Liposomes and Pharmaceutical Compositions

Abstract

The present invention relates to liposomes containing tau peptides, preferably phosphorylated tau peptides, and conjugates containing tau peptides, preferably phosphorylated tau peptides, conjugated to an immunogenic carrier. Pharmaceutical compositions and uses of the liposomes and / or conjugates for the treatment or prevention of a neurodegenerative disease or disorder, such as Alzheimer's disease, are also described.

Description

1 / 96 Liposomes and Pharmaceutical Compositions - Field of the Invention

[001] The present invention relates to the field of medicine. The invention, in particular, relates to liposomes or conjugates of tau peptides and their use to prevent or treat tauopathy, such as Alzheimer's disease. BACKGROUND

[002] Alzheimer's disease (AD) is a progressive, debilitating neurodegenerative disease affecting approximately 44 million people worldwide (Alzheimers.net). Currently available AD therapies in the clinic aim to slow the progression of clinical symptoms, but do not target the underlying pathogenic processes of the disease. Unfortunately, these therapies are minimally effective, and therefore there is an urgent need to develop and test additional preventive and therapeutic measures.

[003] The hallmark pathologies of Alzheimer's disease are an accumulation of extracellular plaques comprising aggregated beta-amyloid protein and intracellular tangles or aggregations of hyperphosphorylated tau protein. The molecular events leading to the accumulation of these proteins are poorly characterized. For amyloid, it is assumed that aberrant cleavage of the amyloid precursor protein leads to an accumulation of the aggregation-prone fragment comprising amino acids 1-42. For tau, it is assumed that dysregulation of kinases, phosphatases, or both leads to aberrant phosphorylation of tau. Once tau becomes hyperphosphorylated, it loses the ability to effectively bind and stabilize microtubules, thus accumulating in the cytoplasm of the affected neuron. Unbound and hyperphosphorylated tau appears to form the first oligomers and then higher-order aggregates, whose presence presumably negatively affects the function of the neuron in which they form, perhaps Petition 870260044022, dated 11 / 05 / 2026, page 9 / 222 2 / 96 through the interruption of normal axonal transport.

[004] In developed countries, individuals diagnosed with Alzheimer's disease or other dementia tauopathies are commonly treated with cholinesterase inhibitors (e.g., Aricept®) or memantines (e.g., Namenda™). These drugs, while reasonably well tolerated, have very modest efficacy. For example, Aricept® delays symptom worsening by 6 to 12 months in approximately 50% of treated individuals. The remainder of the treatment is non-pharmacological and focuses on making patients more able to manage daily tasks as their cognitive abilities decline.

[005] Several published studies (Asuni AA et al., J. Neurosci. August 2007; 27 (34): 9115-29., Theunis C et al., PLoS One. 2013; 8 (8): e72301., Kontsekova E et al., Alzheimers Res Ther. August 1, 2014; 6 (4): 44) demonstrate that active vaccines containing tau peptides can induce anti-tau immune responses in mice or rats; and can reduce the accumulation of pathological tau aggregates in the rodent brain; and can reduce the rate of progression of cognitive decline in animal models of Alzheimer's disease. An active vaccine against pathological tau proteins has been shown to be immunogenic in human patients with Alzheimer's disease (Novak P et al., Lancet Neurology 2017, 16: 123-134). Document WO2010 / 115843 describes an antigenic phosphopeptide mimicking an important pathological phosphoepitope of the tau protein and related compositions for therapeutic and diagnostic use in the treatment of tauopathies, including Alzheimer's disease.However, there are currently no effective vaccines approved on the market to prevent the onset of tau-mediated diseases. Nor are there any effective drugs on the market to intercept or slow the course of the disease once it has begun. Therefore, there is a need. Petition 870260044022, dated 11 / 05 / 2026, page 10 / 222 3 / 96 urgent need to identify new preventive measures (e.g., vaccines) that can prevent these diseases. SUMMARY OF THE INVENTION

[006] In a general aspect, the invention relates to a liposome, comprising:

[007] a. a tau peptide, preferably the tau peptide is a tau phosphopeptide; and

[008] b. a helper T cell epitope,

[009] in which the tau peptide is presented on the surface of the liposome.

[0010] In one embodiment, the liposome further comprises at least one adjuvant comprising a toll-type receptor ligand. Preferably, the liposome further comprises at least one of a toll-type receptor ligand 4 and a toll-type receptor ligand 9.

[0011] In a preferred embodiment, the invention relates to a liposome, comprising:

[0012] a. a tau peptide, preferably the tau peptide is a tau phosphopeptide;

[0013] b. a helper T cell epitope; and

[0014] c. at least one of

[0015] i. a toll-like receptor ligand 9, preferably a lipid-coated CpG oligonucleotide; and

[0016] ii. a toll-like receptor 4 ligand, preferably a toll-like receptor 4 agonist,

[0017] in which the tau peptide is presented on the surface of the liposome.

[0018] In another preferred embodiment, the invention relates to a liposome, comprising:

[0019] a. a tau phosphopeptide; Petition 870260044022, dated 11 / 05 / 2026, page 11 / 222 4 / 96

[0020] b. a helper T cell epitope;

[0021] c. a lipid-encapsulated CpG oligonucleotide; and

[0022] d. an adjuvant containing a toll-type receptor ligand 4;

[0023] in which the phosphopeptide tau is presented on the surface of the liposome.

[0024] In another general aspect, the invention relates to a conjugate comprising a tau peptide, preferably a tau phosphopeptide, and an immunogenic carrier conjugated thereto, wherein the tau peptide is conjugated to the carrier via a linker. The linker may comprise one or more of polyethylene glycol (PEG), succinimidyl 3(bromoacetamide) propionate (SBAP), and m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS). Examples of immunogenic carriers useful for the invention include, among others, California limpet hemocyanin (KLH), tetanus toxoid (TT), CRM197, and a mixture of N. meningitidis outer membrane proteins (OMP), or a derivative thereof.

[0025] In a preferred embodiment, the invention relates to a conjugate having the structure of formula (I): The H2N. JL J 1 H ^-NH Carrier .\ςNH or the structure of formula (II): H / x^ Y Peptide > II tau n OJ Petition 870260044022, dated 11 / 05 / 2026, p. 12 / 222 5 / 96 Carrier H2N s H N. The peptide tau, where x is an integer from 0 to 10, preferably 2 to 6, more preferably 3; en is an integer from 2 to 11, preferably 3 to 11.

[0026] Other aspects of the invention relate to a pharmaceutical composition comprising a liposome or a conjugate of the invention and a pharmaceutically acceptable carrier, methods of preparing the pharmaceutical composition, and the use of the pharmaceutical composition in inducing an immune response against tau, or treatment or prevention of a neurodegenerative disease or disorder in an individual in need of treatment.

[0027] In one embodiment, the invention relates to a method for inducing an immune response in an individual suffering from a neurodegenerative disorder, or for treating or preventing a neurodegenerative disease or disorder in an individual in need of treatment. The method comprises administering to the individual a pharmaceutical composition comprising a liposome of the invention and a pharmaceutically acceptable carrier, or a pharmaceutical composition comprising a conjugate of the invention and a pharmaceutically acceptable carrier. Preferably, the method comprises administering to the individual a pharmaceutical composition of the invention to initiate immunization, and a pharmaceutical composition of the invention to enhance immunization.

[0028] Other aspects, features and advantages of the present invention will be better appreciated by reading the following description. Petition 870260044022, dated 11 / 05 / 2026, page 13 / 222 6 / 96 detailed description of the invention and claims. BRIEF DESCRIPTION OF THE FIGURES

[0029] The preceding summary, as well as the following detailed description of the preferred embodiments of this application, will be better understood when read in conjunction with the attached drawings. It should be understood, however, that the application is not limited to the precise embodiments shown in the drawings.

[0030] Figure 1 illustrates novel vaccines according to embodiments of the invention: a tau liposome according to an embodiment of the invention (above) and a tau conjugate according to an embodiment of the invention (below).

[0031] Figure 2 illustrates that a vaccine comprising a liposome according to an embodiment of the invention (second-generation liposome) containing an encapsulated helper T cell epitope (e.g., tetanus polypeptide (tet)) activates helper T cells.

[0032] Figure 3 illustrates that a vaccine comprising a conjugate according to an embodiment of the invention containing a non-auto or immunogenic carrier protein activates helper T cells.

[0033] Figure 4 shows that tau vaccines according to embodiments of the invention induce sustained high-titer antiphosphorylated tau antibodies in Rhesus monkeys: the geometric mean of the outcome titers per group, as measured by enzyme-linked immunosorbent assay (ELISA), over time, is higher for a vaccine comprising a liposome (Liposome Z) according to an embodiment of the invention or a vaccine comprising a conjugate (Conjugate A) according to an embodiment of the invention, compared with a control liposomal vaccine without the helper T cell epitope. Petition 870260044022, dated 11 / 05 / 2026, page 14 / 222 7 / 96

[0034] Figure 5 shows that serum from Rhesus monkeys immunized with a liposome (liposome Z) according to an embodiment of the invention binds to pathological tau structures in human AD brain sections (left panel) compared with healthy human brain sections (right panel).

[0035] Figure 6 shows that serum from Rhesus monkeys immunized with a conjugate (Conjugate A) according to an embodiment of the invention formulated in a composition containing soluble CpG and alum hydroxide binds to pathological tau structures in human AD brain sections (top row), compared with healthy human brain sections (bottom row).

[0036] Figure 7 shows the antiphosphorylated tau antibody titers in Rhesus monkeys induced by liposomal vaccines according to embodiments of the invention, Liposomes X, Y and Z, each of which contains the encapsulated T-cell epitope T50 and one or more adjuvants; titers were measured by ELISA and presented as outcome titers over time in individual monkeys. In particular:

[0037] Figure 7A shows liposome X-induced antiphosphorylated tau antibody titers with a TLR4 ligand, MPLA (3D (6-acyl) PHAD®) alone as the adjuvant.

[0038] Figure 7B shows titers of antiphosphorylated tau antibodies induced by Y liposome with a TLR9 ligand (lipid-coated CpG oligonucleotide) alone as the adjuvant.

[0039] Figure 7C shows the antiphosphorylated tau antibody titers induced by liposome Z with a combination of a TLR4 ligand, MPLA (3D-(6-acyl)PHAD®) and a TLR9 ligand (lipid-coated CpG oligonucleotide) as adjuvants, which also shows that the combination of two adjuvants induces less variability in antibody titers between individual monkeys. Petition 870260044022, dated 11 / 05 / 2026, p. 15 / 222 8 / 96

[0040] Figure 7D presents the geometric mean of the antibody titers of the immunization groups mentioned above and a control liposomal vaccine with a TLR4 ligand, MPLA, but without a T-cell epitope, and shows that the vaccines according to embodiments of the invention result in higher antibody titers of antiphosphorylated tau antibodies than in a control liposomal vaccine: the titers were measured by ELISA and are presented as geometric mean + / - 95% confidence interval of the outcome titers by group over time.

[0041] Figure 8 shows that immunization using a liposomal vaccine (e.g., liposome X, Y, or Z) or a conjugate vaccine (Conjugate A) according to an embodiment of the invention induces specific IgG antibody titers to enriched paired helical filaments (ePHF) isolated from the brain after death of patients with Alzheimer's disease: antibody titers were measured by Meso Scale Discovery (MSD) technology and are presented as values ​​for individual monkeys on day 50 and the geometric mean + / - 95% CI after the first immunization.

[0042] Figure 9 shows that immunization with a liposomal vaccine according to an embodiment of the invention (Liposome Z) containing encapsulated T50 and a combination of a TLR4 ligand (3D-(6-acyl)PHAD®) and a TLR9 ligand (lipid-coated CpG oligonucleotide) as adjuvants induces antibodies that bind primarily to the N-terminal of the phosphorylated tau peptide of SEQ ID NO: 2 (Figure 9A), while monkeys immunized with a conjugate vaccine according to an embodiment of the invention (Conjugate A) generate IgG antibodies that bind primarily to the C-terminal part of the peptide, both for phosphorylated peptides (left) and for non-phosphorylated peptides (right) (Figure 9B).

[0043] Figures 10 A and B show that vaccination with a Petition 870260044022, dated 11 / 05 / 2026, p. 16 / 222 9 / 96 A liposomal vaccine according to an embodiment of the invention containing encapsulated T50 T-cell epitope and TLR4 ligand (3D-(6acyl)PHAD®) as an adjuvant (S liposome) induces significantly higher antibody titers than the control liposomal vaccine (with a TLR4 ligand, 3D-(6acyl)PHAD®, but without the T50 T-cell epitope, R liposome) and also the liposomal vaccine according to an embodiment of the invention containing T57 surface T-cell epitope (dipalmitoylated T50) and TLR4 ligand (3D-(6acyl)PHAD®, T liposome) in mice: antibody titers on day 21 (Figure 10A) and 35 (Figure 10B) after the first immunization were measured by ELISA and presented as individual values ​​and the geometric mean. by group ± 95% CI; (**: p < 0.01, ***: p < 0.001).

[0044] Figure 11 shows that encapsulation of T48 or T52 T cell peptides in the liposome (M or N liposome, respectively) induces a T cell response specific to the encapsulated peptide in mice: the T cell response was assessed by IFN-γ (Figure 11A) and IL-4 (Figure 11B) ELISPOT.

[0045] Figure 12 shows that the liposomal vaccine containing an encapsulated T-cell epitope (liposome L) and the liposomal vaccine containing an anchored T-cell epitope (liposome O) each induced higher titers of specific antibodies to tau phosphopeptides than the control liposomal vaccine without a T-cell epitope; each of the L liposome, liposome O, and control liposome also contains MPLA as an adjuvant.

[0046] Figure 13 shows that the Tau conjugate (KLH-TAUVACp7.1 or KLH-TAUVAC-p22.1) induces robust TfH cells and Ab titers against the tau peptide in naturally occurring mice, in particular:

[0047] Figure 13A illustrates that the Balb / mouse groups Petition 870260044022, dated 11 / 05 / 2026, p. 17 / 222 10 / 96 Adult females (n = 14 per group) were immunized a total of four times with 100 µg of adjuvanted KLH-tau conjugate vaccine (KLH-TAUVAC-p7.1 or KLH-TAUVAC-p22.1), an active placebo vaccine (KLH plus alum or Ribi), or an inactivated placebo (PBS) according to the scheme shown: four animals from each immunization group were sacrificed seven days after primary immunization, and lymph nodes draining the injection site were collected.

[0048] Figure 13B shows the geometric mean percentage of TfHs by immunization group (n = 4 mice per group analyzed individually) in drainage nodes: all groups that received active vaccines or placebos had measurable TfHs; furthermore, animals that received the KLH-TAUVAC-p7.1, KLH-TAUVACp22.1 vaccine or active KLH placebo plus alum had significantly more TfHs than animals that received inactive placebo (p = 0.0044 for KLH-TAUVAC-p7.1; p = 0.0482 for KLH-TAUVAC-p22.1; p = 0.0063 for KLH, using an ANOVA test followed by Dunnett's adjustment for multiple comparisons).

[0049] Figures 13C-H show changes in serum titers from baseline (day 0) at four time points after immunization (days 14, 28, 56, and 84) to group means (n ​​= 5-10) with a 95% confidence interval: the asterisk indicates the time points at which the antibody response induced by KLH-TAUVAC is significantly greater than that induced by active placebos (p < 0.05, measured using ANOVA followed by Tukey's adjustment for multiple comparisons), more specifically:

[0050] Figure 13C shows binding titrations to the phosphorylated tau peptide p7.1.

[0051] Figure 13D shows binding titrations to the phosphorylated tau peptide p22.1.

[0052] Figure 13E shows tau peptide binding titers Petition 870260044022, dated 11 / 05 / 2026, page 18 / 222 11 / 96 non-phosphorylated 7.1.

[0053] Figure 13F shows binding titrations to the non-phosphorylated tau peptide 22.1.

[0054] Figures 13G and H show binding titers to the carrier protein KLH.

[0055] Figure 14 shows that sera from mice immunized with Tau conjugate also linked pathological tau structures from other tauopathies: combined sera (n = 6) from each vaccination group 84 days after primary immunization were used to stain brain tissue from a case of frontal temporal dementia with MAPT mutation (MAPT P301S, frontal cortex), a case with Pick's disease (frontal cortex), progressive supranuclear palsy (PSP, caudate nucleus) and primary age-related tauopathy (PART, hippocampus); sera from animals that received the active vaccines highlighted the tau-related structures typical of each tauopathy, while sera from animals immunized with an active placebo (KLH-alum or KLH-Ribi) or the inactive one (PBS) did not stain any of them; for reference, an AT8 immunostaining of the corresponding area is shown; scale bar = 50 µm.

[0056] Figure 15 shows that vaccine-induced antibodies reduce aggregated tau in an accelerated tauopathy model, in particular:

[0057] Figure 15A shows that three-month-old P301L transgenic mice (n = 15 per group) received a stereotaxic injection of human ePHF pre-incubated with purified IgG from mice immunized with KLH-TAUVAC-p7.1 plus RIBI or with the active placebo KLH plus RIBI; two months after injection, all mice were sacrificed and the amount of tau aggregated in the mice was determined in total and sarkosil-insoluble fractions. Petition 870260044022, dated 11 / 05 / 2026, page 19 / 222 12 / 96

[0058] Figures 15B and 15C show the total (B) and sarkosil-insoluble (C) fractions collected from the injected hemisphere of each animal: the graphs show the amount of tau measured by MSD; in both the total and insoluble fractions, the brains of mice that received ePHF pre-incubated with IgG from mice immunized with KLH-TAUVAC-p7.1 showed significantly less aggregated tau than mice that received ePHF pre-incubated with control antibodies. (p < 0.0001, using an ANOVA test followed by Holm-Bonferroni adjustment for multiple comparisons.

[0059] Figure 16 shows that the Tau conjugate (Conjugate B) according to an embodiment of the invention induces high antibody titers against phosphorylated Tau and ePHF in non-human primates: Rhesus monkeys were immunized with alum and KLH-TAUVAC-p7 with CpG.1 adjuvant (n = 6) or with KLH (n = 2) on days 1, 29, 85 and 169; blood was collected every 14 days, in particular:

[0060] Figure 16A shows that sera from animals immunized with KLH-TAUVAC-p7.1 were tested for reactivity to the immunizing peptide p7.1 using ELISA.

[0061] Figure 16B shows that sera collected from all animals 50 days after primary immunization showed measurable levels of antibodies against human ePHF using MSD, with 3 of 6 animals showing high reactivity to this antigen.

[0062] Figure 16C shows that sera collected from animals 50 days after primary immunization were applied to human brain sections from healthy individuals or patients with AD, post-immunization sera from pathological tau structures stained by the KLHTAUVAC-p7.1 group, i.e., neurofibrillary tangles, neuropil strands and neuritic plaques in AD brain tissue, while sera from mice immunized with KLH did not show reactivity, and Petition 870260044022, dated 11 / 05 / 2026, page 20 / 222 13 / 96 no coloration was observed in the control tissue.

[0063] Figure 16D shows that, when tested in the tau immunodepletion assay, animals that received KLH-TAUVAC-p7.1 had antibodies capable of binding to and depleting tau seed (p = 0.03 on day 50 using an ANOVA test followed by Dunnett's adjustment for multiple comparisons), while immunization with KLH did not trigger such antibodies.

[0064] Figure 16E shows that pre- and post-immunization sera were also tested in the neutralization assay as serially diluted individual samples; changes from baseline (CFB) were calculated as the difference between FRET counts for readings on day -14 before vaccination (baseline) and after vaccination on days 50, 106, and 190, respectively. The response on a specific post-vaccination day (diai) was calculated as follows: Response = % FRET_diai - % FRET_baseline; a general linear mixed model on the aforementioned responses, with animal random effect, was applied with vaccine groups, variable serum levels treated as categorical variables and all their interactions.

