Method for safe administration of phosphorylated tau peptide vaccine

Abstract

To provide methods for inducing anti-phosphorylated Tau antibodies without inducing a severe adverse event in humans.SOLUTION: The methods include administering to the subject an effective amount of liposomes including a toll-like receptor 4 agonist and a Tau phosphopeptide presented on the surface of the liposome.SELECTED DRAWING: None

Description

[Technical field]

[0001] Reference to Electronically Submitted Sequence Listing The application is filed on February 7, 2020, and has a file size of 22KB. ence Listing 689001_66U1" Contains sequence listings submitted electronically via EFS-Web as a sequence listing in the format The sequence listing submitted via EFS-Web is incorporated herein by reference in its entirety. is incorporated herein by reference.

[0002] The present invention is in the field of medicine. In particular, the present invention relates to phosphorylated tau (pTau) Liposomes of peptides and preventing or treating tauopathies such as Alzheimer's disease Concerning its use for. [Background technology]

[0003] Alzheimer's disease (AD) is a progressive, debilitating neurodegenerative disease that affects an estimated 44 million people worldwide. Alzheimer's disease is a chronic condition that affects the brain and nervous system. Currently commercialized therapies for AD target the clinical symptoms. but do not target the pathogenic process underlying the disease (disease-modifying effect) Unfortunately, current therapies are only minimally effective, and therefore additional prevention is not feasible. And there is an urgent need to develop and test therapeutic interventions.

[0004] The pathology characteristic of Alzheimer's disease involves the formation of highly aggregated amyloid beta proteins. The accumulation of extracellular plaques containing tau protein and intracellular "tangles" of hyperphosphorylated tau protein The molecular events that lead to the accumulation of these proteins are poorly characterized. For amyloids, abnormal cleavage of the amyloid precursor protein leads to It is hypothesized that this leads to the accumulation of aggregation-prone fragments containing acids 1-42. Dysregulation of kinases, phosphatases, or both leads to abnormal phosphorylation of tau It has been hypothesized that hyperphosphorylation of tau effectively binds and binds microtubules. They lose their ability to stabilize and instead accumulate in the cytoplasm of affected neurons. Hyperphosphorylated tau appears to form first oligomers and then higher order aggregates. Their presence may be critical for the function of the neurons in which they are formed, possibly including normal axonal transport. It is believed that this will have a negative impact through interruptions in transport.

[0005] In developed countries, individuals diagnosed with Alzheimer's disease or other dementing tauopathies , commonly referred to as cholinesterase inhibitors (e.g., Aricept®) or memantine. These drugs are used in moderation. It is well tolerated but has very little efficacy. (brand name) delays the progression of symptoms by 6 to 12 months in approximately 50% of treated individuals. The remaining treatments are non-pharmacological and slow the patient's daily activities while decreasing cognitive ability. The focus is on making people more able to handle the challenges of

[0006] Immunotherapies for the prevention and treatment of AD are currently under development. Active immunization using this approach could potentially stimulate both antibody- and cell-mediated immune responses against AD. However, the first widely tested human anti-amyloid beta vaccine Evaluation of Chin was discontinued in 2002. Myelopathy, a type of central nervous system inflammation that can be fatal, Meningoencephalitis is a clinical study of AN-1792, an active immunotherapy targeting Aβ, in AD patients. This was observed in patients exposed to AN-1792 (Orgogozo et al., 2003). The encephaloinflammatory reaction that occurred in 6% of the patients was thought to be induced by unwanted activation of Aβ-specific T cells. can be obtained.

[0007] To date, very little research has been done with drugs that specifically target tau pathology. Tau immunotherapy is now moving into clinical trials, but the field is still in its early stages. The mechanistic understanding of the efficacy and safety of the various approaches is not well established (Sigu rdsson, Neurodegener Dis. 2016; 16(0):34-38). Encephalitis, or inflammation of the brain, is caused by the full-length transcription factor It has also been reported in mice immunized against the protein. The adverse effects were observed in mice immunized with a single injection of phosphorylated tau peptide in a proinflammatory CNS environment. No other studies have reported any side effects from the animals studied (Rosenmann H., 2013. Curr. Alzheimer Res. 10, 217 -228).

[0008] Nonphosphorylated tau peptides in human patients with mild to moderate Alzheimer's disease The long-term safety profile of the vaccine (AADvac1) has been recently published (Novak et al. , Alzheimer's Research & Therapy (2018) 10:108). This vaccine targets N-terminal cysteine Tau sequences coupled to keyhole limpet hemocyanin (KLH) via This vaccine contains a synthetic peptide derived from amino acids 294 to 305. A dose of 40 μg of KLH-coupled peptide (CKD Aluminum hydroxide adjuvant (0.5 mg of Al NIKHVPGGGS) 3 + The patients were enrolled in the study and received the Phase 1 study (FUNDAMANT study). Adverse events observed in 26 patients associated with AADvac1 treatment in the study (AE) was injection site reaction (erythema, swelling, heat, itching, pain, nodule). One or more of the following AEs were observed in 50% of AADvac1-treated patients: Injection site reactions were reversible and primarily mild in severity. Six serious adverse events were reported. Symptoms of acute encephalopathy (SAE) were observed (abdominal strangulation hernia, dehydration, acute psychosis, dementia-like behavior and The SAEs were: psychiatric symptoms, second-degree atrioventricular block, and sinus bradycardia. None of the allergic or anaphylactic reactions were considered to be related to AADvac1 treatment. No adverse drug reactions were observed. Safety signals were determined by clinical laboratory evaluations (coagulation, blood biochemistry, Hematology, hematology, and urinalysis, as well as vital signs evaluation, neurological and psychologic No safety signals were detected by MRI evaluation. No edematous changes occurred. No meningeal changes or meningoencephalitis were observed. Significant microbleeds were observed in one ApoE4 homozygote, and brain surface hemosiderin was increased in one Both events were detected in ApoE4 heterozygotes and were clinically silent. was considered consistent with the background incidence of such lesions in the AD patient population. Summary of the Invention [Problem to be solved by the invention]

[0009] However, the safety profile of the phosphorylated tau peptide vaccine in human patients No reports have been published on the safety and effectiveness of serotonin in treating neurodegenerative diseases such as Alzheimer's disease. There is a need for further treatment. [Means for solving the problem]

[0010] The present invention relates to a liposome comprising a phosphorylated tau peptide displayed on the surface of the liposome. Based on findings from clinical studies of the vaccine. The vaccine appears to be well tolerated in humans. This was higher than the dose used in the tau conjugate vaccine AADvac1 trial. At much higher doses, p-tau peptide liposomal vaccine induced severe adverse events It was shown that anti-phosphorylated tau antibodies were induced in human subjects without the use of tau agonists.

[0011] Thus, in one general aspect, the invention provides a method for treating severe infectious diseases, such as encephalitis, by administering an anti-phosphorylated tau antibody to a patient. A method for inducing tol in a human subject in need thereof without inducing an adverse event, comprising: L-like receptor 4 agonist and SEQ ID NO: 1 to SEQ ID NO: 3 and SEQ ID NO: 5 to SEQ ID NO: 12 and a tau phosphopeptide comprising an amino acid sequence selected from the group consisting of: and administering to the subject a tau phosphopeptide-containing therapeutic agent having a concentration of about 25 nmol per dose. e to about 750 nmole, e.g., about 29.7 nmole to about 742.5 nmole per dose ole, preferably about 90 nmole to about 715 nmole per dose, for example about 89 0.1 nmole to about 712.8 nmole, or about 90 nmole to about 5 nmole per dose 35 nmole, for example about 89.1 nmole to about 534.6 nmole per dose; or from about 90 nmole to about 275 nmole per dose, for example about 89 nmole per dose. The tau phosphopeptide is administered in an amount ranging from 0.1 nmole to approximately 267.3 nmole, and the liposome is In certain embodiments, the tau phosphopeptide is displayed on the surface of the tau protein. The tide is selected from the group consisting of SEQ ID NO: 27 to SEQ ID NO: 29 and SEQ ID NO: 31 to SEQ ID NO: 38. The amino acid sequence of the present invention is selected from the group consisting of the amino acid sequence of SEQ ID NO: 28. In an embodiment, an effective amount of the liposome comprises a toll-like receptor 4 agonist and a liposome comprising SEQ ID NO:28 and a tetrapalmitoylated tau phosphopeptide consisting of the amino acid sequence Mitoylated tau phosphopeptides are presented on the surface of liposomes and are delivered at 2 per dose. 100μg to 2500 per dose, corresponding to 9.7nmole to 742.5nmole 1 μg, preferably 1 μg, corresponding to 89.1 nmole to 712.8 nmole per dose 300μg to 2400μg per dose, e.g. 89.1 nmole per dose, 267 One dose corresponds to 0.3 nmole, 534.6 nmole, or 712.8 nmole Amounts of 300 μg, 900 μg, 1800 μg, or 2400 μg per dose are administered.

[0012] In some embodiments, the present invention provides a method for treating a patient with an anti-phosphorylated tau antibody that induces severe adverse events, such as encephalitis. 20. A method for inducing toll-like receptor 4 (T4) expression in a human subject in need thereof without the need for An effective amount of a liposome comprising an agonist and a tau phosphopeptide displayed on the surface of the liposome. The method includes administering to a subject a peptide having tau phosphopeptides of SEQ ID NO: 1 to SEQ ID NO: 3 and and an amino acid sequence selected from the group consisting of SEQ ID NO:5 to SEQ ID NO:12, The hopeptide is administered in an amount of about 25 nmole to about 750 nmole per dose, preferably For example, the tau phosphopeptide is a tetrapalmitoyl tau phosphopeptide having the amino acid sequence of SEQ ID NO:28. and about 300 μg, about 900 μg, about 1800 μg, or about 2000 μg per dose. 200 μg, or about 2400 μg, or any amount therebetween. do.

[0013] In certain embodiments, the anti-phosphorylated tau antibody can be administered without inducing severe adverse events, such as encephalitis. and a method for inducing in a human subject in need thereof a toll-like receptor 4 agonist, comprising: and an effective amount of liposomes comprising a tau phosphopeptide displayed on the surface of the liposome. The method includes administering to a subject a tau phosphopeptide having a sequence similar to SEQ ID NO:27 to SEQ ID NO:29, and an amino acid sequence selected from the group consisting of SEQ ID NO: 31 to SEQ ID NO: 38, The peptide is about 90 nmole to about 715 nmole per dose, e.g. Approximately 29.7 nmoles, approximately 267.3 nmoles, approximately 534.6 nmoles, or The method comprises administering an amount of about 712.8 nmoles, or any amount therebetween. In this embodiment, an effective amount of liposomes is 265 to 275 nmoles per dose, for example, Winnings: 265, 266, 267, 268, 269, 270, 271, 272, 273, 2 74, or 275 nmoles, or any value therebetween, e.g., about In another embodiment, an effective amount of tau phosphopeptide is The liposomes may contain 530 to 540 nmoles per dose, e.g., 530, 53 1, 532, 533, 534, 535, 536, 537, 538, 539, or 54 0 nmole, or any value therebetween, e.g., 534.6 nmole per dose In another embodiment, an effective amount of liposomes comprises a dose of tau phosphopeptide. or 710-720 nmoles per dose, e.g. 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, or 720 nmoles, or Any value therebetween, for example, tau phosphopeptide in an amount of 712.8 nmoles per dose. A method comprising:

[0014] In certain embodiments, the liposomes are administered subcutaneously.

[0015] In certain embodiments, the liposomes are administered intramuscularly.

[0016] In certain embodiments, the method includes administering an effective amount of a second dose 1 to 24 weeks after the first dose. The method further comprises administering the liposome to a subject.

[0017] In certain embodiments, the liposome comprises a helper T cell epitope and a lipidated Cp In certain embodiments, the oligonucleotide further comprises at least one of: The lipidated CpG oligonucleotide is selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22. and preferably has the nucleotide sequence of SEQ ID NO: 18, The G oligonucleotide has one or more phosphorothioate internucleotide linkages. In general, the CpG oligonucleotide comprises at least one lipophilic group and a linker, preferably a PEG linker. They are covalently linked via an anchor.

[0018] In certain embodiments, the liposome is 1,2-dimyristoyl-sn-glycero- 3-Phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phospho from the group consisting of di-3'-rac-glycerol (DMPG), and cholesterol Further comprising one or more selected lipids.

[0019] In certain embodiments, the helper T cell epitopes are selected from the group consisting of SEQ ID NO: 23 to SEQ ID NO: 26. and preferably SEQ ID NO: 2. It contains a sequence of 3 to 25 amino acids.

[0020] In certain embodiments, the toll-like receptor 4 agonist is monophosphoryl lipid A ( MPLA).

[0021] In certain embodiments, an effective amount of liposomes is between 30 μg and 900 μg per dose. The toll-like receptor 4 agonist is preferably contained in an amount of 100 μg to 585 μg. In certain embodiments, an effective amount of liposomes is between 30 μg and 900 μg per dose, preferably or 100 μg to 585 μg of the toll-like receptor agonist monophosphoryl Contains hexaacyl lipid A, 3-deacylated.

[0022] In certain embodiments, an effective amount of liposomes is between 25 μg and 625 μg per dose. , preferably in an amount of 75 μg to 450 μg of helper T cell epitopes. In an embodiment, an effective amount of liposomes is between 25 μg and 625 μg per dose, preferably A dose of 75 μg to 450 μg of T50 helper T cells consisting of the amino acid sequence of SEQ ID NO: 13 Contains cytoplasmic epitopes.

[0023] In certain embodiments, an effective amount of liposomes is from about 2 nmole to about 1 nmole per dose. 10 nmole, e.g., from about 4.02 nmole to about 100.44 nmole per dose or from about 4 nmole to about 75 nmole per dose, e.g., about 4.0 nmole per dose. 2 nmole to about 72.32 nmole, or about 10 nmole to about 10 per dose 5 nmole, e.g., about 12.06 nmole to about 100.44 nmole per dose or about 70 to about 105 nmole per dose, e.g., about 72.32 nmole per dose. Contains helper T cell epitopes in amounts of about 100.44 nmoles. In one embodiment, an effective amount of liposomes is from about 3 nmole to about 105 nmole per dose. le, preferably about 10 nmole to about 105 nmole per dose, e.g. 13 in an amount of about 12.06 nmoles to about 100.44 nmoles per 1000 of the amino acid sequence of SEQ ID NO:13. In one embodiment, the liposome comprises an effective amount of a T50 helper T cell epitope consisting of the amino acid sequence. The azides may be 2-5 nmole per dose, e.g., 2, 3, 4, or 5 nmole per dose. nmoles, or any value therebetween, e.g., about 3.82, 3.92, The antibody contains helper T cell epitopes in amounts of 4.02 or 4.12 nmoles. In embodiments, an effective amount of liposomes is 10-15 nmoles per dose, e.g., 10, 11, 12, 13, 14, or 15 nmoles per Any value, e.g., 11.86, 11.96, 12.06, 12.16 nmol per dose. In another embodiment, the effective amount of the liposome comprises: 70-75 nmole per dose, e.g. 70, 71, 72, 73, 74 per dose , or 75 nmole, or any value therebetween, e.g., 72.02, 72.1 2, 72.22, 72.32, and 72.42 of the helper T cell epitope. In another embodiment, the effective amount of liposomes is between 98 and 103 nmoles per dose, e.g. For example, 98, 99, 100, 101, 102, or 103 nmoles per dose, or or any value therebetween, e.g., 100.24, 100.34, 100. Helper T cell epitopes in amounts of 44, 100.54, or 100.64 nmoles Includes.

[0024] In certain embodiments, an effective amount of liposomes is between 50 μg and 1250 μg per dose. The lipidated CpG oligonucleotide comprises an amount of 150 μg to 800 μg. In certain embodiments, an effective amount of liposomes is between 50 μg and 1250 μg per dose. , preferably in an amount of 150 μg to 800 μg, consisting of the nucleotide sequence of SEQ ID NO: 18 Contains CpG oligonucleotides.

[0025] In certain embodiments, the liposome comprises: (1) a tau phosphopeptide having the amino acid sequence of SEQ ID NO: 28; (2) Toll-like receptor 4 amyloid containing monophosphoryl hexaacyl lipid A, 3-deacylated Gonist, (3) a helper T cell epitope comprising the amino acid sequence of SEQ ID NO: 39; (4) a lipidated CpG oligonucleotide comprising the nucleotide sequence of SEQ ID NO: 18, and To (5) 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1, 2-Dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol ( DMPG), and cholesterol. Includes.

[0026] In certain embodiments, the subject has Alzheimer's disease, e.g., early stage Alzheimer's disease. , mild cognitive impairment (MCI) due to Alzheimer's disease, mild Alzheimer's disease, or In another embodiment, the subject is in need of treatment for mild to moderate Alzheimer's disease. Although they are amyloid positive in the MRI, they have not yet demonstrated significant cognitive impairment.

