvement of cognitive function.

[0128] Item 15. The method according to any one of Items 1 to 14, wherein the SCO-spondin-derived peptide is selected from the group consisting of peptides having the sequence of SEQ ID NO: 1 or 2.

[0129] Item 16. The method according to any one of Items 1 to 14, wherein the SCO-spondin-derived peptide is selected from the group consisting of peptides having sequences set forth in SEQ ID NOs: 3 to 63.

[0130] Item 17. The method according to any one of Items 1 to 16, further comprising administering a sufficient amount of an acetylcholinesterase inhibitor to the subject.

[0131] Item 18. The method according to Item 17, wherein the acetylcholinesterase inhibitor is DPZ.

[0132] Section 19. - selecting patients who have received therapeutic treatment with an acetylcholinesterase inhibitor, preferably DPZ, and then - optionally determining that the patient is in a phase in which the patient no longer responds to the treatment and / or shows intolerance to said treatment, - administering to said patient a therapeutic treatment comprising administering to the subject via a systemic route a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient, or a combination as defined above. (1) treating a tauopathy in a subject; (2) reducing or disrupting tau aggregation in a subject; or (3) reducing tau protein in a subject, comprising:

[0133] Section 20. - administering to a patient in need thereof a therapeutic treatment with an acetylcholinesterase inhibitor, preferably DPZ, and then - optionally determining that the patient is in a phase in which the patient no longer responds to the treatment and / or shows intolerance to said treatment, - administering to said patient a therapeutic treatment comprising administering to the subject via a systemic route a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient, or a combination as defined above. (1) treating a tauopathy in a subject; (2) reducing or disrupting tau aggregation in a subject; or (3) reducing tau protein in a subject, comprising:

[0134] Section 21. - determining the initial cognitive status of the patient, for example using the MMSE and / or ADAS-cog methods, - administering to said patient a therapeutic treatment comprising administering to the subject via a systemic route a therapeutic amount of a peptide derived from SCO-Spondin and a pharmaceutically acceptable vehicle or excipient, or a combination as defined above; - assessing the patient's cognitive status during the course of the therapeutic treatment using the same method, i.e., the MMSE and / or ADAS-cog method, - comparing the cognitive function treatment status during the treatment course with the initial cognitive function status of the patient; - optionally deciding to stop treatment in cases where cognitive function has partially or completely improved, or to continue treatment in cases where cognitive function has not improved or is insufficient (neither partial nor complete improvement has been achieved); Item 19. The method according to any one of items 1 to 18, comprising:

[0135] Item 22. If a decision is made to proceed with treatment: - assessing the patient's cognitive status during the course of the therapeutic treatment using the same method, i.e., the MMSE and / or ADAS-cog method, - comparing the cognitive function treatment status during the treatment course with the initial cognitive function status of the patient; - optionally deciding to stop treatment in cases where cognitive function has partially or completely improved, or to continue treatment in cases where cognitive function has not improved or is insufficient (neither partial nor complete improvement has been achieved); Item 22. The method of item 21, further comprising:

[0136] Item 23. If a decision is made to stop treatment: - determining the further cognitive status of said patient after a treatment-free period, for example using the MMSE and / or ADAS-cog instruments, - comparing the further cognitive treatment status with the patient's initial cognitive status and / or with the patient's cognitive status during the treatment course, - optionally deciding to do nothing if the situation is still better than the initial situation or is not significantly different from the situation at the time the treatment was stopped, or, if this is not the case, to decide on a further period of treatment according to the invention. Item 23. The method of item 21 or 22, further comprising:

[0137] Item 24. The method according to any one of Items 1 to 23, wherein the tauopathy is selected from the group consisting of Alzheimer's disease (AD), progressive supranuclear palsy (PSP), tau-positive frontotemporal dementia such as Pick's disease, dementia with Lewy bodies, corticobasal degeneration, Niemann-Pick disease type C, chronic traumatic encephalopathy including dementia pugilistica, and postencephalitic parkinsonism.

[0138] Item 25. The method according to any one of Items 1 to 24, wherein the tauopathy is Alzheimer's disease (AD).

[0139] Item 26. The method according to any one of Items 1 to 25, wherein the SCO-spondin-derived peptide is administered to the subject intravenously, intraarterially, or intraperitoneally.

[0140] Item 27. The method according to any one of Items 1 to 26, wherein the formation of amyloid beta plaques is inhibited or reduced, the aggregation of amyloid beta is prevented or reduced, and / or already formed amyloid beta accumulations or amyloid beta aggregates are depolymerized.

[0141] Item 28. Amyloid beta 1-42 Item 28. The method according to any one of Items 1 to 27, wherein hyperphosphorylation of tau protein is inhibited or reduced.

[0142] Item 29. The method according to any one of Items 1 to 28, wherein the peptide is NX210, NX218, or a mixture of NX210 and NX218.