[0065] Figure 17 shows that mice immunized with a conjugate vaccine (Conjugate A) according to an embodiment of the invention and a combination with alum hydroxide (alum) and CpG oligo adjuvant (CpG) result in higher titer antibody responses to the vaccine peptide: adult female C57BL / 6 mice (n = 5-6 per group) were immunized intramuscularly with 2 or 0.2 pg of Conjugate A vaccine, and the conjugate vaccine was administered alone, with alum, with CpG, or with alum and CpG combined; all mice received a primary immunization on day 0 of the study, followed by a single booster immunization on day 28; doses for the alum adjuvant were 500 µg per mouse per injection, and doses for the CpG adjuvant were Petition 870260044022, dated 11 / 05 / 2026, p. 21 / 222 14 / 96 of 20 µg / mouse per injection; the graphs show the results of ELISA binding using serum collected from mice immunized with the T3.5 vaccine peptide as a coating antigen, with mean outcome-specific T3.5 titers per group plotted, before immunization (day 0) and at two time points after immunization (days 28 and 42), and with error bars representing the standard error; the tables show the statistical analysis of the results, where antibody titers were compared by the non-parametric Kruskal-Wallis test, and pairwise group comparisons were assessed by the Wilcoxon Signed Rank test as post-hoc to the Kruskal-Wallis test; in particular:

[0066] Figure 17A shows that the mice were immunized with 2 ug of the Conjugate A vaccine.

[0067] Figure 17B shows that the mice were immunized with 0.2 ug of the Conjugate A vaccine.

[0068] Figure 18 shows that tau vaccines according to embodiments of the invention with different ratios of tau peptide to T cell epitope induce sustained high titer antiphosphorylated tau antibodies in Rhesus monkeys. DETAILED DESCRIPTION OF THE INVENTION

[0069] Various publications, articles, and patents are cited or described in the background and throughout the specification; each of these references is incorporated herein by reference in its entirety. The discussion of documents, acts, materials, devices, articles, or the like that have been included in this specification is for the purpose of providing context for the invention. Such discussion is not an admission that any or all of these matters are part of the prior art with respect to any inventions described or claimed.

[0070] Unless otherwise defined, all terms Petition 870260044022, dated 11 / 05 / 2026, page 22 / 222 15 / 96 technical and scientific terms used herein have the same meaning as is commonly understood by one skilled in the art to which this invention relates. Otherwise, certain terms used in this document have the meanings set forth in the specification.

[0071] It should be noted that, as used here and in the attached claims, the singular forms a, an and the, a include plural reference, unless the context clearly indicates otherwise.

[0072] Unless otherwise indicated, any numerical values, such as a concentration or a range of concentrations described herein, should be understood as modified in all occurrences by the term approximately. Thus, a numerical value normally includes ± 10% of the quoted value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (weight / volume) includes 0.9% (weight / volume) to 11% (weight / volume). As used herein, the use of a numerical range expressly includes all possible subranges, all individual numerical values ​​within that range, including whole numbers within those ranges and fractions of the values, unless the context clearly indicates otherwise.

[0073] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood as referring to each element in the series. Those skilled in the art will recognize, or will be able to verify, using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be covered by the invention.

[0074] As used herein, the terms comprise, including, includes, has, containing, contains or including, or any other variation thereof, shall be understood Petition 870260044022, dated 11 / 05 / 2026, p. 23 / 222 16 / 96 as implying the inclusion of a stated integer or group of integers, but not the exclusion of any other integer or group of integers, and is intended to be non-exclusive or open. For example, a composition, a mixture, a process, a method, an article, or an apparatus comprising a list of elements is not necessarily limited to those elements alone, but may include other elements not expressly listed or inherent in such composition, mixture, process, method, article, or apparatus. Furthermore, unless expressly stated otherwise, "or" refers to an inclusive "or" and not an exclusive "or". For example, a condition A or B is satisfied by any of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).

[0075] It should also be understood that the terms about, approximately, generally, substantially, and similar terms, used herein when referring to a dimension or characteristic of a component of the preferred invention, indicate that the described dimension / characteristic is not a strict limit or parameter and does not exclude small variations that are functionally equal or similar, as would be understood by one skilled in the art. At a minimum, such references that include a numerical parameter would include variations that, using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.), would not vary the least significant digit.

[0076] As used in this document, the term tau or tau protein, also known as microtubule-associated tau protein, MAPT, neurofibrillary tangle protein, paired helical filament-tau, PHF-tau, MAPTL, MTBT1, refers to a Petition 870260044022, dated 11 / 05 / 2026, page 24 / 222 17 / 96 Abundant protein of the central and peripheral nervous system having multiple isoforms. In the human central nervous system (CNS), there are six major tau isoforms with sizes ranging from 352 to 441 amino acids in length due to alternative splicing (Hanger et al., Trends Mol Med. 15: 112-9, 2009). Examples of tau include, but are not limited to, tau isoforms in the CNS, such as the longer 441-amino acid tau isoform (4R2N) which has four repeats and two insertions, and the shorter (fetal) 352-amino acid isoform (3R0N) which has three repeats and no insertions. Examples of tau also include the big tau isoform expressed in peripheral nerves which contains 300 additional residues (exon 4a). Friedhoff et al., Biochimica et Biophysica Acta 1502 (2000) 122 to 132. Examples of tau include a human big tau which is a 758 amino acid protein encoded by a 6762 nucleotide-long mRNA transcript (NM_016835).4), or isoforms thereof. The amino acid sequence of the human big tau exemplified is represented in GenBank accession number NP_058519.3. As used herein, the term tau includes tau homologs from other non-human species, such as Macaca Fascicularis (cynomolgus apes) or Pan troglodyta (chimpanzee). As used herein, the term tau includes proteins comprising mutations, for example, point mutations, fragments, insertions, deletions, and splice variants of naturally occurring full-length tau. The term tau also encompasses post-translational modifications of the tau amino acid sequence. Post-translational modifications include, but are not limited to, phosphorylation.

[0077] As used herein, the term peptide or polypeptide refers to a polymer composed of amino acid residues, related naturally occurring structural variants and their non-naturally occurring synthetic analogues, linked through Petition 870260044022, dated 11 / 05 / 2026, p. 25 / 222 18 / 96 peptide bonds. The term refers to a peptide of any size, structure, or function. Typically, a peptide has at least three amino acids. A peptide may be naturally occurring, recombinant, or synthetic, or any combination thereof. Synthetic peptides may be synthesized, for example, using an automated polypeptide synthesizer. Examples of tau peptides include any tau protein peptide of about 5 to about 30 amino acids in length, preferably about 10 to about 25 amino acids in length, more preferably about 16 to about 21 amino acids in length. In the present description, peptides are listed from the N-terminal to the C-terminal using the standard three-letter or one-letter amino acid abbreviation, wherein phosphoresidues are indicated with p.Examples of tau peptides useful in the invention include, but are not limited to, tau peptides comprising the amino acid sequence of any of the SEQ ID NOs: 1-12, or tau peptides with an amino acid sequence that is at least 75%, 80%, 85%, 90%, or 95% identical to the amino acid sequence of any of the SEQ ID NOs: 1-12.

[0078] As used herein, the term phosphopeptide or phospho-epitope refers to a peptide that is phosphorylated at one or more amino acid residues. Examples of tau phosphopeptides include any tau peptide comprising one or more phosphorylated amino acid residues. Examples of tau phosphopeptides useful in the invention include, but are not limited to, tau phosphopeptides comprising the amino acid sequence of any of the SEQ ID NOs: 1-3 or 5-12, or tau phosphopeptides with an amino acid sequence that is at least 75%, 80%, 85%, 90%, or 95% identical to the amino acid sequence of any of the SEQ ID NOs: 1-3 or 5-12.

[0079] The tau peptides of the present invention can be synthesized by solid-phase peptide synthesis or by systems of Petition 870260044022, dated 11 / 05 / 2026, page 26 / 222 19 / 96 Recombinant expression. Automated peptide synthesizers are commercially available from various suppliers, such as Applied Biosystems (Foster City, California). Recombinant expression systems can include bacteria, such as E. coli, yeast, insect cells, or mammalian cells. Procedures for recombinant expression are described by Sambrook et al., Molecular Cloning: A Laboratory Manual (CSHP Press, NY 2nd ed., 1989).

[0080] Tau is a human autoprotein. This means that, in principle, all lymphocytes carrying a specific receptor for tau should have been deleted during development (central tolerance) or rendered unresponsive by a peripheral tolerance mechanism. This problem has proven to be a significant obstacle to the development of vaccines against autoproteins or altered autoproteins (e.g., tumor antigens).

[0081] The generation of high-quality antibodies against an antigen (auto or infectious) requires the action not only of B lymphocytes, which produce the antibody, but also of CD4+ helper T lymphocytes. CD4+ T cells provide critical survival and maturation signals for B lymphocytes, and animals deficient in CD4+ T cells are profoundly immunosuppressed. CD4+ T cells are also subject to tolerance mechanisms, and an additional obstacle to generating strong anti-autoantibody responses (e.g., anti-tau) is that tau-reactive CD4+ T cells are also likely rare or nonexistent in the human / animal repertoire.

[0082] Although not wishing to be bound by the theory, it is believed, but in no way limiting the scope of the present invention, that this problem is circumvented by the vaccine compositions of the present invention.

[0083] In one embodiment, a liposome comprising a Petition 870260044022, dated 11 / 05 / 2026, p. 27 / 222 20 / 96 tau peptide (an example is shown in Figure 1; top) is produced which also comprises a T cell epitope capable of binding most or all HLA DR (human leukocyte antigen - D-related antigen) molecules. The T cell epitope is then able to activate CD4+ T cells and provides essential maturation and survival signals to tau-specific B cells (Figure 2). In another embodiment, a conjugate of a tau peptide with a carrier protein is produced (an example is shown in Figure 1; bottom), which generates a strong helper T cell response (Figure 3). In this embodiment, unbound recognition is used, in which vehicle-specific T cells provide survival and maturation signals to autoreactive B cells.Therefore, tau-specific B cells receive crucial signals to trigger affinity maturation, immunoglobulin class switching, and to establish a long-term memory cluster. Tau liposomes and tau conjugates can be used to generate high-quality antibodies against tau antigen in homologous or heterologous immunization schedules, with liposome or conjugate used at the beginning and / or booster. Liposomes

[0084] In a general aspect, the invention relates to a liposome, comprising:

[0085] a. a tau peptide, preferably the tau peptide is a tau phosphopeptide; and

[0086] b. a helper T cell epitope,

[0087] in which the tau peptide is presented on the surface of the liposome.

[0088] Liposomes according to embodiments of the invention are also referred to herein as improved liposomes, improved liposomal vaccines, or liposomal vaccines according to Petition 870260044022, dated 11 / 05 / 2026, page 28 / 222 21 / 96 embodiments of the invention or Tau liposomes or optimized liposomal vaccines of 2nd generation liposomes.

[0089] As used herein, the term liposome generally refers to a lipid vesicle that is made of materials with a high lipid content, for example, phospholipids, cholesterol. The lipids in these vesicles are usually arranged in the form of lipid bilayers. Lipid bilayers generally encapsulate a volume that is sandwiched between several onion-like shells of lipid bilayers, forming multilamellar lipid vesicles (MLVs) or contained within an amorphous central cavity. Lipid vesicles with an amorphous central cavity are unilamellar lipid vesicles, that is, those with a single peripheral bilayer surrounding the cavity. Large unilamellar vesicles (LUVs) typically have a diameter of 100 nm to a few micrometers, such as 100 to 200 nm or more, while small unilamellar lipid vesicles (SUVs) typically have a diameter of less than 100 nm, such as 20 to 100 nm, typically 15 to 30 mm.

[0090] According to particular embodiments, the liposome comprises one or more tau peptides. According to particular embodiments, the tau peptides in the liposome may be the same or different.

[0091] Any suitable tau peptide known to those skilled in the art may be used in the invention in view of the present description. According to particular embodiments, one or more of the tau peptides comprise the amino acid sequence of one of the SEQ ID NOs: 1-12. In other embodiments, one or more of the tau peptides comprise an amino acid sequence that is at least 75%, 80%, 85%, 90% or 95% identical to the amino acid sequence of one of the SEQ ID NOs: 1-12, wherein none of the amino acid residues are phosphorylated or one or more amino acid residues are phosphorylated. Petition 870260044022, dated 11 / 05 / 2026, p. 29 / 222 22 / 96

[0092] According to particular embodiments, one or more of the tau peptides is a tau phosphopeptide. According to particular embodiments, the one or more tau phosphopeptides comprise the amino acid sequence of one of the SEQ ID Nos: 1-3 or 5-12, or an amino acid sequence that is at least 75%, 80%, 85%, 90% or 95% identical to the amino acid sequence of one of the SEQ ID Nos: 1-3 or 5-12, wherein one or more of the indicated amino acid residues are phosphorylated. Preferably, the tau phosphopeptide comprises the amino acid sequence of one of the SEQ ID Nos: 1-3. The tau peptide may have an amidated C-terminal.

[0093] According to embodiments of the application, a tau peptide is presented on the liposome surface. A tau peptide, preferably a tau phosphopeptide, may be presented on the liposome surface using methods known in the art, in view of the present description. See, for example, the relevant description in U.S. Patents Nos. 8,647,631 and 9,687,447, the contents of which are incorporated herein by reference. According to particular embodiments, the one or more tau peptides, including phosphopeptides, further comprise one or more modifications, such as palmitoylation or dodecyl modification, to enable the tau peptides to be presented on the liposome surface. Additional amino acid residues, such as Lys, Cys, or sometimes Ser or Thr, may be added to the tau peptide to facilitate the modification. It has been reported that the position of lipid anchors induces different conformations of the peptide sequence (Hickman et al., J. Biol. Chem. Vol. 286, NO. 16, p.13966 to 13976, April 22, 2011). Although not wishing to be limited by theory, it is believed that the addition of hydrophobic portions at both ends may increase the pathological beta-sheet conformation of the tau peptide. Thus, one or more tau peptides further comprise hydrophobic portions at both ends. O. Petition 870260044022, dated 11 / 05 / 2026, page 30 / 222 23 / 96 modified tau peptide may have the C-terminal amidated. Preferably, a tau peptide presented on the liposome surface consists of the amino acid sequence from one of SEQ ID NO: 27 to SEQ ID NO: 38.

[0094] As used herein, the term helper T cell epitope refers to a polypeptide comprising an epitope that is capable of recognition by a helper T cell. Examples of helper T cell epitopes include, but are not limited to, tetanus toxoid (e.g., P2 and P30 epitopes, also referred to as T2 and T30, respectively), hepatitis B surface antigen, cholera B toxin, toxoid, diphtheria toxoid, measles virus F protein, Chlamydia trachomatis outer membrane major protein, Plasmodium falciparum circumsporozita T, falciparum CS antigen, Schistosoma mansoni triose phosphate isomerase, Bordetella pertussis, Clostridium tetani, Pertusaria trachythallina, Escherichia coli TraT, and influenza virus hemagglutinin (HA).

[0095] Any suitable helper T cell epitope known to those skilled in the art may be used in the invention in view of the present description. According to particular embodiments, the helper T cell epitope comprises 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 comprises two or more of the amino acid sequences from SEQ ID NO: 23 to SEQ ID NO: 26 fused through a linker, such as a peptide linker comprising one or more amino acids, for example, Val (V), Ala (A), Arg (R), Gly (G), Ser (S), Lys (K). The length of the linker may vary, preferably from 1 to 5 amino acids. Preferably, the helper T cell epitope comprises three or more of the amino acid sequences from SEQ ID NO: 23 to SEQ ID NO: 26 fused through one or more linkers selected from Petition 870260044022, dated 11 / 05 / 2026, p. 31 / 222 24 / 96 of the group consisting of VVR, GS, RR, RK. The helper T cell epitope may have its C-terminal amidated.

[0096] According to embodiments of the application, helper T cell epitopes can be incorporated into the liposomal surface, for example, anchored by a covalently linked hydrophobic moiety, wherein said hydrophobic moiety is an alkyl group, a fatty acid, a triglyceride, diglyceride, steroid, sphingolipid, glycolipid or phospholipid, particularly an alkyl group or a fatty acid, particularly with a carbon backbone of at least 3 carbon atoms, particularly of at least 4 carbon atoms, particularly of at least 6 carbon atoms, particularly of at least 8 carbon atoms, particularly of at least 12 carbon atoms, particularly of at least 16 carbon atoms. In one embodiment of the invention, the hydrophobic moiety is palmitic acid. Alternatively, helper T cell epitopes can be encapsulated in liposomes.According to particular designs, the helper T cell epitope is encapsulated in the liposome.

[0097] The helper T cell epitope can be modified to its desired location on liposomes using methods known in the art in view of the present description. According to particular embodiments, the helper T cell epitope useful for the invention comprises an amino acid sequence from one of SEQ ID NO: 39 to SEQ ID NO: 44. Preferably, the helper T cell epitope consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17.

[0098] According to particular embodiments, the liposome comprises a tau peptide and a helper T cell epitope in a weight ratio of 1:1.2:1.3:1.4:1.5:1 or 6:1.

[0099] In one embodiment, the liposome also comprises Petition 870260044022, dated 11 / 05 / 2026, page 32 / 222 25 / 96 less an adjuvant comprising a toll-type receptor ligand. Thus, in another general aspect, the invention relates to a liposome comprising:

[00100] a. a tau peptide, preferably a tau phosphopeptide;

[00101] b. a helper T cell epitope; and

[00102] c. at least one of

[00103] i. a toll-type receptor ligand 9, and

[00104] ii. a toll-type receptor ligand 4.

[00105] As used herein, the term toll-like receptor or TLR refers to a class of pattern recognition receptor (PRR) proteins that play a fundamental role in the innate immune response. TLRs recognize pathogen-associated molecular patterns (PAMPs) of microbial pathogens, such as bacteria, fungi, parasites, and viruses, which can be distinguished from host molecules. TLRs are membrane-spanning proteins that typically function as dimers and are expressed by cells involved in the innate immune response, including antigen-presenting dendritic cells and phagocytic macrophages. There are at least ten members of the human TLR family, TLR1 to TLR10, and at least twelve members of the murine TLR family, TLR1 to TLR9 and TLR11 to TLR13, and they differ in the types of antigens they recognize.For example, TLR4 recognizes lipopolysaccharides (LPS), a component present in many Gram-negative bacteria, as well as viral proteins, polysaccharides, and endogenous proteins such as low-density lipoprotein, beta-defensins, and heat shock proteins; and TLR9 is a nucleotide-sensing TLR that is activated by unmethylated or double-stranded cytosine phosphate-guanine (CpG) dinucleotides, which are abundant in prokaryotic genomes but rare in vertebrate genomes. TLR activation leads to a series of events. Petition 870260044022, dated 11 / 05 / 2026, page 33 / 222 26 / 96 signaling, resulting in the production of type I interferons (IFNs), inflammatory cytokines and chemokines, and the induction of immune responses. Eventually, this inflammation also activates the adaptive immune system, which results in the elimination of invading pathogens and infected cells.

[00106] As used herein, the term ligand refers to a molecule that forms a complex with a biomolecule (e.g., a receptor) to serve a biological purpose. According to particular embodiments, the toll-like receptor ligand is a toll-like receptor agonist.