[0027] The present invention also provides a method for administering an anti-phosphorylated tau antibody to a patient without inducing severe adverse events such as encephalitis. A vaccine combination for use in inducing in a human subject of need thereof. , vaccines, including priming and booster vaccines of the present invention; The present invention also relates to a combination of an anti-phosphorylated tau antibody and a method for treating a patient suffering from a severe adverse event such as encephalitis. In the manufacture of a pharmaceutical product for induction in a human subject in need thereof, Use of a vaccine combination, the vaccine combination being a priming vaccine according to an embodiment of the present invention. The present invention provides a method for administering an anti-phosphorylated tau antibody to a patient suffering from encephalitis or the like, including administering the antibody to a patient suffering from encephalitis or the like, and providing a booster vaccine. The present invention relates to a method for inducing a cellular endothelial cell death in a human subject without inducing severe adverse events. All aspects and embodiments of the present invention provide for the use of anti-phosphorylated tau antibodies to treat severe adverse events, such as encephalitis. Vaccine for use in induction in human subjects in need thereof without inducing The combination of phospho-tau and / or anti-phospho-tau antibody was administered without inducing severe adverse events such as encephalitis. , in the manufacture of a pharmaceutical product for induction in a human subject in need thereof, The present invention can be applied to the use of the entire body.

[0028] Further aspects, features, and advantages of the present invention are set forth in the following detailed description of the invention and the claims. This will be better understood by reading the scope of the requirements.

[0029] The foregoing summary and the following detailed description of preferred embodiments of the present application should be taken in conjunction with the accompanying drawings. However, the present application is not to be construed as limiting the present invention, and It should be understood that the present invention is not limited to the precise embodiment. [Brief description of the drawings]

[0030] [Figure 1] Graph showing pTau IgG titers following three intramuscular administrations of 35 and 80 pg / dose of ACI-35.030 to female C57BL / 6 mice, once every two weeks. Geometric mean + 95% CI is represented per group of 10 mice. [Diagram 2] 1 is a graph showing that 1200 μg / dose and 2400 μg / dose of ACI-35.030 induced durable titers of anti-phosphorylated tau antibodies in rhesus monkeys. Geometric mean antibody titers per group expressed in AU / mL are presented as measured by ELISA. [Diagram 3] FIG. 1 shows that three injections of 2400 μg / dose of ACI-35.030 induced antibodies that bound to tau tangles in AD brain sections (panel B) with no cross-reactivity with colon sections (panel D). Pre-treatment samples did not bind to AD brain sections (panel A) or colon sections (panel C). [Figure 4] 1 is a graph showing that intramuscular injection of 1800 μg / dose of ACI-35.030 induced anti-ePHF IgG antibody titers with lower within-group variability compared to subcutaneous injection of the same vaccine at the same dose. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] Various publications, articles, and patents are cited or referenced in the background and throughout this specification. Each of these references is incorporated herein by reference in its entirety. Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is expressly incorporated by reference in its entirety. The purpose of this discussion is to provide a context for the invention. Any or all of these matters may be part of the prior art with respect to any invention sought. This does not constitute an authorization for the formation of a

[0032] Unless otherwise defined, all technical and scientific terms used herein are intended to be understood as meaning the terminology of the present invention. has the same meaning as commonly understood by a person skilled in the art to which it pertains. As used herein, certain terms have the meanings set forth herein.

[0033] As used in this specification and the appended claims, the singular forms "a," "an," "an" and "an" are used interchangeably. "An" and "the" refer to the plural unless the context clearly indicates otherwise. It should be noted that this includes the referent of

[0034] Unless otherwise stated, any numerical value, e.g., a concentration or concentration range, described herein The range should be understood to be modified in all instances by the term "about." Therefore, numerical values ​​typically include ±10% of the indicated value. For example, 1 mg / m The concentration of L includes 0.9 mg / mL to 1.1 mg / mL. Similarly, the concentration includes 1% to 10% (w / v ) in a concentration range of 0.9% (w / v) to 11% (w / v), inclusive. The use of numerical ranges includes all possible subranges, i.e., unless the context clearly dictates otherwise. That is, all individual numbers within such ranges, including the integers within such ranges and any fractional parts of that value. Explicitly includes the value.

[0035] Unless otherwise indicated, the term "at least" preceding a series of elements refers to all elements in that series. It should be understood that this refers to any element of the above. Those of ordinary skill in the art can determine this using no more than routine experimentation. It will be appreciated that those skilled in the art will recognize or have discovered many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by this invention. .

[0036] As used herein, "comprises," "comprising," "including "includes," "including," "has," "having," The term "contains" or "containing" or its equivalent Any other variation of these includes the stated integer or group of integers but any other integer. It can be understood to imply that several groups are not excluded, and non-exclusive or The lists are intended to be non-limiting. For example, compositions, mixtures, processes, methods, etc. that include the elements of the list may be used. The laws, articles, or devices are not necessarily limited to those elements only, but may be modified in any way without prior notice. or other substances inherent in such composition, mixture, process, method, article, or apparatus. Furthermore, unless expressly stated to the contrary, "Or" refers to a non-exclusive or, not an exclusive or. For example, if condition A or B is the following: A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and both A and B Satisfied by either one of them both being true (or existing).

[0037] As used herein, "about" when referring to dimensions or characteristics of components of the preferred invention The terms "approximately," "generally," "substantially," and similar terms are used herein to refer to the The dimensions / characteristics listed are not intended to be precise boundaries or parameters, as will be appreciated by those of ordinary skill in the art. indicates that the invention does not exclude minor variations therefrom that are functionally the same or similar. It should also be understood that the references include, at a minimum, numerical parameters. The subject matter described herein is based on accepted mathematical and industrial principles (e.g., rounding, measurement, etc.) in the art. including variations that do not change the least significant digit, such as deviations from the least significant digit, or other systematic errors, manufacturing tolerances, etc. It can be seen.

[0038] The present invention provides an anti-phosphorylated tau antibody that can be administered to patients who need it without inducing severe adverse events such as encephalitis. In certain embodiments, the method comprises inducing in a human subject a liposome-derived marker for inflammatory bowel disease. The antibody contains a tau phosphopeptide displayed on the surface of the tau phosphopeptide and a toll-like receptor 4 agonist. The method includes administering to the subject an effective amount of liposomes.

[0039] As used herein, the term "anti-phosphorylated tau antibody" refers to an antibody that binds to the amino acid sequence of tau. The present invention refers to an antibody that binds to tau that is phosphorylated at one or more amino acid residues in the The amino acid residues to be phosphorylated include, for example, serine (Ser), threonine (Thr), or or tyrosine (Tyr). The site of phosphorylated tau to which anti-phosphorylated tau antibodies bind is preferably a site that is specifically phosphorylated in neurodegenerative diseases such as Alzheimer's disease Examples of sites of phosphorylated tau to which anti-phosphorylated tau antibodies bind include, for example, Ty r18, Ser199, Ser202, Thr205, Thr212, Ser214, S These include Er396, Ser404, Ser409, Ser422, and Thr427. As used throughout this application, amino acid positions are those amino acid sequences deposited in GenBank. Human microtubule-associated protein tau isoform 2, designated by the number NP_005901.2 This is obtained by referring to the array.

[0040] Whether or not anti-phosphorylated tau antibodies can be induced upon administration is determined using a biological sample derived from the subject. (e.g., blood, plasma, serum, PBMCs, urine, saliva, feces, CSF, or lymphatic fluid) , antibodies directed against immunogenic tau peptides administered in pharmaceutical compositions, e.g., IgG Alternatively, it can be determined by testing for the presence of IgM antibodies (e.g., Ha (See, for example, R. Low, 1989, Antibodies, Cold Spring Harbor Press). The titers of antibodies produced in response to administration of the provided compositions can be determined using an enzyme-linked immunosorbent assay (ELISA). ELISA), other ELISA-based assays (e.g., MSD-Meso Scal e Discovery), dot blot, SDS-PAGE gel, ELISPOT, Alternatively, it can be measured by antibody-dependent cellular phagocytosis (ADCP) assay.

[0041] As used herein, the term "adverse event" (AE) refers to an adverse event that occurs after administration of a pharmaceutical formulation. Any untoward medical occurrence in a patient that does not necessarily have a causal relationship to the treatment In the embodiments of the present invention, AE refers to an increase in severity using the following definitions: It is rated on a three-point scale: mild (Grade 1), easily tolerated by the subject, and Refers to AEs that cause minor discomfort and do not interfere with daily activities; moderate (Grade 2), positive Refers to an AE that is sufficiently bothersome to interfere with normal daily activities and may require intervention. Severe (Grade 3), interfering with normal daily activities and usually requiring treatment or other intervention A A severe AE (SAE) is any event at any dose that results in any of the following outcomes: Any AE occurring at the time of onset of the event: Death, which is an outcome, not an event; Refers to an event in which the patient is at risk of death; life-threatening; life-threatening does not refer to an event that, if it had been more severe, might have caused death; Hospitalization for treatment, i.e. unplanned overnight hospitalization or extension of an existing hospitalization; Permanent or significant impairment or substantial destruction of the ability to perform vital functions; congenital anomaly / Congenital defects; potentially putting the patient at risk or any of the other outcomes listed above A significant medical event (research) that may require medical or surgical intervention to prevent one of the following: (e.g., emergency room or hospital admission for allergic bronchospasm) Hospitalization means intensive care at home, or blood disease or convulsions that do not require hospitalization. Hospitalization or prolonged hospitalization constitutes a criterion for an AE to be serious, but , which in itself is not considered an SAE. In the absence of an AE, hospitalization or prolonged hospitalization is should not be reported as an SAE by the participating investigators. This applies in the following circumstances: This may be the case in cases where hospitalization or prolonged hospitalization is required by protocol. or the hospitalization or extension of hospitalization is necessary in accordance with routine procedures followed by the central facility (e.g., This should be recorded in the study file. Hospitalization for elective procedures for pre-existing conditions that did not worsen during the study were not considered AEs. It is not done.

[0042] Complications occurring during hospitalization are AEs. Complications that prolong hospitalization or result in other SAEs If any of the criteria are met, the event is an SAE.

[0043] As used herein, the term "encephalitis" refers to encephalitis caused by infectious and non-infectious causes. As used herein, the term "meningoencephalitis" refers to inflammation of the brain that can cause inflammation of the meninges and It refers to a condition characterized by infection or inflammation of the brain. Diagnosis may be made by techniques known to those of skill in the art in light of the present disclosure, e.g., clinical, neurological, and psychiatric examinations, biological sampling including blood and CSF sampling, MRI scanning, as well as by electroencephalography (EEG).

[0044] As used herein, the term "liposome" generally refers to a liposome having a high lipid content. Refers to lipid vesicles made from materials such as phospholipids and cholesterol. These vesicles The lipids in the membrane are generally organized in the form of lipid bilayers. Interspersed among multiple onion-like frameworks of lipid bilayers that form small lipid vesicles (MLVs) or encapsulates the volume contained within the central amorphous cavity. The lipid vesicles having the structure are unilamellar lipid vesicles, i.e., they have a single peripheral bilayer surrounding a cavity. Large unilamellar vesicles (LUVs) are generally lipid vesicles ranging in size from 100 nm to several microns. vesicles, e.g., small unilamellar lipid vesicles ( SUVs are generally less than 100 nm, e.g., between 20 and 100 nm, typically between 15 and 30 nm in diameter.

[0045] As used herein, microtubule-associated protein tau, MAPT, neurofibrillary tangle protein tau Also known as protein, paired helical filament-tau, PHF-tau, MAPTL, and MTBT1 The commonly known term "tau" or "tau protein" refers to a protein that has multiple isoforms. It refers to abundant central and peripheral nervous system proteins that are involved in the regulation of the human central nervous system (CNS). Six major tau isoforms were selected, ranging in size from 352 to 441 amino acids in length. It exists due to alternative splicing (Hanger et al., Trends Mol Med. 15:112-9, 200 9) Examples of tau include, but are not limited to, tau isoforms in the CNS: For example, the microtubule-associated protein tau isoform has four repeats and two inserts. The longest tau isoform, 441 amino acids long (4R2N), also named form 2; For example, the amino acid sequence of the human Other examples of tau include isoform 2, which has three repeats and an insertion The 352 amino acid sequence of the microtubule-associated protein tau isoform 4, also known as tau isoform 4, does not contain the fragment. The shortest (fetal) isoform (3R0N), e.g., the amino acid sequence G Human tau isoform 4, represented by enBank accession number NP_058525.1, was identified. Examples of tau include the terminal tau gene, which contains 300 additional residues (exon 4a). One example is the "large tau" isoform expressed in peripheral nerves. Friedhoff et al., Biochi mica et Biophysica Acta 1502 (2000) 122-132. An example of tau is the 6762 nucleoside. The 758 alpha-nucleotide sequence encoded by the 10-kDa mRNA transcript (NM_016835.4) was Human large tau, a 1-amino acid long protein, or its isoforms The amino acid sequence of an exemplary human large tau is located under GenBank accession number NP_058519. As used herein, the term "tau" includes tau-specific tau expression in species other than humans, e.g. For example, Macaca Fascicularis (Macaca fascicularis), Rhesus monkey, or It contains a homologue of tau from chimpanzees (Pan troglodytes). As used herein, the term "tau" includes mutations of full-length wild-type tau, such as point mutations. The term "tau" includes proteins containing fragments, insertions, deletions, and splice variants. The term also encompasses post-translational modifications of the tau amino acid sequence. Examples of cleavage include, but are not limited to, phosphorylation.

[0046] As used herein, the term "peptide" or "polypeptide" refers to a peptide. Polymers composed of amino acid residues linked via bond bonds, related naturally occurring The term refers to structural variants of any of the above-mentioned compounds, as well as non-naturally occurring synthetic analogues thereof. Typically, a peptide is at least three amino acids in length. The peptide may be a naturally occurring peptide, a recombinant peptide, or The synthetic peptides may be, for example, a fully automated peptide synthesis. An example of a tau peptide can be synthesized using a polypeptide synthesizer. 5 to about 30 amino acids in length, preferably about 10 to about 25 amino acids in length, more preferably about 16 to about 30 amino acids in length Any peptide of the tau protein of about 21 amino acids in length is included in the present disclosure. The amino acids are listed using the standard three- or one-letter amino acid abbreviations, where the phosphate residue is indicated by "p." Examples of tau peptides useful in the present invention include those listed below from N-terminus to C-terminus. Tau peptides including, but not limited to, any of the amino acid sequences of SEQ ID NOs: 1 to 12, has at least 75%, 80%, 85%, or 90% identical amino acid sequence to any one of SEQ ID NOs: 1 to 12. Examples of tau peptides include those having 0% or 95% identical amino acid sequence.

[0047] As used herein, the term "phosphopeptide" or "phosphoepitope" refers to a peptide that is phosphorylated at one or more amino acid residues. An example of a phosphopeptide is any taupe peptide that contains one or more phosphorylated amino acid residues. Petite is one example.

[0048] The tau peptides of the present invention can be synthesized by solid-phase peptide synthesis or recombinant expression systems. Automated peptide synthesizers are available from a number of suppliers, including Applied Bio Systems (Foster City, Calif.). Expression systems include bacteria, such as E. coli, yeast, insect cells, or mammalian cells. Recombinant expression procedures are described in Sambrook et al., Molecular Cloning: A Laboratory Manual (CSHP Press, NY 2d ed., 1989).

[0049] In certain embodiments, the liposomes comprise one or more tau peptides. In embodiments, the tau peptides in the liposomes may be the same or different. In light of the present disclosure, any suitable tau peptide known to those of skill in the art may be used in the present invention. In certain embodiments, the one or more tau peptides can be selected from the group consisting of SEQ ID NOs: 1 to 5. In another embodiment, the one or more taupeptides include one of the amino acid sequences of 12. The nucleotide sequence is at least 75%, 80%, 80%, or 90% identical to one of the amino acid sequences of SEQ ID NOs: 1 to 12. Contains 5%, 90%, or 95% identical amino acid sequence, and all amino acid residues are phosphate The polypeptides are either unmodified or phosphorylated at one or more amino acid residues.

[0050] In certain embodiments, the one or more tau peptides are tau phosphopeptides. In certain embodiments, the one or more tau phosphopeptides are selected from the group consisting of SEQ ID NOs: 1-3. or one of the amino acid sequences of SEQ ID NOs: 1 to 3 or 5 to 12. One of the amino acid sequences has at least 75%, 80%, 85%, 90%, or 95% % identical amino acid sequence, where one or more of the indicated amino acid residues are phosphorylated Preferably, the tau phosphopeptide is one of the amino acid sequences of SEQ ID NOs: 1 to 3. The tau peptide may be amidated at the C-terminus.