[0143] The invention will now be described in more detail, by way of non-limiting example, with reference to the drawings. [Brief explanation of the drawings]

[0144] [Figure 1] Figure 1 shows the effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on Aβ25-35-induced spatial working memory impairment in mice. Latency to reach the platform was determined during 5 days of training. Doses are expressed in mg per kg. n equals 11-12 per group. Data are expressed as time to find the platform (seconds). ns (not significant), ***, ###, or □□□ p<0.001 versus the Aβ25-35 / Vhc group by two-way ANOVA followed by Tukey's multiple comparison test. [Figure 2]Figure 1 shows the effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on Aβ25-35-induced spatial working memory impairment in mice. Time spent in each quadrant was determined during the probe trial. Doses are expressed in mg per kg. n equals 11-12 per group. Data are expressed as the percentage of time spent in the target quadrant compared to the mean percentage of time spent in the three other quadrants. ns (not significant), ***p<0.001 vs. Aβ25-35 / Vhc group by two-way ANOVA followed by Tukey's multiple comparison test. [Figure 3] Figure 1 shows the effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on biochemical changes (Aβ1-42 and phosphorylated tau levels) in the mouse brain induced by Aβ25-35. Doses are expressed in mg per kg. n equals 5-6 per group. Data are expressed as percentage of the control group (Sc.Aβ / Vhc group). ns (not significant), ***p<0.001 vs. Aβ25-35 / Vhc group by one-way ANOVA followed by Dunnett's test. [Figure 4] Figure 1 shows the effect of administering active or subactive doses of NX210 or NX218 (0.1-2 mg / kg once daily IP), or active or subactive doses of DPZ (0.25-1 mg / kg once daily PO), or a combination of subactive doses of NX210 or NX218 with subactive doses of DPZ (NX210 and NX218 at 0.1 mg / kg IP, and DPZ at 0.25 mg / kg PO) on Aβ25-35-induced short-term memory impairment in mice. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed as the percentage of alternation. ns (not significant), ***p<0.001 vs. Aβ25-35 / Vhc group by one-way ANOVA followed by Dunnett's test. [Figure 5]Figure 1 shows the effect of administration of active or subactive doses of NX210 or NX218 (0.1-2 mg / kg once daily, IP), or active or subactive doses of DPZ (0.25-1 mg / kg once daily, PO), or a combination of subactive doses of NX210 or NX218 with subactive doses of DPZ (NX210 and NX218 at 0.1 mg / kg, IP, and DPZ at 0.25 mg / kg, PO) on long-term memory impairment in mice induced by Aβ25-35, i.e., step-through latency (STL) measured during the retention session. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed in seconds. By Kruskal-Wallis test followed by Dunn's multiple comparison test, ns (not significant), **p<0.01, and ***p<0.001 relative to the Aβ25-35 / Vhc group. [Figure 6] Figure 1 shows the effect of administration of NX210 or NX218 (2 mg / kg once daily starting on D11) on Aβ25-35-induced short-term memory impairment in mice. Doses are expressed in mg per kg. n equals 6 per group. Data are expressed as the percentage of alternation behavior over time (D08-15-22-29). One-way ANOVA followed by Dunnett's test: ns (not significant), ***p<0.001 for the Aβ25-35 / D11 vhc group (* for Sc. Aβ / D11 vhc, $ for Aβ25-35 / D11 NX210 IP2, and □ for Aβ25-35 / D11 NX218 IP2). [Figure 7]Administration of NX218 (2 mg / kg IP once daily for 120 days (Group 4)) or escalating doses of NX218 (2 mg / kg IP once daily from days 44 to 78, then 4 mg / kg once daily from days 79 to 99, then 8 mg / kg once daily from days 100 to 113) on Aβ25-35-induced short-term memory impairment in mice. Figure 1 shows the long-term effects of administering PZ (1 mg / kg orally once daily for 43 days) (Group 5), or a combination of subactive doses of NX218 (0.1 mg / kg IP once daily for 120 days) and DPZ (0.25 mg / kg orally once daily for 120 days) (Group 6), or transient treatment with NX218 (2 mg / kg IP once daily for 28 days from days 11 to 38 (Group 3)). Data are expressed as the percentage of alternation behavior over time. n equals 5-6 per group. ns (not significant), **p<0.01, ***p<0.001 vs. Aβ25-35 vhc group (Group 2) by one-way ANOVA followed by Dunnett's test. [Figure 8] Figure 1 shows the PK profile after IV administration of NX210 in monkeys (bolus IV injection of 10 mg / kg NX 210). Mean monkey plasma concentrations (ng / mL) of NX218 (cyclic form of NX210) are plotted on the y-axis on a decimal log scale. [Example]

[0145] Introduction: Alzheimer's disease (AD) and related tauopathies are the most common age-related neurodegenerative disorders, yet no curative treatment exists. Current drugs only temporarily slow the symptoms of dementia but cannot alter the course of the disease. As a result, there is an urgent need to identify novel treatments.

[0146] The examples use a tauopathy model in which mice are injected with Aβ on day 01 (D01). 25-35The peptide (or scrambled Aβ peptide (Sc.Aβ) in sham animals) was injected intracerebroventricularly (ICV). This severe animal model elicits specific biomarkers (cerebral amyloid beta) involved in the physiopathology and progression of human tauopathies. 1-42 These findings mimic the changes in the brain (including accumulation of phospholipids and hyperphosphorylated tau protein) and the profound cognitive impairment (memory loss) experienced by patients (Maurice et al., 1996, 1998, 2013; Meunier et al., 2006).

[0147] In this dose-response study with peptides derived from SCO-spondin, treatment reduced Aβ 25-35 Peptides were injected with NX210, NX218, or vehicle (Vhc, HO) starting 1 hour (as a model of early disease) or 10 days later (as a model of late disease). NX210, NX218, and vehicle were administered intraperitoneally (IP) once daily or every other day (once every two days). The synthesis of NX210 and NX218 is described in Examples 1 and 2.