[00107] As used herein, the term agonist refers to a molecule that binds to one or more TLRs and induces a receptor-mediated response. For example, an agonist may induce, stimulate, augment, activate, facilitate, enhance, or regulate receptor activity. Such activities are termed agonistic activities. For example, a TLR4 or TLR9 agonist may activate or augment cell signaling through the bound receptor. Agonists include, but are not limited to, nucleic acids, small molecules, proteins, carbohydrates, lipids, or any other molecules that bind to or interact with receptors. Agonists may resemble the activity of a natural receptor ligand. Agonists may be homologous to these natural receptor ligands with respect to sequence, conformation, charge, or other characteristics, so that they can be recognized by the receptors.This recognition can result in physiological and / or biochemical changes within the cell, such that the cell reacts to the presence of the agonist in the same way as if the natural receptor ligand were present. According to particular modalities, the toll-like receptor agonist is at least one of a toll-like receptor agonist 4 and a toll-like receptor agonist 9. Petition 870260044022, dated 11 / 05 / 2026, p. 34 / 222 27 / 96

[00108] As used herein, the term toll-type receptor agonist 4 refers to any compound that acts as a TLR4 agonist. Any suitable toll-type receptor 4 agonist known to those skilled in the art in view of the present description may be used in the invention. Examples of toll-type receptor 4 ligands useful for the invention include TLR4 agonists, including but not limited to monophosphoryl lipid A (MPLA). As used herein, the term monophosphoryl lipid A or MPLA refers to a modified form of lipid A, which is the biologically active part of Gram-negative bacterial lipopolysaccharide (LPS) endotoxin. MPLA is less toxic than LPS while retaining immunostimulatory activity. As a vaccine adjuvant, MPLA stimulates cellular and humoral responses to vaccine antigen.Examples of MPLA include, but are not limited to, 3-O-desacyl-4'-monophosphoryl lipid A, hexaacyl monophosphoryl lipid A, 3-desacyl, 3-desacyl monophosphoryl lipid A, and structurally related variants thereof. MPLA useful for the invention can be obtained using methods known in the art or from a commercial source, such as 3D-(6-acyl)PHAD®, PHAD®, PHAD®, PHAD®-504, 3D-PHAD® from Avanti Polar Lipids (Alabaster, Alabama, USA) or MPLTM from various commercial sources. According to particular embodiments, the toll-like receptor 4 agonist is MPLA. As used herein, the term toll-like receptor 9 agonist refers to any compound that acts as a TLR9 agonist. Any suitable toll-like receptor 9 agonist known to those skilled in the art in view of the present description may be used in the invention.Examples of toll-like receptor 9 ligands useful for the invention include TLR9 agonists including, but not limited to, CpG oligonucleotides.

[00109] As used herein, the term CpG oligonucleotide, CpG oligodeoxynucleotide, or CpG ODN refers to a Petition 870260044022, dated 11 / 05 / 2026, p. 35 / 222 28 / 96 oligonucleotide comprising at least one CpG motif. As used herein, oligonucleotide, oligodeoxynucleotide or ODN refers to a polynucleotide formed from a plurality of linked nucleotide units. Such oligonucleotides may be obtained from existing nucleic acid sources or may be produced by synthetic methods. As used herein, the term CpG motif refers to a nucleotide sequence containing unmethylated cytosine-phosphate-guanine (CpG) dinucleotides (i.e., a cytosine (C) followed by a guanine (G)) linked by a phosphate bond or a phosphodiester backbone or other internucleotide linkages.

[00110] According to particular embodiments, the CpG oligonucleotide is lipidized, that is, conjugated (covalently linked) to a lipid moiety.

[00111] As used herein, a lipid moiety refers to a moiety containing a lipophilic structure. Lipid moieties, such as an alkyl group, a fatty acid, a triglyceride, diglyceride, steroid, sphingolipid, glycolipid, or phospholipid, particularly a sterol, such as cholesterol or fatty acids, when linked to highly hydrophilic molecules, such as nucleic acids, can substantially improve plasma protein binding and, consequently, the circulating half-life of the hydrophilic molecules. Furthermore, binding to certain plasma proteins, such as lipoproteins, has been shown to increase uptake in specific tissues expressing the corresponding lipoprotein receptors (e.g., LDL receptor-HDL receptor or SR-B1 scavenger receptor).In particular, a lipid portion conjugated with phosphopeptides and / or CpG oligonucleotides allows said peptides and / or oligonucleotides to be anchored to the membrane of a liposome through a hydrophobic portion. Petition 870260044022, dated 11 / 05 / 2026, page 36 / 222 29 / 96

[00112] According to particular embodiments, in view of the present description, the CpG oligonucleotide may comprise any suitable internucleotide linkages.

[00113] As used herein, the term internucleotide linkage refers to a chemical bond linking two nucleotides through their sugars, consisting of a phosphorus atom and a charged or neutral group between adjacent nucleosides. Examples of internucleotide linkages include phosphodiester (po), phosphorothioate (ps), phosphorodithioate (ps2), methylphosphonate (mp), and methylphosphorothioate (rp). Phosphorothioate, phosphorodithioate, methylphosphonate, and methylphosphorothioate are stabilizing internucleotide linkages, while phosphodiester is a naturally occurring internucleotide linkage. Phosphorothioate oligonucleotides are typically synthesized as a random racemic mixture of Rp and Sp phosphorothioate linkages.

[00114] Any suitable CpG oligonucleotide known to those skilled in the art may be used in the invention in view of the present description. Examples of such CpG oligonucleotides include, but are not limited to, CpG2006 (also known as CpG 7909), CpG 1018, CpG2395, CpG2216, or CpG2336.

[00115] A CpG oligonucleotide can be lipidized using methods known in the art in view of the present description. In some embodiments, the 3' terminus of a CpG oligonucleotide is covalently linked to a cholesterol molecule via a phosphate linkage, optionally via a PEG linker. Another lipophilic portion can also be covalently linked to the 3' terminus of a CpG oligonucleotide. For example, a CpG oligonucleotide can be covalently linked to a lipid anchor of the same length as the liposome phospholipids: a palmitic acid chain (using Pal-OH or similar, activated for coupling) or two palmitic acids (e.g., using 1,2-dipalmitoyl-sn-glycero-3- Petition 870260044022, dated 11 / 05 / 2026, page 37 / 222 30 / 96 phosphoethanolamine-N-(succinyl) or similar, activated for coupling), optionally via a PEG linker. See, for example, the relevant description in U.S. Patent No. 7,741,297, the contents of which are incorporated herein by reference. The PEG length may vary, for example, from 1 to 5 PEG units.

[00116] Other linkers can also be used to covalently link a CpG oligonucleotide to a lipophilic moiety (such as a cholesterol molecule), examples of which include, but are not limited to, an alkyl spacer with 3 to 12 carbons. A short linker compatible with the chemistry of oligonucleotides such as aminodiol is required. In some embodiments, no linker is used for covalent linking. See, for example, Ries et al., Convenient synthesis and application of versatile nucleic acid lipid membrane anchors in liposome assembly and fusion, Org. Biomol. Chem., 2015, 13, 9673, the relevant description of which is incorporated herein by reference.

[00117] According to particular embodiments, the lipid-enhanced CpG oligonucleotide useful for the invention comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22, wherein the nucleotide sequence comprises one or more phosphorothioate internucleotide linkages, and the nucleotide sequence is covalently linked to at least one cholesterol molecule by means of a linker. Any suitable linkers may be used to covalently link a CpG oligonucleotide to a cholesterol molecule. Preferably, the linker comprises polyethylene glycol (PEG).

[00118] According to particular modalities, the liposome comprises:

[00119] a. a tau phosphopeptide;

[00120] b. a helper T cell epitope; Petition 870260044022, dated 11 / 05 / 2026, page 38 / 222 31 / 96

[00121] c. a lipid-encapsulated CpG oligonucleotide; and

[00122] d. a toll-type 4 receptor ligand;

[00123] in which the tau phosphopeptide is presented on the surface of the liposome and the helper T cell epitope is encapsulated in the liposome.

[00124] According to particular embodiments, the liposome comprises:

[00125] a. a tau peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 27 to SEQ ID NO: 38;

[00126] b. a helper T cell epitope having an amino acid sequence selected from the group consisting of SEQ ID NO: 39 to SEQ ID NO: 44, preferably the helper T cell epitope consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17;

[00127] c. a lipid-encapsulated CpG oligonucleotide with a nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22, wherein the CpG oligonucleotide comprises one or more phosphorothioate internucleotide linkages, and the CpG oligonucleotide is covalently linked to at least one cholesterol via a ligand; and

[00128] d. monophosphoryl lipid A (MPLA).

[00129] According to particular embodiments, the liposome further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG), and cholesterol.

[00130] According to particular embodiments, the liposome further comprises a buffer. Any suitable buffer known to those skilled in the art, in view of the present description, may be used in the invention. In one embodiment, the liposome comprises a phosphate-buffered saline solution. According to embodiments Petition 870260044022, dated 11 / 05 / 2026, page 39 / 222 32 / 96 particular cases, the tampon comprises histidine and sucrose.

[00131] According to particular embodiments, the liposome comprises DMPC, DMPG, cholesterol, tau phosphopeptide and helper T cell epitope in the molar ratio of 9:1:7:0.07:0.04.

[00132] The liposomes of the invention can be produced using methods known in the art, taking into account the present description.

[00133] An exemplary liposome of the present application is illustrated in Figure 1. More specifically, a tetrapalmitoylated tau phosphopeptide (pTau T3 peptide, SEQ ID NO: 28) is presented on the liposome surface via two palmitic acids at each end of the tau peptide. A TLR-9 ligand comprising lipidated CpG (CpG7909-Chol Adjuvant) is incorporated into the liposomal membrane via covalently bound cholesterol. A TLR-4 ligand (3D-(6-acyl)PHAD® Adjuvant) is also incorporated into the membrane. A helper T cell epitope (PAN-DR T50 ligand) is encapsulated. Conjugates

[00134] In a general aspect, the invention relates to a conjugate comprising a tau peptide and an immunogenic carrier conjugated to it.

[00135] According to particular aspects, the conjugate has the following structure: VEHICLE Peptide

[00136] or the structure of formula (II): Petition 870260044022, dated 11 / 05 / 2026, p. 40 / 222 33 / 96 Tau peptide XlNH Carrier

[00137] in which

[00138] x is an integer from 0 to 10; and

[00139] n is an integer from 2 to 15, preferably 3 to 11.

[00140] According to particular modalities, x is an integer from 1 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. According to particular modalities, x is 3.

[00141] According to particular modalities, n is 2 to 15, 3 to 11, 3 to 9, 3 to 8 or 3 to 7.

[00142] According to particular embodiments, the conjugate comprises one or more tau peptides. According to particular embodiments, the tau peptides of the conjugate may be the same or different.

[00143] According to particular embodiments, in view of the present description, any suitable tau peptides may be used in the invention. According to particular embodiments, one or more of the tau peptides comprise the amino acid sequence of one of the SEQ ID NOs: 1-12 or an amino acid sequence that is at least 75%, 80%, 85%, 90% or 95% identical to the amino acid sequence of one of the SEQ ID NOs: 1-12, where none, one or more of the amino acid residues are phosphorylated.

[00144] According to particular embodiments, one or more of the tau peptides is a tau phosphopeptide. According to particular embodiments, the one or more tau phosphopeptides comprise the amino acid sequence of one of the SEQ ID NOs: 1-3 or 5-12, or an amino acid sequence that is at least 75%, 80%, 85%, 90% Petition 870260044022, dated 11 / 05 / 2026, p. 41 / 222 34 / 96 or 95% identical to the amino acid sequence of one of the SEQ ID NOs: 1-3 or 5-12, where one or more of the indicated amino acid residues are phosphorylated.

[00145] According to particular embodiments, the phosphopeptide tau consists of the amino acid sequence of one of the SEQ ID NOs: 13.

[00146] As used herein, the term immunogenic vehicle refers to an immunogenic substance that can be coupled to a tau peptide. An immunogenic moiety coupled to a tau peptide can induce an immune response and elicit the production of antibodies that can specifically bind to the tau peptide. Immunogenic moieties are operative moieties that include proteins, polypeptides, glycoproteins, complex polysaccharides, particles, nucleic acids, polynucleotides, and the like that are recognized as foreign and thus elicit an immune response from the host. Any suitable immunogenic vehicle known to those skilled in the art in view of the present description may be used in the invention. According to particular embodiments, the immunogenic vehicle is California limpet hemocyanin (KLH), tetanus toxoid, CRM197 (a non-toxic form of diphtheria toxin), a mixture of N. meningitidis outer membrane proteins (OMP), or a derivative thereof.Depending on the specific modalities, the immunogenic vehicle is KLH or CRM197.

[00147] According to particular embodiments, the tau peptide is conjugated to the carrier via a linker. As used herein, the term linker refers to a chemical moiety that links an immunogenic carrier to a tau peptide. Any suitable linker known to those skilled in the art in view of the present description may be used in the invention. Linkers may be, for example, a single covalent bond, a substituted or unsubstituted alkyl group, a moiety Petition 870260044022, dated 11 / 05 / 2026, p. 42 / 222 35 / 96 substituted or unsubstituted heteroalkyl, a polyethylene glycol (PEG) linker, a peptide linker, a sugar-based linker, or a cleavable linker, such as a disulfide bond or a protease cleavage site, or an amino acid, or a combination thereof. Examples of the linker may include one or more polyethylene glycol (PEG), succinimidyl 3-(bromoacetamide) propionate (SBAP), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), or one or more amino acids such as Cys, Lys, or sometimes Ser or Thr, or a combination thereof.

[00148] According to particular embodiments, the ligand comprises (C2H4O)x-cysteine-acetamidopropionamide or mmaleimidobenzoyl-N-hydroxysuccinimide ester - cysteine ​​- (C2H4O)x, where x is an integer from 0 to 10, such as 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[00149] According to particular embodiments, the vehicle is covalently linked to the N-terminal of the tau peptide, through a ligand.

[00150] According to other particular embodiments, the vehicle is covalently linked to the C-terminal of the tau peptide, through a ligand.

[00151] According to particular modalities, the conjugate has the structure of: VY KS(p)PV VSG DT S( p )PRH L-CON HJ You

[00152] where n is an integer from 2 to 15, preferably 3 to 11, more preferably 3 to 7.

[00153] The conjugates of the invention can be made by methods Petition 870260044022, dated 11 / 05 / 2026, p. 43 / 222 36 / 96 known in the art, in view of the present description. For example, the above conjugate can be formed by the reaction succinimidyl 3-(bromoacetamide)propionate (SBAP): ___,Br OO NH o

[00154] with an amino group of CRM197 to form an amide linkage. This CRM197 precursor can be subsequently reacted with the tau peptide (e.g., the phosphorylated tau peptide of SEQ ID NO: 2) conjugated at its N-terminus or at its C-terminus to a PEG-cysteine ​​ligand with a free nucleophilic thiol group to form the tau phosphopeptide conjugate.

[00155] An exemplary conjugate according to an embodiment of the present application is illustrated in Figure 1. More specifically, multiple tau phosphopeptides (pTau Peptide T3.76) are covalently linked to a CRM197 carrier protein.

[00156] Pharmaceutical Compositions

[00157] In a general aspect, the invention relates to pharmaceutical compositions comprising a therapeutically effective amount of liposome or conjugate of the invention, together with a pharmaceutically acceptable excipient and / or vehicle. Pharmaceutically acceptable excipients and / or vehicles are well known in the art (see Remington's Pharmaceutical Science (15th ed.), Mack Publishing Company, Easton, Pa., 1980). The preferred formulation of the pharmaceutical composition depends on the intended mode of administration and therapeutic application. The compositions may include pharmaceutically acceptable non-toxic vehicles or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for administration to animals or humans. The diluent is selected so as not to affect the biological activity of the combination. Petition 870260044022, dated 11 / 05 / 2026, page 44 / 222 37 / 96 Examples of such diluents are distilled water, phosphate-buffered physiological saline solution, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include other non-toxic, non-therapeutic, non-immunogenic, and similar vehicles, adjuvants, or stabilizers. It is understood that the characteristics of the vehicle, excipient, or diluent will depend on the route of administration for a particular application.

[00158] The pharmaceutical composition may contain a mixture of the same immunogenic tau peptide. Alternatively, the pharmaceutical composition may contain a mixture of different immunogenic tau peptides of the present invention.

[00159] Another problem associated with vaccines against neuronal diseases is that exceptionally high antibody titers are likely to be required to ensure efficacy. This is because the target antigen for the vaccine is located in the brain. The brain is separated from the circulation by a specialized cellular structure called the blood-brain barrier (BBB). The BBB restricts the passage of substances from the circulation into the brain. This prevents toxins, microbes, etc., from entering the central nervous system. The BBB also has the potentially less desirable effect of preventing the efficient entry of immune mediators (such as antibodies) into the interstitial and cerebrospinal fluid surrounding the brain.

[00160] Approximately 0.1% of antibodies present in systemic circulation cross the BBB and enter the brain. This means that systemic titers induced by a vaccine targeting a CNS antigen must be at least 1000 times greater than the minimum effective titer to be effective in the brain.

[00161] According to particular embodiments, the pharmaceutical compositions of the present invention further comprise one or more suitable adjuvants. Thus, the tau peptides of the present invention, Petition 870260044022, dated 11 / 05 / 2026, page 45 / 222 38 / 96 present in the liposome or conjugate can be administered in combination with a suitable adjuvant to achieve the desired immune response in the individual. Suitable adjuvants can be administered before, after, or concomitantly with the administration of the liposome or conjugate of the present invention. Preferred adjuvants enhance the intrinsic response to an immunogene without causing conformational changes in the immunogene that affect the qualitative form of the response. Examples of adjuvants are aluminum salts (alum), such as aluminum hydroxide, aluminum phosphate, and aluminum sulfate. These adjuvants can be used with or without other specific immunostimulatory agents, such as MPLA Class (3 De-O-acylated monophosphoryl lipid A (MPLTM), monophosphoryl hexaacyl 3-desacyl-synthetic lipid A (3D-(6-acyl)PHAD®), PHAD™, PHAD®-504, 3D-PHAD®) lipid A), polymeric or monomeric amino acids, such as polyglutamic acid or polylysine.These adjuvants can be used with or without other specific immunostimulating agents, such as muramyl peptides (e.g., N-acetylmuramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetylnormuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-Lalanyl-D-isoglutaminyl-L-alanine-2-(1'-2' dipalmitoyl-sn-glycero-3hydroxyphosphoryloxy)-ethylamine (MTP-PE), N-acetylglucsaminyl-Nacetylmuramyl-L-Al-D-isoglu-L-Ala-dipalmitoxy propylamide (DTP-DPP) Theramide™) or other components of the bacterial cell wall. Oil-in-water emulsions include MF59 (see WO 90 / 14837), containing 5% squalene, 0.5% Tween 80, and 0.5% Span 85 (optionally containing varying amounts of MTP-PE) formulated into submicron particles using a microfluidizer; SAF, containing 10% squalene, 0.4% Tween 80, 5% pluronic-blocked polymer L121, and thrMDP, either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion; and the... Petition 870260044022, dated 11 / 05 / 2026, page 46 / 222 39 / 96 Ribi™ Adjuvant System (RAS) (Ribi ImmunoChem, Hamilton, Mont.) Tween 80 at 0.2% and one or more bacterial cell wall components selected from the group consisting of monophosphoryl lipid A (MPLT™), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL™+ CWS (Detox™). Other adjuvants include Freund's complete adjuvant (CFA) and cytokines, such as interleukins (IL-1, IL-2, and IL-12), macrophage colony-stimulating factor (M-CSF), and tumor necrosis factor (TNF).

[00162] As used herein, the term in combination, in the context of administering two or more therapies to an individual, refers to the use of more than one therapy. The use of the term in combination does not restrict the order in which the therapies are administered to an individual. For example, a first therapy (e.g., a composition described herein) may be administered before (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 before), concomitantly with, 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 after) the administration of a second therapy to a individual.

[00163] The pharmaceutical compositions of the present invention can be formulated according to methods well known in the art. The optimum ratios of each component in the compositions can be determined by techniques well known to those skilled in the art, in view of the present description. Methods of Use

[00164] Another general aspect of the invention relates to methods for Petition 870260044022, dated 11 / 05 / 2026, page 47 / 222 40 / 96 induce an immune response against tau protein in an individual suffering from a neurodegenerative disease, disorder or condition, comprising administering to the individual a pharmaceutical composition according to an embodiment of the invention. According to particular aspects, the immune response is induced against phosphorylated tau protein, preferably ePHF.

[00165] Another general aspect of the invention relates to methods for treating or preventing a neurodegenerative disease, disorder or condition, comprising administering to the individual a pharmaceutical composition according to an embodiment of the invention.