[0051] In an embodiment of the present application, the tau peptide is presented on the surface of a liposome. Peptides, preferably tau phosphopeptides, are known in the art in view of the present disclosure. The present invention can be presented on the surface of liposomes using methods such as those described in, for example, US Pat. Nos. 8,647,631 and 9,687,444, which are incorporated herein by reference. No. 7, and related disclosures in International Patent Application No. PCT / US18 / 57286. In certain embodiments, one or more tau peptides, including phosphopeptides, are The tau peptide may be modified in one or more ways to present the tau peptide on the surface of the liposome, e.g., Further include lumi-ylation or dodecyl modifications. Additional amino acid residues, such as Lys, Cys, or optionally Ser or Thr, is added to the tau peptide to facilitate modification. The position of the lipid anchor induces different conformations of the peptide sequence. It was reported (Hickman et al., J. Biol. Chem. vol. 286, NO. 16, pp. 13966-139 76, April 22, 2011). Without wishing to be bound by theory, Addition of the end of the tau peptide to the β-sheet structure may increase the pathological beta-sheet conformation of the tau peptide. Thus, the tau peptide or peptides further comprise hydrophobic moieties at both termini. The modified tau peptide may be amidated at the C-terminus. The tau peptide displayed on the surface of the antibody is one of the peptides represented by SEQ ID NO: 27 to SEQ ID NO: 38. It consists of an amino acid sequence.

[0052] Examples of tau liposomes useful in the present invention include, but are not limited to, those described in U.S. Pat. ,647,631 and 9,687,447, and International Patent Application Nos. PCT / The tau liposomes described in US18 / 57286 are included, the disclosures of which are: , the entirety of which is incorporated herein by reference.

[0053] As used herein, the term "effective amount" refers to a desired biological or physiological effect in a subject. refers to the amount of an active ingredient or component that induces a medical response. Selection of a specific effective dose depends on: The disease to be treated or prevented, the symptoms involved, the patient's weight, the patient's immune status, and the It is believed that the present invention can be performed by one of ordinary skill in the art based on the consideration of several factors, including other factors known to those skilled in the art. The exact dosage to be used in the formulation can be determined by the individual administering the formulation. The method, route of administration, target site, physiological state of the patient, other medications administered, and severity of the disease. The dose may also depend on the individual patient's condition and should be determined according to the practitioner's judgment and the circumstances of each patient. The effective amount of tau phosphopeptide also depends on whether an adjuvant is also administered. Higher dosages are required in the absence of adjuvant. Effective doses are determined in vitro. o or can be extrapolated from dose-response curves derived from animal model test systems.

[0054] In an embodiment of the present application, an effective amount of liposomes is administered without inducing severe adverse events such as encephalitis. comprising a tau phosphopeptide in an amount sufficient to increase the level of anti-phosphorylated tau antibodies. In certain embodiments, an effective amount of liposomes is from about 25 nmole to about 750 nmole per dose. nmoles, for example about 29.7 nmoles to about 742.5 nmoles per dose, preferably or about 90 nmole to about 715 nmole per dose, for example about 89 nmole per dose 0.1 nmole to about 712.8 nmole, or about 90 nmole to about 5 nmole per dose 35 nmole, for example about 89.1 nmole to about 534.6 nmole per dose; or from about 90 nmole to about 275 nmole per dose, for example about 89 nmole per dose. The tau phosphopeptide is administered in an amount of about 0.1 nmole to about 267.3 nmole. The amount of tau phosphopeptide can also be expressed by weight. For example, 29.7 nM per dose. Each mole contains 100 μg of tetrapalmitochondrial consisting of the amino acid sequence of SEQ ID NO: 28. 742.5 nmoles per dose corresponds to a toylated tau phosphopeptide 2500 μg of tetrapalmitoylated tau phospholipase A consisting of the amino acid sequence of SEQ ID NO: 28 89.1 nmole per dose corresponds to peptide sequence no. 300 μg per dose It corresponds to the tetrapalmitoylated tau phosphopeptide consisting of the amino acid sequence of No. 28, and is used for 1 712.8 nmoles per dose is 2400 μg of the amino acid sequence of SEQ ID NO:28 per dose 534.6 per dose, corresponding to a tetrapalmitoylated tau phosphopeptide consisting of nmole contains 1800 μg of tetracycline consisting of the amino acid sequence of SEQ ID NO: 28 per dose. Corresponding to lumitoylated tau phosphopeptide. Tetrapalmitoylated tau phosphopeptide has four lipid chains that allow for the presentation of tau phosphopeptides to the surface of liposomes Tetrapezil consisting of the amino acid sequence of SEQ ID NO: 28 at doses of 300, 900, and 1800 μg The lumi-toylated tau phosphopeptides were either 169, 508, or 1016 μg, respectively. The corresponding "naked" peptide does not have any lipid chains.

[0055] In an embodiment of the present application, an effective amount of liposomes is about 25 nmole to about 25 nmole per dose. Tau phosphopeptide in an amount of about 750 nmole, e.g., about 25 nmole per dose; Approximately 30 nmole, approximately 35 nmole, approximately 40 nmole, approximately 45 nmole, approximately 50 nmole mole, about 55 nmole, about 60 nmole, about 65 nmole, about 70 nmole , about 75 nmole, about 80 nmole, about 85 nmole, about 90 nmole, about 95 nmole, about 100 nmole, about 125 nmole, about 150 nmole, about 175 nmole, about 200 nmole, about 225 nmole, about 250 nmole, about 275 nmole, about 300 nmole, about 325 nmole, about 350 nmole, about 375 nmole, about 400 nmole, about 425 nmole, about 450 nmole, about 475 nmole, about 500 nmole, about 525 nmole, about 550 nmole, about 575 nmole, about 600 nmole, about 625 nmole, about 650 nmole, about 675 nmole, about 700 nmole, about 725 nmole, about 750 nmole of sequence number The tau phosphopeptide includes one of the amino acid sequences of 1 to 3 or 5 to 12. Preferably, the tau phosphopeptide has one of the amino acid sequences of SEQ ID NO: 27 to SEQ ID NO: 38. More preferably, the tau phosphopeptide comprises the amino acid sequence of SEQ ID NO:28. do.

[0056] In an embodiment of the present application, an effective amount of liposomes is between 100 μg and 250 μg per dose. 0μg, 300μg to 2400μg, 300μg to 1800μg, or 300μg to 9 00μg, e.g. 100μg, 150μg, 200μg, 250μg, 30 0μg, 400μg, 500μg, 600μg, 700μg, 800μg, 900μg, 1000μg, 1100μg, 1200μg, 1300μg, 1400μg, 1500μg g, 1600μg, 1700μg, 1800μg, 1900μg, 2000μg, 210 Tetra in amounts of 0 μg, 2200 μg, 2300 μg, 2400 μg, or 2500 μg Contains palmitoylated tau phosphopeptides.

[0057] In an embodiment of the present application, the tau phosphopeptide is presented on the surface of a liposome. In an embodiment of the present application, the tau phosphopeptide is a tau phosphopeptide having any of SEQ ID NOs: 1 to 3 or 5 to 12. Preferably, the tau phosphopeptide comprises one of the amino acid sequences of SEQ ID NO: 27 to SEQ ID NO: 38. More preferably, the tau phosphopeptide comprises one of the amino acid sequences listed in the table below. It consists of the amino acid sequence of SEQ ID NO:28.

[0058] In another embodiment of the present application, the effective amount of liposomes is from 30 μg to 90 μg per dose. 0 μg, preferably 100 μg to 585 μg of a toll-like receptor 4 agonist. For example, effective amounts of liposomes include 30 μg, 50 μg, 100 μg per dose. , 150μg, 200μg, 250μg, 300μg, 330μg, 360μg, 390 μg, 420μg, 450μg, 480μg, 500μg, 520μg, 540μg, 5 in amounts of 60 μg, 580 μg, 600 μg, 700 μg, 800 μg, or 900 μg It may include a toll-like receptor 4 agonist.

[0059] In an embodiment of the present application, toll-like receptor 4 is 3D-(6-acyl)PHAD( Registered trademarks.

[0060] In another embodiment of the present application, the effective amount of liposomes is from 25 μg to 62 μg per dose. Further comprising a helper T cell epitope in an amount of 5 μg, preferably 75 μg to 450 μg. For example, effective amounts of liposomes may be 25 μg, 50 μg, 75 μg, 100 μg, or 250 μg per dose. μg, 125μg, 150μg, 175μg, 200μg, 225μg, 250μg, 2 75μg, 300μg, 325μg, 350μg, 375μg, 400μg, 425μg , 450μg, 475μg, 500μg, 525μg, 550μg, 575μg, 600 The antibody may contain helper T cell epitopes in an amount of 625 μg, or 625 μg.

[0061] In another embodiment of the present application, the effective amount of liposomes is about 3 nmoles per dose. ~ about 105 nmole, e.g., about 4 nmole, about 5 nmole, about 6 nmole, per dose ole, about 7 nmole, about 8 nmole, about 9 nmole, about 10 nmole, about 15 nmole nmole, about 20 nmole, about 25 nmole, about 30 nmole, about 35 nmole e, about 40 nmole, about 45 nmole, about 50 nmole, about 55 nmole, about 6 0 nmole, approx. 65 nmole, approx. 70 nmole, approx. 75 nmole, approx. 80 nmole le, about 85 nmole, about 90 nmole, about 95 nmole, about 100 nmole, or further comprising a helper T cell epitope in an amount of about 105 nmole.

[0062] In an embodiment of the present application, the helper T cell epitope has the amino acid sequence of SEQ ID NO: 13. T50 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 14; 6 helper T cell epitope, T48 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 15 a T51 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 16; or a T52 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 17, Preferably, the helper T cell epitope is a T50 epitope consisting of the amino acid sequence of SEQ ID NO: 13. It is a triple T cell epitope.

[0063] In certain embodiments, an effective amount of liposomes is between 50 μg and 1250 μg per dose. and a lipidated CpG oligonucleotide in an amount of 150 μg to 800 μg. For example, an effective amount of liposomes may be 50 μg, 100 μg, 150 μg, or , 200μg, 250μg, 300μg, 350μg, 400μg, 450μg, 500 μg, 550μg, 600μg, 650μg, 700μg, 750μg, 800μg, 8 50μg, 900μg, 950μg, 1000μg, 1050μg, 1100μg, 12 The lipidated CpG oligonucleotide may be present in an amount of 100 μg, 1250 μg, or 1,250 μg. do.

[0064] In an embodiment of the present application, the lipidated CpG oligonucleotide is selected from the group consisting of SEQ ID NOs: 18 to 22. and preferably a CpG oligonucleotide comprising one of the nucleotide sequences The modified CpG oligonucleotide is a CpG oligonucleotide comprising the nucleotide sequence of SEQ ID NO:18. In an embodiment of the present application, the lipidated CpG oligonucleotide is or more phosphorothioate internucleotide linkages, (PEG)-containing linker is covalently linked to cholesterol, It is a CpG oligonucleotide comprising the nucleotide sequence of number 18.

[0065] In embodiments, the effective amount of liposomes is 50 μg, 100 μg, 15 μg, or 20 μg per dose. 0μg, 200μg, 250μg, 300μg, 350μg, 400μg, 450μg, 500μg, 550μg, 600μg, 650μg, 700μg, 750μg, 800μg g, 850μg, 900μg, 950μg, 1000μg, 1050μg, 1100μg , 1,200 μg, or 1,250 μg, shared via a PEG linker with cholesterol It comprises CpG oligonucleotides linked by bonds.

[0066] In certain embodiments, the human subject is in need of treatment for a neurodegenerative disease, disorder, or condition. Let us assume that.

[0067] As used herein, a "neurodegenerative disease, disorder, or condition" includes any disease, disorder, or condition that is contemplated by the present disclosure. The term "neurodegenerative disease" includes any neurodegenerative disease, disorder, or condition known to those of skill in the art. Examples of diseases, disorders, or conditions that may be caused by the formation of neurofibrillary lesions include or a neurodegenerative disease or disorder associated therewith, such as a tau-related disease or disorder called a tauopathy. In certain embodiments, the present invention relates to a neurodegenerative disease, disorder, or condition. or conditions such as Alzheimer's disease, Parkinson's disease, Creutzfeldt-Jakob disease, Boxer dementia, Down syndrome, Gerstmann-Straussler-Scheinker disease, Inclusion Myositis, prion protein cerebral amyloid angiopathy, traumatic brain injury, amyotrophic lateral sclerosis, Parkinson-dementia complex in Guam, non-Guam type motor neuron with neurofibrillary tangles Disease, argyrophilic grain dementia, corticobasal degeneration, amyotrophic lateral sclerosis with Lewy dementia Dementia Lewy Amyotrophic Lateral Sclerosis, diffuse neurofibrillary fibrosis with calcification Fibrinogen tangles, frontotemporal dementia, and Parkinsonism, preferably linked to chromosome 17. Frontotemporal Dementia with Dysfunction (FTDP-17), Frontotemporal Dementia, Hallerhorden Spa Zucker's disease, multiple system atrophy, Niemann-Pick disease type C, Pick's disease, progressive subcortical gliosis , progressive supranuclear palsy, subacute sclerosing panencephalitis, tangle only dementia, Postencephalitic Parkinsonism, Myotonic Dystrophy, Chronic Traumatic Encephalopathy (CTE), Primary Sex-Age-Related Tauopathy (PART), cerebrovascular disease, or Lewy Body Dementia (LBD) Any disease exhibiting coexistence of tau and amyloid pathology, including, but not limited to, In certain embodiments, the neurodegenerative disease, disorder, or condition is an arachidonic acid disorder. In a preferred embodiment, the neurodegenerative disease is Zheimer's disease or another tauopathy. The disease, disorder, or condition is Alzheimer's disease.

[0068] The clinical course of Alzheimer's disease progresses through multiple stages, with the pattern of progression of cognitive and functional impairment. The stages can be divided into steps as known in the art, including, for example, the NIA-AA research framework. For example, each of the contents can be defined using a rating scale. Dubois et al., Alzheimer's & Dementia 12 (2016) 292- 323, Dubois et al., Lancet Neurol 2014; 13: 614-29, Jack et al., Alzheimer's & D See ementia 14 (2018) 535-562.

[0069] In a preferred embodiment, the neurodegenerative disease, disorder, or condition is early Alzheimer's disease Alzheimer's disease, mild cognitive impairment (MCI) due to Alzheimer's disease, mild Alzheimer's disease, and has mild to moderate Alzheimer's disease.

[0070] In some embodiments, the subject in need of treatment is amyloid positive in the brain, They have not yet shown significant cognitive impairment. Amyloid deposition in the brain is a condition known in the art. Detecting using methods such as PET scanning, immunoprecipitation mass spectrometry, or other methods. can be done.

[0071] As used herein, the term "toll-like receptor" or "TLR" refers to a naturally occurring A class of pattern recognition receptor (PRR) proteins that play important roles in immune responses TLRs are molecules that can distinguish microbial pathogens, e.g., bacteria, fungi, and fungi, from host molecules. Recognizes pathogen-associated molecular patterns (PAMPs) derived from fungi, parasites, and viruses. LRs typically function as dimers and mediate antigen-presenting dendritic cells and phagocytic macrophages. It is a transmembrane protein expressed by cells involved in the innate immune response, including at least The ten human TLR family members, namely TLR1 to TLR10, as well as a few At least 12 mouse TLR family members, namely TLR1-TLR9 and TLR1-TLR2, are identified. There are three types of TLRs: TLR11 to TLR13, which differ in the types of antigens they recognize. R4 binds lipopolysaccharide (LPS), a component of many gram-negative bacteria, as well as cytoplasmic endothelial cells. Viral proteins, polysaccharides, and integral proteins, e.g., low density lipoproteins, vein proteins, TLR9 recognizes tadefensins and heat shock proteins in the prokaryotic genome. Unmethylated cytosine-phosphoryl esterase (UCP) is abundant in mammalian genomes but rare in vertebrate genomes. Nucleotides activated by cytosine-guanine (CpG) single- or double-stranded dinucleotides TLR activation triggers a series of signaling events, resulting in type I TLRs. Production of interferons (IFNs), inflammatory cytokines, and chemokines, and This leads to the induction of an immune response. Ultimately, this inflammation also activates the adaptive immune system, which then attacks the invader. This results in the elimination of pathogens and infected cells.

[0072] As used herein, the term "agonist" refers to an agent that binds to one or more TLRs. An agonist is a molecule that inhibits the activity of a receptor and induces a receptor-mediated response. induce, activate, elevate, activate, facilitate, enhance Such activity can be referred to as "agonist activity." For example, TLR4 or TLR9 agonists inhibit cell signaling through the bound receptors. Agonists can activate or increase the activity of the receptors. binds to or interacts with nucleic acids, small molecules, proteins, carbohydrates, lipids, or receptors An agonist is any molecule that mimics the activity of a natural receptor ligand. Agonists can be classified into two types: by sequence, by conformation, by charge, or by the mechanism by which the agonist acts on the receptor. These natural receptor ligands are identical in terms of other characteristics that enable them to be recognized. This recognition allows cells to recognize agonists in the same way as they would if the natural receptor ligand were present. Physiological and / or biochemical changes may occur within the cell in response to its presence. In one embodiment, the toll-like receptor agonist is a toll-like receptor 4 agonist and and / or toll-like receptor 9 agonists.