[0148] Furthermore, NX210 or NX218 were combined with standard therapy (an inhibitor of acetylcholinesterase, here donepezil, DPZ). To this end, subactive doses of the NX210 or NX218 peptides were combined with subactive doses of DPZ to study potential synergistic therapeutic effects.

[0149] Finally, a study was conducted to evaluate the long-term effects of NX218 and / or DPZ-based treatment.

[0150] Example 1 Synthesis of NX peptide The process for preparing peptides of sequences SEQ ID NO: 1, 2, or any of the peptides of sequences 3 to 63, particularly the peptide used in the Examples section, e.g., NX210 (SEQ ID NO: 3), is based on solid-phase peptide synthesis, applying N-α-Fmoc (side chain) protected amino acids as building blocks in the assembly of peptides. The protocol used consists of coupling an N-α-Fmoc-protected amino acid at its C-terminal glycine attached to an MPPA linker onto MBHA resin, followed by an Fmoc coupling / deprotection sequence. Assembly of the peptide on the resin is followed by a step of simultaneous peptide cleavage from the resin and deprotection of the amino acid side chains.

[0151] The crude peptide is precipitated, filtered, and dried. Prior to purification by preparative reverse-phase chromatography, the peptide is dissolved in an aqueous solution containing acetonitrile. The purified peptide in solution is concentrated, followed by an ion-exchange step to obtain the peptide in its acetate salt form.

[0152] Those skilled in the art can refer to US 6,995,140 and WO2018146283 for further synthetic details, and WO2017 / 051135 for oxidized forms of the peptides disclosed herein, all of which are incorporated herein by reference.

[0153] Those skilled in the art can further utilize standard methods for producing any of the disclosed peptides of the present invention, including peptides modified or protected at the N-terminus and C-terminus. For acetylation and / or amidation of peptides at the N-terminus and C-terminus, respectively, those skilled in the art can refer to standard techniques, such as those described in Biophysical Journal, Vol. 95, November 2008, pp. 4879-4889, which is also incorporated by reference.

[0154] Example 2 Synthesis of cyclic NX peptide A polypeptide with the sequence WSGWSSCSRSCG was added to human serum albumin (HSA) in a 1:1 ratio and incubated for 1-3 hours at room temperature in air with stirring. Using HPLC, we observed the formation of a peak corresponding to the polypeptide sequence WSGWSSCSRSCG, in which two cysteines are linked by a disulfide bridge. After removing the albumin by precipitation, the product was then purified and analyzed by HPLC. While it was found that smaller amounts of albumin were easier to remove, using different ratios of albumin and polypeptide with the sequence WSGWSSCSRSCG allowed for the influence of the cyclization rate and final cyclization yield. The cyclized compound was NX218.

[0155] Those skilled in the art can refer to WO2017051135 for further synthetic details, which is incorporated herein by reference.

[0156] Example 3 Dose-response efficacy of NX210 and NX218 Cognitive assessment: a. Spontaneous alternation behavior (Y-maze) Aβ 25-35 Seven days after ICV injection of the peptide (D08), all animals were tested for spontaneous alternation performance in a Y-maze (short-term memory test), an index of spatial working memory. Each mouse was placed at the end of one arm and allowed to move freely through the maze during an 8-minute session. An alternation was defined as consecutive entries into all three arms. The maximum number of alternations was therefore the total number of arm entries minus 2, and the percentage of alternations was calculated as (actual number of alternations / maximum number of alternations) × 100. This parameter includes the percentage of alternations (an index of memory) (Maurice et al., 1996, 1998; Meunier et al., 2006).

[0157] [Table 3] Table 1. Aβ 25-35 Effect of IP administration of NX210 or NX218 peptides (0.1-1-2-3.75 mg / kg once a day or every other day (1 / 2d)) on short-term memory impairment induced by Aβ in mice. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed as percentage of alternation behavior. One-way ANOVA followed by Dunnett's test showed that Aβ 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0158] Administration of NX210 (1, 2, and 3.75 mg / kg) or NX218 (1, 2, and 3.75 mg / kg) once daily or every other day significantly reversed spatial short-term working memory impairment. Complete recovery was achieved at 2 and 3.75 mg / kg for NX210 and at 1, 2, and 3.75 mg / kg for NX218, levels similar to those observed in uninjured sham animals (Sc.Aβ / Vhc). This demonstrates the potent efficacy of both the NX210 and NX218 peptides in countering the cognitive dysfunction (short-term memory loss) observed in this AD / tauopathy animal model (Table 1).

[0159] Once-daily administration of NX210 (0.1 mg / kg) or NX218 (0.1 mg / kg) significantly reduced Aβ 25-35 Since these compounds do not interfere with the short-term memory impairment induced by α-glucan, 0.1 mg / kg is set as the subactive dose of these two compounds (Table 1), allowing for combination studies with standard treatments (Example 4).

[0160] b. Step-Through Passive Avoidance Test (STPA) After the Y-maze test, the same animals also underwent a step-through passive avoidance (STPA) test, which is an index of contextual long-term memory (Aβ 25-35Eight and nine days after the ICV injection of the peptide (D09 and D10, respectively), the apparatus consisted of a box with two compartments (light and dark) separated by a guillotine door. A shock generator scrambler was used to deliver foot shock to the dark compartment. On the first day (training session), each mouse was placed in the light compartment with the guillotine door initially closed. After 5 seconds, the door was raised. Once the mouse entered the dark compartment and placed all of its paws on the grid floor, the door was closed and a foot shock was delivered for 3 seconds. 24 hours after this training session, a retention test (in which no foot shock was administered) was conducted. Specifically, each mouse was again placed in the light compartment with the door closed. After 5 seconds, the door was raised. The time it took for the mouse to enter the dark compartment, called the step-through latency (STL), was recorded up to 300 seconds. The time it takes the mouse to exit the dark compartment, called the escape latency (EL), is recorded up to 300 seconds ( Maurice et al., 1996 , 1998 ; Meunier et al., 2006 ).