[00166] As used herein, the terms induce and stimulate and variations thereof refer to any measurable increase in cellular activity. Induction of an immune response may include, for example, activation, proliferation or maturation of a population of immune cells, increasing the production of a cytokine and / or other indicator of enhanced immune function. In certain embodiments, induction of an immune response may include increasing B cell proliferation, producing antigen-specific antibodies, increasing antigen-specific T cell proliferation, enhancing dendritic cell antigen presentation and / or increasing the expression of certain cytokines, chemokines and co-stimulatory markers.

[00167] The ability to induce or stimulate an anti-tau immune response after administration in an animal or human organism can be evaluated in vitro or in vivo using a variety of assays that are standard in the art. For a general description of the techniques available for evaluating the initiation and activation of an immune response, see, for example, Coligan et al. (1992 and 1994, Current Protocols in Immunology; ed. J. Wiley & Sons Inc., National Institutes of Health). Measurement of cellular immunity can be performed by methods Petition 870260044022, dated 11 / 05 / 2026, page 48 / 222 41 / 96 easily recognized in the art, for example, by measuring cytokine profiles secreted by activated effector cells, including those derived from CD4+ and CD8+ T cells (e.g., quantification of IL-4 or IFN gamma-producing cells by ELISPOT), by determining the activation state of effector immune cells (e.g., T cell proliferation assays by a classical [3H]thymidine uptake), testing antigen-specific T lymphocytes in a sensitized individual (e.g., peptide-specific lysis in a cytotoxicity assay, etc.).

[00168] The ability to stimulate a cellular and / or humoral response can be determined by testing a biological sample (e.g., blood, plasma, serum, PBMCs, urine, saliva, feces, CSF, or lymphatic fluid) from the individual for the presence of antibodies directed to the immunogenic tau peptide(s) administered in the pharmaceutical composition (see, for example, Harlow, 1989, Antibodies, Cold Spring Harbor Press). For example, antibody titers produced in response to administration of a composition that provides an immunogene can be measured by enzyme-linked immunosorbent assay (ELISA), dot blots, SDS-PAGE gels, ELISPOT, or antibody-dependent cellular phagocytosis (ADCP) assay.

[00169] As used herein, the term individual refers to an animal. According to particular modalities, the individual is a mammal including a non-primate (e.g., a camel, donkey, zebra, cow, pig, horse, goat, sheep, cat, dog, rat, rabbit, guinea pig, or mouse) or a primate (e.g., a monkey, chimpanzee, or human). According to particular modalities, the individual is a human.

[00170] As used herein, the term therapeutically effective amount refers to an amount of an active ingredient or component that elicits the desired biological or medicinal response. Petition 870260044022, dated 11 / 05 / 2026, p. 49 / 222 42 / 96 in an individual. A therapeutically effective amount can be determined empirically and routinely in relation to the stated objective. For example, in vitro assays may optionally be employed to help identify ideal dosage ranges. The selection of a specific effective dose can be determined (e.g., through clinical trials) by those skilled in the art, based on consideration of several factors, including the disease to be treated or prevented, the symptoms involved, the patient's body mass, the patient's immune status, and other factors known to those skilled in the art. The exact dose to be employed in the formulation will also depend on the route of administration and the severity of the disease, and should be decided according to the physician's judgment and the circumstances of each patient. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[00171] As used in this document, the terms treat, treating, and treatment are all intended to refer to an improvement or reversal of at least one measurable physical parameter related to a neurodegenerative disease, disorder, or condition, which is not necessarily discernible in the individual, but which may be discernible in the individual. The terms treat, treat, and treatment may also refer to causing regression, preventing progression, or at least slowing the progression of the disease, disorder, or condition. In a specific modality, treat, treat, and treatment refer to a relief, prevention of the development or onset, or reduction in the duration of one or more symptoms associated with the neurodegenerative disease, disorder, or condition. In a specific modality, treat, treat, and treatment refer to the prevention of recurrence of the disease, disorder, or condition. In a specific modality, treat, treat, and treatment refer to an increase Petition 870260044022, dated 11 / 05 / 2026, page 50 / 222 43 / 96 in the survival rate of an individual with the disease, disorder, or condition. In a specific modality, treat, treating, and treatment refer to the elimination of the disease, disorder, or condition in the individual.

[00172] According to particular modalities, a therapeutically effective amount refers to the amount of therapy that is sufficient to achieve one, two, three, four, or more of the following effects: (i) reducing or improving the severity of the disease, disorder, or condition being treated or a symptom associated with it; (ii) reducing the duration of the disease, disorder, or condition being treated or a symptom associated with it; (iii) preventing the progression of the disease, disorder, or condition being treated, or a symptom associated with it; (iv) causing regression of the disease, disorder, or condition being treated, or a symptom associated with it; (v) preventing the development or onset of the disease, disorder, or condition being treated, or a symptom associated with it; (vi) preventing the recurrence of the disease, disorder, or condition being treated, or a symptom associated with it; (vii) reducing hospitalization of an individual having the disease, disorder, or condition being treated, or a symptom associated with it;(viii) reduce the length of hospital stay of an individual with the disease, disorder or condition to be treated or a symptom associated with it; (ix) increase the survival of an individual with the disease, disorder or condition to be treated, or a symptom associated with it; (x) inhibit or reduce the disease, disorder or condition to be treated, or a symptom associated with it in an individual; and / or (xi) increase or improve the prophylactic or therapeutic effect of another therapy.

[00173] As used herein, a neurodegenerative disease, disorder, or condition includes any neurodegenerative disease, disorder, or condition known to those skilled in the art, with a view to the present description. Examples of neurodegenerative diseases, disorders, or conditions include neurodegenerative diseases or disorders caused by or associated with the formation of neurofibrillary fibrillation lesions, such as Petition 870260044022, dated 11 / 05 / 2026, p. 51 / 222 44 / 96 diseases, disorders or conditions associated with tau, known as tauopathies. According to particular modalities, neurodegenerative disease, disorder, or condition includes any of the diseases or disorders that show the coexistence of tau and amyloid pathologies, including, but not limited to, Alzheimer's disease, Parkinson's disease, Creutzfeldt-Jakob disease, Dementia pugilistica, Down syndrome, Gerstmann-Straussler-Scheinker disease, inclusion body myositis, cerebral amyloid angiopathy due to prion protein, traumatic brain injury, amyotrophic lateral sclerosis, Guam complex parkinsonism-dementia, non-Guamian motor neuron disease with neurofibrillary tangles, argyrophilic granule dementia, corticobasal degeneration, amyotrophic lateral sclerosis - Lewy body dementia, diffuse neurofibrillary tangles with calcification, frontotemporal dementia,Preferably frontotemporal dementia with parkinsonism associated with chromosome 17 (FTDP-17), frontotemporal lobar dementia, Hallevorden-Spatz disease, multiple system atrophy, Niemann-Pick disease type C, Pick's disease, progressive subcortical gliosis, progressive supranuclear palsy, subacute sclerosing panencephalitis, entanglement-only dementia, postencephalitic parkinsonism, myotonic dystrophy, chronic traumatic encephalopathy (CTE), primary age-related tauopathy (PART), or Lewy body dementia (LBD). Depending on the particular modalities, the neurodegenerative disease, disorder, or condition is Alzheimer's disease or another tauopathy.

[00174] The present invention also provides a method for promoting the clearance of tau aggregates from the brain of an individual, said method comprising administering to the individual a pharmaceutical composition according to an embodiment of the invention, under conditions effective for promoting the clearance of tau aggregates. Petition 870260044022, dated 11 / 05 / 2026, page 52 / 222 45 / 96 tau originates from the individual's brain. According to particular modalities, tau aggregates are neurofibrillary tangles or their precursors of pathological tau.

[00175] The present invention also provides a method for delaying the progression of a behavioral phenotype related to tau pathology in an individual, said method comprising administering to the individual a pharmaceutical composition according to an embodiment of the invention, under conditions effective in delaying the progression of the behavioral phenotype related to tau pathology in the individual.

[00176] In a preferred embodiment of the present invention, the administration of a tau peptide, via administration of a pharmaceutical composition according to an embodiment of the invention, induces an active immune response in the individual to the tau peptide and the pathological form of tau, thereby facilitating the clearance of related tau aggregates, delaying the progression of tau pathology-related behavior and / or treating the underlying tauopathy. According to this aspect of the present invention, an immune response involves the development of a beneficial humoral response (antibody-mediated) directed against the tau peptide and a cellular response (mediated by antigen-specific T cells or their secretory products) directed against the T cell epitope or the immunogenic vehicle.

[00177] As used herein, a behavioral phenotype related to tau pathology includes, without limitation, cognitive impairments, early personality alteration and disinhibition, apathy, abulia, mutism, apraxia, perseveration, stereotyped movements / behaviors, hyperorality, disorganization, inability to plan or organize sequential tasks, selfishness / insensitivity, antisocial traits, lack of empathy, interruption, agrammatic speech with Petition 870260044022, dated 11 / 05 / 2026, page 53 / 222 46 / 96 frequent paraphasic errors, but relatively preserved comprehension, impaired comprehension and word-finding deficits, slowly progressive gait instability, retropulsions, freezing, frequent falls, axial rigidity unresponsive to levodopa, supranuclear gaze palsy, square wave spasms, slow vertical saccades, pseudobulbar palsy, limb apraxia, dystonia, cortical sensory loss, and tremor.

[00178] When performing the methods of the present invention, it is preferable to select an individual having or at risk of having Alzheimer's disease or another tauopathy, an individual with tau aggregates in the brain, or an individual exhibiting a entanglement-related behavioral phenotype prior to administration of the immunogenic peptides or antibodies of the present invention. Treatable individuals include individuals at risk of disease but who do not exhibit symptoms, as well as patients currently exhibiting symptoms. In the case of Alzheimer's disease, virtually anyone is at risk of developing Alzheimer's disease. Therefore, the present methods can be administered prophylactically to the general population without the need for any assessment of the individual patient's risk. The present methods are especially useful for individuals who have a known genetic risk for Alzheimer's disease.These individuals include those who have relatives who have suffered from the disease and those whose risk is determined by analysis of genetic or biochemical markers.

[00179] In asymptomatic patients, treatment can begin at any age (e.g., 10, 20, 30 years). However, it is usually not necessary to start treatment until a patient reaches 40, 50, 60, or 70 years of age. Treatment typically involves multiple doses over a period of time. Treatment can be monitored through antibody analysis or cell responses. Petition 870260044022, dated 11 / 05 / 2026, page 54 / 222 47 / 96 T or B cells are activated in response to the therapeutic agent over time. If the response diminishes, a booster dose is indicated.

[00180] In prophylactic applications, pharmaceutical compositions containing tau peptides are administered to a patient susceptible to, or otherwise at risk of, developing Alzheimer's disease or another tauopathy, in a quantity sufficient to eliminate or reduce the risk, decrease the severity, or delay the onset of the disease, including biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes presented during the development of the disease. In therapeutic applications, pharmaceutical compositions containing a tau peptide are administered to a patient suspected of or already suffering from this disease in a quantity sufficient to cure or at least partially interrupt the symptoms of the disease (biochemical, histological, and / or behavioral), including its complications and intermediate pathological phenotypes in the development of the disease.

[00181] The effective doses of a pharmaceutical composition of the invention, for the prevention and / or treatment of neurodegenerative disease, disorder or condition, vary according to many different factors, including mode of administration, target site, physiological state of the patient, other medications administered, and whether the treatment is prophylactic or therapeutic. The amount of peptides depends on whether an adjuvant is also administered, with higher doses being required in the absence of an adjuvant. The timing of injections can vary significantly from once daily, once a year, and once a decade. A typical regimen consists of an immunization followed by booster injections at time intervals, such as 6-week intervals. Another regimen consists of an immunization followed by booster injections 1, 2, 6, 9, and 12 months later. Yet another regimen involves an injection every two months for life. Petition 870260044022, dated 11 / 05 / 2026, p. 55 / 222 48 / 96 Alternatively, booster injections may be irregular, as indicated by monitoring of the immune response.

[00182] It is readily apparent to those skilled in the art that the regimen for initial and booster administrations may be adjusted based on the immune responses measured after administration. For example, booster compositions are generally administered weeks or months after administration of the initial composition, for example, approximately 2 to 3 weeks or 4 weeks, or 8 weeks, or 16 weeks, or 20 weeks, or 24 weeks, or 26 weeks, or 28 weeks, or 30 weeks, or 32 weeks, or 36 weeks, or one to two years after administration of the initial composition.

[00183] Peptides can be administered parenterally, topically, intravenously, orally, subcutaneously, intra-arterially, intracranially, intraperitoneally, intradermally, intranasally, or intramuscularly for prophylactic and / or therapeutic treatment. The most typical route of administration for an immunogenic agent is subcutaneous or intramuscular injection. The latter type of injection is most typically performed in the muscles of the arm or leg.

[00184] Depending on particular aspects, one or more booster immunizations may be administered. The antigens in the respective initial and booster compositions, although many booster compositions are used, need not be identical, but must share antigenic determinants or be substantially similar to each other.

[00185] The composition may, if desired, be presented in a kit, package or dispenser, which may contain one or more unit dosage forms containing the active ingredient. The kit, for example, may comprise a metal or plastic foil, such as a blister pack. The kit, package or dispenser may be accompanied by instructions for administration. Petition 870260044022, dated 11 / 05 / 2026, page 56 / 222 49 / 96

[00186] According to particular embodiments, the kit comprises at least one pharmaceutical composition comprising a liposome according to an embodiment of the invention and one pharmaceutical composition comprising a conjugate according to an embodiment of the invention. Modalities

[00187] The invention also provides the following non-limiting embodiments.

[00188] Modality 1 is a liposome, which comprises:

[00189] a. a tau peptide; and

[00190] b. a helper T cell epitope;

[00191] in which the tau peptide is presented on the surface of the liposome.

[00192] Mode 2 is the liposome of Mode 1, where the tau peptide is a tau phosphopeptide.

[00193] Mode 3 is the liposome of Mode 1 or 2, further comprising a toll-like receptor ligand.

[00194] Modality 4 is the liposome of Modality 3, wherein the toll-like receptor ligand comprises at least one of a toll-like receptor ligand 4 and a toll-like receptor ligand 9.

[00195] Mode 5 is the liposome of Mode 3 or 4, where the toll-type receptor ligand is a toll-type receptor ligand 4.

[00196] Mode 6 is the liposome of Mode 5, in which the toll-like receptor ligand 4 comprises monophosphoryl lipid A (MPLA).

[00197] Mode 7 is the liposome of Mode 3 or 4, where the toll-like receptor ligand is a toll-like receptor ligand 9.

[00198] Mode 8 is the liposome of Mode 7, in which the toll-like receptor ligand 9 comprises a lipid-coated CpG oligonucleotide. Petition 870260044022, dated 11 / 05 / 2026, page 57 / 222 50 / 96

[00199] Modality 9 is the liposome of Modality 1, comprising:

[00200] a. a tau peptide;

[00201] b. a helper T cell epitope; and

[00202] c. at least one of

[00203] i. a toll-type receptor ligand 9, and

[00204] ii. a toll-type receptor ligand 4.

[00205] Modality 10 is the liposome of Modality 9, in which the tau peptide is a tau phosphopeptide.

[00206] Mode 11 is the liposome of Mode 9 or 10, in which the toll-like receptor ligand 9 is a lipid-coated CpG oligonucleotide.

[00207] Modality 12 is the liposome of any of the Modalities 9 to 11, wherein the liposome comprises toll-type receptor ligand 4 and toll-type receptor ligand 9.

[00208] Modality 13 is the liposome of Modality 12, in which the toll-like receptor ligand 4 comprises monophosphoryl lipid A (MPLA).

[00209] Modality 14 is a liposome, which comprises:

[00210] a. a tau phosphopeptide;

[00211] b. a helper T cell epitope;

[00212] c. a lipid-encapsulated CpG oligonucleotide; and

[00213] d. an adjuvant containing a toll-type receptor ligand 4;

[00214] in which the tau phosphopeptide is presented on the surface of the liposome.

[00215] Modality 15 is the liposome of Modality 14, in which the toll-like receptor ligand 4 comprises monophosphoryl lipid A (MPLA).

[00216] Modality 16 is the liposome of any of the Petition 870260044022, dated 11 / 05 / 2026, page 58 / 222 51 / 96 Modalities 1 to 15, in which the helper T cell epitope is encapsulated in the liposome.

[00217] Modality 16a is the liposome of any of the Modalities 1 to 15, in which the helper T cell epitope is incorporated into the liposome membrane.

[00218] Modality 16b is the liposome of any of the Modalities 1 to 15, in which the helper T cell epitope is presented on the liposome surface.

[00219] Modality 17 is a liposome composition, comprising:

[00220] a. a tau phosphopeptide;

[00221] b. a helper T cell epitope;

[00222] c. a lipid-encapsulated CpG oligonucleotide; and

[00223] d. a monophosphoryl lipid A (MPLA);

[00224] in which the phosphopeptide tau is presented on the surface of the liposome, and

[00225] the T cell epitope is encapsulated in the liposome.

[00226] Modality 17a is the liposome of Modality 17, wherein MPLA is 3-O-desacyl-4'-monophosphoryl-lipid A, preferably MPLTM.

[00227] Modality 17b is the liposome of Modality 17, in which MPLA is monophosphoryl hexaacyl lipid A, 3-desacyl, preferably 3D-(6-acyl)PHAD®.

[00228] Modality 17c is the liposome of Modality 17, in which MPLA is monophosphoryl-3-desacyl-lipid A, preferably 3D-PHAD®.

[00229] Modality 18 is the liposome of any of the Modalities 1 to 17c, further comprising one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-racglycerol (DMPG), and cholesterol.

[00230] Modality 19 is the liposome of any of the Petition 870260044022, dated 11 / 05 / 2026, page 59 / 222 52 / 96 Embodiments 1 to 18, wherein the tau peptide has an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12, or at least 85%, 90% or 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12.

[00231] Modality 19-1 is the liposome of Modality 19, in which the tau peptide is a phosphopeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-3 and 5-12.

[00232] Modality 19-2 is the liposome of Modality 19-1, where the tau phosphopeptide comprises the amino acid sequence of SEQ ID NO: 1.

[00233] Modality 19-3 is the liposome of Modality 19-1, where the tau phosphopeptide comprises the amino acid sequence of SEQ ID NO: 2.

[00234] Modality 19-4 is the liposome of Modality 19-1, where the tau phosphopeptide comprises the amino acid sequence of SEQ ID NO: 3.

[00235] Modality 19a is the liposome of any of the Modalities 19, 19-1, 19-2, 19-3 and 19-4, in which the amino acid sequence further comprises one or more modifications to allow the tau peptide to be presented on the liposome surface.

[00236] Modality 19b is the liposome of Modality 19a, in which one or more modifications comprise at least one palmitoylation and dodecyl modification.

[00237] Modality 19c is the liposome of Modality 19a or 19b, in which the tau peptide is modified at its N-terminus by one or more modifications.

[00238] Modality 19d is the liposome of any of the Petition 870260044022, dated 11 / 05 / 2026, p. 60 / 222 53 / 96 Modalities 19a to 19c, in which the tau peptide is modified at its C-terminal by one or more modifications.

[00239] Modality 19e is the liposome of Modality 19d, in which the tau peptide is palmitoylated at its N-terminus and C-terminus.

[00240] Modality 19f is the liposome of any of the Modalities 19a-19e, in which the tau peptide further comprises one or more additional amino acids to facilitate one or more modifications.

[00241] Modality 19g is the liposome of Modality 19f, in which one or more additional amino acids are selected from the group consisting of Lys, Cys, Ser, and Thr.

[00242] Modality 19h is the liposome of any of the Modalities 19 to 19g, in which the tau peptide is amidated at its C-terminal.

[00243] Modality 19i is the liposome of any of the Modalities 19 to 19h, in which the tau peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 27 to SEQ ID NO: 38.