[0073] As used herein, the terms "induce" and "activate" and the associated These variations refer to any measurable increase in cellular activity. Induction of an immune response includes, for example, For example, it may increase the activation, proliferation, or maturation of immune cell populations and cytokine production. and / or other indicators of increased immune function. In this case, the induction of immune responses is achieved by increasing the proliferation of B cells and producing antigen-specific antibodies. enhance the proliferation of antigen-specific T cells, and improve dendritic cell antigen presentation. and / or increase the expression of certain cytokines, chemokines, and costimulatory markers. Examples of adding

[0074] As used herein, the term "toll-like receptor 4 agonist" refers to a TLR4 In light of this disclosure, those of skill in the art will recognize that Any suitable toll-like receptor 4 agonist can be used in the present invention. Examples of toll-like receptor 4 ligands useful for As used herein, the term "TLR4 agonist" includes, but is not limited to, TLR4 agonists. In this case, the term "monophosphoryl lipid A" or "MPLA" refers to the endotoxin Gram-positive It refers to a modified form of lipid A, which is the biologically active portion of lipopolysaccharide (LPS) of M. PLA is less toxic than LPS but maintains its immunostimulatory activity. Vaccine adjuvant As such, MPLA stimulates both cellular and humoral responses to vaccine antigens. Examples of MPLA include, but are not limited to, 3-O-desacyl-4'-monophos Phosphoryl lipid A, monophosphoryl hexaacyl lipid A, 3-deacylated (synthetic) (3D- (6-acyl)PHAD®), monophosphoryl 3-deacylated aryl The MPLAs useful in the present invention include: , which can be obtained using methods known in the art or from commercial sources; For example, Avanti Polar Lipids (Alabaster, Alabama a, 3D-(6-acyl) PHAD®, PHAD® manufactured by USA , PHAD®-504, 3D-PHAD®, or various commercially available In certain embodiments, the toll-like receptor 4 agonist is MPL™ from a source. In certain embodiments, the tau phosphopeptide and toll-like receptor agonist are Liposomes containing the agonist 4 bind to most or all of the HLA DR (human leukocyte antigen receptor 4) receptors. -antigen D related) molecules. The T cell epitopes are then expressed as CD4 + It can activate T cells and promote their essential maturation and Tau liposomes provide tau-specific B cells with apoptosis and survival signals. In an allogeneic or heterologous immunization scheme used in the immunization and / or boost , can be used to generate high quality antibodies against the p-tau antigen.

[0075] As used herein, the term "helper T cell epitope" refers to a T helper cell Helper T cell epitopes are polypeptides that contain epitopes capable of recognition by T cell helper Examples of peptides include, but are not limited to, tetanus toxoid (e.g., T2 and T30), hepatitis B surface antigen, cholera Toxin B, toxoid, diphtheria toxoid, measles virus F protein, chlamydia Chlamydia trachomatis major outer membrane protein, Plasmodium falciparum smodium falciparum circumsporozoite T, Plasmodium falciparum (P. falciparum) CS Antigen, Schistosoma mansoni triosephosphate isomerase, Pertussis Bordetella pertussis, Clostridium tetani, Pertusaria trachy Pertusaria trachythallina, Escherichia coli TraT, Nara and influenza virus hemagglutinin (HA).

[0076] In view of the present disclosure, any suitable helper T cell epitope known to those of skill in the art may be used in the present invention. In a particular embodiment, the helper T cell epitope is of the sequence Contains at least one amino acid sequence selected from the group consisting of SEQ ID NO: 23 to SEQ ID NO: 26 Preferably, the helper T cell epitope is linked to a linker, e.g., one or more amino acids. Acids, e.g., Val(V), Ala(A), Arg(R), Gly(G), Ser(S) 23 to SEQ ID NO: 24, fused together via a peptide linker containing Lys(K). The linker may comprise two or more of the amino acid sequences of SEQ ID NO:26. Preferably, the helper T cell epitope is 1 to 5 amino acids. fused together via one or more linkers selected from the group consisting of S, RR, RK The helper T comprises three or more of the amino acid sequences of SEQ ID NO: 23 to SEQ ID NO: 26. The cellular epitope may be amidated at its C-terminus.

[0077] In an embodiment of the present application, the helper T cell epitope is incorporated on the surface of the liposome. can be immobilized, for example, by a covalently attached hydrophobic moiety, where the hydrophobic moiety , in particular at least 3 carbon atoms, in particular at least 4 carbon atoms, in particular at least 6 at least 8 carbon atoms, in particular at least 12 carbon atoms, in particular at least Alkyl groups, fatty acids, triglycerides having a carbon skeleton of at least 16 carbon atoms; Diglycerides, steroids, sphingolipids, glycolipids, or phospholipids, especially alkyl groups or a fatty acid. In one embodiment of the invention, the hydrophobic moiety is palmitic acid. Alternatively, the helper T cell epitope can be encapsulated in a liposome. In this, helper T cell epitopes are encapsulated in liposomes.

[0078] In view of the present disclosure, the helper T cell epitopes can be expressed in a desired position in the liposome. In certain embodiments, the nucleic acid may be modified using methods known in the art. The helper T cell epitope useful in the present invention is one of SEQ ID NO: 39 to SEQ ID NO: 44. Preferably, the helper T cell epitope comprises one amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 14. It consists of an amino acid sequence selected from the group consisting of numbers 17.

[0079] In certain embodiments, a method for treating tau phosphopeptides comprising administering toll-like receptor 4 agonists to a subject .... The liposomes also contain a toll-like receptor 9 agonist. The term "TLR9 agonist" refers to any compound that acts as an agonist of TLR9. In view of the present disclosure, any suitable toll-like receptor agonist known to those of skill in the art may be used. Toll-like receptor 9 agonists may also be used in the present invention. Examples of the target gene include, but are not limited to, TLR9 oligonucleotides. Examples include Nist.

[0080] As used herein, "CpG oligonucleotide," "CpG oligodeoxynucleotide," The term "nucleotide sequence of a nucleic acid molecule," or "CpG ODN," refers to a nucleic acid sequence that contains at least one CpG motif. As used herein, "oligonucleotide" refers to an oligonucleotide that contains "Oligodeoxynucleotide" or "ODN" means a set of multiple linked nucleotides. Such oligonucleotides are known as polynucleotides formed from existing units. The nucleic acid may be obtained from any suitable nucleic acid source or may be produced by synthetic methods. As used herein, the term "CpG motif" refers to a phosphate bond or phosphodiesterase (Pd) bond. Unmethylated cytosine-phosphate linked by a steric backbone or other internucleotide linkage -CpG dinucleotide (i.e., cytosine (C) followed by guanine (G) "A" refers to a nucleotide sequence containing the nucleotide sequence

[0081] In certain embodiments, the CpG oligonucleotide is lipidated, i.e., coupled with a lipid moiety. They are conjugated (linked by a covalent bond).

[0082] As used herein, a "lipid moiety" refers to a moiety that contains a lipophilic structure. Moieties such as alkyl groups, fatty acids, triglycerides, diglycerides, steroids, sulfur Ingolipids, glycolipids, or phospholipids, especially sterols such as cholesterol or fatty acids When binding to highly hydrophilic molecules, e.g. nucleic acids, plasma protein binding and the resulting This can substantially increase the circulating half-life of hydrophilic molecules. Binding to plasma proteins, such as lipoproteins, binds to the corresponding lipoprotein receptors. (e.g., LDL receptor, HDL receptor, or scavenger receptor SR-B1) It has been shown that phosphopeptides and phosphopeptides increase uptake into certain tissues. and / or the lipid moiety conjugated to the CpG oligonucleotide is and / or anchoring the oligonucleotide to the membrane of the liposome via a hydrophobic moiety. This makes it possible.

[0083] In certain embodiments, the CpG oligonucleotide may be any Suitable internucleotide linkages may also be included.

[0084] As used herein, the term "internucleotide linkage" refers to the linkage between two nucleotides. via their sugars, phosphorus atoms between adjacent nucleosides and charged or neutral An example of an internucleotide linkage is the phosphodiester (p o), phosphorothioate (ps), phosphorodithioate (ps2), methylphosphonate Phosphorothioate (mp), and methyl phosphorothioate (rp). Phosphorothioates, phosphorodithioates, methylphosphonates, and methylphosphorothioates are A stabilized internucleotide linkage, phosphodiester is a naturally occurring internucleotide linkage. Oligonucleotide phosphorothioates are typically Rp and Sp phosphorothioates. It is synthesized as a random racemic mixture of thioate linkages.

[0085] In light of the present disclosure, any suitable CpG oligonucleotide known to those of skill in the art may be used in the present invention. Examples of such CpG oligonucleotides include Including, but not limited to, CpG2006 (also known as CpG7909) (SEQ ID NO: 18), CpG1018 (SEQ ID NO: 19), CpG2395 (SEQ ID NO: 20), CpG2216 ( CpG2336 (SEQ ID NO: 21), or CpG2336 (SEQ ID NO: 22).

[0086] CpG oligonucleotides can be synthesized using methods known in the art in light of the present disclosure. In some embodiments, the CpG oligonucleotide can be lipidated by cholesterol. In some embodiments, the CpG oligonucleotide is directly linked to a nucleic acid molecule via a covalent bond. The 3' end of the tide is bound to a cholesterol molecule via a phosphate bond, and optionally to a PEG linker. In some embodiments, the CpG oligonucleotide is covalently linked via The 5' end is linked to a cholesterol molecule by a phosphate bond, optionally via a PEG linker. Other lipophilic moieties may also be attached to the 5' or 6' ends of CpG oligonucleotides. or covalently linked to the 3' end. The lipid anchor is the same length as the phospholipid derived from the liposome, e.g., one palmitic acid chain (using activated Pal-OH or analogues for coupling), and is two palmitic acids (e.g., 1,2-dipalmitate) that have been activated for coupling. yl-sn-glycero-3-phosphoethanolamine-N-(succinyl) or similar and optionally via a PEG linker. See, for example, U.S. Patent No. 7,741, the contents of which are incorporated herein by reference. See the related disclosure in U.S. Pat. No. 5,297,513. The length of the PEG can vary, e.g. For example, 1 to 5 PEG units.

[0087] Other linkers also link CpG oligonucleotides to lipophilic moieties, such as cholesterol. Examples include, but are not limited to, Examples of the spacer include, but are not limited to, alkyl spacers having 3 to 12 carbon atoms. A short linker compatible with leutidine chemistry is required as the aminodiol. In this case, the linker is not used for covalent attachment. Ries et al., "Convenient synthesis and application of vers atile nucleic acid lipid membrane anchors in the assembly and fusion of liposome s, Org. Biomol. Chem., 2015, 13, 9673.

[0088] In a particular embodiment, the lipidated CpG oligonucleotide useful in the present invention is The nucleotide sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:18 to SEQ ID NO:22. The string contains one or more phosphorothioate internucleotide linkages and has the nucleotide sequence is covalently linked to at least one cholesterol via a linker. In a preferred embodiment, the lipidated CpG oligonucleotide is the nucleotide sequence of SEQ ID NO:18. a nucleic acid sequence comprising one or more phosphorothioate internucleotide linkages; Any suitable linker may be used to link a CpG oligonucleotide to a sterol. Can be used to covalently link nucleotides to cholesterol molecules. Preferably, the linker comprises polyethylene glycol (PEG).

[0089] In certain embodiments, the liposome is 1,2-dimyristoyl-sn-glycero- 3-Phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phospho from the group consisting of di-3'-rac-glycerol (DMPG), and cholesterol Further comprising one or more selected lipids.

[0090] In certain embodiments, the liposome further comprises a buffer. Any suitable buffer known to one skilled in the art can be used in the present invention. The liposomes comprise phosphate buffered saline. In certain embodiments, the buffer contains histidine and and sucrose.

[0091] An exemplary liposome for use in the present invention is a liposome comprising tau tetrapalmitoylated phosphopeptide. (pTau peptide T3, SEQ ID NO: 28), comprising two palladium ions at each end of the tau peptide Tau tetrapalmitoylated phospholipids displayed on the surface of liposomes via mitotic acid. peptide; lipidated CpG ( TLR-9 ligand containing the adjuvant CpG7909-Chol (SEQ ID NO: 18) Thus, the cholesterol molecule is covalently linked via a CpG and PEG linker. TLR-9 ligand; TLR-4 ligand (monophosphoryl ribosomal ligand) that is integrated into the membrane PAD A (e.g., 3D-(6-acyl)PHAD®); encapsulated helper T cell epitope (PAN-DR binding site T50, SEQ ID NO: 13); and liposomes ,2-Dimyristoyl-sn-glycero-3-phosphocholine as a lipid component of the (DMPC), 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1 -glycerol)] sodium salt (DMPG), and cholesterol.

[0092] The liposomes of the present invention can be made using methods known in the art in light of this disclosure. The optimal ratio of each of the liposome components can be determined by one of skill in the art in light of this disclosure. The amount of the ion exchange reaction can be determined by known techniques.

[0093] The liposomes can be administered by any means suitable for prophylactic and / or therapeutic treatment. In a preferred embodiment, the liposomes are administered by subcutaneous or intramuscular injection. Intramuscular injections are most typically given into the arm or leg muscles.

[0094] In one general aspect, the present invention provides a method for administering a therapeutically effective amount of liposomes in a pharma- ceutically acceptable vehicle. and / or a carrier. and / or carriers are well known in the art (Remington's Pharmaceutical Sciences (15th edition) d.), Mack Publishing Company, Easton, Pa., 1980. The preferred formulation depends on the intended method of administration and therapeutic application. is defined as a vehicle commonly used to formulate pharmaceutical compositions for The combination may include a non-toxic pharma- ceutically acceptable carrier or diluent. Examples of such diluents are distilled water, These are physicochemical solutions: phosphate-buffered saline, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also contain other carriers, adjuvants, or non-toxic, non-therapeutic agents. The characteristics of the carrier, excipient, or diluent may include, but are not limited to, non-immunogenic, non-specific, and non-immunogenic stabilizers. It will be appreciated that this may depend on the route of administration for a particular application.

[0095] The vaccine's target antigens are located in the brain, which is protected by a special barrier called the blood-brain barrier (BBB). It is separated from the circulation by a cellular structure. The BBB limits the transfer of substances from the circulation to the brain. This prevents toxins, microorganisms, etc. from entering the central nervous system. The BBB also functions as a barrier to immune responses. This prevents mediators (e.g., antibodies) from efficiently entering the interstitial and cerebrospinal fluids that surround the brain. This has the potentially more undesirable effect of

[0096] Approximately 0.1% of antibodies present in the systemic circulation cross the BBB and enter the brain. Therefore, systemic titers induced by vaccines targeting CNS antigens are effective in the brain. This suggests that the effective titer must be at least 1000 times greater than the minimum effective titer. The minimum serum antibody titer required to produce efficacy is not readily apparent. Therefore, not only the quantity but also the quality (e.g., avidity) of the immune response may affect CNS disorders, e.g., neurodegeneration. Must-consider for safe and effective immunotherapy to target a disease, disorder, or condition It won't happen.

[0097] Thus, in certain embodiments, the pharmaceutical compositions of the present invention are used to ameliorate the desired immune response in a subject. The composition further comprises one or more suitable adjuvants to achieve an immune response. The adjuvant can be administered before, after, or simultaneously with administration of the liposomes. Adjuvants induce conformational changes in immunogens that affect the qualitative form of the response. Adjuvants enhance the intrinsic response to immunogens without providing any protective effect against the immune system. Examples of adjuvants include aluminum salts ( Alum), e.g., aluminum hydroxide, aluminum phosphate, and aluminum sulfate Such adjuvants may be other specific immunostimulants, such as MPLAs (3-deoxyglucose -O-acylated monophosphoryl lipid A (MPL™), monophosphoryl hexaacyl Lulipid A 3-Deacylation Synthesis (3D-(6-acyl)PHAD®, PHA D™, PHAD®-504, 3D-PHAD®) Lipid A , in combination with polymeric or monomeric amino acids, such as polyglutamic acid or polylysine. Such adjuvants may be used with or without other specific immunostimulants, such as muramyl peptides (e.g., N-acetyl-muramyl-L-threo N-acetylglutamyl-D-isoglutamine (thr-MDP), N-acetyl-normuramyl-L-aramin N-acetylglutamyl-D-isoglutamine (nor-MDP), N-acetylglutamyl-L-alanyl- D-Isoglutaminyl-L-alanine-2-(1'-2'-dipalmitoyl-sn-glyceryl) 3-Hydroxyphosphoryloxy)-ethylamine (MTP-PE), N-acetyl Glucosaminyl-N-acetylmuramyl-L-Al-D-isoglu-L-Ala-di Palmitoxypropylamide (DTP-DPP) Theramide™, or can be used with or without other bacterial cell wall components. For emulsions, microfluidizers are used to produce particles smaller than a micrometer. Formulated with 5% squalene, 0.5% Tween 80, and 0.5% Span 85 MF59 (International Publication No. 99 / 023663) containing 0 / 14837 brochure); for submicron emulsions Microfluidized or vortexed to produce larger particle size emulsions. Mixed with 10% squalene, 0.4% Tween 80, 5% pluronic blocker SAF containing copolymer L121, and thr-MDP; and 0.2% Twe en80, and monophosphoryl lipid A (MPL™), trehalose diglyceride triglyceride (TDM), and cell wall skeleton (CWS), preferably MPL™+CWS (D etox™) Ribi™ Adjuvant System (RAS) (Ribi ImmunoCh em, Hamilton, Mont. Other adjuvants include complete Freund's adjuvant (CFA), as well as cytokines such as interleukins (IL-1, IL-2, and IL-12), macrophage colony-stimulating factor (MCSF) SF), and tumor necrosis factor (TNF).