[0161] [Table 4] Table 2. Aβ 25-35 Effect of IP administration of NX210 or NX218 (0.1-1-2-3.75 mg / kg once a day or every other day (1 / 2d)) on long-term memory impairment in mice induced by Aβ: step-through latency (STL) measured during the retention session. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed in seconds. Kruskal-Wallis test followed by Dunn's multiple comparison test showed no significant difference in Aβ 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0162] [Table 5] Table 3. Aβ 25-35Effect of IP administration of NX210 or NX218 (0.1-1-2-3.75 mg / kg once a day or every other day (1 / 2d)) on long-term memory impairment induced by Aβ in mice: escape latency (EL) measured during the retention session. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed in seconds. Kruskal-Wallis test followed by Dunn's multiple comparison test showed significant differences in Aβ 25-35 ns (not significant), **p<0.01, and ***p<0.001 relative to the / Vhc group.

[0163] Administration of NX210 (2 and 3.75 mg / kg) or NX218 (1, 2, and 3.75 mg / kg) once daily or every other day completely reversed contextual long-term memory impairment (STL and EL), reaching levels similar to those in uninjured sham animals (Sc.Aβ / Vhc). This dose-effect relationship demonstrates the potent efficacy of both NX210 and NX218 in counteracting the cognitive dysfunction (long-term memory impairment) seen in this tauopathy model (Tables 2-5).

[0164] Once-daily administration of NX210 or NX218 (0.1 mg / kg) did not prevent contextual long-term memory impairment (STL and EL), confirming that 0.1 mg / kg is a subactive dose for both compounds (Tables 2-3).

[0165] c. Morris Water Maze Place Learning-Reference Memory Test (MWM) After the Y-maze and instead of STPA testing, some animals underwent the Morris Water Maze (MWM) test to assess spatial working memory impairment (six days of testing from D09 to D14). This gold-standard test consisted of a circular pool filled with turbid water placed in an externally cued room (sink, contrasting wallpaper, shelf). The platform was submerged during acquisition. Training consisted of three swims per day over five days, between D09 and D13, with a 20-minute intertrial interval. Each mouse had 90 seconds to swim and 20 seconds to find the platform.

[0166] The probe trial (PT) is performed 24 hours after the last swim on D14. In the PT, the platform is removed and each animal is allowed to swim for 60 seconds. The starting position of each mouse corresponds to one of two positions away from the platform location in a counterbalanced order. During the PT, one quadrant of the platform is designated "target," and the other quadrants (opposite, adjacent right, and adjacent left) are designated "other." The time spent in each quadrant is recorded. Results are expressed as the percentage of time spent in the target quadrant compared to the average percentage of time spent in the three other quadrants.

[0167] Each day, the latency to find the platform during the first (Swim 1), second (Swim 2), third (Swim 3), fourth (Swim 4), and fifth (Swim 5) trials was monitored and averaged over the 6 days of the experiment (Maurice et al., 2013).

[0168] [Table 6] Table 4. Aβ 25-35Effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on spatial working memory impairment induced by Aβ in mice: Latency to reach the platform was determined during 5 days of training. Doses are expressed in mg per kg. n equals 11–12 per group. Data are expressed as time to find the platform (seconds). Two-way ANOVA followed by Tukey's multiple comparison test was used to evaluate Aβ levels. 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0169] [Table 7] Table 5. Aβ 25-35 Effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on spatial working memory impairment induced by Aβ in mice: time spent in each quadrant was determined during the probe trial. Doses are expressed in mg per kg. n equals 11–12 per group. Data are expressed as the percentage of time spent in the target quadrant (target) compared with the mean percentage of time spent in the three other quadrants (other). Two-way ANOVA followed by Tukey's multiple comparison test was used to evaluate the effect of Aβ on the spatial working memory impairment in mice. 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0170] Administration of NX210 or NX218 significantly reversed the learning profile and spatial long-term memory impairment in this model, as shown by the Morris water maze results (Tables 4-5 and Figures 1-2). On the second day of training (D10), mice treated with NX210 or NX218 memorized the location of the hidden platform as efficiently as uninjured sham mice (Sc.Aβ / Vhc). The beneficial effects were maintained throughout the 5-day training trial (Table 4 and Figure 1).

[0171] The beneficial effects on spatial memory were confirmed by the probe test (D14), in which the platform was removed. In this test, mice treated with NX210 or NX218 spent approximately 40% of their total swimming time in the target quadrant (i.e., the location of the removed platform), which was comparable to intact sham mice (Sc.Aβ / Vhc) in terms of memory for the platform location. Conversely, Aβ 25-35 / Vhc mice did not remember the location of the platform and therefore spent approximately 25% of their swim time in each quadrant (Table 5 and Figure 2).

[0172] This again demonstrates the strong efficacy of both NX210 and NX218 in countering the cognitive impairment (spatial working memory impairment) seen in this tauopathy model.