[00244] Modality 19j is the liposome of any of the Modalities 19-19i, where the tau peptide consists of the amino acid sequence of SEQ ID NO: 27.

[00245] Modality 19k is the liposome of any of the Modalities 19-19i, in which the tau peptide consists of the amino acid sequence of SEQ ID NO: 28.

[00246] Modality 19l is the liposome of any of the Modalities 19-19i, in which the tau peptide consists of the amino acid sequence of SEQ ID NO: 29.

[00247] Modality 20 is the liposome of any of the Modalities 1 to 19l, in which the helper T cell epitope comprises at least one amino acid sequence selected from the group consisting of: SEQ ID NO: 23 to SEQ ID NO: 26. Petition 870260044022, dated 11 / 05 / 2026, p. 61 / 222 54 / 96

[00248] Modality 20a is the liposome of Modality 20, in which the helper T cell epitope comprises at least two amino acid sequences selected from the group consisting of: SEQ ID NO: 23 to SEQ ID NO: 26.

[00249] Modality 20b is the liposome of Modality 20, in which the helper T cell epitope comprises at least three amino acid sequences selected from the group consisting of: SEQ ID NO: 23 to SEQ ID NO: 26.

[00250] Modality 20c is the liposome of Modality 20, where the helper T cell epitope comprises the four amino acid sequences from: SEQ ID NO: 23 to SEQ ID NO: 26.

[00251] Modality 20d is the liposome of any of the Modalities 20a to 20c, in which two or more amino acid sequences selected from the group consisting of SEQ ID NO: 23 to SEQ ID NO: 26 are covalently linked by a ligand.

[00252] Modality 20e is the liposome of Modality 20d, wherein the linker comprises one or more amino acids selected from the group consisting of Val (V), Ala (A), Arg (R), Gly (G), Ser (S), Lys (K).

[00253] Modality 20f is the liposome of Modality 20e, wherein the linker comprises an amino acid sequence selected from the group consisting of VVR, GS, RR, and RK.

[00254] Modality 20g is the liposome of any of the Modalities 20 to 20f, in which the helper T cell epitope is amidated at its C-terminus.

[00255] Modality 20h is the liposome of any of the Modalities 20 to 20g, in which the helper T cell epitope is modified for insertion into the liposome membrane, presentation on the liposome surface, or encapsulation within the liposome, depending on the intended location of the helper T cell epitope.

[00256] Modality 20i is the liposome of any of the Petition 870260044022, dated 11 / 05 / 2026, page 62 / 222 55 / 96 Modalities 20 to 20h, in which the helper T cell epitope consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17.

[00257] Modality 20j is the liposome of any of the Modalities 1 to 20i, in which the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 6:1.

[00258] Modality 20k is the liposome of any of the Modalities 1 to 20i, in which the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 5:1.

[00259] Modality 20l is the liposome of any of the Modalities 1 to 20i, wherein the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 4:1.

[00260] Modality 20m is the liposome of any of the Modalities 1 to 20i, in which the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 3:1.

[00261] Modality 20n is the liposome of any of the Modalities 1 to 20i, wherein the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 2:1.

[00262] Modality 20o is the liposome of any of the Modalities 1 to 20i, in which the liposome comprises the tau peptide and the helper T cell epitope in a weight ratio of 1:1.

[00263] Modality 21 is the liposome of any of the Modalities 1 to 20, in which the lipid-encapsulated CpG oligonucleotide comprises the nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22.

[00264] Modality 21a is the liposome of Modality 21, in which the CpG oligonucleotide has one or more phosphorothioate internucleotide linkages.

[00265] Modality 21b is the liposome of Modality 21a, in which the CpG oligonucleotide has all the internucleotide bonds. Petition 870260044022, dated 11 / 05 / 2026, p. 63 / 222 56 / 96 phosphorothioate.

[00266] Modality 21c is the liposome of any of the Modalities 21 to 21b, in which the lipid-encapsulated CpG oligonucleotide comprises the CpG oligonucleotide covalently linked to at least one lipophilic group via a linker.

[00267] Modality 21d is the liposome of Modality 21c, where the ligand comprises (C2H4O)n, where n is an integer from 0 to 10.

[00268] Modality 21e is the liposome of Modality 21c, where the ligand comprises an alkyl spacer with 3 to 12 carbons.

[00269] Modality 21f is the liposome of any of the Modalities 21 to 21e, where at least one lipophilic group is cholesterol.

[00270] Modality 21g is the liposome of either Modalities 21 to 21f, wherein the lipid-encapsulated CpG oligonucleotide comprises the nucleotide sequence SEQ ID NO: 18 or SEQ ID NO: 19 covalently linked to a cholesterol molecule via a ligand comprising (C2H4O)n, wherein n is an integer from 3 to 5.

[00271] Modality 22 is a liposome, which comprises:

[00272] a. a tau peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 27 to SEQ ID NO: 38;

[00273] b. a helper T cell epitope having an amino acid sequence selected from the group consisting of SEQ ID NO: 39 to SEQ ID NO: 44, preferably the helper T cell epitope consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17;

[00274] c. a lipid-encapsulated CpG oligonucleotide with a nucleotide sequence selected from the group consisting of SEQ ID NO: Petition 870260044022, dated 11 / 05 / 2026, p. 64 / 222 57 / 96 a SEQ ID NO: 22, wherein the CpG oligonucleotide comprises one or more phosphorothioate internucleotide linkages, and the CpG oligonucleotide is covalently linked to at least one cholesterol via a ligand; and

[00275] d. monophosphoryl lipid A (MPLA).

[00276] Modality 22a is a liposome of Modality 22, comprising:

[00277] a. a tau phosphopeptide consisting of the amino acid sequence SEQ ID NO: 27, SEQ ID NO: 28 or SEQ ID NO: 29;

[00278] b. a helper T cell epitope consisting of the amino acid sequence SEQ ID NO: 13

[00279] c. a lipid-containing CpG oligonucleotide consisting of the nucleotide sequence SEQ ID NO: 18 or SEQ ID NO: 19 covalently linked to a cholesterol via a linker comprising (C2H4O)n, where n is an integer from 3 to 7; and

[00280] d. monophosphoryl lipid A (MPLA).

[00281] Modality 22b is the liposome of Modality 22 or 22a, where MPLA is 3-O-desacyl-4'-monophosphoryl-lipid A, preferably MPLTM.

[00282] Modality 22c is the liposome of Modality 22 or 22a, where MPLA is preferentially 3D-(6-acyl)PHAD®.

[00283] Modality 22d is the liposome of Modality 22 or 22a, in which MPLA is preferably 3D-PHAD®.

[00284] Modality 23 is the liposome of any of the Modalities 22 to 22d, in which the helper T cell epitope is encapsulated in the liposome.

[00285] Embodiment 24 is a pharmaceutical composition comprising the liposome of any of Embodiments 1 to 23 and a pharmaceutically acceptable carrier.

[00286] Modality 25 is a combination that includes a Petition 870260044022, dated 11 / 05 / 2026, page 65 / 222 58 / 96 phosphopeptide tau and an immunogenic carrier conjugated to it via a ligand, having the following structure: Carrier Tau peptide

[00287] where x is an integer from 0 to 10; and

[00288] n is an integer from 2 to 15.

[00289] Modality 25a is a conjugate comprising a tau phosphopeptide and an immunogenic carrier conjugated to it via a ligand, having the structure of formula (II): Carrier H2N s H N-Tau peptide

[00290] in which

[00291] x is an integer from 0 to 10; and

[00292] n is an integer from 2 to 15.

[00293] Modality 26 is the conjugate of Modality 25 or 25a, where x is an integer from 2 to 6.

[00294] Modality 27 is the conjugate of Modality 25 or 25a, where x is 3.

[00295] Modality 28 is the combination of any of the Modalities 25 to 25a, where n is 3 to 7.

[00296] Modality 29 is the combination of any of the Modalities 25 to 28, in which the vehicle is an immunogenic vehicle. Petition 870260044022, dated 11 / 05 / 2026, p. 66 / 222 59 / 96 selected from the group consisting of California limpet hemocyanin (KLH), tetanus toxoid, CRM197 and a mixture of N. meningitidis outer membrane proteins (OMP) or a derivative thereof.

[00297] Modality 30 is the conjugate of any of the Modalities 25 to 29, wherein the tau phosphopeptide consists of the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 12.

[00298] Modality 30a is the conjugate of Modality 30, in which the tau phosphopeptide consists of the amino acid sequence SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[00299] Modality 31 is the combination of any of the Modalities 25 to 30, where the vehicle is CRM197.

[00300] Modality 32 is the conjugate of Modality 25, with the structure of: VY KS(p)PV VSG DT S( p )PRH L-CON HJ You

[00301] where n is 3-7.

[00302] Modality 32a is the conjugate of Modality 25, where KLH- [m-maleimidobenzoyl-N-hydroxysuccinimide ester - cysteine ​​(C2H4O)x - Tau peptide]n Petition 870260044022, dated 11 / 05 / 2026, p. 67 / 222 60 / 96 Tau peptide

[00303] in which

[00304] the Tau peptide consists of SEQ ID NO: 1 or SEQ ID NO: 3;

[00305] x is an integer from 0 to 10; and

[00306] n is an integer from 2 to 15.

[00307] Modality 33 is a pharmaceutical composition comprising the combination of any of the Modalities 25 to 32a and a pharmaceutically acceptable carrier.

[00308] Possibility 33a is the pharmaceutical composition of claim 33, further comprising an adjuvant.

[00309] Embodiment 33b is the pharmaceutical composition of claim 33a, wherein the adjuvant comprises at least one of a TLR-4 ligand and a TLR-9 ligand.

[00310] Modality 34 is a method for inducing an immune response in an individual suffering from a neurodegenerative disorder, comprising administering to the individual at least one of the pharmaceutical compositions of Modalities 24 and 33 to 33b.

[00311] Modality 35 is the method of Modality 34, comprising administering to the individual at least one of the pharmaceutical compositions of Modalities 24 and 33 to 33b to initiate immunization, and administering to the individual at least one of the pharmaceutical compositions of Modalities 24 and 33 to 33b to boost immunization.

[00312] Modality 36 is a method for the treatment or Petition 870260044022, dated 11 / 05 / 2026, page 68 / 222 61 / 96 prevention of a neurodegenerative disease or disorder in an individual in need of treatment, comprising administering to the individual at least one of the pharmaceutical compositions of Modality 24 or 33.

[00313] Modality 37 is the method of Modality 36, comprising administering to the individual at least one of the pharmaceutical compositions of Modalities 24 and 33 to 33b to initiate immunization, and administering to the individual at least one of the pharmaceutical compositions of Modalities 24 and 33 to 33b to boost immunization.

[00314] Modality 38 is the method of any of the Modalities 34 to 37, in which the neurodegenerative disease or disorder is caused by or associated with the formation of neurofibrillary lesions.

[00315] Modality 39 is the method of any of the Modalities 34 to 38, in which the neurodegenerative disease or disorder is Alzheimer's Disease, Parkinson's Disease, Creutzfeldt-Jakob Disease, pugilistic dementia, Down Syndrome, Gerstmann-Straussler-Scheinker Disease, inclusion body myositis, cerebral amyloid angiopathy due to prion protein, traumatic brain injury, amyotrophic lateral sclerosis, Guam complex dementia, non-Guamian neurological disease with neurofibrillary tangles, argyrophilic grain dementia, corticobasal degeneration, Lewy body dementia - amyotrophic lateral sclerosis, diffuse neurofibrillary tangles with calcification, frontotemporal dementia, preferably frontotemporal dementia with parkinsonism associated with chromosome 17 (FTDP-17), frontotemporal lobar dementia, Hallevorden-Spatz disease, multiple system atrophy, Niemann-Pick disease type C, Pick's disease,progressive subcortical gliosis, progressive supranuclear palsy, subacute sclerosing panencephalitis, dementia only, Petition 870260044022, dated 11 / 05 / 2026, page 69 / 222 62 / 96 due to entanglement, post-encephalitic parkinsonism, myotonic dystrophy, chronic traumatic encephalopathy (CTE), primary age-related tauopathy (PART), or Lewy body dementia (LBD).

[00316] Modality 40 is the method of any of the Modalities 34 to 39, in which the neurodegenerative disease or disorder is Alzheimer's disease, Parkinson's disease, Down syndrome, progressive supranuclear palsy (PSP), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Pick's disease, corticobasal degeneration, Lewy body dementia, amyotrophic lateral sclerosis, myotonic dysplasia, chronic traumatic encephalopathy (CTE), cerebral angiopathy, primary age-related tauopathy (PART), or Lewy body dementia (LBD).

[00317] Modality 40b is the method of any of the Modalities 34 to 39, in which the neurodegenerative disease or disorder is Alzheimer's disease, progressive supranuclear palsy (PSP), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17) or Pick's disease and PART (primary age-related tauopathy).

[00318] Modality 40c is the method of any of the Modalities 34 to 39, in which the neurodegenerative disease or disorder is Alzheimer's disease, Parkinson's disease, Down syndrome, frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), corticobasal degeneration, Lewy body dementia - amyotrophic lateral sclerosis, myotonic dysplasia, chronic traumatic encephalopathy (CTE), cerebral angiopathy, primary age-related tauopathy (PART) or Lewy body dementia (LBD).

[00319] Modality 41 is a kit comprising at least one of the pharmaceutical compositions of Modality 24 and the pharmaceutical composition of Modality 33, 33a or 33b.

[00320] Modality 42 is a helper T cell epitope that Petition 870260044022, dated 11 / 05 / 2026, page 70 / 222 63 / 96 consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17.

[00321] Modality 43 is a pharmaceutical composition comprising the helper T-cell epitope of Modality 42.

[00322] Modality 44 is a method for enhancing an immune response to an antigen in an individual in need of treatment, comprising administering the antigen to the individual together with the pharmaceutical composition of Modality 43. EXAMPLES

[00323] The following examples of the invention are to further illustrate the nature of the invention. It should be understood that the following examples do not limit the invention and that the scope of the invention should be determined by the appended claims.

[00324] The experimental methods used in the following examples, unless otherwise indicated, are all common methods. The reagents used in the following embodiments, unless otherwise indicated, are all purchased from common reagent suppliers. Example 1: Preparation of Liposomal Vaccines Preparation of the control liposomal vaccine (ethanol injection technique)

[00325] The liposomal control vaccine was produced by the ethanol (EtOH) injection technique followed by extrusion. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands) and MPLA (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of 9:1:7:0.05 in a 20:1 (by volume) mixture of EtOH and terbutanol (t-BuOH) at 60°C. The lipid / ethanol solution was diluted in phosphate-buffered saline (PBS) pH 7.4 at 60°C to maintain the EtOH concentration at 10%, resulting in the formation of Petition 870260044022, dated 11 / 05 / 2026, page 71 / 222 64 / 96 multilamellar liposomal vesicles (MLVs). The MLVs were then subjected to 5 sequential extrusion passages through three 0.08 µm pore size polycarbonate filters (Whatman) in series using Emulsiflex-C5 (Avestin, Canada). The resulting liposomes were diluted in PBS pH 7.4 and heated to 60°C to obtain a liposome solution before the addition of the tau peptide.

[00326] A phosphorylated tau peptide tetrapalmitoylated acetate of SEQ ID NO: 2 (Bachem AG, Switzerland), herein referred to as the active pharmaceutical ingredient (API), was dissolved in PBS at pH 11.4 with 2.0% octyl βD-glucopyranoside (Sigma-Aldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was injected into the liposomal solution at 60°C, followed by agitation for 30 minutes at 60°C. Concentration was achieved by ultrafiltration to a final target volume, and buffer changes were performed 10 times with PBS pH 7.4 during diafiltration. The resulting liposomes, with the API presented on the surface of the liposomes, were then filtered by sterilization by passing through two 0.2 µm polycarbonate syringe filters in series, and the final product was stored at 5°C. Preparation of X, Y, Z and Z+ Liposome Vaccines

[00327] Liposome X and Y vaccines were produced using thin lipid film technology, followed by homogenization and extrusion.

[00328] Z+ liposome vaccines, with a final API concentration of 1200 pg / mL and a final T50 concentration of 1200 pg / mL, were produced by the ethanol injection followed by extrusion technique, and Z liposome vaccines, with a final API concentration of 400 pg / mL and a final T50 concentration of 100 pg / mL, were produced by thin lipid film technology followed by homogenization and extrusion.

[00329] The Z++ liposome vaccine, with a final concentration of Petition 870260044022, dated 11 / 05 / 2026, page 72 / 222 65 / 96 An API of 400 pg / mL and a final T50 concentration of 400 pg / mL were produced using thin lipid film technology followed by homogenization and extrusion.

[00330] Z+++ liposome vaccines, with a final API concentration of 1200 pg / mL and a final T50 concentration of 300 pg / mL, were produced by the ethanol injection followed by extrusion technique.

[00331] Preparation of X, Y, Z and Z++ Liposome Vaccines by the thin lipid film technique

[00332] Liposomal vaccines X, Y, Z, and Z++ were produced using thin lipid film technology followed by homogenization and extrusion. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands), and monophosphoryl hexaacyl 3-desacyl-synthetic Lipid A (3D-(6acyl)PHAD®) (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of 9:1:7:0.05 in EtOH at 60°C, except for liposome Y, which did not contain 3D-(6acyl)PHAD®. The ethanol was evaporated under vacuum rotary evaporator to obtain a thin lipid film.

[00333] The lipid film was rehydrated with PBS at pH 7.4, 5% DMSO (all from Sigma-Aldrich) containing 0.15 mg / mL of T50 peptide (Peptides & Elephants, Germany). The sample was gently agitated for 15 min and vigorously vortexed to dissolve the thin lipid film. The resulting multilamellar vesicles were subjected to 10 freeze-thaw cycles (liquid N2 and water bath at 37°C) and homogenized followed by sequential extrusion through polycarbonate membranes (Whatman, UK) with a pore size of 0.08 µm. The homogenization and extrusion steps were performed in an EmulsiFlex-C5 (Avestin, Canada). Liposomes extruded with encapsulated T50 peptide were concentrated by ultrafiltration, and the buffer was changed to PBS pH 7.4 by diafiltration. The resulting liposomes were diluted in PBS pH 7.4 and heated. Petition 870260044022, dated 11 / 05 / 2026, page 73 / 222 66 / 96 at 60°C to obtain a liposome solution before adding tau peptide and adjuvant.

[00334] CpG2006-cholesterol (CpG2006-Chol) (Microsynth, Switzerland) is a DNA oligonucleotide with all internucleotide linkages as thiophosphate that is modified at the 5' terminus with a cholesterol molecule via a phosphate linkage using a PEG spacer. CpG2006-Cholesterol (CpG2006-Chol) (Microsynth, Switzerland) was dissolved in PBS pH 7.4 at 1 mg / mL and injected into liposome solutions (except for liposome X, which does not contain CpG2006Chol) followed by incubation for 15 minutes before API insertion.

[00335] The API (Bachem AG, Switzerland) was dissolved in PBS pH 11.4 with 2% Octyl-εD-glucopyranoside (Sigma-Aldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was injected into the liposome solution at 60°C followed by stirring for 30 minutes at 60°C.Concentration was performed via ultrafiltration to obtain the target value (400 pg / mL API and 100 pg / mL T50 for liposomes X, Y, and Z; and 400 pg / mL API and 400 pg / mL T50 for liposome Z++), and buffer changes were performed 10 times with PBS pH 7.4 during diafiltration. The resulting liposomes with the API present on their surface were then sterilely filtered through 0.2 µm polycarbonate syringe filters, and the final product was stored at 5°C. Liposome O Preparation by Ethanol Injection Technique

[00336] The liposome vaccine O was produced by the ethanol (EtOH) injection technique followed by extrusion. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands) and MPLA (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of 9:1:7:0.05 in a 20:1 (by volume) mixture of EtOH and tert-butanol (t-BuOH) at 60°C. The lipid / ethanol solution was diluted in buffered saline solution. Petition 870260044022, dated 11 / 05 / 2026, page 74 / 222 67 / 96 with phosphate (PBS) pH 7.4 at 60°C to maintain the EtOH concentration at 10%, resulting in the formation of multilamellar liposomal vesicles (MLVs). The MLVs were then subjected to 5 sequential extrusion passes through three 0.08 µm pore size polycarbonate filters (Whatman) in series using the Emulsiflex-C5 (Avestin, Canada). The resulting liposomes were diluted in PBS pH 7.4 and heated to 60°C to obtain a liposome solution before the addition of the tau peptide.