[0098] As used herein, the term "in combination" refers to the administration of two or more therapies to a subject. In the context of administration of a compound, the term "in combination" refers to the use of more than one therapy. The use of does not limit the order in which the therapies are administered to a subject. For example, a first therapy (e.g., The compositions described herein may be administered within 5 minutes, 15 minutes, 30 minutes, or 60 minutes prior to administration of the second therapy. 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), at the same time, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 Hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours , 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks later) It can be administered to elephants.

[0099] The timing of dosing can vary greatly from once a day to once a year to once every 10 years. A typical regimen would consist of an immunization followed by a time interval, e.g., 1–24 weeks. Another regimen consists of an immunization followed by 1, 2, 4, and 6 booster injections at intervals. 10, 20, and 30 months after vaccination, followed by booster injections at 6, 8, 10, and 12 months. Another regimen is lifelong Alternatively, booster injections can be given to monitor the immune response. If indicated by a scrub, it may be on an irregular basis.

[0100] The regimen for priming and boosting doses was designed to determine the immune response measured after administration. It will be readily understood by those skilled in the art that the results can be adjusted based on the answer. The priming composition generally is administered several weeks or months after administration of the priming composition, e.g., about 1 week, or 2 weeks, or 3 weeks, or 4 weeks, or 8 weeks, or 16 weeks, 20 weeks, 24 weeks, 28 weeks, or 32 weeks , or 36 weeks, or 40 weeks, or 44 weeks, or 48 weeks, or is 52 weeks, 56 weeks, 60 weeks, 64 weeks, or 68 weeks or 72 weeks, or 76 weeks, or 1 to 2 years after administration of the priming composition. It will be administered later.

[0101] In certain embodiments, one or more booster immunizations may be administered. The antigens in the boosting and boosting compositions depend on how many boosting compositions are used. are used, they do not have to be identical, but may share a common antigenic determinant or may be mutually exclusive. It should be substantially similar to

[0102] The pharmaceutical compositions of the present invention can be formulated according to methods known in the art in light of this disclosure. The optimal ratio of each component in the composition can be determined by one of ordinary skill in the art in light of this disclosure. This can be determined by techniques known to those skilled in the art.

[0103] In a preferred embodiment of the invention, the method comprises administering a pharmaceutical composition according to an embodiment of the invention. Administration of targeted tau peptides stimulates an active immune response against tau peptides and pathological forms of tau. Induce tau-related aggregate clearance in mice, thereby preventing tau pathology-related behavior slow the progression of the disease and / or treat the underlying tauopathy.

[0104] Tau is a human “self” protein. This means that, in principle, there is no specific receptor for tau. Either all lymphocytes present in the body are eliminated during development (central tolerance) or peripheral This means that the immune system would have been refractory to the vaccine due to the mechanism of immune tolerance. This is a major milestone in the development of vaccines against "altered self" proteins (e.g., tumor antigens). It has been shown to be a major obstacle to the development of the immune system. The goal is to stimulate not only antibody-producing B lymphocytes, but also CD4 + of T "helper" lymphocytes It also requires CD4 + T cells transmit vital survival and maturation signals to B lymphocytes. and CD4 + T cell-deficient animals are severely immunosuppressed. + T cells also play a role in immunity. Susceptible to tolerance mechanisms and the induction of strong anti-self (e.g., anti-tau) antibody responses A further obstacle in the treatment of tau-reactive CD4 + T cells also enter the human / animal repertoire The challenge is that it is likely to be rare or nonexistent in the

[0105] In this aspect of the invention, the immune response is a beneficial humoral (antibody) directed against tau peptides. body-mediated responses, and cell-mediated (antigen) responses directed against T cell epitopes or immunogenic carriers It involves the development of a response (mediated by specific T cells or their secreted products).

[0106] As used herein, tau pathology associated behavioral phenotypes include, but are not limited to, cognitive impairment. Harm, early personality change and disinhibition, apathy, akinetic mutism, apraxia, perseveration, stereotyped movements / behaviors , lip-syncopation, disorganization, inability to plan or organize sequential tasks, selfish behavior / callousness, Antisocial traits, lack of empathy, lack of consistency, frequent erroneous errors, but relatively good understanding Slowly progressing agrammatic speech, impaired comprehension and word finding, slowly progressive gait instability, retropulsion , freezing, frequent falls, non-levodopa-responsive axial rigidity, supranuclear gaze palsy, square wave eye movements, Slow vertical saccadic eye movements, pseudobulbar palsy, limb apraxia, dystonia, cortical sensory loss, etc. Other symptoms include tremors.

[0107] In carrying out the methods of the invention, in certain embodiments of the invention, the immunogenic peptides of the invention are Patients with Alzheimer's disease or other tauopathies prior to administration of the peptide or antibody subjects with or at risk of having tau aggregates in the brain, or subjects with high levels of tau aggregates in the brain It is preferable to select subjects exhibiting a deficiency-associated behavioral phenotype. is a method for identifying individuals at risk for disease but who are not showing symptoms, and for patients who are currently showing symptoms. In the case of Alzheimer's disease, virtually everyone has a risk of developing the disease. Therefore, the method does not require any assessment of the risk of the subject patient. The method can be administered prophylactically to the general population without the need for any known treatment for Alzheimer's disease. It is particularly useful for individuals at genetic risk. Such individuals include those who have undergone Individuals with relatives who have been tested and those who have been identified through analysis of genetic or biochemical markers In a preferred embodiment, the subject is an individual whose risk is determined by Alzheimer's disease, preferably early Alzheimer's disease, mild cognitive impairment due to Alzheimer's disease ( MCI), mild Alzheimer's disease, or mild to moderate Alzheimer's disease requiring treatment It is necessary.

[0108] In asymptomatic patients, treatment can begin at any age (e.g., 10, 20, 30 years). However, it is usually not until patients reach age 40, 50, 60, or 70 years. There is no need to initiate treatment. Treatment typically involves multiple dosages over a period of time. The method comprises assaying activated T cell or B cell responses to an antibody or therapeutic agent. If the response drops, booster doses can be administered. The amount is appropriate.

[0109] In prophylactic applications, pharmaceutical compositions containing tau peptides can be used to treat Alzheimer's disease or other conditions. Patients susceptible to or otherwise at risk for a tauopathy are treated with The biochemical, histological, and / or behavioral manifestations of, and complications from, the disease, and its occurrence Eliminate or reduce the risk of disease, including pathological endophenotypes present in It is administered in an amount sufficient to reduce the severity or delay the onset of the condition. Pharmaceutical compositions containing tau peptides may be used in patients suspected of or already suffering from such diseases. The patient is then assessed for disease progression, including disease complications and pathological endophenotypes in disease development. Treat or lessen the symptoms (biochemical, histological, and / or behavioral) of the disease The drug is administered in an amount sufficient to partially halt both the

[0110] The composition may, if desired, contain one or more unit dosage forms containing the active ingredient. The kit may be provided in a kit, container, or dispensing device that can For example, it may contain metal or plastic foil, such as a blister pack. The packet, container, or dispensing device can be accompanied by instructions for administration.

[0111] Embodiment The present invention also provides the following non-limiting embodiments.

[0112] Embodiment 1 provides an anti-phosphorylated tau antibody that can be administered to patients who require it without inducing severe adverse events. A method of inducing in a subject, comprising administering to said subject from about 25 nmole to about 750 nmole per dose e., preferably about 90 nmole to about 715 nmole per dose, about 9 0 nmole to about 535 nmole, or about 90 nmole to about 275 nmole per dose mole, e.g., about 25 nmole, about 30 nmole, about 35 nmole per dose , about 40 nmole, about 45 nmole, about 50 nmole, about 55 nmole, about 60 nmole, approx. 65 nmole, approx. 70 nmole, approx. 75 nmole, approx. 80 nmole e, about 85 nmole, about 90 nmole, about 95 nmole, about 100 nmole, about 125 nmole, about 150 nmole, about 175 nmole, about 200 nmole, about 225 nmole, about 250 nmole, about 275 nmole, about 300 nmole, about 325 nmole, approx. 350 nmole, approx. 375 nmole, approx. 400 nmole, approx. 425 nmole, approx. 450 nmole, approx. 475 nmole, approx. 500 nmole, approx. 525 nmole, about 550 nmole, about 575 nmole, about 600 nmole, about 625 nmole, approx. 650 nmole, approx. 675 nmole, approx. 700 nmole, approx. 725 nmoles, about 750 nmoles, or any value therebetween. An amino acid sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:3 and SEQ ID NO:5 to SEQ ID NO:12 and an effective amount of a liposome comprising a tau phosphopeptide comprising a toll-like receptor 4 agonist. The method includes administering to a subject a tau phosphopeptide that is presented on the surface of a liposome. This is a method.

[0113] Embodiment 1a provides a method for administering an anti-phosphorylated tau antibody to a subject in need thereof without inducing encephalitis. The method for inducing the above-mentioned IL-16 receptor agonist in a patient is described below. 0μg~2400μg, 300μg~1800μg, or 300μg~900μg, e.g. For example, 100 μg, 150 μg, 200 μg, 250 μg, 300 μg, 40 0μg, 500μg, 600μg, 700μg, 800μg, 900μg, 1000μg , 1100μg, 1200μg, 1300μg, 1400μg, 1500μg, 1600 μg, 1700μg, 1800μg, 1900μg, 2000μg, 2100μg, 22 00μg, 2300μg, 2400μg, 2500μg, or any value between them , tetrapalmitoylated tau phosphopeptide displayed on the surface of liposomes, and and administering to a subject an effective amount of a liposome comprising an LL-like receptor 4 agonist. The phosphopeptides consist of SEQ ID NO:1 to SEQ ID NO:3 and SEQ ID NO:5 to SEQ ID NO:12. The method comprises the step of:

[0114] In a second embodiment, the tau phosphopeptide is selected from SEQ ID NO: 27 to SEQ ID NO: 29 and SEQ ID NO: 31. The method of any one of the preceding claims, wherein the amino acid sequence is selected from the group consisting of SEQ ID NO: 38. This is a method according to the present invention.

[0115] Embodiment 3a is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 27. The method according to any one of modes 1 and 2.

[0116] Embodiment 3b is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 28. The method according to any one of modes 1 and 2.

[0117] Embodiment 3c is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 29. The method according to any one of modes 1 and 2.

[0118] Embodiment 3d is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 31. The method according to any one of modes 1 and 2.

[0119] Embodiment 3e is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 32. The method according to any one of modes 1 and 2.

[0120] Embodiment 3f is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 33. The method according to any one of modes 1 and 2.

[0121] Embodiment 3g is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 34. The method according to any one of modes 1 and 2.

[0122] Embodiment 3h is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 35. The method according to any one of modes 1 and 2.

[0123] Embodiment 3i relates to an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 36. The method according to any one of modes 1 and 2.

[0124] Embodiment 3j is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 37. The method according to any one of modes 1 and 2.

[0125] Embodiment 3k is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 38. The method according to any one of modes 1 and 2.

[0126] Embodiment 4 is the method of any one of embodiments 1 to 3k, wherein the liposomes are administered subcutaneously. This is the method.

[0127] Embodiment 5 is the method of any one of embodiments 1 to 3k, wherein the liposomes are administered intramuscularly. This is the method.

[0128] In embodiment 6, a second dose of an effective amount of liposomes is administered 1 to 24 weeks after the first dose. 6. The method of any one of embodiments 1 to 5, further comprising administering

[0129] In embodiment 7, the liposome comprises a helper T cell epitope and a lipidated CpG oligonucleotide. At least one of the peptides, preferably a helper T cell epitope and a lipidated C 7. The method of any one of claims 1 to 6, further comprising a pG oligonucleotide. be.

[0130] In embodiment 7a, the effective amount of liposomes is between 30 μg and 900 μg per dose, preferably 100μg to 585μg, for example 30μg, 50μg, 100μg, 150μg, 2 00μg, 250μg, 300μg, 330μg, 360μg, 390μg, 420μg , 450μg, 480μg, 500μg, 520μg, 540μg, 560μg, 580 μg, 600 μg, 700 μg, 800 μg, or 900 μg, or any interval therebetween 8. The method of embodiment 7, comprising any amount of the toll-like receptor 4 agonist.

[0131] Embodiment 7b is a method for treating toll-like receptor 4 agonists comprising administering toll-like receptor 4 agonists a monophosphoryl hexaacyl lipid. The method according to embodiment 7 or 7a, wherein A is 3-deacylation.

[0132] In embodiment 7c, the effective amount of liposomes is between 25 μg and 625 μg per dose, preferably 75μg to 450μg, for example 25μg, 50μg, 75μg, 100μg, 125μg μg, 150μg, 175μg, 200μg, 225μg, 250μg, 275μg, 3 00μg, 325μg, 350μg, 375μg, 400μg, 425μg, 450μg , 475μg, 500μg, 525μg, 550μg, 575μg, 600μg, or 625 μg, 630 μg, or any value therebetween. The method according to any one of embodiments 7 to 7b.

[0133] Embodiment 7d is a variant in which the helper T cell epitope consists of the amino acid sequence of SEQ ID NO: 13. T50 helper T cell epitope, T46 helper consisting of the amino acid sequence of SEQ ID NO: 14 T cell epitope, T48 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 15 a T51 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO: 16, or the sequence 17. The T52 helper T cell epitope of embodiment 7 or 7c et al.

[0134] Embodiment 7e is a T helper T cell epitope comprising the amino acid sequence of SEQ ID NO: 13. 50 helper T cell epitope.

[0135] In embodiment 7f, the effective amount of liposomes is between 50 μg and 1250 μg per dose, preferably or 150μg to 800μg, for example 50μg, 100μg, 150μg, 200μg , 250μg, 300μg, 350μg, 400μg, 450μg, 500μg, 550 μg, 600μg, 650μg, 700μg, 750μg, 800μg, 850μg, 9 00μg, 950μg, 1000μg, 1050μg, 1100μg, 1200μg, or 1250 μg of lipidated CpG oligonucleotide, or any value therebetween. The method of any one of embodiments 7 to 7e, comprising:

[0136] In an eighth embodiment, the lipidated CpG oligonucleotide comprises SEQ ID NO: 18 to SEQ ID NO: 22. The method according to any one of embodiments 7 to 7f, having a nucleotide sequence selected from the group consisting of: This is the method described above.

[0137] Embodiment 9a is a method for preparing a CpG oligonucleotide having the nucleotide sequence of SEQ ID NO: 18. 9. The method according to embodiment 8,

[0138] Embodiment 9b is a method for preparing a CpG oligonucleotide having the nucleotide sequence of SEQ ID NO: 19. 9. The method according to embodiment 8,

[0139] Embodiment 9c is a method for preparing a CpG oligonucleotide comprising the steps of: 9. The method according to embodiment 8,

[0140] Embodiment 9d is a method for preparing a CpG oligonucleotide comprising the steps of: 9. The method according to embodiment 8,

[0141] Embodiment 9e is a variant in which the CpG oligonucleotide has the nucleotide sequence of SEQ ID NO:22. 9. The method according to embodiment 8,

[0142] Embodiment 10 is a method for preparing a CpG oligonucleotide comprising the steps of: The method of any one of embodiments 8 to 9e, wherein the nucleic acid has an internucleotide linkage.

[0143] Embodiment 11 is a method for preparing a CpG oligonucleotide comprising the steps of: 11. The method according to any one of embodiments 8 to 10, wherein said first and second nucleotide sequences are linked together.

[0144] Embodiment 11a is a CpG oligonucleotide comprising at least one lipophilic group and a PEG-linked nucleotide. 11. The method according to any one of embodiments 8 to 10, wherein the carboxyl group is covalently linked via a carboxyl group. It is.