[0173] Biochemical and histological evaluation: a. Evaluation of brain biomarkers To evaluate the potential biological effects of NX210 or NX218 treatment on pathological biomarkers, the Sc.Aβ / Vhc group (n=6), Aβ 25-35 / Vhc group (n=6), Aβ 25-35 / NX210 IP 2mg / kg group (n=5), and Aβ 25-35 Mice in the / NX218 IP 2mg / kg group (n=5) were treated with daily compound administration (once a day) and Aβ 25-35 Ten days after ICV injection of the peptide (D11), the mice were sacrificed under anesthesia.

[0174] For each mouse, the brain was quickly removed and dissected on an ice-cold metal plate into two hemihippocampi, two hemifrontal cortices, and the remaining brain. Immediately after collection, the brain samples were frozen on dry ice and stored at -80°C. After thawing, the brain structures were dissociated in 50 mM Tris-150 mM NaCl buffer, pH 7.5, and sonicated for 20 seconds. After centrifugation, the protein-containing supernatant was used to perform an ELISA assay according to the manufacturer's instructions (see below). - Aβ 1-42 (Amyloid-beta 1-42) in the left hippocampus, using an ELISA kit from Cloud-Clone Corp., reference number: CEA946Mu, batch: L190107385 - For pTau (phosphorylated tau protein), left hippocampus, use Fisher Scientific ELISA kit, reference number: 10591255, batch: 187860001

[0175] For all assays, absorbance was read at 450 nm and the concentration of each sample was calculated using a specific standard curve. All samples were run in duplicate and the average of these duplicates was used for calculations.

[0176] [Table 8] Table 6. Aβ 25-35 Biochemical changes in the mouse brain induced by Aβ 1-42 Effect of IP administration of NX210 or NX218 (2 mg / kg once daily) on Aβ levels (and phosphorylated tau levels). Doses are expressed in mg per kg. n equals 5-6 per group. Data are expressed as percentage of the control group (Sc.Aβ / Vhc group). One-way ANOVA followed by Dunnett's test showed that Aβ 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0177] In this tauopathy model, Aβ 25-35 Compared to the / Vhc group, once-daily administration of NX210 (2 mg / kg) or NX218 (2 mg / kg) for 10 days surprisingly and significantly reduced several brain biomarkers typically associated with the progression of AD and other tauopathies, such as amyloid beta 1-42 (Aβ 1-42 ) and hyperphosphorylated tau protein (pTau) (Table 6 and Figure 3). 1-42and pTau levels were similar to those in healthy sham animals (for both the NX210- and NX218-treated groups, and the NX218-treated group, respectively).

[0178] b.Histological study To measure neuronal loss in this model (described by Maurice et al., 2013), a well-known pathological event occurring in patients with AD / tauopathy, hippocampal neurons were cultured in sham mice, Aβ-treated mice, and then cultured in mice. 25-35 / vehicle mice, and Aβ 25-35 Counts were made in the brains of / NX210-treated mice.

[0179] To do this, daily compound administration of NX210 (IP) or vehicle (IP) and Aβ 25-35 Fourteen days after ICV injection of the α- or scrambled peptide (D15), mice were anesthetized with ketamine / xylazine and then transcardially perfused with phosphate-buffered saline (PBS, 15 ml at 5 ml / min, pH 7.4) until fluid flow became clear, followed by 4% paraformaldehyde solution (PFA, 25 ml at 5 ml / min) until fixational eye movements disappeared. The brains were carefully removed and kept in 4% PFA at 4°C for 24–48 hours. The brains were then dehydrated through a series of gradient ethanol baths to remove water and then infiltrated with wax. Finally, the infiltrated brains were embedded in wax blocks and sectioned at 5 ± 1 μm. Nine brain sections per animal were stained with cresyl violet (CV), and CA1 pyramidal neurons were counted in coronal sections spaced 100 μm apart.

[0180] [Table 9] Table 7. Aβ 25-35 Effect of IP administration of NX210 (2 mg / kg once daily) on the neuronal loss observed in mice injected with Aβ. Data are expressed as the ratio of the number of nuclei over the length of the analyzed area and as a percentage of the control (Sc.Aβ / Vhc). One-way ANOVA followed by Dunnett's test showed that Aβ25-35 *p<0.05 and ***p<0.001 versus the / Vhc group.

[0181] NX210 (2 mg / kg) inhibits Aβ 25-35 It significantly prevents the neuronal loss observed in the hippocampus of mice injected with the peptide (Table 7). 25-35 Only 82 percent of pyramidal neurons remained in / Vhc mice (compared to sham mice), compared with 93 percent after treatment with NX210.

[0182] Example 4 Standard treatment in combination with NX210 or NX218 Acetylcholinesterase inhibitors, such as donepezil (DPZ), are currently the standard treatment for patients suffering from Alzheimer's disease or tauopathy. The efficacy response to these compounds is unpredictable, and loss of efficacy occurs over time. Increasing the dose is only a temporary option, because the side effects associated with this increased dosage are poorly tolerated by patients (Homma et al., 2009; Jackson et al., 2004). Ultimately, patients will therefore discontinue treatment, leaving them without a therapeutic solution.

[0183] Cognitive assessment: a. Spontaneous alternation behavior (Y-maze) Aβ 25-35 Seven days after ICV injection of the peptides (D08), all animals were tested for spontaneous alternation performance in the Y-maze, an index of spatial working memory (short-term memory test, see Example 3 for methodological details).