[00337] The T46 peptide (Pepscan, Netherlands) was dissolved in PBS pH 7.4 at 1 mg / mL and injected into the liposome solutions, followed by incubation for 15 minutes before API insertion.

[00338] The API (Bachem, Switzerland) was dissolved in PBS pH 11.4 with 2% Octyl-D-glucopyranoside (Sigma-Aldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was injected into the liposome solution at 60°C followed by agitation for 30 min at 60°C. Concentration was achieved by ultrafiltration to obtain the target value (400 pg / mL of API and 100 pg / mL of T46), and buffer changes were performed 10 times with PBS pH 7.4 during diafiltration. The resulting liposomes with the API presented on the surface of the liposomes were then sterilely filtered through 0.2 µm polycarbonate syringe filters, and the final product was stored at 5°C. Preparation of Z+ and Z+++ Liposome Vaccines by Ethanol Injection

[00339] Z+ and Z+++ liposome vaccines were produced using an ethanol injection-based process. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands) and 3D-(6-acyl)PHAD® (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of approximately 9:1:7:0.04 in Petition 870260044022, dated 11 / 05 / 2026, page 75 / 222 68 / 96 EtOH at 60°C. The T50 peptide (Bachem AG, Switzerland) was dissolved in 10 mM His / 270 mM sucrose (pH 5.8 to 6.0). Then, the lipid-ethanol solution was injected into the solution containing the T50 peptide and gently stirred for 15 min, resulting in multilamellar vesicles (MLVs). The MLVs were homogenized (6 times for the Z+ liposome, and without homogenization for the Z+++ liposome), followed by sequential extrusion through polycarbonate membranes (Whatman, UK) with a pore size of 0.08 µm (5 passes for the Z+ liposome, 3 to 5 times for the Z+++ liposome). The homogenization and extrusion steps were performed in an EmulsiFlex-C5 (Avestin, Canada) for the Z+ liposome. The Z+++ liposome extrusion was performed using a LIPEX filter extruder. The extruded liposomes were concentrated by ultrafiltration, and the buffer was changed to 20 mM His / 145 mM NaCl pH 7.4 by diafiltration.The resulting liposomes with encapsulated T50 peptide were diluted in 20 mM His / 145 mM NaCl, pH 7.4, and heated to 60°C to obtain a liposome solution before the addition of the API and adjuvant.

[00340] CpG2006-Chol (Microsynth, Switzerland for the Z+ liposome; Avecia, USA for the Z+++ liposome) was dissolved in 20 mM His / 145 mM NaCl pH 7.4 at 1 mg / mL and injected into the liposomal solution followed by incubation for 15 minutes before API insertion.

[00341] The API (Bachem AG, Switzerland) was dissolved in carbonate buffer pH 10.2 with 1% octyl β-D-glucopyranoside (SigmaAldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was injected into the Z+ liposome solution at 60°C, followed by stirring for 30 min at 60°C. The peptide solution was mixed into the Z+ liposome solution using in-line mixing at 60°C, followed by stirring for 30 min at 60°C. Concentration was achieved by ultrafiltration to obtain the target value (1200 pg / mL of API and 1200 pg / mL of T50 for Z+ liposome; and 1200 pg / mL of API and 300 pg / mL of T50). Petition 870260044022, dated 11 / 05 / 2026, page 76 / 222 69 / 96 for Z+++ liposomes), and buffer changes were performed 10 times with 10 mM His / 270 mM Sucrose, pH 6.5 during diafiltration. The resulting Z+ liposomes with the API presented on the surface of the liposomes and the resulting Z+++ liposomes with the API presented on the surface of the liposomes were then sterilely filtered through 0.2 µm polycarbonate syringe / capsule filters, and the final product was stored at 5°C. Preparation of L, M and N Liposome Vaccines

[00342] L, M, and N liposome vaccines were produced using thin lipid film technology, followed by homogenization and extrusion. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands), and MPLA (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of 9:1:7:0.05 in EtOH at 60°C. The ethanol was evaporated under vacuum rotary evaporator to obtain a thin lipid film.

[00343] The lipid film was rehydrated with PBS pH 7.4, 5% DMSO (all from Sigma-Aldrich) containing either:

[00344] · 0.15 mg / mL of T48 peptide (Peptides & Elephants, Germany) - for M liposome; or

[00345] · 0.13 mg / mL of T50 peptide (Peptides & Elephants, Germany) - for L liposomes; or

[00346] · 0.15 mg / mL of T52 peptide (Peptides & Elephants, Germany) - for liposome N.

[00347] The sample was gently agitated for 15 minutes and vigorously vortexed to dissolve the thin lipid film. The resulting multilamellar vesicles were subjected to 10 freeze-thaw cycles (liquid N2 and water bath at 37°C) and then homogenized followed by sequential extrusion through polycarbonate membranes (Whatman, UK) with a pore size of 0.08 µm. The homogenization and extrusion steps were Petition 870260044022, dated 11 / 05 / 2026, page 77 / 222 70 / 96 liposomes were performed on an EmulsiFlex-C5 (Avestin, Canada). The extruded liposomes were concentrated by ultrafiltration and the buffer was changed to PBS pH 7.4 by diafiltration. The resulting liposomes with encapsulated T48, T50, or T52 peptide were diluted in PBS pH 7.4 and heated to 60°C to obtain a liposome solution before the addition of the tau peptide.

[00348] The API (Bachem AG, Switzerland) was dissolved in PBS pH 11.4 with 2% Octyl-εD-glucopyranoside (Sigma-Aldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was injected into the liposome solution at 60°C followed by agitation for 30 min at 60°C. Concentration was achieved by ultrafiltration to obtain a target value (400 pg / mL of API and 100 pg / mL of T48, T50, or T52), and buffer changes were performed 10 times with PBS pH 7.4 during diafiltration. The resulting liposomes with the API present on the surface of the liposomes were then sterilely filtered through 0.2 µm polycarbonate syringe filters, and the final product was stored at 5°C. Preparation of R, S and T Liposome Vaccines

[00349] Liposome vaccines R, S, and T were produced using a process based on ethanol injection followed by extrusion. First, DMPC (Lipoid GmbH, Ludwigshafen, Germany), DMPG (Lipoid GmbH, Ludwigshafen, Germany), cholesterol (Dishman, Netherlands), and 3D-(6-acyl)PHAD® (Avanti Polar Lipids, AL, USA) were solubilized in a molar ratio of 9:1:7:0.04 in EtOH at 60°C. For liposomes R and T, the lipid-ethanol solution above was mixed with 10 mM histidine, pH 5.8, supplemented with 270 mM sucrose to achieve 10% solvent (EtOH), and then incubated for 30 minutes at 60°C. For the S liposome, the T50 peptide (Bachem AG, Switzerland) was dissolved in 10 mM His / 270 mM sucrose (pH 5.8 to 6.0). The lipid-buffer mixture related to the R, S, and T liposomes was gently stirred for 15 min, resulting in multilamellar vesicles. Petition 870260044022, dated 11 / 05 / 2026, page 78 / 222 71 / 96 (MLVs). The resulting multilamellar vesicles were extruded through polycarbonate membranes (Whatman, UK) with a pore size of 0.08 µm (5X) performed in a high-pressure EmulsiFlex-C5 system (Avestin, Canada).

[00350] The extruded liposomes were concentrated by ultrafiltration and the buffer was changed to 20 mM His / 145 mM NaCl pH 7.4 by diafiltration. The resulting liposomes with T50 encapsulated for liposome S and the resulting liposomes R and T were further diluted in 20 mM His / 145 mM NaCl pH 7.4 and heated to 60°C to obtain a liposomal solution before the additions of API and T57 for liposome T.

[00351] For liposome T, T57 was dissolved at 1 mg / mL in 1% Octylβ-D-glucopyranoside (Sigma-Aldrich, USA) in deionized distilled water and inserted into the liposome, followed by incubation for 15 minutes at 60°C before API insertion.

[00352] The API (Bachem AG, Switzerland) was dissolved in carbonate buffer pH 10.2 with 1% octyl-β-D-glucopyranoside (SigmaAldrich, USA) at a concentration of 1 mg / mL, and the peptide solution was mixed into the liposome solution at 60°C, followed by stirring for 30 min at 60°C. Concentration was performed by ultrafiltration to obtain the following target value:

[00353] - 1200 pg / mL of API for liposome R;

[00354] - 1200 pg / mL of API and 300 pg / mL of T50 for the liposome S; and

[00355] - 1200 pg / mL of API and 300 pg / mL of T57 for the liposome T.

[00356] Buffer exchange was performed 10 times with 10 mM His / 270 mM sucrose pH 6.5 during diafiltration. The resulting liposomes with the API presented on the surface of the liposomes were then sterilely filtered by passing through syringe filters. Petition 870260044022, dated 11 / 05 / 2026, page 79 / 222 72 / 96 polycarbonate of 0.2 µm and the final product was stored at 5°C. Example 2: Preparation of Conjugate Vaccine Peptides and Adjuvants

[00357] The sequences of two multiphosphorylated peptide epitopes (TAUVAC-p7.1 and TAUVAC-p22.1, which have three and two phosphorylated amino acids, respectively) were refined by optimizing length to better bind to B cell surface immunoglobulin and to ensure that the sequences do not contain epitopes predicted to bind to human HLA class IA, B, and C molecules with high affinity. The latter criterion was important to avoid inducing a cytotoxic CD8+ T cell response against tau that could potentially cause significant neuronal damage. Using the T cell epitope prediction tool from the Immune Epitope Database and Analysis Resources, the TAUVAC-p7.1 peptide did not show predicted epitopes capable of binding to human HLA class IA, B, C, and HLA class II DQ and DR molecules with high affinity, while the TAUVAC-p22 peptide was predicted.1 contained epitopes that bound to HLA class II DQ and DR molecules with intermediate / high affinity (data not shown).

[00358] The phosphorylated tau peptides (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3) used in this study were synthetically produced (Pepscan, NL) with the phosphoresidues added during synthesis. A conjugate comprising phosphorylated tau peptide having the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 covalently linked to a KLH carrier via a ligand is hereby referred to as Conjugate B or Conjugate C, respectively. A conjugate comprising phosphorylated tau peptide having the amino acid sequence of SEQ ID NO: 2 covalently linked to a CRM carrier via a ligand is hereby referred to as Conjugate A.

[00359] To manufacture Conjugates B and C, vaccine peptides Petition 870260044022, dated 11 / 05 / 2026, p. 80 / 222 73 / 96 were conjugated to the KLH carrier protein via a melaminemidobenzoyl-N-hydroxysuccinimide ester (MBS) ligand and an extra cysteine ​​at the N-terminus of the peptide. Unbound peptide was removed using a Sephadex G25 column before concentrating the conjugate. The conjugates were mixed prior to injection into a potent multicomponent adjuvant (Sigma Adjuvant System, Sigma-Aldrich) or a single-component depot adjuvant (aluminum hydroxide, Alhydrogel®, Invivogen) following the manufacturer's instructions.

[00360] Vaccine peptides were conjugated to the CRM197 carrier protein via a polyethylene glycol (PEG) cysteine-acetamidopropionamide linker. The phosphorylated tau peptide having the amino acid sequence SEQ ID NO: 2 was synthetically produced (Polypeptide Laboratories SAS), with phosphorus residues and the PEG3 spacer added during synthesis. Conjugate A was fabricated by conjugating the CRM197 carrier protein via a succinimidyl 3-(bromoacetamide) propionate (SBAP) linker to a cysteine ​​at the N-terminal of the peptide. The SBAP was linked to primary amines of the CRM197 protein (-NH2) via NHS ester reaction chemistry. Excess SBAP linker was removed using ultrafiltration and diafiltration (UF / DF). The intermediate CRM197-SBAP was conjugated with the phosphorylated tau peptide and, once the reaction was complete, the conjugation reaction was stopped by adding an excessive amount of L-cystine to cool the reaction.The crude product conjugated with CRM197 peptide was purified using a Capto Q ImpRes chromatography column (GE Healthcare) and eluted using an isocratic salt method. The purified CRM197 peptide product was then formulated in 20 mM Tris, 250 mM sucrose, pH 8.1 at a concentration of 0.5 mg / mL using UF / DF. The CRM197 tau peptide drug substance (DS) was generated by adding a 10% PS80 stock buffer to achieve a final PS80 concentration of 0.01%. Petition 870260044022, dated 11 / 05 / 2026, page 81 / 222 74 / 96 was thoroughly mixed before filtration. Example 3: Vaccine-Induced IgG Antibodies Specific for Tau Phosphopeptide

[00361] All animal experiments were approved and conducted in accordance with local legislation on animal experimentation. Rhesus monkeys (Macaca mulatta) were obtained from Kunming Biomed International Ltd, China, Yunnan Yinmore Bio-Tech Co. LTD, China, and Yunnan Laboratory Primates Inc., China. The animals were two to five years old at the start of immunization and had a minimum weight of 2.5 kg. A detailed clinical examination was performed before the start of treatment and weekly thereafter. In addition, the monkeys were observed twice daily and clinical signs were recorded.

[00362] Adult Rhesus monkeys (n = 3 males and 3 females per group) were immunized subcutaneously with 1800 pg of tetrapalmitoylated phosphorylated tau peptide with SEQ ID NO: 2 acetate per dose of the control liposomal vaccine (liposome with tetrapalmitoylated phosphorylated tau peptide of SEQ ID NO: 2 and MPLA) or a liposomal vaccine according to the application modality, for example, Liposome Z (liposome with tetrapalmitoylated phosphorylated tau peptide of SEQ ID NO: 2, 3D-(6-acyl)PHAD®, CpG 2006 lipid oligonucleotide and T50 T cell peptide) or 15 pg per dose of a conjugate vaccine according to an embodiment of the invention (for example, Conjugate A, phosphorylated tau peptide of SEQ ID NO: 2 linked to CRM197) co-injected with alum and CpG oligonucleotide CpG 2006 on days 1, 29, and 85. Bleeding was performed before immunization and on days 8, 22, 36, 50, 64, 78, 92, 106, 120, 134, and 148, and sera were isolated.

[00363] Specific IgG antibody titers were determined by ELISA using the phosphorylated tau peptide of SEQ ID Petition 870260044022, dated 11 / 05 / 2026, page 82 / 222 75 / 96 NO: 2 as a coating antigen. Serum from an individual immunized monkey was serially diluted in assay buffer (PBS, 0.05% Tween20, 1% BSA) and applied to 96-well plates that were coated with the relevant peptide. After two hours of incubation, samples were removed and plates washed in PBST (PBS, 0.05% Tween20). Antibodies were detected using an HRP-conjugated anti-monkey IgG (KPL), followed by ABTS substrate (Roche). All samples were run at eight two-fold dilutions, with positive and negative control samples included in each plate. Data were expressed as the geometric mean of outcome titers (last serum dilution inducing a positive response) per group.

[00364] As shown in Figure 4, the liposome Z vaccine and conjugate A induced higher titers of phosphopeptide-specific IgG compared to the control liposomal vaccine. Example 4: Vaccine-Induced Antibodies Specific to Pathological Tau Structures in the Human Brain

[00365] All brain tissues were obtained from the Netherlands Brain Bank (NBB) and were collected from donors after signing an informed consent form for a brain autopsy and the use of the samples, as well as their clinical information, for research purposes. Paraffin sections from non-demented (healthy) controls, Alzheimer's disease (AD), frontal temporal lobe dementia with tau pathology (FTD-tau), Pick's disease, primary age-related tauopathy (PART), and progressive supranuclear palsy (PSP) were used. Brain regions included parietal cortex, middle frontal gyrus, hippocampus, or caudate nucleus.

[00366] In particular, formalin-fixed paraffin-embedded sections from parietal cortices of a control (healthy) human subject and a human subject suffering from the disease of Petition 870260044022, dated 11 / 05 / 2026, page 83 / 222 76 / 96 Alzheimer's disease (AD Braak V / VI) sections were stained with postimmune monkey serum diluted 1:100 in normal antibody diluent (Immunological). The sections were then washed and stained with an anti-goat monkey HRP (Abcam). The staining was finally visualized using 3,3'-diaminobenzidine (ADB; Dako), which deposits a specific brown stain in the presence of horseradish peroxidase (HRP). The slides were counterstained with hematoxylin, dehydrated, and mounted with Quick D mounting medium (Klinipath). Images were captured with a Leica DC500 microscope.

[00367] The results in Figure 5 show that post-immunization sera from Rhesus monkeys immunized with liposome Z (liposome with tetrapalmitoylated phosphorylated tau peptide of SEQ ID NO: 2, 3D-(6-acyl)PHAD®, lipidated CpG oligonucleotide CpG 2006 and T cell peptide T50) stained pathological tau structures in human brain sections. Serum was collected from Rhesus monkeys on day 106 after primary immunization with the enhanced liposomes. This monkey received immunizations at months 0, 1, and 3 prior to serum collection. The left panel (AD Braak V / VI) shows staining of the parietal cortex from a Stage V Braak donor. Arrows indicate the staining of tau tangles. The right panel (healthy) shows the parietal cortex staining of a Stage 0 Braak donor. Serum was applied to the sections at a 1:100 dilution, followed by goat anti-monkey antibody at 1:100, and the staining was visualized using a DAB developer.

[00368] The results in Figure 6 show that serum from Rhesus monkeys immunized with Conjugate A containing phosphorylated tau peptide from SEQ ID NO:2 plus soluble CpG and alum hydroxide binds to pathological tau structures in human AD brain sections. Serum was collected on day 106 after primary immunization with the CRM conjugate vaccine. These monkeys received immunizations on Petition 870260044022, dated 11 / 05 / 2026, page 84 / 222 77 / 96 months 0, 1, and 3 before serum collection. The upper panels (AD) show parietal cortex staining, including tau tangles, from a Stage V Braak donor. The lower panels (CTRL) show parietal cortex staining from a Stage 0 Braak donor. Serum was applied to the sections at a 1:100 dilution, followed by 1:100 goat monkey anti-antibody, and staining was visualized using a DAB developer. Example 5: Liposomal Vaccines with One or Two Adjuvants

[00369] The addition of two adjuvants in the enhanced liposomal vaccine increases the level of titers of specific IgG antibodies to tau phosphopeptides, as well as the consistency of the antibody response among individuals.