[0145] In embodiment 12, the CpG oligonucleotide is covalently linked to a cholesterol group. The method according to embodiment 11 or 11a,

[0146] Embodiment 12a is a CpG oligonucleotide comprising a cholesterol group and a PEG linker. 13. The method of embodiment 12, wherein the covalent linkage is via a

[0147] Embodiment 13 is a method for preparing a liposome comprising administering to a subject a subject the liposome is 1,2-dimyristoyl-sn-glycero-3-phosphorylated. Phosphoryl-3, 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3 '-rac-glycerol (DMPG), and cholesterol. The method according to any one of embodiments 1 to 12a, further comprising one or more lipids. It is law.

[0148] In embodiment 14, the helper T cell epitope is a group consisting of SEQ ID NO: 23 to SEQ ID NO: 26. Any one of embodiments 1 to 13, comprising at least one amino acid sequence selected from The present invention relates to a method for

[0149] Embodiment 15 is a method for the preparation of a peptide comprising the steps of: 15. The method of embodiment 14, comprising the amino acid sequence of SEQ ID NO:25.

[0150] In embodiment 16, the helper T cell epitope is selected from SEQ ID NOs: 13 to 17 and 39 to 44. 16. The method of embodiment 14 or 15, comprising an amino acid sequence selected from the group consisting of: do.

[0151] Embodiment 17 is a method for treating toll-like receptor 4 agonists comprising administering toll-like receptor 4 agonists to a subject, comprising administering to the subject a toll-like receptor 4 agonist, ... 17. The method of any one of the preceding claims, wherein

[0152] Embodiment 18 provides a method for administering an anti-phosphorylated tau antibody to a patient in need thereof without inducing severe adverse events. The method of inducing in a subject, the method comprising administering to said subject from about 25 nmole to about 750 nmole per dose. le, preferably about 90 nmole to about 715 nmole, or about 90 nmole per dose nmole to about 535 nmole, or about 90 nmole to about 275 nmole per dose mole, e.g., about 25 nmole, about 30 nmole, about 35 nmole per dose , about 40 nmole, about 45 nmole, about 50 nmole, about 55 nmole, about 60 nmole, approx. 65 nmole, approx. 70 nmole, approx. 75 nmole, approx. 80 nmole e, about 85 nmole, about 90 nmole, about 95 nmole, about 100 nmole, about 125 nmole, about 150 nmole, about 175 nmole, about 200 nmole, about 225 nmole, about 250 nmole, about 275 nmole, about 300 nmole, about 325 nmole, approx. 350 nmole, approx. 375 nmole, approx. 400 nmole, approx. 425 nmole, approx. 450 nmole, approx. 475 nmole, approx. 500 nmole, approx. 525 nmole, about 550 nmole, about 575 nmole, about 600 nmole, about 625 nmole, approx. 650 nmole, approx. 675 nmole, approx. 700 nmole, approx. 725 nmole, about 750 nmole, or any value therebetween, or one dose 300μg to 2400μg per serving, for example 300μg to 1800μg, or 300μ g to 900 μg, e.g. 100 μg, 150 μg, 200 μg, 250 μg per dose , 300μg, 400μg, 500μg, 600μg, 700μg, 800μg, 900 μg, 1000μg, 1100μg, 1200μg, 1300μg, 1400μg, 15 00μg, 1600μg, 1700μg, 1800μg, 1900μg, 2000μg, 2100μg, 2200μg, 2300μg, 2400μg, 2500μg, or Tetrapalmitoylated tauphos displayed on the surface of liposomes with any value between these values. administering to a subject an effective amount of liposomes containing a tau phosphopeptide; is selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3 and SEQ ID NO: 5 to SEQ ID NO: 12 The liposomes are composed of monophosphoryl lipid A (MPLA), 1,2-diamino acid sequence, Ristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl -sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG), cholesterol a terol, and a buffer.

[0153] Embodiment 18a provides a method for administering an anti-phosphorylated tau antibody to a subject in need thereof without inducing encephalitis. A method for inducing an inflammatory response in a patient with an inflammatory bowel disorder, comprising administering to the patient an inflammatory bowel disorder comprising administering to the patient an inflammatory bowel disorder comprising administering to the patient a therapeutically effective amount of an inflammatory bowel disorder, the method ... Preferably, about 90 nmole to about 715 nmole, or about 90 nmole per dose. le to about 535 nmole, or about 90 nmole to about 275 nmol per dose e.g., about 25 nmole, about 30 nmole, about 35 nmole, about 4 nmole, 0 nmole, approx. 45 nmole, approx. 50 nmole, approx. 55 nmole, approx. 60 nmole le, about 65 nmole, about 70 nmole, about 75 nmole, about 80 nmole, about 85 nmole, approx. 90 nmole, approx. 95 nmole, approx. 100 nmole, approx. 125 nmole, about 150 nmole, about 175 nmole, about 200 nmole, about 225 nmole, about 250 nmole, about 275 nmole, about 300 nmole, about 325 nmole, about 350 nmole, about 375 nmole, about 400 nmole, about 425 nmole, about 450 nmole, about 475 nmole, about 500 nmole, about 525 nmole, about 550 nmole, about 575 nmole, about 600 nmole, about 625 nmole, approx. 650 nmole, approx. 675 nmole, approx. 700 nmole, approx. 725 nmole, about 750 nmole, or any value therebetween, or per dose 100μg to 2500μg or 300μg to 2400μg, for example 300μg to 18 00μg or 300μg to 900μg, e.g. 100μg, 150μg per dose , 200μg, 250μg, 300μg, 400μg, 500μg, 600μg, 700 μg, 800μg, 900μg, 1000μg, 1100μg, 1200μg, 1300 μg, 1400μg, 1500μg, 1600μg, 1700μg, 1800μg, 19 00μg, 2000μg, 2100μg, 2200μg, 2300μg, 2400μg, 2500 μg of tetritol displayed on the surface of the liposomes, or any value therebetween. administering to the subject an effective amount of liposomes comprising lapalmitoylated tau phosphopeptide; and the tau phosphopeptides are selected from SEQ ID NO:1 to SEQ ID NO:3 and SEQ ID NO:5 to SEQ ID NO:12. and wherein the liposome comprises an amino acid sequence selected from the group consisting of monophosphoryl lipid A ( MPLA), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) , 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycero The method further comprises adding dichloromethane (DMPG), cholesterol, and a buffer.

[0154] Embodiment 19 is a method for preparing a liposome comprising administering to a patient a helper T cell epitope and a cholesterol group. 19. The method of claim 18, further comprising covalently linking a CpG oligonucleotide to 18a.

[0155] Embodiment 19a is a CpG oligonucleotide comprising a cholesterol group and a PEG linker. 20. The method of embodiment 19, wherein the covalent linkage is via a

[0156] Embodiment 19b is a variant in which the helper T cell epitope is selected from SEQ ID NO:23, SEQ ID NO:24, and and the amino acid sequence of SEQ ID NO: 25.

[0157] Embodiment 19c is a method for the preparation of a peptide comprising the helper T cell epitope of SEQ ID NOs: 13-17 and 39-44. The method according to embodiment 19 or 19a, comprising an amino acid sequence selected from the group consisting of: It is.

[0158] Embodiment 19d is an embodiment in which the helper T cell epitope comprises the amino acid sequence of SEQ ID NO: 39. The method according to embodiment 19 or 19a.

[0159] Embodiment 19e is an embodiment in which the helper T cell epitope comprises the amino acid sequence of SEQ ID NO: 13. The method according to embodiment 19 or 19a.

[0160] Embodiment 19f relates to a method for the preparation of a CpG oligonucleotide having the nucleotide sequence of SEQ ID NO:18. The method of any one of embodiments 19 to 19e,

[0161] Embodiment 19g is a method for preparing a CpG oligonucleotide comprising the steps of: The method of any one of embodiments 19 to 19e,

[0162] Embodiment 20a is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 27. The method according to any one of forms 18 to 19g.

[0163] Embodiment 20b is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 28. The method according to any one of forms 18 to 19g.

[0164] Embodiment 20c is an embodiment in which the tau phosphopeptide consists of the amino acid sequence of SEQ ID NO: 29. The method according to any one of forms 18 to 19g.

[0165] Embodiment 21a is any of embodiments 18 to 20c, wherein the buffer comprises a phosphate buffer. This is one method.

[0166] Embodiment 21b is a method for preparing a medicament comprising the steps of: Included are methods according to any one of embodiments 18 to 20c.

[0167] Embodiment 22a is a method for preparing a phospholipid comprising administering to a subject a subject, the subject being satisfied, in which the MPLA is monophosphoryl lipid A (e.g., 3D-(6-acyl The method according to any one of embodiments 18 to 21b, comprising administering to said patient a therapeutically effective amount of PHAD®. It is.

[0168] Embodiment 23 is a method according to any one of embodiments 1 to 22a, wherein the subject is in need of treatment for Alzheimer's disease. The method according to any one of the above.

[0169] Embodiment 24 is a method according to any one of embodiments 1 to 22a, wherein the subject is in need of prevention of Alzheimer's disease. The method according to any one of the above.

[0170] Embodiment 25 is a method for treating Alzheimer's disease, including administering to a subject a therapeutically effective amount of serotonin to a subject suffering from Alzheimer's disease, mild cognitive impairment due to Alzheimer's disease, or a combination of these conditions. cognitive impairment (MCI), mild Alzheimer's disease, or mild to moderate Alzheimer's disease 24. The method of embodiment 23 in need of treatment.

[0171] Embodiment 26 is any one of embodiments 1 to 25, wherein the subject is a human subject. This is the method. EXAMPLES

[0172] The following examples of the present invention are intended to further illustrate the principles of the present invention. The examples do not limit the invention, the scope of which should be determined by the appended claims. It should be understood that there is.

[0173] All experimental methods used in the following examples are routine unless otherwise indicated. The reagents used in the following embodiments are all available from standard reagent suppliers unless otherwise specified. are purchased from suppliers.

[0174] In all the following examples, ACI-35.030 refers to 1200 μg / mL of SEQ ID NO: A phosphorylated tau peptide with a sequence of 28 amino acids, MPLA (3D-(6-acyl) PHAD®), DMPC, DMPG, cholesterol, 300 μg / mL The helper T cell epitope in column number 13 is covalently linked to a cholesterol group via a PEG linker. The present invention relates to a method for the preparation of a CpG oligonucleotide comprising the steps of: In the liposome preparation of the embodiment, ACI-35 is a liposome preparation having the amino acid sequence of SEQ ID NO:28. The phosphorylated tau peptide, MPLA, DMPC, DMPG, cholesterol, and 1 is a liposome formulation according to an embodiment of the present invention containing a buffer solution.

[0175] [Example 1] Dose response of ACI-35.030 in C57BL / 6J mice Objective: AC when administered to C57BL / 6 female mice by intramuscular (im) injection. To evaluate the immunogenicity of the I-35.030 vaccine.

[0176] Methods: The study design is described in Table 1 .

[0177] [Table 1]

[0178] Specific IgG antibody responses directed against phosphorylated tau peptides (determined by ELISA) ) was measured.

[0179] Results / Conclusion: Target doses of 35 and 80 μg in the ACI-35.030 (1200:300) vaccine Immunization with T3 of the present invention was directed to the T3.5 peptide having the amino acid sequence of SEQ ID NO:2. The antibody induced the IL-1 induced ...

[0180] [Example 2] ACI-35.030 (tetrapalmitoylated T3 on the liposome surface) in rhesus monkeys Phospho-tau peptide, MPLA (3D-(6-acyl)PHAD®), and and CpG7909-Chol, and encapsulated T50 vaccines Assessment of IgG responses Objective: ACI-35 administered to male and female rhesus monkeys by subcutaneous or intramuscular injection. 030 (Tetrapalmitoylated T3 phospho-tau peptide on the liposome surface, MPLA( 3D-(6-acyl)PHAD (registered trademark), and CpG7909-Chol, etc. To evaluate the immunogenicity of the T50 vaccine encapsulated in porcine mice and thereby to determine the optimal vaccine. Determine route of administration and concentration.

[0181] Methods: The study design is described in Table 2.

[0182] [Table 2]

[0183] The theoretical dose of liposomal vaccine is 1800 μg of tetrapalmitoyl per animal. The total amount of the peptides was 1800 μg of phosphopeptide (T3) and 1800 μg of T50 peptide.

[0184] Four doses were administered at the following intervals: days 1, 29, 85, and 169. Blood samples for the study were collected at the following intervals: -14, 8, 22, 36, 50, 64, 78, 9 Days 2, 106, 120, 134, 148, 162, 176, and 190.

[0185] Phosphorylated and non-phosphorylated tau peptides (T3.5 and T3.6, respectively; ELISA A), full-length phospho-tau (p-tau) protein (determined by ELISA), We also identified pathological forms of tau extracted from human brains of Alzheimer's disease patients (human P Specific IgG antibody responses to HF (determined by MSD) were measured. The animals were monitored for symptoms (health status, behavioral changes, etc.) twice daily from the day of delivery through the entire study. Body weights were recorded for all animals at least once before the start of treatment and weekly thereafter. Recorded at least once.

[0186] Results / Conclusion: Overall, neither systemic adverse effects nor persistent skin sensitivity at the injection site were reported with either vaccine. No inflammatory bowel disease could be observed even after immunization with the chin regimen.

[0187] Concentrated (1200:1200) and non-concentrated (400:400) ACI-35.030 Cutin induced similar T3.5- and human PHF-specific IgG titers. The difference between allogeneic IgG titers and human PHF-specific IgG titers was observed after sc administration of the same vaccine formulation. No significant difference was detected between the im and postnatal doses.

[0188] [Example 3] Eight subcutaneous injections of ACI-35 every 2 weeks, followed by a 2-week treatment-free period (GLP) Subsequent repeat-dose toxicity study in C57BL / 6 mice Objective: AC in C57BL / 6 mice following subcutaneous injection every 2 weeks for 14 weeks. To evaluate the potential toxicity of I-35. After completion of the treatment period, designated animals were placed in a 2-week free The treatment period was followed to assess the reversibility of any findings. Gender was assessed during the study.

[0189] Methods: The study design is described in Table 3.

[0190] [Table 3]

[0191] Results / Conclusion: ACI-35-free or ACI-35 at the highest dose level of 390 μg / injection Repeated injections of induced a decrease in the albumin / globulin (A / G) ratio after eight injections. This change, generally observed in immunized animals, was due to a decrease in the globulin (antibody) plasma percentage. This is consistent with the increase in antibody production in all ACI-35 treated animals (IgM and This is explained by the fact that the IgG titers increased repeatedly (in the IgG and IgG classes). After injection, IgM titers were higher and females tended to produce more antibodies than males.

[0192] At the site of injection, a localized and transient reaction (thickening) was observed after 98 or 390 μg of peptide. At these doses, the effect was observed in both sexes in the groups treated with ACI-35. Increased spleen and liver weights and decreased thymus weights in the rats, and decreased kidney weights in the females only. At the injection site, macroscopic findings were predominantly observed in the 98 and 390 μg / injection groups. Microscopically, PBS control, ACI-35-free, Administration of ACI-35 at 24 μg / injection was well tolerated locally. Foamy macrophage and mononuclear inflammatory infiltrates associated with loaded liposomes were dose-dependent. were seen at all dose levels with significant adverse events and severity, but not in the PBS control group. A subacute inflammatory reaction accompanied by fibrosis and possibly degeneration / necrosis of the subcutaneous tissue occurred at 390 μm. There was a slight increase in the case of ACI-35 at 100 g / injection.

[0193] Administration of ACI-35 without or at 390 μg / injection induced extramedullary hematopoiesis. Induced in the spleen and liver and increased myeloid cell numbers in bone marrow, which were not liposomal In addition, mononuclear inflammatory cell infiltrates in multiple systemic organs were observed primarily in females. This was seen with ACI-35 at 390 μg / injection in the 100-mL mice, whereas the 100-mL mice without ACI-35 showed better response. This finding is consistent with previous findings in mice, which are known to have an easily inducible immune system. This was the kind of response that could be expected in these animals because they were strains of and therefore were not considered clinically significant. The only systemic finding after administration was extramedullary hematopoiesis in the spleen and liver. There were no systemic findings following administration of ACI-35.

[0194] None of these findings were associated with mild to moderate severity, complete or partial recovery after 2 weeks. Given the partial recovery and the absence of significant clinical or clinicopathologic correlates, As a result, under the experimental conditions of this study, was administered to C57BL / 6J male and female mice every 2 weeks for 3 months. The ACI-35 dose level of 0 μg peptide was the NOAEL (no signs of systemic toxicity, no local (There were no adverse events at these levels.)

[0195] [Example 4] 6-week GLP recovery period in cynomolgus monkeys with ACI-35 Month-long subcutaneous toxicity study Objective: To evaluate the efficacy and safety of ACI-35 when administered subcutaneously for seven doses in cynomolgus monkeys once every four weeks. To assess cumulative toxicity of treatment-related changes, including reversibility or progression, or any delayed toxicity. was evaluated during a 4-week recovery period for some animals following the treatment period.

[0196] Methods: The study design is described in Table 4.