[0184] [Table 10] Table 8. Aβ 25-35Effects of administration of active or subactive doses of NX210 or NX218 (0.1-2 mg / kg once daily IP), or active or subactive doses of DPZ (0.25-1 mg / kg once daily PO), or a combination of subactive doses of NX210 or NX218 with subactive doses of DPZ (NX210 and NX218 at 0.1 mg / kg IP, and DPZ at 0.25 mg / kg PO) on short-term memory impairment induced by NF-κB in mice. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed as percentage of alternation. One-way ANOVA followed by Dunnett's test showed significant differences in Aβ levels. 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0185] At subactive doses, once-daily administration of NX210 (0.1 mg / kg), NX218 (0.1 mg / kg), or DPZ (0.25 mg / kg) significantly reduced Aβ 25-35 (Table 8 and Figure 4, see also Example 3).

[0186] Surprisingly, the inventors found that a subactive dose of NX210 or NX218 (0.1 mg / kg, IP) in combination with a subactive dose of DPZ (0.25 mg / kg, PO) significantly reduced Aβ 25-35 The results showed that tauopathy-associated cognitive impairment was significantly and completely reversed by tauopathy-induced spatial short-term working memory impairment (Table 8 and Figure 4), demonstrating a synergistic therapeutic effect between the drugs on cognitive impairment associated with tauopathy.

[0187] b. Step-Through Passive Avoidance Test (STPA) Aβ 25-35 Eight and nine days after ICV injection of the peptides (D09 and D10, respectively), animals were tested for contextual long-term memory performance in the step-through passive avoidance (STPA) test (see Example 3 for method details).

[0188] [Table 11] Table 9. Aβ 25-35 Effects of administration of active or subactive doses of NX210 or NX218 (0.1–2 mg / kg once daily IP), or active or subactive doses of DPZ (0.25–1 mg / kg once daily PO), or a combination of subactive doses of NX210 or NX218 with subactive doses of DPZ (NX210 and NX218 at 0.1 mg / kg IP, and DPZ at 0.25 mg / kg PO) on long-term memory impairment induced by NF-κB in mice: step-through latency (STL) measured during the retention session. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed in seconds. A Kruskal-Wallis test followed by Dunn's multiple comparison test was used to measure Aβ levels. 25-35 ns (not significant), **p<0.01, and ***p<0.001 relative to the / Vhc group.

[0189] [Table 12] Table 10. Aβ 25-35 Effects of administration of active or subactive doses of NX210 or NX218 (0.1–2 mg / kg once daily IP), or active or subactive doses of DPZ (0.25–1 mg / kg once daily PO), or a combination of subactive doses of NX210 or NX218 with subactive doses of DPZ (NX210 and NX218 at 0.1 mg / kg IP, and DPZ at 0.25 mg / kg PO) on long-term memory impairment induced by Aβ in mice: escape latency (EL) measured during the retention session. Doses are expressed in mg per kg. n equals 12 per group. Data are expressed in seconds. Kruskal-Wallis test followed by Dunn's multiple comparison test showed significant differences in Aβ levels. 25-35 ns (not significant), *p<0.05 and ***p<0.001 compared to the / Vhc group.

[0190] At subactive doses, once-daily administration of NX210 (0.1 mg / kg), NX218 (0.1 mg / kg), or DPZ (0.25 mg / kg) significantly reduced Aβ 25-35 (STL and EL, Tables 9-12 and Figure 5, see also Example 3).

[0191] Surprisingly, the inventors have demonstrated in their studies that a subactive dose of NX210 or NX218 (0.1 mg / kg, IP) in combination with a subactive dose of DPZ (0.25 mg / kg, PO) significantly inhibits Aβ 25-35 The results show that NX210 or NX218 significantly and completely reversed the long-term memory impairment induced by tauopathy (STL and EL, Tables 9-12 and Figure 5). This result supports the synergistic beneficial effect of combining NX210 or NX218 with an acetylcholinesterase inhibitor (here, donepezil) on cognitive dysfunction associated with tauopathy.

[0192] Example 5 Late-stage efficacy of NX210 or NX218 Cognitive assessment: a. Spontaneous alternation behavior (Y-maze) Aβ in tauopathy 25-35 In mouse models, Aβ 25-35 Ten days after ICV injection of the peptide (D11), the physiopathology was characterized by high cognitive impairment seen in the Y-maze and STPA tests (Example 3, Tables 1-5, vehicle group), as well as by the expression of two pathological brain biomarkers, amyloid beta 1-42 (Aβ 1-42 ) and the large changes in hyperphosphorylated tau protein (pTau) (Example 3, Table 6 and Figure 3, vehicle group) are already at a very late stage.

[0193] The efficacy of the SCO-Spondin-derived peptides NX210 and NX218 was therefore evaluated during this late physiopathological situation. Specifically, administration of NX210, NX218, or vehicle was initiated on D11 and repeated once daily until the end of the experiment.

[0194] All animals were tested for spontaneous alternation performance in the Y-maze (see Example 3 for method details), an index of short-term memory, starting on D08 (i.e., pre-treatment values) and once a week for 3 weeks (D15, D22, D29).

[0195] [Table 13A] [Table 13B] Table 11. Aβ 25-35 Effect of administration of NX210 or NX218 (2 mg / kg once daily starting on D11) on short-term memory impairment induced by Aβ in mice. Doses are expressed in mg per kg. n equals 6 per group. Data are expressed as percentage of alternation over time (D08-15-22-29). One-way ANOVA followed by Dunnett's test showed that Aβ 25-35 ns (not significant), ***p<0.001 compared to the / Vhc group.

[0196] On D08 (value before treatment), all Aβ 25-35 Strong memory impairment was also observed in both groups (Table 11 and Figure 6).