[00370] Adult Rhesus monkeys (n = 3 males and 3 females per group) were immunized subcutaneously on days 1, 29, 85, and 169 with 1800 pg of tetrapalmitoylated phosphorylated tau peptide with SEQ ID NO: 2 acetate per dose of the control liposomal vaccine or the enhanced liposomal vaccine with encapsulated T50 cell epitope, containing 3D-(6-acyl)PHAD® adjuvant alone (liposome X, Figure 7A), CpG 2006 lipid oligonucleotide adjuvant alone (liposome Y, Figure 7B), or both 3D-(6-acyl)PHAD® adjuvants and CpG 2006 lipid oligonucleotide (liposome Z, Figure 7C). Bleeding samples were collected before immunization and on days 8, 22, 36, 50, 64, 78, 92, 106, 120, 134, 148, 162, 176, and 190, and sera were isolated. Specific IgG antibody titers in the sera were determined by ELISA using phosphorylated tau peptide of SEQ ID NO: 2 as a coating antigen and an anti-monkey IgG secondary antibody.The resulting antibody levels are presented as outcome titers (last serum dilution inducing a positive response) for each individual monkey over time. Each immunization group is represented in a panel (Figures 7A-C). A. Petition 870260044022, dated 11 / 05 / 2026, page 85 / 222 The geometric mean of outcome titers per group (78 / 96) ± 95% confidence interval is presented in Figure 7D. In summary, Figures 7A-D show that the inclusion of two adjuvants in the liposomal vaccine containing encapsulated T50 improved the level and consistency of the antibody response to tau phosphopeptide, resulting in less variability in antibody response among individual monkeys. More specifically, as shown in Figure 7D, liposomal vaccines enhanced with phosphorylated tau peptide of SEQ ID NO: 2, T50 cell epitope (liposome X, Y, and Z), and 1 or 2 adjuvants induced higher titers against tau phosphopeptide than the control liposomal vaccine without T cell epitope. All monkeys responded when injected with each of the enhanced liposomal vaccines, while 4 of 6 animals responded with the control liposomal vaccine. Example 6: Vaccine-Induced Antibodies Specific for Enriched Paired Helical Filaments (ePHF)

[00371] Groups of Rhesus monkeys (n = 3 males and 3 females per group) were immunized subcutaneously by vaccination on day 1 and day 29 with (i) the enhanced liposomal vaccine containing the T50 cell epitope and 3D-(6-acyl) PHAD® adjuvant alone (liposome X), (ii) the enhanced liposomal vaccine containing the T50 cell epitope and CpG 2006 lipid adjuvant alone (liposome Y), (iii) the enhanced liposomal vaccine containing the T50 cell epitope and two adjuvants (3D-(6-acyl) PHAD® and CpG2006 lipid, liposome Z) or (iv) the conjugate vaccine (phosphorylated tau peptide of SEQ ID NO: 2 linked to CRM197) co-injected with alum and CpG oligonucleotide CpG 2006 (Conjugate A).

[00372] Enriched paired helical filament (ePHF) preparations were obtained from brain tissue after death from individuals with histologically confirmed AD by extraction. Petition 870260044022, dated 11 / 05 / 2026, page 86 / 222 79 / 96 sarcosyl insoluble tau, using a modified method of Greenberg and Davies (Greenberg and Davies, 1991, Proc Natl Acad Sci USA, 87 (15): 5827-31). Titrations of specific antibodies to enriched paired helical filaments (ePHF) were assessed using the Mesoscale Discovery (MSD) platform. MSD streptavidin plates were coated with biotinylated anti-tau antibody (HT7-biotin, ThermoScientific) prior to incubation with ePHF isolated from Alzheimer's disease patients, while ePHF-specific IgG antibodies were further detected using a SulfoTag-labeled anti-human IgG antibody that reacts with monkey IgG antibodies. More specifically, ePHF was added to 96-well small streptavidin plates MSD Gold (MSD) previously saturated with 1% BSA and coated with biotinylated HT-7 (Thermo Scientific).After one hour of incubation, the plates were washed with PBST and serial dilutions of the sera were added and incubated for two hours. Bound antibodies were detected using a SulfoTag-labeled anti-human IgG antibody followed by a fixation step in 1% PFA before the addition of T-reading buffer. The plates were analyzed using a Sector Imager (MSD). Results were expressed in arbitrary units per milliliter (AU / mL) for each individual monkey, along with the geometric mean per group. Titrations of ePHF-specific antibodies on day 50 after the first immunization are shown.

[00373] Figure 8 shows that all vaccines induced high titers of specific IgG antibodies to ePHF.

[00374] Similar results with high titers of specific IgG antibodies for ePHF were also obtained with other liposomes, such as the Z+ liposome, administered to Rhesus monkeys by intramuscular administration. Example 7: The Specific Antibody Amplitude for Petition 870260044022, dated 11 / 05 / 2026, page 87 / 222 80 / 96 Tau Phosphopeptide Induced by Liposomal Vaccine and Conjugate Vaccine in Rhesus Monkeys

[00375] Groups of Rhesus monkeys (n = 3 males and 3 females per group) were immunized subcutaneously on days 1 and 29 with (i) the enhanced liposomal vaccine containing encapsulated T50 and two adjuvants: TLR4 ligand (3D-(6-acyl)PHAD®) and lipid-coated CpG 2006 oligonucleotide (liposome Z) and (ii) the conjugate vaccine (phosphorylated tau peptide of SEQ ID NO:2 linked to CRM) (conjugate A) co-injected with alum and CpG 2006 oligonucleotide. The antibody epitope recognition profile was determined by epitope mapping ELISA three weeks after the second immunization (day 50) using a library of N-terminal biotinylated 8-mer peptides, displaced by one amino acid and covering the tau peptide sequence. phosphorylated from SEQ ID NO: 2, as well as the sequence from SEQ ID NO: 4 (VYKSPVVSGDTSPRHL, non-phosphorylated tau peptide having the same amino acid sequence as SEQ ID NO: 2) and the corresponding full-length biotinylated peptides.

[00376] Figure 9 shows that monkeys immunized with liposome Z produced IgG antibodies that bind primarily to the N-terminal portion of the phosphorylated peptide of SEQ ID NO: 2 (Figure 9A), while monkeys immunized with the conjugate vaccine (phosphorylated tau peptide of SEQ ID NO: 2 linked to CRM) generated IgG antibodies that bind primarily to the C-terminal portion of the tau peptide of SEQ ID NO: 2 (Figure 9B). Example 8: Increased Titers of Tau Phosphopeptide-Specific IgG Antibodies Induced by Liposomal Vaccine with Encapsulated T-Cell Epitope

[00377] Three groups of C57BL / 6J mice (n = 10 per group) were immunized subcutaneously on days 0 and 14 with i) a liposomal vaccine containing a TLR4 agonist (3D-(6-acyl)PHAD®), Petition 870260044022, dated 11 / 05 / 2026, page 88 / 222 81 / 96 (liposome R), ii) liposomal vaccine containing encapsulated T50 T-cell epitope and TLR4 ligand (3D-(6-acyl)PHAD®) as adjuvant (liposome S), or iii) liposomal vaccine containing T57 T-cell epitope anchored to the liposomal surface (i.e., dipalmitoylated T50) and TLR4 ligand (3D-(6-acyl)PHAD®) as adjuvant (liposome T). The level of IgG antibodies specific for the phosphorylated tau peptide of SEQ ID NO: 2 was measured 21 and 35 days after the first injection into mouse plasma by ELISA; results were presented as individual mouse values, along with the geometric mean per group ± 95% CI expressed in arbitrary units (AU) per mL.As shown in Figure 10A, vaccination with the liposomal vaccine containing encapsulated T50 (Liposome S) induced significantly higher antibody titers than the control liposomal vaccine (Liposome R) and the liposomal vaccine containing anchored T-cell epitope (Liposome T) 21 days after the first immunization (Kruskal-Wallis test: p = 0.0089 and p = 0.002, respectively) and also higher antibody titers than the control liposomal vaccine and significantly higher antibody titers than the liposomal vaccine containing anchored T-cell epitope 35 days after the first immunization (Kruskal-Wallis test: p = 0.7591 and p = 0.053, respectively) (Figure 10B). Example 9: T-Cell Response Induced by Liposomal Vaccines Specific for the Incorporated T-Cell Epitope

[00378] Three groups of C57BL / 6J mice (n = 5 per group) were immunized subcutaneously on days 0, 14, and 28 with (i) the liposomal vaccine enhanced with encapsulated T48 T-cell peptide (containing T-cell epitopes PADRE, T2, T30, and T17 separated with the GS ligand) and a TLR4 agonist as an adjuvant (MPLA) (Liposome M), (ii) the liposomal vaccine enhanced with T52 Petition 870260044022, dated 11 / 05 / 2026, page 89 / 222 82 / 96 encapsulated (containing T cell epitopes PADRE, T2, and T30 separated with the RK ligand) and a TLR4 agonist (MPLA) as an adjuvant (Liposome N) or (iii) PBS. Mouse spleens were harvested 42 days after the first immunization for analysis of T cell responses to IL-4 and IFN-γ ELISPOT. Single-cell suspensions were incubated with medium, T48 or T52 peptide at 10 pg / mL for 48 hours. Plates were incubated with biotinylated anti-mouse IL-4 or IFN-γ monoclonal antibodies and with streptavidin alkaline phosphatase (AP). Staining was revealed by adding the AP substrate. Figure 11 shows that re-stimulation of mouse splenocytes with the same peptide as that encapsulated in the liposome induced IL-4 spot-forming cells (Figure 11B) and IFN-γ (Figure 11A), while splenocytes from mice injected with PBS did not.This confirmed that adding a T-cell epitope to the vaccine induced the activation of specific T cells, allowing them to provide even more assistance in producing antibodies for tau-specific B cells. Example 10: Liposomal Vaccines Containing Encapsulated T-Cell Epitope and Anchored T-Cell Epitope

[00379] Groups of Rhesus monkeys (n = 6 per group) were immunized subcutaneously on days 1, 29, 85, and 169 with (i) a liposomal vaccine containing the encapsulated T50 T-cell epitope and TLR4 ligand (MPLA) as an adjuvant (liposome L), (ii) a liposomal vaccine containing the anchored T46 T-cell epitope and TLR4 ligand (MPLA) as an adjuvant (liposome O), and (iii) a control liposomal vaccine containing a TLR4 ligand (MPLA) as an adjuvant and no T-cell epitope. Bleeding was performed before immunization (on day -14) and on days 8, 22, 36, 50, 64, 78, 92, 106, 120, 134, 148, 148, Sera 162, 176, and 190 were isolated. Specific IgG antibody titers were determined by ELISA using the tau peptide. Petition 870260044022, dated 11 / 05 / 2026, page 90 / 222 83 / 96 phosphorylated SEQ ID NO: 2 antigen as a coating antigen and an anti-monkey IgG secondary antibody. The resulting antibody levels were calculated as outcome titers (last serum dilution inducing a positive response) and the data were expressed as geometric mean per group. As shown in Figure 12, the liposomal vaccine containing an encapsulated T-cell epitope (L liposome) and the liposomal vaccine containing an anchored T-cell epitope (O liposome) each induced higher titers of tau phosphopeptide-specific antibodies than the control liposomal vaccine without a T-cell epitope. Example 11: Antibody Response in Mice Induced by a Conjugate Vaccine

[00380] Female BALB / c mice (14 mice per group) were immunized with Conjugate B or Conjugate C (containing SEQ ID NO: 1 or SEQ ID NO: 3 covalently linked to KLH) following the scheme shown in Figure 13A and using vaccine candidates adjuvanted with a potent multicomponent adjuvant (Sigma Adjuvant System®, Sigma-Aldrich, then called Ribi) or a single-component depot adjuvant (Alhydrogel® 2% adjuvant or aluminum hydroxide gel, InvivoGen, then called alum). The amino acid sequence of SEQ ID NO: 1 contains only one amino acid difference compared to the mouse protein, while the sequence of SEQ ID NO: 3 is 100% conserved between humans and mice. Thus, the selected epitopes can reasonably be considered autoproteins for mice, and mice should be a relevant model for investigating the limitations that immune tolerance may place on immunogenicity.

[00381] As a first measure of vaccine immunogenicity, flow cytometry was used to measure T cell induction. Petition 870260044022, dated 11 / 05 / 2026, page 91 / 222 84 / 96 follicular helper cells (TfHs) in cervical lymph nodes draining the vaccine injection site (four mice per group). TfHs are a specialized population of CD4+ T cells characterized by the expression of CXCR5, PD-1, and ICOS, among other molecules. TfHs expand after exposure to a vaccine or other immune stimuli and support affinity maturation of B cells in the germinal center (Crotty, 2011, Annual Reviews of Immunology. Vol. 29: p. 621-663). The number of induced TfHs correlates positively with the protective efficacy of vaccines in humans (Bentebibel et al., 2013, Sci Transl Med., 5 (176): 176ra32; Spensieri et al., 2013, Proc Natl Acad Sci USA., 110 (35): 14330-5) and small animals. As shown in Figure 13B, both vaccines, as well as the KLH control immunization plus adjuvant, induced measurable TfHs in vaccinated mice.Furthermore, all animals that received active vaccine (Conjugate B and Conjugate C groups) or active placebo (KLH) plus alum had significantly higher TfHs than animals that received an inactive placebo (PBS group) when cervical lymph node drainage was collected seven days after the first immunization (P = 0.0044 for KLH-TAUVAC-p7.1; P = 0.0482 for KLH-TAUVAC-p22.1; P = 0.0063 for KLH, using an ANOVA test followed by Dunnett's adjustment for multiple comparisons).

[00382] ELISA was performed to determine the serum titer of antibodies binding to tau phosphopeptides and KLH on day 0 and at four additional time points after immunization (days 14, 28, 56, and 84, see Figures 13C, D, G, and H). As shown in Figure 13C, immunization with Conjugate B induced binding antibodies reactive against the corresponding vaccine peptide.For animals immunized with Conjugate B and Ribi adjuvant, binding titers against the vaccine peptide were significantly higher than binding titers induced by the active placebo (compare the... Petition 870260044022, dated 11 / 05 / 2026, p. 92 / 222 85 / 96 conjugated B plus Ribi to KLH plus Ribi) at all time points measured (P < 0.001 using an ANOVA test followed by Tukey's adjustment for multiple comparisons). For the group with alum adjuvant, the difference was significant only on days 56 and 84 (P = 0.001 and 0.012, respectively).

[00383] The tau-specific antibody response to Conjugate C (Figure 13D) was of a generally lower magnitude than the response to Conjugate B, although assay differences (different coating peptide) preclude direct statistical comparison between the two vaccines. However, antibody titers against Conjugate C were significantly higher in mice vaccinated with Conjugate C plus Ribi than in mice that received active placebo KLH-Ribi on days 28 and 84 (P = 0.001 and 0.008, respectively) after immunization; titers in the alum adjuvant group were not significantly different from those in the active placebo group.

[00384] Although the carrier protein protects the phosphopeptide from degradation in vivo to some extent, it was likely that phosphatase digestion of the peptide antigens in vivo would expose some unphosphorylated peptide to the immune system. To determine whether this exposure resulted in the generation of antibodies capable of binding to the unphosphorylated peptide in Conjugate B and Conjugate C, ELISA was performed using unphosphorylated peptides as the coating antigen. As shown in Figures 13E-F, the response to unphosphorylated tau peptides was low compared to the response of active placebos to the same unphosphorylated peptide. Furthermore, in animals immunized with Conjugate B and Ribi, the titers of binding to the phosphorylated peptide were significantly higher than the titers of binding to the non-phosphorylated peptide at all time points measured (P = 0.009 on day 14; P < 0.0001 on days 28, 56, and 84 using a Petition 870260044022, dated 11 / 05 / 2026, p. 93 / 222 86 / 96 ANOVA test). For the group with alum adjuvant, the difference was significant only on days 56 and 84 (P = 0.0002 and 0.001, respectively). For animals immunized with Conjugate C, responses to the phosphorylated peptide were greater only when Ribi adjuvant was used (P < 0.0001 on day 28; P = 0.0001 on days 56 and 84). Example 12: Antibodies Induced by Conjugate Vaccines Bind to Physiologically Relevant Forms of Altered Tau

[00385] To further determine whether vaccine-induced antibodies could bind to physiologically relevant forms of altered tau, post-immunization sera from vaccinated mice were used to stain post-mortem human brain sections collected from patients with Alzheimer's disease (5 AD cases), patients affected by other tauopathies (3 PART, FTD, PICK, and PSP cases), or healthy age-matched controls (5 control cases, CTRL). As expected, sera from control animals (PBS and active placebo groups) did not bind to brain sections, while AT8, a monoclonal antibody that binds to pTau [pSer202, pThr205] obtained from a murine clone, showed strong immunoreactivity of the tau pathology in an adjacent tissue section from the corresponding area (Figure 14).Sera from animals immunized with the active Conjugate B and Conjugate C vaccines bind to pathological tau structures not only in AD sections (data not shown), but also in those of other tauopathies (Figure 14). Conjugate B-induced antibodies reacted with (pre-)tangles, neuropil strands, and neuritic plaques in AD cases. These post-immunization sera were also able to immunoreact with neurofibrillary tangles and neuropil strands in PART brain tissue, neuronal inclusions and neuropil strands in FTD-tau tissue (MAPT P301S), inclusions and astrocytes in some cases of Pick's disease, and finally, Petition 870260044022, dated 11 / 05 / 2026, p. 94 / 222 87 / 96 as neuronal inclusions, neuropil threads, and astrocytes typical of PSP. Polyclonal sera induced by the C-conjugate also reacted to the characteristics of the pathological tau structures of each tauopathy. In AD cases, staining was primarily focused on neurofibrillary tangles and, to a lesser extent, on neuritic plaques and neuropil threads. Lower magnification of the corresponding areas showed similar results (unknown data). Example 13: Vaccine-Induced Antibodies Are Functional in Mice

[00386] The protective efficacy of the Conjugate B vaccine was tested in a tauopathy injection model (Peeraer et al., 2015, Neurobiol Dis., 73: 83-95). In this model, mice susceptible to tauopathy due to a genetic mutation (P301L) receive an intracerebral injection of enriched PHF isolated from the human AD brain, following the schedules indicated in Figure 15A. The injection, performed before the onset of transgene-induced tauopathy, accelerates the development of tauopathy in these animals. On the other hand, when the ePHF seed is premixed with an antibody capable of suppressing tau seeding activity such as AT8, the induction of tauopathy is reduced (unpublished data, not shown).

[00387] Following the scheme in Figure 15A, the development of tauopathy was evaluated after stereotaxic injection of human PHF enriched premixed with IgG purified from serum of animals immunized with Conjugate B, Ribi, or the active control KLH-Ribi. Two months after injection, the brains of these mice were harvested, and the amount of tau aggregated in the total and sarkosil-insoluble fractions was determined using standard biochemical analysis. The data obtained showed that when mice were injected with ePHF that had been premixed with IgG from mice vaccinated with Conjugate B, there was Petition 870260044022, dated 11 / 05 / 2026, page 95 / 222 88 / 96 significantly less phospho-tau aggregated in the total (Figure 15B) and sarkosil-insoluble (Figure 15C) fractions compared to animals that received the control injection (p < 0.0001 KLH Ribi vs KLHTAUVAC-p7.1 Ribi using an ANOVA test followed by Holm-Bonferroni adjustment for multiple comparisons). As sarkosil-insoluble tau is well accepted to correlate with the pathological characteristics of tauopathy, this result demonstrates that antibodies induced by vaccination with KLH-TAUVAC-p7.1 are protective in vivo. Example 14: Vaccine-Induced Antibodies Are Functional in Non-Human Primates

[00388] Rhesus monkeys were immunized with alum-adjuvanted CpG oligonucleotide conjugate B (n = 6) or with KLH (n = 2) on days 1, 29, 85, and 169. Blood was collected every 14 days, and sera from animals immunized with conjugate B were tested for reactivity to the immunization peptide using ELISA (Figure 16A) and human ePHF using MSD (Figure 16B). Immunization with conjugate B resulted in a sustained and consistent antibody response against the vaccine phosphopeptide. In addition, all animals showed measurable levels of antibodies against human ePHF, with 3 of 6 animals showing high reactivity to this antigen. Sera collected from animals 50 days after primary immunization were applied to human brain sections from healthy individuals or patients with AD (Figure 16C).Post-immunization sera from the Conjugate B group stained pathological tau structures, i.e., neurofibrillary tangles, neuropil strands, and neuritic plaques in AD brain tissue, while sera from KLH-immunized mice showed no reactivity. No staining was observed in control tissue. When tested in the tau immunodepletion assay, animals that received Conjugate B had antibodies capable of binding to and depleting tau seed (p = 0.03 on day 50). Petition 870260044022, dated 11 / 05 / 2026, page 96 / 222 89 / 96 using an ANOVA test followed by Dunnett's adjustment for multiple comparisons), while KLH immunization did not trigger such antibodies (Figure 16D). Pre- and post-immunization sera were also tested in the neutralization assay as serially diluted individual samples (Figure 16E). Changes from baseline (CFB) were calculated as the difference between FRET counts for readings on day -14 before vaccination (baseline) and after vaccination on days 50, 106, and 190, respectively. The response on a specific post-vaccination day (diai) was calculated as follows:

[00389] Response = %FRET_diai — %FRET_baseline

[00390] A general linear mixed model was applied to the responses mentioned above, with animal as a random effect, with vaccine groups, days, and variable serum levels treated as categorical variables and all their interactions. Given the exploratory nature of the study, no multiple test adjustment was considered. Hypothesis testing was performed at the 5% significance level. Example 15: Mice immunized with the conjugate vaccine in combination with alum and CpG oligonucleotide adjuvants resulted in higher titer antibody responses to the vaccine peptide.