[0197] [Table 4]

[0198] Results / Conclusion: 358, 716, and 1431 μg / injection once every 28 days for a 6-month period Subcutaneous administration of ACI-35 to cynomolgus monkeys at 4 °C was well tolerated in all animals. did.

[0199] Although a relationship to treatment could not be excluded, a slight increase in neutrophil counts and glomerular filtration rate were observed. The slight decrease in course level was due to the fact that all values ​​were within the range of historical background data and were consistent with the preliminary results. No relevant variation in values ​​was recorded and therefore not considered to be harmful.

[0200] All test article treatment groups developed antibodies. Two weeks after the first dose, the immune response was The mean anti-p-tau antibody titers were similar across the test article treatment groups. After 4 weeks of recovery, animals were treated with the seventh dose. There was a clear decrease in antibody levels recorded two weeks after administration.

[0201] Based on the results obtained, under the conditions of these studies, 1 The dose of 1431 μg / injection administered once was considered the NOAEL (no observed adverse effect level) for this study. It was thought.

[0202] [Example 5] Rhesus monkeys were administered ACI-35.030, followed by a 4-week recovery period (GLP). Three-Month Intramuscular Toxicity Study in Objective: The objective of this study was to administer muscle stimulation to naive male and female rhesus monkeys three times a month. The objective of this study was to evaluate the potential toxicity of ACI-35.030 when administered intramuscularly. The reversibility of possible treatment-related changes was assessed using a 4-week recovery period for two animals / sex in the control and high-dose groups. was evaluated during the period.

[0203] DESIGN: The study protocol applied was as follows (Table 5).

[0204] [Table 5]

[0205] Test and reference substances were administered by single intramuscular injection into three thigh sites on days 1, 29, and 85. All animals were monitored throughout the study for survival / death and clinical signs. On each dosing day, animals were observed at least twice daily for pre-dose and post-dose Evaluation of local reactions at the administration site (Draizus method) was performed 6, 24, and 48 hours after administration. The pieces were then subjected to core processing.

[0206] Food consumption was estimated qualitatively for each cage throughout the study. Body weight was calculated from the start of the acclimation period. Patients were evaluated weekly until the end of the study. Fundus examinations were performed once during the pre-study period and 3 / 4 days after the final dose. Electrocardiogram (limb leads I, II, and III, and augmented lead aVR (augmented lead aVR) was performed. aVL (augmented potential right), aVL (augmented potential left), and aVF (augmented potential foot) were measured during the preliminary and final Each animal was recorded once approximately 24 hours after the last dose.

[0207] Clinical pathology evaluations (hematology, clinical chemistry, coagulation, and urinalysis) were performed exploratory on all animals. During the study, once 2 days after the last injection (day 87) and at recovery for all surviving animals. The test was conducted once towards the end of the period (day 125).

[0208] Blood samples of serum for anti-p-tau and anti-T50 ELISA determination were collected from -14 , 8, 22, 36, 50, 64, 78, 92, 99, 106, 120, and 127 days (Days 106, 120, and 127 were collected only for recovery animals).

[0209] PBMCs were collected from all groups on day -14, 2 weeks after the final immunization, and in the recovery group. IL-4 and IFN-γ ELISPOT analysis was performed to detect tau Specific T cell responses were determined.

[0210] Immunophenotyping was performed on all animals, sampled during pre-study and once at necropsy. The evaluation was based on blood.

[0211] Blood was collected from the PMB on days -14 and 99 (main and recovery) and on day 127 (recovery). C (ELISpot for T cell responses to T3.5).

[0212] Cerebrospinal fluid (CSF) samples were obtained from all animals under sedation, during pre-study and before each post-mortem examination. , and were taken for cytological evaluation.

[0213] After completion of the scheduled treatment or recovery period, all animals were necropsied and various organs were weighed. Histopathological examination was performed on the brain, injection sites, and lymph nodes of all study animals. carried out.

[0214] Immunohistochemistry (IHC) technique was used to detect the antibodies induced by ACI-35.030. Potential binding activity to various human tissues was evaluated in a dedicated study, where The cross-reactivity of ACI-35.030-induced monkey antibodies to mouse tissues was Serum sampled on day 99 of treated animals was used to compare the IL-1 expression levels of IL-1 from three unrelated individuals. The antibody was evaluated in a panel of 42 frozen human tissues and blood smears.

[0215] result: Mortality, weight, physical changes, appetite, ophthalmology, electrocardiogram, clinical pathology (hematology) , coagulation, clinical chemistry, and urinalysis), or ACI-35.0 for immunophenotyping 30 There were also no related effects.

[0216] Transient mild to moderate signs of skin irritation (erythema and edema) at the injection site , was observed in several animals across all groups following immunization. Transient severe local edema was observed. One of five females received 2400 μg T3 / injection in the treated hind limb at 48 hours. Edema was observed on days 29 and 87. However, edema was observed 1 day later (days 30 and 8 It disappeared on the 8th day.

[0217] Several dose-independent differences in mean organ weights were observed between the control and test article-treated groups. Macroscopically, inguinal, iliac, and pelvic lymph nodes were found in all groups, including controls. Randomly described as enlarged over a wide area, often associated with microscopic lymphoid hyperplasia Most recovered after a 4-week recovery period, and the vaccine / vaccine adjuvant activation Microscopically, minimal to mild inflammation was observed at the injection site. It was found in skeletal muscle and in the dermis and / or subcutaneous tissue, with Minor degeneration and / or regeneration was observed across all groups, including the control. Inflammation was observed at the site of the needle puncture. This may be due to a combination of the injection technique associated with the needle and a local immune response to the injected material. These changes were not considered adverse and were reversible after a 4-week recovery period. It was predicted that.

[0218] There were no microscopic brain changes associated with ACI-35.030 administration.

[0219] ACI-35.030 was sustained at both 1200 and 2400 μg / dose. The antibody induced persistent anti-pTau IgG titers (Figure 2).

[0220] Tau-specific T cell responses were not observed at the end of treatment, and T cell activation-related toxicities, e.g., myelopathy, This suggests a low risk of meningoencephalitis.

[0221] Immunotherapy was performed based on 42 frozen human tissue and blood smears from three unrelated individuals. Histochemistry showed that ACI-35.030 inducers tested at 1 / 300 and 1 / 100 The derived monkey antibody specifically stained tau tangles in AD brain sections (Figure 3, Panels A and B). Before treatment with CI-35.030, Panel B: ACI-35 at 2400 μg / dose. 030 after three injections, serum diluted 1 / 100) was not detected in any of the control tissues tested. They emphasized that the presence of IgG1 did not result in on- or off-target staining. As an example, colon section staining with pre- and post-treatment serum (panels C and D, respectively) The structure of the cascade ...

[0222] Conclusion: Overall, the absence of ACI-35-associated changes after immunization on days 1, 29, and 85 Therefore, the highest dose level of 2400 μg T3 was considered the no observed effect level (NOEL). .

[0223] [Example 6] EpiScreen™ Immunogenicity Analysis of Peptides the purpose: Two 16-mer peptides, namely tau 393-408 [pS396 / pS404] ( T3.5 having the amino acid sequence of SEQ ID NO: 2) and Tau393-408 (Tau3.5 having the amino acid sequence of SEQ ID NO: 4) The amino acid sequence of T3.6 was detected in 51 human donors and expressed on CD4 + T cell response The ability to induce .

[0224] design: EpiScreen™ Time-Course T Cell Proliferation Assay and IL-2 ELISPOT Assay PBMCs from 51 human donors were cultured with either peptide at a final concentration of 5 μM. For each donor, a reproducibility control (incubated with 100 μg / mL KLH) was (cells incubated with 0.3 μM humanized A33), and benchmark clinical control ( The proliferation and IL-2 production were measured at various time points. was later evaluated.

[0225] EpiScreen™ Data Analysis For proliferation assays and IL-2 ELISPOT assays, a stimulation index of 2 or greater ( An empirical threshold for the index of indetermination (SI) (SI ≥ 2.00) has been established, which allows Samples inducing a response above this threshold were considered positive, and SI ≥ 1.90 was considered a borderline response sample. I thought of it as a fee.

[0226] Previous EpiScreen™ time-course T cell assays with a range of biologics The percentage of donor T cell responses in the EpiScreen™ assay and clinical Overall, the results show a clear correlation with the level of immunogenicity observed in EpiSc. Protein therapeutics that induce greater than 10% positive responses in the reen™ assay are Associated with a significant risk of immunogenicity in clinical practice.

[0227] result: The overall correlation between the proliferation assay and the IL-2 ELISPOT assay was high (KLH In this case, 98% of the donors were positive for IL-2 EL A donor was defined as an initiating donor in both the ISPOT assay and the proliferation assay. Analysis of the combined data set from these two assays revealed that 1 of 51 (2%) patients had Ner responded to tau 393–408 [pS396 / pS404], 2 of 51 (4%) We found that 10 donors responded to tau 393–408. Considering donors responding to only one of the assays, 4 of 51 donors (8% ) responded to tau 393–408 [pS396 / pS404], and 4 of 51 donors ( 8%) responded to tau 393–408. Thus, the overall frequency and magnitude of responses was For both peptides, responses were low, with a maximum of 8% of donors responding.

[0228] Conclusion: The test peptide (Tau 393-408 [pS 396 / pS404] and tau 393–408) CD4 + Ability to induce T cell responses The study was conducted on a cohort of 51 HLA-typed donors. + The overall relative risk of inducing a T cell response was low for both peptides (2-8% ) was shown.

[0229] In comparison to protein therapeutics tested in the EpiScreen™ assay The data from this study show that both peptides inhibit CD4 + Induction of T cell responses in humans This indicates that the substance may be considered to have a low potential risk of

[0230] [Example 7] Safety and Efficacy of ACI-35 in Humans The safety, tolerability, and immunogenicity of the ACI-35 vaccine (ACI-35) were evaluated in the UK and Phase Ib clinical study in patients with mild to moderate AD in Japan and Finland The study was conducted in patients with mild to moderate Alzheimer's disease (ACI-35-1201). A Phase Ib Multicenter Study of the Safety, Tolerability, and Immunogenicity of ACI-35 in Patients with (Double-blind, randomized, placebo-controlled study).

[0231] Objective: To evaluate the efficacy and safety of 3 doses and 3 dosing regimens in patients with mild to moderate AD. ACI-35 was evaluated for safety and tolerability, as well as anti-phospho-tau (p-tau) immunoglobulins. To preliminary evaluate the induction of serum IgG titers.

[0232] Methods: Each of the five cohorts received various doses (30 0 μg, 900 μg, or 1800 μg of tetrapalmitoylated phosphopeptide (TPAP) peptide T3, SEQ ID NO: 28), and / or a dosing regimen lasting for 6 months (2, 3, or 5 doses administered) and then 12 to 16 months after the final injection (Cohort 1). or a booster injection 6 months later (cohorts 2–5).

[0233] [Table 6]

[0234] Twenty-four patients were randomized and received at least one dose of the study vaccine. The age was 73.6 ± 5.88 years, and the mean MMSE score was 2. The mean mean ± SD was 3.3 ± 2.71. Fifteen women and nine men were randomized. Twenty-two patients were Two patients in cohort 2 (one 900 μg, one placebo) completed the study. A serious medical condition unrelated to ACI-35 that significantly impeded or made impossible the completion of the study. The study was discontinued due to the condition.

[0235] The inclusion criteria were: 1. National Institute of Neurological and Communicative Disorders and Stroke – Alzheimer’s Disease and Related Likely to have Alzheimer's disease (AD) according to disability association criteria 2. Age 60 or older and 85 or younger 3.Mini-Mental State Examination (MMSE) score of 18-28 at screening 4. Patients must be taking a stable dose of an acetylcholinesterase inhibitor at least prior to screening. had to take the test for three months. 5. A trusted spouse or guardian to provide informed consent, assist with clinical evaluation, and report safety issues. are patients who are cared for by other home caregivers 6. In the opinion of the researcher, has understood and signed a written informed consent and is fully Patients who were able to comply with all study procedures (consent must be obtained prior to any study-related procedures). (Note that 7. Women must be at least 1 year postmenopausal and / or surgically Had to be infertile 8. Female partners of male patients who are neither postmenopausal nor surgically infertile should be High-quality contraceptive methods, such as dual barrier or hormonal contraception, must be used. There was none

[0236] Results / Conclusion: Overall, ACI-35 demonstrated efficacy at all doses and treatment regimens tested in human subjects. The study was safe and well tolerated in patients with bronchitis, and injections were administered once in all study cohorts. Immediately after injection into the eye, it caused rapid induction of anti-p-tau antibodies. Injection site erythema, injection site reaction , and fatigue were the most common treatment-emergent adverse events (TEAEs) associated with the study drug. Evidence suggesting the development of CNS inflammation was provided clinically, laboratory, and radiologically. With the exception of injection site reactions, there were no significant differences in the pattern of AEs compared to placebo. No findings suggested a relationship to the study medication. Injection site symptoms were generally mild and self-limited. Reactions were more frequent at higher doses and in cohorts 4 and 5, both of which received active medication. Asymptomatic hypoglycemia was reported by all patients. Given that the phenomenon is known to occur naturally, it is not likely that There was an uncertain association with 5 SAEs reported, all in patients from cohort 2. One patient was treated with placebo and two SAEs (urosepsis) occurred. These 2 SAEs were not related to the study drug. Possibly related SAEs to the study drug (acute pyelonephritis, sinus node dysfunction, and dizziness) ) was reported in two patients treated with ACI-35, and these were the only observations. Adverse reactions were reported in this study. No deaths were reported.

[0237] [Example 7] Safety and Efficacy of ACI-35.030 in Humans Safety, Tolerability, and Immunity of ACI-35.030 Vaccine (ACI-35.030) Epidemiology of early AD (mild cognitive impairment (MCI) due to AD and mild AD) in Europe A Phase Ib / IIa, multicenter, double-blind, randomized, placebo-controlled clinical study in patients with The results will be evaluated in a study (ACI-35-1802).

[0238] Objective: To evaluate patients with early AD (e.g., mild cognitive impairment (MCI) due to AD and mild AD) ACI-35.030 at up to three doses in participants with early AD safety and tolerability in patients with tau-resistant tau, as well as induction of anti-phospho-tau (anti-p-tau) in serum Preliminary study conducted over a 74-week time frame to determine induction of an immune response against abnormal forms of tau protein To evaluate.

[0239] Secondary objectives: Over a 74 week time frame, e.g., IgG titers to tau and p-tau and tau-tau The immunogenicity of the investigational vaccine was further evaluated by evaluating the induction of IgM titers against the vaccine in serum. Further evaluation of the avidity of antibodies induced by immunization will be performed. and.

[0240] Exploratory objective: To identify putative biomarkers of AD progression, all of which are The study included studies on total tau and p-tau protein concentrations in blood and / or CSF. To explore the effect of cloning; to explore the effect of the investigational vaccine on the activation of T cells in the blood inflammatory cytokines in the blood (e.g., IL-1β, IL-2, IL-6, IL- 8, IL-10, IFN-γ, and TNF-α) to explore the effects of the investigational vaccine on To explore the effects of the investigational vaccine on behavioral, cognitive and functional activities.

[0241] Methods: Each of the three cohorts will be followed for 48 weeks (0, 8, 24, and Dosage at week 48), amount of p-tau peptide T3 in the composition (300 μg, 900 μg g, or 1800 μg of the tetrapalmitoylated phosphopeptide pTau peptide T3, Various doses of ACI-35.030 as referred to in column number 28) followed by 24 weeks The study consisted of patients undergoing treatment with a safety follow-up period of 6 months.

[0242] 24 patients were randomized into 3 subcohorts, with 2 patients in each subcohort 10 patients will receive placebo and 6 patients will receive ACI-35.030. The dose will be administered intramuscularly. do.

[0243] Safety assessments were conducted by telephone in all study patients immediately after each dose and 48-72 hours after. In each subcohort, the first dose of the first 4 patients was administered A safety evaluation was conducted over a 48-72 hour period and the study vaccine and This will be carried out after confirming that there are no relevant clinically relevant safety issues.

[0244] All treated patients will have a 24 week (6 month) safety follow-up period after the end of the treatment period. During this period, patients were followed up at the first follow-up visit 19 weeks after the last dose and at the end of the follow-up period. Participants will be asked to attend the final visit (26 weeks after the last dose). Regular review of safety data by the Data Safety Monitoring Board (DSMB) Will be monitored throughout the study.

[0245] Interim analyses will be performed as follows:

[0246] The first interim analysis was conducted after all Cohort 1 patients completed Visit 4 (Week 10). , i.e., 2 to 4 weeks after the second injection. The objective is to assess safety, tolerability, and efficacy up to this point. and immunogenicity data to determine whether to initiate cohort 2. do.

[0247] The second interim analysis was conducted after all Cohort 2 patients completed Visit 4 (Week 10). , i.e., 2 to 4 weeks after the second injection. The objective is to assess safety, tolerability, and efficacy up to this point. and immunogenicity data to determine whether to initiate cohort 3. do.