[0197] On D15, five daily doses of NX210 or NX218 (2 mg / kg) reduced Aβ 25-35The results already showed a tendency to reverse the spatial short-term working memory impairment induced by NX210 (p-values ​​= 0.19 and 0.09 for NX210 and NX218, respectively, Table 11 and Figure 6). At D22, after 12 days of treatment, administration of NX210 or NX218 significantly reversed the working memory impairment (Table 11 and Figure 6). Finally, at D29, after 19 days of treatment, administration of NX210 or NX218 completely reversed the memory impairment seen in this tauopathy model (97% and 91%, respectively, compared to the sham group, Table 11 and Figure 6).

[0198] In conclusion, NX210 and NX218 were able to completely reverse cognitive and memory impairments in this tauopathy model, even when treatment was initiated at a very advanced pathophysiological stage (late in the pathogenesis).

[0199] Example 6 Long-term effects of NX218 and / or DPZ-based treatments A long-term study (120 days) was conducted to examine the ability of NX218 treatment to maintain cognitive recovery over time and to serve as a second-line treatment when DPZ loses its activity. 25-35 This was done in a mouse model.

[0200] Mice were assigned to six treatment groups with 5-6 animals per group, as described below.

[0201] [Table 14]

[0202] On day 1 (D01), amyloid β (Aβ) was administered to induce amyloid toxicity. 25-35 ) or not cause (Sc.Aβ) Sc.Aβ or Aβ 25-35 The peptide was injected ICV.

[0203] Control group: From D01 to D120, mice in groups 1 and 2 were administered IP once a day (od) with vehicle (water for injection).

[0204] Test compound NX218: - From D11 to D38, mice of group 3 were administered 2 mg / kg NX218 IP od (mice were not treated with vehicle from D01 to D10). - From D01 to D120, mice in group 4 were administered 2 mg / kg NX218 IP od.

[0205] DPZ rescued by test compound NX218: From D01, mice in group 5 were administered an active dose (1 mg / kg) of DPZ PO by gavage at od until its effectiveness disappeared, which disappeared on D36. DPZ was administered until D43, and then from D44 to D78, DPZ was replaced by an active dose (2 mg / kg) of NX218 via IP administration once a day. - From D79 to D99, a higher dose (4 mg / kg) of NX218 was administered IP od. - From D100 to D113, a higher dose (8 mg / kg) of NX218 was administered IP od. - NX218 administration was stopped from D114 until the animals were sacrificed.

[0206] Test compound NX218 in combination with DPZ: From D01 to D120, mice of group 6 were co-administered with NX218 and DPZ at their respective subactive doses: NX218:IP, od, 0.1 mg / kg DPZ: PO, od, 0.25 mg / kg

[0207] Cognitive assessment: All groups received D08 (Aβ 25-35 Behavioral testing was performed once a week starting 7 days after peptide injection to monitor the effects of the compounds.

[0208] Spontaneous alternation procedure in the Y-maze (YM, assessment of spatial short-term memory / working memory as described in Example 3) at D08, D15, D22, D29, D36, D43, D50, D57, D64, D71, D78, D85, D92, D99, D106, D113, and D120. The results are shown in Table 12 and Figure 7.

[0209] [Table 15A] [Table 15B] [Table 15C] [Table 15D] Table 12. Aβ 25-35 Administration of NX218 (2 mg / kg IP once daily for 120 days (Group 4)) or escalating doses of NX218 (2 mg / kg IP once daily from days 44 to 78, then 4 mg / kg IP once daily from days 79 to 99, then 8 mg / kg once daily from days 100 to 113) for short-term memory impairment in mice induced by DP Long-term effects of administration of NX218 (1 mg / kg orally once daily for 43 days) (Group 5), or a combination of subactive doses of NX218 (0.1 mg / kg IP once daily for 120 days) and DPZ (0.25 mg / kg orally once daily for 120 days) (Group 6), or transient treatment with NX218 (2 mg / kg IP once daily for 28 days from day 11 to day 38 (Group 3)). Data are expressed as the percentage of alternation behavior over time. n equals 5-6 per group. One-way ANOVA followed by Dunnett's test revealed significant differences in Aβ levels. 25-35 ns (not significant), **p<0.01, ***p<0.001 compared to the vhc group (group 2).

[0210] At 120 days follow-up, mice treated with 2 mg / kg NX218 (Groups 3 and 4) showed complete and sustained recovery without any loss of efficacy, in contrast to DPZ (Group 5), which was only effective until D36.

[0211] Mice with known pathology and cognitive impairment transiently treated with NX218 peptide from D11 to D38 (group 3) completely reversed the memory changes seen in YM. Furthermore, the benefit was maintained until D120 without re-administration of NX peptide, highlighting its disease-modifying effect (targeting disease pathogenic pathways to significantly modify or reverse the progression of pathology (i.e., cognitive impairment)) rather than a temporary symptomatic effect (as with commercially available drugs such as DPZ).

[0212] After DPZ resistance after 4 weeks of treatment (37% change), NX218 treatment (group 5) completely reversed memory loss for up to 17 weeks (variation between 53% and 80% depending on the time and dose of NX peptide administration).

[0213] Mice treated with NX218+DPZ (Group 6) showed complete and sustained reversal of spatial short-term working memory impairment (71%-80% change over the course of the study compared with 35-49% in vehicle-treated mice and 73-80% in the scrambled Aβ group) for up to 17 weeks without loss of efficacy.