[00391] Adult female C57BL / 6 mice (n = 5 to 6 per group) were immunized intramuscularly with 2 µg (Figure 17A) or 0.2 µg (Figure 17B) of Conjugate A vaccine. The conjugate vaccine was administered alone (without adjuvant), with alum hydroxide, with CpG oligonucleotide, or with alum and CpG oligonucleotide combined. All mice received a primary immunization on day 0 of the study, followed by a single booster immunization on day 28. The dose for the alum adjuvant was 500 µg per mouse per injection and the dose for the CpG oligonucleotide adjuvant was 20 µg per mouse per injection. The graphs in Figure 17 Petition 870260044022, dated 11 / 05 / 2026, p. 97 / 222 Figures 90 / 96 show the results of linkage ELISA using serum collected from mice before immunization (day 0) and at two time points after immunization (days 28 and 42) with the T3.5 vaccine peptide as the coating antigen. Mean outcome-specific titers of T3.5 per group are represented, with error bars representing standard error. The tables show the statistical analysis of the results, in which antibody titers were compared using the non-parametric Kruskal-Wallis test, and comparisons between groups were assessed by the Wilcoxon Signed Rank test as a post-hoc test to the Kruskal-Wallis test.

[00392] The results shown in Figure 17 illustrate that at both doses, the unadjuvanted vaccine failed to induce a strong immune response. The use of alum or CpG oligonucleotide or a combination of both improved the magnitude of the antibody response (p < 0.0152). Furthermore, for animals immunized with 2 µg of vaccine, the adjuvant combination provided significantly higher antibody titers than the adjuvants alone on day 28 (p = 0.0028). The alum-CpG oligonucleotide combination also performed better than CpG oligonucleotide alone for animals immunized with 0.2 pg of vaccine on day 42 (p = 0.0497). These data support the use of the alum and CpG oligonucleotide adjuvant combination. Example 16: Liposomal vaccines with different Tau peptide ratios: T-cell epitopes induce high and sustained levels of specific IgG antibody titers for tau phosphopeptides.

[00393] Adult Rhesus monkeys (n = 6 per group) were immunized subcutaneously on days 1, 29, 85, and 169 with 1800 pg of tetrapalmitoylated phosphorylated tau peptide with SEQ ID NO: 2 acetate per dose in the T-cell epitope-enhanced liposomal vaccine. Petition 870260044022, dated 11 / 05 / 2026, p. 98 / 222 91 / 96 encapsulated T50, containing 3D-(6-acyl) PHAD® adjuvant and lipid-coated CpG 2006 oligonucleotide with: i) 400 pg / mL of phosphorylated tau peptide of SEQ ID NO: 2 and 100 pg / mL of T50 (Z liposome), ii) 1200 pg / mL of phosphorylated tau peptide of SEQ ID NO: 2 and 1200 pg / mL of T50 (Z+ liposome), iii) 400 pg / mL of phosphorylated tau peptide of SEQ ID NO: 2 and 400 pg / mL of T50 (Z++ liposome), iv) 1200 pg / mL of phosphorylated tau peptide of SEQ ID NO: 2 and 300 pg / mL of T50 (liposome Z+++). Bleeding was performed before immunization and on days 8, 22, 36, 50, 64, 78, 92, 106, 120, 134, 148, 162, 176, and 190, and sera were isolated. Specific IgG antibody titers in the sera were determined by ELISA, using phosphorylated tau peptide of SEQ ID NO: 2 as a coating antigen and an anti-monkey IgG secondary antibody.The resulting antibody levels were calculated as the outcome titers (last serum dilution inducing a positive response) for each individual monkey over time. The geometric mean of the outcome titers per group ± 95% confidence interval is presented in Figure 18, showing that all four liposomal vaccines tested induced high and sustained titers against the tau peptide. Sequence Listing

[00394] SEQ ID NO: 1 - phospho-tau (7.1) peptide

[00395] GDRSGYS [pS] PG [pS] PG [pT] PGSRSRT

[00396] SEQ ID NO: 2 - phospho-tau peptide (T3.5)

[00397] VYK [pS] PVVSGDT [pS] PRHL

[00398] SEQ ID NO: 3 - phospho-tau peptide (22.1)

[00399] SSTGSIDMVD [pS] PQLA [pT] LA

[00400] SEQ ID NO: tau peptide 4

[00401] VYKSPVVSGDTSPRHL

[00402] SEQ ID NO: 5 - phospho-tau peptide Petition 870260044022, dated 11 / 05 / 2026, page 99 / 222 92 / 96

[00403] RENAKACTDHGAEIVYK [pS] PVVSGDT [pS] PRHL

[00404] SEQ ID NO: 6 - phospho-tau peptide

[00405] RQEFEVMEDHAGT [pY] GL

[00406] SEQ ID NO: 7 - phospho-tau peptide

[00407] PGSRSR [pT] P [pS] LPTPPTR

[00408] SEQ ID NO: 8 - phospho-tau peptide

[00409] GYSSPG [pS] PG [pT] PGSRSR

[00410] SEQ ID NO: 9 - phospho-tau peptide

[00411] GDT [pS] PRHL [pS] NVSSTGSID

[00412] SEQ ID NO: 10 - phospho-tau peptide

[00413] PG [pS] PG [pT] PGSRSR [pT] P [pS] LP

[00414] SEQ ID NO: 11 - phospho-tau peptide

[00415] HL [pS] NVSSTGSID

[00416] SEQ ID NO: 12 - phospho-tau peptide

[00417] VSGDT [pS] PRHL

[00418] SEQ ID NO: 13 - T50 T Cell Epitope

[00419] AKFVAAWTLKAAVVRQYIKANSKFIGITELVVRFNNFTVS FWLRVPKVSASHLE-NH2

[00420] SEQ ID NO: 14 - T46 T Cell Epitope

[00421] AKFVAAWTLKAAAGSQYIKANSKFIGITELGSFNNFTVSFW LRVPKVSASHLEK (Pal) K (Pal)-NH2

[00422] SEQ ID NO: 15 - T48 helper T cell epitope

[00423] AKFVAAWTLKAAAGSQYIKANSKFIGITELGSFNNFTVSFW LRVPKVSASHLEGSLINSTKIYSYFPSVISKVNQ-NH2

[00424] SEQ ID NO: 16 - T51 helper T cell epitope

[00425] AKFVAAWTLKAAARRQYIKANSKFIGITELRRFNNFTVSFW LRVPKVSASHLE-NH2

[00426] SEQ ID NO: 17 - T52 helper T cell epitope

[00427] AKFVAAWTLKAAARKQYIKANSKFIGITELRKFNNFTVSFW LRVPKVSASHLE-NH2 Petition 870260044022, dated 11 / 05 / 2026, pages 100 / 222 93 / 96

[00428] SEQ ID NO: 18 - CpG 2006 (also known as CpG 7909)

[00429] 5'-tcgtcgttttgtcgttttgtcgtt-3'

[00430] where lowercase letters stand for internucleotide bonds phosphorothioate (ps)

[00431] SEQ ID NO: 19 - CpG 1018

[00432] 5'-tgactgtgaacgttcgagatga-3'

[00433] where lowercase letters signify phosphorothioate internucleotide bonds

[00434] SEQ ID NO: 20 - CpG2395

[00435] 5'-tcgtcgttttcggcgcgcgccg-3'

[00436] where lowercase letters signify phosphorothioate internucleotide bonds

[00437] SEQ ID NO: 21 - CpG2216

[00438] 5'-ggGGGACGATCGTCgggggg-3'

[00439] where lowercase letters mean phosphorothioate internucleotide bonds and uppercase letters mean phosphodiester (po) bonds

[00440] SEQ ID NO: 22 - CpG2336

[00441] 5'- gggGACGACGTCGTGgggggg -3',

[00442] where lowercase letters mean phosphorothioate internucleotide bonds and uppercase letters mean phosphodiester bonds

[00443] SEQ ID NO: 23 - Pan DR epitope peptide (PADRE)

[00444] AKFVAAWTLKAAA

[00445] SEQ ID NO: 24 - P2

[00446] QYIKANSKFIGITEL

[00447] SEQ ID NO: 25 - P30

[00448] FNFTVSFWLRVPKVSASHLE

[00449] SEQ ID NO: 26 - TT586-605 Petition 870260044022, of 11 / 05 / 2026, p. 101 / 222 94 / 96

[00450] LINSTKIYSYFPSVISKVNQ

[00451] SEQ ID NO: 27 - palmitoylated phospho-tau peptide (palmitoylated 7.1)

[00452] K (pal) K (pal) GDRSGYS [pS] PG [pS] PG [pT] PGSRSRTK (pal) K (pal)

[00453] SEQ ID NO: 28 - palmitoylated phospho-tau peptide (T3, palmitoylated T3.5)

[00454] K (pal) K (pal) VYK [pS] PVVSGDT [pS] PRHLK (pal) K (pal)

[00455] SEQ ID NO: 29 - palmitoylated phospho-tau peptide (palmitoylated 22,1)

[00456] K (pal) K (pal) SSTGSIDMVD [pS] PQLA [pT] LAK (pal) K (pal)

[00457] SEQ ID NO: 30 - palmitoylated tau peptide

[00458] K (pal) K (pal) VYKSPVVSGDTSPRHLK (pal) K (pal)

[00459] SEQ ID NO: 31 - palmitoylated phospho-tau peptide

[00460] K (pal) K (pal) RENAKACTDHGAEIVYK [pS] PVVSGDT [pS] PRHLK (pal) K (pal)

[00461] SEQ ID NO: 32 - palmitoylated phospho-tau peptide

[00462] K (pal) K (pal) RQEFEVMEDHAGT [pY] GLC (pal) K (pal)

[00463] SEQ ID NO: 33 - palmitoylated phospho-tau peptide

[00464] K (pal) K (pal) PGSRSR [pT] P [pS] LPTPPTRK (pal) K (pal)

[00465] SEQ ID NO: 34 - palmitoylated phospho-tau peptide

[00466] K (pal) K (pal) GYSSPG [pS] PG [pT] PGSRSRK (pal) K (pal)

[00467] SEQ ID NO: 35 - palmitoylated phospho-tau peptide

[00468] K (pal) K (pal) GDT [pS] PRHL [pS] NVSSTGSIDK (pal) K (pal)

[00469] SEQ ID NO: 36 - palmitoylated phospho-tau peptide

[00470] K (pal) K (pal) PG [pS] PG [pT] PGSRSR [pT] P [pS] LPK (pal) K (pal)

[00471] SEQ ID NO: 37 - palmitoylated phospho-tau peptide Petition 870260044022, of 11 / 05 / 2026, p. 102 / 222 95 / 96

[00472] K (pal) K (pal) HL [pS] NVSSTGSIDK (pal) K (pal)

[00473] SEQ ID NO: 38 - palmitoylated phospho-tau peptide

[00474] K (pal) K (pal) VSGDT [pS] PRHLK (pal) K (pal)

[00475] SEQ ID NO: 39 - T50 without C-terminal amide

[00476] AKFVAAWTLKAAAVVRQYIKANSKFIGITELVVRFNNFTVS FWLRVPKVSASHLE

[00477] SEQ ID NO: 40 - T46 without -Lys (Pal) -Lys (Pal) -NH2 at the C terminal

[00478] AKFVAAWTLKAAAGSQYIKANSKFIGITELGSFNNFTVSFW LRVPKVSASHLE

[00479] SEQ ID NO: 41 - T48 without the C terminal amide

[00480] AKFVAAWTLKAAAGSQYIKANSKFIGITELGSFNNFTVSFW LRVPKVSASHLEGSLINSTKIYSYFPSVISKVNQ

[00481] SEQ ID NO: 42 - T51 without terminal C-amide

[00482] AKFVAAWTLKAAARRQYIKANSKFIGITELRRFNNFTVSFW LRVPKVSASHLE

[00483] SEQ ID NO: 43 - T52 without terminal C-amide

[00484] AKFVAAWTLKAAARKQYIKANSKFIGITELRKFNNFTVSFW LRVPKVSASHLE

[00485] SEQ ID NO: 44 - T57

[00486] AKFVAAWTLKAAAVVRQYIKANSKFIGITELVVRFNNFTVS FWLRVPKVSASHLE-K (Pal) K (Pal) -NH2 References Asuni AA et al., J Neurosci. August 22, 2007;27(34):9115-29 Bentebibel et al., 2013, Sci Transl Med., 5(176):176ra32 Crotty, 2011, Annual Reviews of Immunology. Vol 29: pp. 621-663 Friedhoff et al., Biochimica et Biophysica Acta 1502 (2000) 122 to 132 Petition 870260044022, dated 11 / 05 / 2026, p. 103 / 222 96 / 96 Greenberg and Davies, 1991, Proc Natl Acad Sci USA, 87(15):5827-31 Hanger et al., Trends Mol Med. 15:112-9, 2009 Hickman et al., J. Biol. Chem. vol. 286, NO. 16, pp. 13966–13976, April 22, 2011 Kontsekova E et al., Alzheimers Res Ther. August 1, 2014;6(4):44 Novak P et al., Lancet Neurology 2017, 16: 123 to 134 Peeraer et al., 2015, Neurobiol Dis., 73: 83–95 Ries et al., 2015, Org. Biomol. Chem., 13: 9673 Spensieri et al., 2013, Proc Natl Acad Sci USA., 110(35):14330-5 Theunis C et al., PLoS One. 2013; 8(8): e72301 US 7,741,297 US 8,647,631 US 9,687,447 WO 90 / 14837 WO 2010 / 115843 Petition 870260044022, dated 11 / 05 / 2026, page 104 / 222

Claims

1 / 6 CLAIMS 1. Liposome, characterized in that it comprises: a. a tau peptide having the amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 5, 9 and 12; b. a T helper cell epitope comprising at least one amino acid sequence selected from the group consisting of: SEQ ID NOs: 23, 24, 25 and 26; c. a lipid-coated CpG oligonucleotide having the nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22, wherein the CpG oligonucleotide comprises one or more phosphorothioate internucleotide linkages, and wherein the CpG oligonucleotide is covalently linked to at least one cholesterol through a ligand; and d. a monophosphoryl lipid A (MPLA); wherein the tau peptide is presented on the surface of the liposome.

2. Liposome, according to claim 1, characterized in that: a. the tau peptide has an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 31, 35 and 38; b. a lipid-coated CpG oligonucleotide has the nucleotide sequence of SEQ ID NO: 18; and c. the helper T cell epitope comprises the amino acid sequences of SEQ ID NO: 23, SEQ ID NO: 24 and SEQ ID NO: 25, wherein the amino acid sequences are covalently linked to each other, optionally by means of one or more ligands.

3. Liposome, according to claim 2, characterized in that the helper T cell epitope comprises an amino acid sequence selected from the group consisting of Petition 870260044022, dated 11 / 05 / 2026, page 105 / 222 2 / 6 SEQ ID Nos: 39, 40, 41, 42 and 43.

4. Liposome, according to claim 2, characterized in that the helper T cell epitope comprises an amino acid sequence selected from the group consisting of the SEQ ID NOs: 13, 14, 15, 16, 17 and 44.

5. Liposome, according to claim 1, characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol.

6. Liposome, characterized in that it comprises: a. a tau peptide comprising the amino acid sequence of SEQ ID NO: 2; b. a helper T cell epitope presenting an amino acid sequence selected from the group consisting of SEQ ID NOs: 39, 40, 41, 42 and 43; c. a lipid-coated CpG oligonucleotide presenting the nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO: 22, wherein the CpG oligonucleotide is covalently linked to at least one cholesterol through a ligand; and d. monophosphoryl-lipid A (MPLA); wherein the tau peptide is presented on the surface of the liposome.

7. Liposome, according to claim 6, characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol, wherein: a. the tau peptide has the amino acid sequence of Petition 870260044022, dated 11 / 05 / 2026, page 106 / 222 3 / 6 SEQ ID NO: 28; b. the helper T cell epitope has the amino acid sequence of SEQ ID NO: 39; and c. the lipid-coated CpG oligonucleotide has the nucleotide sequence of SEQ ID NO:

18.

8. Liposome, according to claim 7, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

13.

9. Liposome, according to claim 6, characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol, wherein: a. the tau peptide has the amino acid sequence of SEQ ID NO: 28; b. the helper T cell epitope has the amino acid sequence of SEQ ID NO: 40; and c. the lipid-coated CpG oligonucleotide has the nucleotide sequence of SEQ ID NO:

18.

10. Liposome, according to claim 9, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

14.

11. Liposome, according to claim 6, characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol, wherein: a. the tau peptide has the amino acid sequence of SEQ ID NO: 28; b. the T helper cell epitope has the amino acid sequence of SEQ ID NO: 41; and c. the lipid-coated CpG oligonucleotide has the nucleotide sequence of SEQ ID NO:

18.

12. Liposome, according to claim 11, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

15.

13. Pharmaceutical composition, characterized in that it comprises the liposome, as defined in claim 6, and a pharmaceutically acceptable carrier.

14. Pharmaceutical composition, characterized in that it comprises the liposome, as defined in claim 1, and a pharmaceutically acceptable carrier.

15. Liposome, according to claim 1, characterized in that: the tau peptide has an amino acid sequence selected from the group consisting of the following sequence numbers: 28, 31, 35 and 38.

16. Liposome, according to claim 15, characterized in that the helper T cell epitope comprises an amino acid sequence selected from the group consisting of the following sequences: 39, 40, 41, 42 and 43.

17. Liposome, according to claim 15, characterized in that the helper T cell epitope comprises an amino acid sequence selected from the group consisting of the following SEQ ID NOs: 13, 14, 15, 16, 17 and 44.

18. Liposome, according to claim 6, characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol.

19. Liposome, according to claim 6, Petition 870260044022, dated 11 / 05 / 2026, page 108 / 222 5 / 6, characterized in that the helper T cell epitope has the amino acid sequence of SEQ ID NO:

39.

20. Liposome, according to claim 6, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

40.

21. Liposome, according to claim 6, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

41.

22. Liposome, according to claim 6, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

42.

23. Liposome, according to claim 6, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

43.

24. Liposome, according to claim 6, characterized in that the tau peptide has the amino acid sequence of SEQ ID NO:

28.

25. Liposome, characterized in that it comprises: a. a tau peptide comprising the amino acid sequence of SEQ ID NO: 28, wherein the tau peptide is present on the surface of the liposome; b. a helper T cell epitope with an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 14 or 15; c. a lipid-linked CpG oligonucleotide with the nucleotide sequence selected from the group consisting of SEQ ID NO: 18, wherein the CpG oligonucleotide is covalently linked to at least one cholesterol by means of a ligand; and d. monophosphoryl lipid A (MPLA).

26. Liposome, according to claim 25, Petition 870260044022, dated 11 / 05 / 2026, page 109 / 222 6 / 6 characterized in that it further comprises one or more lipids selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phosphoryl-3'-rac-glycerol (DMPG) and cholesterol.

27. Liposome, according to claim 25, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

13.

28. Liposome, according to claim 25, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

14.

29. Liposome, according to claim 25, characterized in that the helper T cell epitope has the amino acid sequence SEQ ID NO:

15.

30. Pharmaceutical composition, characterized in that it comprises liposomes, as defined in claim 25, and a pharmaceutically acceptable carrier. Petition 870260044022, dated 11 / 05 / 2026, pp. 110 / 222

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