[0248] The third interim analysis was conducted after all Cohort 3 patients completed Visit 6 (Week 26). The objective is to determine whether cohort 1, 2, or 3 Either of these will be extended to provide the most favorable profile in terms of immunogenicity, safety, and tolerability. A decision will be made to collect additional safety / tolerability data at the proposed doses.

[0249] The fourth interim analysis will be conducted at the end of the treatment period (i.e., 2–4 weeks after the fourth injection). The objective is to review the data to date, including data from patients in subcohort expansions, if applicable. The objective of this study is to consider safety / tolerability and immunogenicity data. Biomarker results are The results can be subsequently taken up on other cohorts. The results obtained were compared across all study cohorts to identify the best strategy for further clinical development. Select from.

[0250] The fifth interim analysis was at the end of the safety follow-up period, i.e., all cohort 1 patients had 11 The study will be conducted after completion of the first visit (week 74). The objectives are the same as the fourth interim analysis: Results are then compared across all cohorts.

[0251] The study population consisted of subjects with a diagnosis of mild AD or MCI due to AD according to the NIA-AA criteria. Patients aged 50-75 years (male and female) with

[0252] The selection criteria were as follows: 1. Male or female, between 50 and 75 years old. 2. Mild cognitive impairment (MCI) due to AD or mild AD according to NIA-AA criteria, and a Clinical Dementia Rating (CDR) total score of 0.5 or 1. Mini-Mental State Examination (MMSE) score of 3.22 or higher. 4. CSF at screening consistent with NIA-AA2018 criteria for AD pathology Amyloid beta 42 (Aβ42) and phosphorylated tau levels. CSF Aβ42 levels In borderline cases, other findings can help determine amyloid positivity, e.g. Aβ42 / Aβ40 ratio and positive amyloid PET scan or positive CSF per case A history of Aβ42 levels may be considered. CSF performed within 3 months prior to screening The results of the sampling were consistent with the presence of amyloid pathology and the corresponding CSF samples is allowed on a case-by-case basis, provided that it can be used in research for testing. 5. Any marketed treatment for AD in at least 3 months prior to baseline or stable doses of acetylcholinesterase inhibitors and / or were patients receiving memantine. 6. Trusted informant to ensure adherence, assist with clinical evaluations, and report safety issues or patients being cared for by a caregiver. 7. Women must be at least 1 year postmenopausal and / or surgically Women who are not of childbearing potential or postmenopausal must be infertile. Negative pregnancy test result at screening and termination of participation from the screening visit Women must be willing to use a highly effective method of contraception until the end of pregnancy. will be performed throughout the treatment period to ensure that subjects are able to continue receiving the study vaccine. Male patients with partners of childbearing potential should take appropriate precautions during the study. They must be willing to use contraception. 8. In the opinion of the researcher, understands and agrees to the written informed consent Patients who can do this. 9. Patients and informants or caregivers must be fluent in one of the study languages ​​and have lumbar Must be able to comply with all study procedures including punctures.

[0253] The exclusion criteria were as follows: 1. Patients are treated with placebo only and the placebo formulation does not induce any specific immune response. Use active immunization unless there is documented evidence that it is not expected to lead to Participation in previous clinical trials for AD and / or neurological disorders. 2. Subjects are treated with placebo only, to prevent the placebo from inducing any specific immune response. Any passive immunization should be avoided unless there is documented evidence that it is not expected to have an adverse effect on the Prior screening for previous clinical trials of AD and / or neurological disorders Participation within the last 12 months. 3.Treatment for AD and / or neurological disorders using any small molecule drug, including BACE-1 inhibitors Participation in a previous clinical trial within the past 3 months prior to screening. 4. Concurrent participation in any other clinical trial using an experimental or approved drug or therapy. . 5. At a dilution of at least 1 / 160 in patients without clinical symptoms of autoimmune disease Presence of positive antinuclear antibody (ANA) titers. 6. Current or past history of autoimmune disease or clinical symptoms consistent with the presence of an autoimmune disease Medical history. 7. Immunosuppressant medications unless temporarily prescribed for >3 months prior to screening or immunosuppression, including but not limited to the use of systemic steroids. 8. Severe allergic reactions, including but not limited to, to previous vaccines and / or medications. History of undetermined severe allergic reactions (e.g. anaphylaxis). 9. Prior history of clinically significant hypoglycemic episodes. 10. Currently met Diagnostic and Statistical Manual of Mental Disorders V (DSM-V) criteria Drug or alcohol abuse or dependence currently occurring or having occurred within the past 5 years. 11. To assess the safety and tolerability of study procedures and / or adherence to all scheduled study visits. Any clinically significant medical condition likely to interfere. 12. Patients receiving any medication likely to affect the immune system and / or investigational vaccines Any clinically significant medical condition that is expected to potentially impair the immunization potential of the any history of acquired or innate immune suppression disorder). 13. Hydralazine, procainamide, quinidine, isoniazid, TNF inhibitors, mino Use of cyclins within the 12 months immediately prior to screening. 14. Use of diltiazem for at least 3 months prior to screening, unless on a stable dose use. 15. Using the Columbia Suicide Severity Scale, subject answers "No" to questions 4 or 5 regarding suicidal ideation. "Yes" to suicidal behavior in the past 12 months Overt suicide risk, defined. 16. Concurrent psychiatric or neurological disorders other than those considered related to AD (e.g. For example, head injury with loss of consciousness, symptomatic stroke, Parkinson's disease, severe carotid artery obstructive disease. disease, TIA). 17. History or presence of uncontrolled epileptic seizures. In case of history of epileptic seizures, the epileptic episode The operation must be well controlled so that no outbreaks were present within 2 years prior to the baseline. Antiepileptic medication use must be on a stable dose for at least 3 months prior to screening. In some cases it is acceptable. 18. History of meningoencephalitis within the past 10 years prior to screening. 19. Patients with a history of hemorrhagic and / or non-hemorrhagic stroke. 20. Presence or history of peripheral neuropathy. 21. History of inflammatory neuropathy with possible CNS involvement. 22. Screening for structural evidence of alternative pathologies inconsistent with AD that may be causing patient symptoms MRI scan. Benign meningiomas less than 1 cm in diameter, 3 or more lacunar infarcts or A single infarct >1 cm or any single area of ​​superficial hemosiderosis other than the cerebrospinal fluid Evidence of space-occupying disease or evidence of prior major hemorrhage ≥10 mm. T2 * In MRI Microbleeds may occur up to 10 times regardless of location. 23. MRI examination was not performed for patients with metal implants and / or severe occlusions that contraindicate MRI studies Unable to do so for any reason, including but not limited to, phobia. 24. Significant hearing or vision impairment or inability to comply with protocol and outcome measures Any other issues judged by the researcher to be relevant in preventing the implementation of the study. 25. Clinically significant infection or major surgery within 3 months prior to screening. Planned surgical procedures anticipated to be performed during participation in the study should be reported to the medical Must be reviewed and approved by the Supervisor. 26. Any medication administered within the past 2 months prior to baseline, including influenza vaccine vaccine. 27. Clinically significant arrhythmias on ECG at screening or other clinically significant An abnormality. 28. Myocardial infarction, unstable angina, or significant coronary artery disease within 1 year prior to baseline. 29. Treated squamous cell carcinoma, basal cell carcinoma, and intraepidermal melanoma or completely removed History of cancer other than noninvasive prostate cancer or noninvasive breast cancer that may have been present and treated Patients who have had this condition within the past 5 years. 30. Hematological parameters, liver function tests, and other biochemical parameters in the opinion of the field investigator. Clinically significant deviation of a biological measurement from normal values ​​that are judged to be clinically significant. 31. Pregnant or pregnant as confirmed by serological testing at screening Intended for women who are planning to undergo chemotherapy or are breastfeeding. 32. Any anticoagulant or antiplatelet drug except aspirin at a dose of less than 100 mg daily Patients undergoing lumbar puncture (risk of bleeding during planned or unplanned lumbar puncture) to avoid). 33. Patients receiving antipsychotics, except for stable low doses for the treatment of insomnia. 34. Have donated blood or blood products or participated in the study within 30 days prior to screening Patients who plan to donate blood while participating. 35. Positive VDRL (Venereal Disease Laboratory) consistent with active syphilis at screening. 36. Patients with a positive HIV test result at screening. 37. Active hepatitis B and / or C as measured by laboratory testing at screening Patients who have: 38. Creatinine >1.5 times the upper limit of normal, abnormal thyroid function test results, or serum Patients with clinically significant decreases in B12 or folate levels (Note: All thyroid replacement medications Oral doses of folate, B12, or B12 must be stable for at least 3 months prior to screening. must be).

[0254] Results / Conclusion: The following primary endpoints will be assessed: Safety and tolerability - adverse events, immediate and delayed reactogenicity (e.g. anaphylaxis, local and systemic reactogenicity, including pain, redness, immune complex disease, swelling, and fever; well-tolerated Global assessment; suicidal ideation (C-SSRS); behavior (NPI); cognition and and functional assessment (RBANS, CDR-SB); vital signs; MRI imaging; electrocardiogram routine hematology and biochemistry evaluation of blood and urine; anti-DNA antibodies in blood Assessment of autoimmune antibodies, including; inflammatory markers in blood and CSF. Immune response - Anti-p-tau IgG titers in serum (geometric mean, change from baseline, responder rate) , peak, and area under the curve).

[0255] The following secondary endpoints will be evaluated: Immune response - serum anti-tau IgG, anti-p-tau and anti-tau IgM titers (geometric mean, basal change from line, responder rate, peak, and area under the curve), and I Determination of gG response profiles.

[0256] The following exploratory endpoints will be assessed: Change from baseline in blood and / or CSF biomarker titers (e.g., total tau and p-tau proteins), change from baseline in blood T cell activation levels, Inflammatory cytokines in the blood (e.g., IL-1B, IL-2, IL-6, IL-8, IL -10, IFN-γ, and TNF-α) titers, and suicidal ideation (C -SSRS), behavioral (NPI), cognitive and functional activities (RBANS, CDR-SB) Change from baseline in score.

[0257] [Example 8] Intramuscular injection induced a greater alloimmune response than subcutaneous injection Groups of rhesus monkeys (n=3 males and 3 females per group) were treated with 1800 μg / dose Vaccination with ACI-35.030 on days 1, 29, 85, and 169 The mice were immunized intramuscularly or subcutaneously by immunization with chlorine.

[0258] Preparation of enriched paired helical fibrils (ePHFs) was performed according to a modified version of Greenberg and Using the method of Avies (1991, Proc Natl Acad Sci USA, 87(15):5827-31), Sarkosyl extraction of soluble tau obtained from postmortem brain tissue of histologically confirmed AD subjects The antibody titers specific for enriched helical fibrils (ePHFs) were analyzed using Mesoscale D The MSD Streptavidin Virus Discovery platform was used to evaluate the The viridin plate was incubated with a biotinylated anti-tau capture antibody (HT7-biotin, ThermoSci After coating with entific, ePHFs isolated from AD patients were used. The ePHF-specific IgG antibody was cross-reacted with the monkey IgG antibody. Further detection was performed using a SulfoTag-labeled anti-human IgG antibody. ePHFs were pre-saturated with 1% BSA and incubated with biotinylated HT-7 (Thermo Sci MSD Gold small spot streptavidin coated with entific After 1 hour of incubation, the plates were The plates were washed with PBST, serial dilutions of serum were added, and incubated for 2 hours. The bound antibodies were then stained using SulfoTag-labeled anti-human IgG antibody followed by 1% PFA. After a fixation step on 0.5% CO, detection was performed by adding read buffer T. The plates were then incubated at 4°C for 1 h at 37°C for 2 h at 37°C. The results were analyzed using a 3D tor Imager (MSD). The data are expressed in arbitrary units per milliliter (AU / mL) along with the geometric mean per unit volume (Figure 1 4) ePHF-specific antibodies on days 50, 106, and 190 after the first immunization. Represents antibody titer.

[0259] FIG. 4 shows that the liposomal vaccine induced high ePHF-specific IgG titers; 4 shows that intramuscular injection induced a greater alloantibody response than subcutaneous injection. [Sequence table]

[0260] [ka]

[0261] [ka]

[0262] [ka]

Claims

1. Anti-phosphorylated tau antibodies can be administered to human subjects in need of them without inducing severe adverse events. The method comprises administering toll-like receptor 4 agonists and SEQ ID NO: 1 to SEQ ID NO: 3 and and a tau phosphoprotein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:5 to SEQ ID NO:

12. and administering to the subject an effective amount of a liposome comprising a tau phospholipase C peptide, The peptide is administered in an amount of about 25 nmoles to about 750 nmoles per dose, The method, wherein a phosphopeptide is displayed on the surface of said liposome.

2. The tau phosphopeptides are SEQ ID NOs:27 to 29 and SEQ ID NOs:31 to 39. 38, and preferably, said tau phosphopeptide The method of claim 1, wherein the peptide consists of the amino acid sequence of SEQ ID NO:

28.

3. The effective amount of liposomes is from 100 μg to 2500 μg of the tau phospholipase per dose. Peptide, preferably 300 μg to 2400 μg per dose, 300 μg per dose to 1800 μg, or 300 μg to 900 μg of said tau phosphopeptide per dose The method of claim 1 or 2, comprising:

4. The effective amount of liposomes is 300 μg per dose, 900 μg per dose, 1800 μg per dose, or 2400 μg of said tau phosphopeptide per dose. The method of claim 3 .

5. The method of claim 1 , wherein the liposomes are administered subcutaneously.

6. 5. The method of claim 1, wherein the liposomes are administered intramuscularly.

7. administering a second dose of said effective amount of liposomes to said subject 1 to 24 weeks after the first dose. The method of claim 1 , further comprising:

8. The liposome comprises a helper T cell epitope and a lipidated CpG oligonucleotide. The method of claim 1 , further comprising:

9. The lipidated CpG oligonucleotide is selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO:

22. and the CpG oligonucleotide has a nucleotide sequence selected from one or more nucleotides. having phosphorothioate internucleotide linkages, said CpG oligonucleotide comprising at least covalently linked to another lipophilic group, optionally via a PEG linker; The method according to claim 8.

10. The CpG oligonucleotide is linked to at least one lipophilic group via a PEG linker. The method of claim 9 , wherein the linker is a bond.

11. The liposome is a 1,2-dimyristoyl-sn-glycero-3-phosphocholine (D MPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac- glycerol (DMPG), and cholesterol; or 11. The method of claim 1, further comprising a plurality of lipids.

12. The helper T cell epitopes are selected from the group consisting of SEQ ID NOs: 13 to 17, SEQ ID NOs: 23 to 29, No. 26, and at least one selected from the group consisting of SEQ ID NO: 39 to SEQ ID NO: 44 The method according to any one of claims 8 to 11, comprising the amino acid sequence of:

13. The effective amount of liposomes is in an amount of about 2 nmole to about 110 nmole per dose. The method according to any one of claims 8 to 12, comprising said helper T cell epitope.

14. The effective amount of liposomes is between 25 μg and 620 μg per dose, preferably SEQ ID NO:13 to SEQ ID NO:17, SEQ ID NO:23 to SEQ ID NO:26, No. 26, and an amino acid sequence selected from the group consisting of SEQ ID NO: 39 to SEQ ID NO:

44. and preferably said helper T cell epitope 9. A T50 helper T cell epitope consisting of the amino acid sequence of SEQ ID NO:

13.

14. The method according to any one of claims 1 to 13.

15. The effective amount of liposomes is between 30 μg and 900 μg per dose, preferably and preferably contains the toll-like receptor 4 agonist in an amount of 100 μg to 585 μg per The toll-like receptor 4 agonist is monophosphoryl hexaacyl lipid A, 3-deoxyribonucleic acid.

15. The method of claim 1, wherein the compound is a sil.

16. The effective amount of liposomes is between 50 μg and 1250 μg per dose, preferably Selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22 in an amount of 150 μg to 800 μg per and preferably said lipidated CpG oligonucleotide having a nucleotide sequence The lipidated CpG oligonucleotide comprises the nucleotide sequence of SEQ ID NO:

18.

16. The method according to any one of claims 8 to 15.

17. The liposome is (1) The tau phosphopeptide having the amino acid sequence of SEQ ID NO:

28. (2) The toll-like receptor containing monophosphoryl hexaacyl lipid A, 3-deacylated 4 agonists, (3) The helper T cell epitope comprising the amino acid sequence of SEQ ID NO:

39. (4) The lipidated CpG oligonucleotide comprising the nucleotide sequence of SEQ ID NO:

18. Rabi (5) 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1, 2-Dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol ( At least one lipid selected from the group consisting of:

17. The method of any one of claims 1 to 16, comprising:

18. The subject is diagnosed with Alzheimer's disease, preferably early Alzheimer's disease, Treatment of mild cognitive impairment (MCI) due to 18. The method of any one of claims 1 to 17, comprising:

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