[0214] Example 7 Pharmacokinetics in Animals Preliminary in vitro experiments showed that NX210 was rapidly converted to NX218 by oxidation in rat plasma. Therefore, NX210 PK in animals was followed by measuring its cyclic form, NX218. First, preliminary PK studies in rats were performed to validate the method for detecting NX218 in plasma, and then transferred to monkeys by repeating the experiments to perform more robust PK studies. All PK studies were performed using 0.9% NaCl as a vehicle.

[0215] Preliminary PK studies in rats: In the four rats tested, NX218 concentrations declined rapidly and were no longer quantifiable 3 hours after a slow bolus IV injection of 49 mg / kg NX210 (data not shown). This study demonstrated that the identification of NX218 in animal plasma is feasible.

[0216] PK studies in monkeys: In the three monkeys tested, data were reproducible after repeat studies performed on different days (D22, D37, and D51) following a bolus IV injection of 10 mg / kg NX210. NX210 concentrations declined rapidly until 30 minutes after injection (Figure 8). The half-life was estimated to be approximately 12 minutes (Table 13), and therefore was within the same range as that observed in rats and dogs (data not shown), with high consistency across repeated doses.

[0217] [Table 16] AUC: area under the curve, CL: total clearance, Cmax: maximum concentration, t1 / 2: terminal elimination half-life, Tmax: time to reach Cmax, Vss: volume of distribution at steady state Summary statistics of mean monkey plasma PK parameters for NX218 (±standard deviation where available)

[0218] Example 8 Pharmacokinetics in healthy human subjects As in animals, PK studies of NX210 in healthy human subjects were conducted following measurements of its cyclic form, NX218. Six healthy subjects were administered a 10 mg / kg dose via 12-minute IV infusion. 1 / 2 was estimated based on data from three patients (see table below). The half-life was estimated to be approximately 19 minutes (Table 14), and therefore within the same range as that seen in animals.

[0219] [Table 17] AM=arithmetic mean, SD=standard deviation, Min=minimum, Max=maximum, GM=geometric mean; CV: Coefficient of variation Kel: clearance rate constant AUC: Area under the curve Cmax: maximum concentration t1 / 2: terminal elimination half-life Tmax: Time to reach Cmax CV% = arithmetic CV, which is equal to SD / AM × 100 a: There were 3 subjects with an AUClast / AUC∞ ratio <0.80 or adjusted R2 value <0.80, and therefore their Kel, t 1 / 2 , and AUC∞ are not reportable and are not included in the summary statistics.

[0220] (References) TIFF0007784378000022.tif148170

Claims

1. Amino acid sequence X1-WS-A1-WS-A2-CS-A3-A4-CG-X2 (SEQ ID NO: 1) 1. A composition for use in the treatment of a tauopathy comprising a peptide of the formula: - A1 and A2 are independently selected from G and S, and A3-A4 are selected from RS, VS, VT, and RT; - X1 and X2 consist of an amino acid sequence consisting of 1 to 6 amino acids, or X1 and X2 are absent; - the N-terminal amino acid can be acetylated, the C-terminal amino acid can be amidated, or the N-terminal amino acid can be acetylated and the C-terminal amino acid can be amidated; The peptide is administered to the subject via a systemic route; composition.

2. X1 is a hydrogen atom, P, AP, LAP, or VLAP, and / or The composition of claim 1, wherein X2 is a hydrogen atom, L, LG, LGL, LGLI, or LGLIF.

3. The peptide has an amino acid sequence WS-A1-WS-A2-CS-A3-A4-CG (SEQ ID NO: 2) The composition according to claim 1 or 2,

4. 4. The composition according to any one of claims 1 to 3, wherein the peptide is a linear peptide or an oxidized peptide in which the cysteines present in the peptide formulae of SEQ ID NOs: 1 and 2 form disulfide bridges, or a mixture of both linear and oxidized peptides.

5. The composition according to any one of claims 1 to 4, wherein the peptide is a peptide of a sequence selected from the group consisting of the sequences SEQ ID NO: 3 to 63.

6. 6. The composition of claim 5, wherein the peptide is a peptide of sequence SEQ ID NO: 3 in linearized form, in cyclized form, or a mixture of both.

7. The composition according to any one of claims 1 to 6, wherein the tauopathy is selected from the group consisting of Alzheimer's disease (AD), progressive supranuclear palsy (PSP), tau-positive frontotemporal dementia such as Pick's disease, dementia with Lewy bodies, corticobasal degeneration, Niemann-Pick disease type C, chronic traumatic encephalopathy including dementia pugilistica, and post-encephalitic parkinsonism.

8. The composition of any one of claims 1 to 6, wherein the tauopathy is Alzheimer's disease (AD).

9. 9. The composition of any one of claims 1 to 8, which induces a reduction or disruption of tau aggregation in a subject, a reduction in tau protein in a subject, and / or a reduction in the level of phosphorylated tau protein.

10. 10. The composition of any one of claims 1 to 9, wherein the composition is administered to a patient via intravenous, intraperitoneal, intranasal, subcutaneous, intramuscular, sublingual, or oral routes.

11. The composition of any one of claims 1 to 10, wherein the subject is also treated with a sufficient amount of an acetylcholinesterase inhibitor.

12. 11. The composition of any one of claims 1 to 10, further comprising an acetylcholinesterase inhibitor.

13. 13. The composition of claim 12, wherein the acetylcholinesterase inhibitor is DPZ.

14. Use of the composition of any one of claims 1 to 6 for the manufacture of a medicament for treating a tauopathy through the systemic route.

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