Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
  • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/0005—Vertebrate antigens
  • A61K39/0007—Nervous system antigens; Prions
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/0005—Vertebrate antigens
  • A61K39/0008—Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K40/00—Cellular immunotherapy
  • A61K40/10—Cellular immunotherapy characterised by the cell type used
  • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K40/00—Cellular immunotherapy
  • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
  • A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
  • A61K47/6911—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
  • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
  • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
  • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
  • A61K2039/55511—Organic adjuvants
  • A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
  • A61K2039/55511—Organic adjuvants
  • A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing adjuvants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
  • A61K2039/55511—Organic adjuvants
  • A61K2039/55572—Lipopolysaccharides; Lipid A; Monophosphoryl lipid A
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
  • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/62—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier
  • A61K2039/627—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier characterised by the linker

Definitions

• the present invention is in the field of medicine.
• the invention in particular relates to a liposome that contains a phosphorylated Tau peptide for inducing a sustained antibody response against phosphorylated Tau protein (pTau) in a subject in need of preventing or treating Tauopathy, such as Alzheimer’s Disease.
• pTau phosphorylated Tau protein
• AD Alzheimer’ s Disease
• Alzheimers.net Alzheimer’ s Disease
• AD therapies that are currently commercialized aim to act on the clinical symptoms, but do not target the pathogenic processes that underlie the disease (disease-modifying effect).
• the current therapies are only minimally efficacious, and there is therefore an urgent need to develop and test additional preventive and therapeutic measures.
• Alzheimer’ s Disease The hallmark pathologies for Alzheimer’ s Disease are an accumulation of extracellular plaques comprising notably aggregated amyloid beta protein and intracellular “tangles” or aggregations of hyperphosphorylated Tau protein.
• the molecular events that lead to accumulation of these proteins are poorly characterized.
• amyloid it is hypothesized 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 hypothesized that dysregulation of either kinases, phosphatases, or both, leads to aberrant phosphorylation of Tau.
• Tau becomes hyperphosphorylated it loses the ability to effectively bind and stabilize microtubules, and instead accumulates in the cytoplasm of the affected neuron.
• the unbound and hyperphosphorylated Tau appears to form first oligomers and then higher order aggregates, the presence of which presumably negatively affects the function of the neuron in which they form, perhaps via interruption of normal axonal transport.
• Alzheimer’s Disease or other dementing Tauopathies are commonly treated with cholinesterase inhibitors (e.g., Aricept®) or memantine (e.g., NamendaTM). These drugs, although reasonably well tolerated, have very modest efficacy. For example, Aricept® delays the worsening of symptoms for 6-12 months in approximately 50% of the treated individuals. The remainder of treatment is non- pharmacologic, and focuses on making patients more capable of managing day to day tasks as their cognitive ability declines.
• cholinesterase inhibitors e.g., Aricept®
• memantine e.g., NamendaTM
• AADvacl an active peptide vaccine designed to target pathological Tau in Alzheimer’s Disease
• the AADvacl contains a synthetic peptide derived from amino acids 294 to 305 of the Tau sequence coupled to keyhole limpet hemocyanin (KLH) through an N-terminal cysteine.
• KLH keyhole limpet hemocyanin
• ACI-35 a vaccine using a synthetic peptide based on human p-Tau396/404 was show'n to have improved motor abilities and extended survival of mice carrying a P301L mutation (Theunis et al., PLOS ONE. 2013. 8(8): e72301).
• ACI-35 was well-tolerated and elicited an antibody response, with a limited boostability.
• the invention is based on findings from clinical studies of an improved liposomal vaccine comprising a phosphorylated Tau peptide presented on the surface of the liposome.
• the vaccine induced potent and sustained immune response, such as sustained antibody responses against pTau and the antibody responses could be boosted by booster shots.
• the invention provides a method of inducing an antibody response against a phosphorylated Tau protein (pTau) in a human subject in need thereof, comprising administering to the subject an effective amount of a liposome containing:
• a Tau phosphopeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:27 to SEQ ID NO:29 and SEQ ID NO:31 to SEQ ID NO:38 at an amount of 25-750 nmoles, such as 300 pg to 1800 pg, per dose;
• a toll-like receptor 4 agonist comprising monophosphoryl lipid A
• helper T-cell epitope having an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17, SEQ ID NO:23 to SEQ ID NO:26, and SEQ ID NO:39 to SEQ ID NO:44;
• the effective amount of the liposome comprises:
• Tau phosphopeptide consisting of the amino acid sequence of SEQ ID NO: 28 at the amount of 300 pg to 1800 pg per dose;
• helper T-cell epitope at an amount of 75 pg to 550 pg per dose
• the CpG oligonucleotide has one or more phosphorothioate internucleotide linkages, and the CpG oligonucleotide is covalently linked to at least one lipophilic group, optionally via a PEG linker.
• the Tau phosphopeptide is administered at an amount of about 25 nmoles to about 750 nmoles per dose, such as about 29.7 nmoles to about 742.5 nmoles per dose, preferably about 90 nmoles to about 715 nmoles, such as about 89.1 nmoles to about 712.8 nmoles per dose, or about 90 nmoles to about 535 nmoles per dose, such as about 89.1 nmoles to about 534.6 nmoles per dose, or about 90 nmoles to about 275 nmoles per dose, such as about 89.1 nmoles to about 267.3 nmoles per dose.
• the Tau phosphopeptide consists of an amino acid sequence selected from the group consisting of SEQ ID NO:27 to SEQ ID NO:29 and SEQ ID NO:31 to SEQ ID NO:38, preferably consists of an amino acid sequence of SEQ ID NO:28.
• the tetrapalmitoylated Tau phosphopeptide is administered at an amount of 100 pg to 2500 pg per dose, corresponding to 29.7 nmoles to 742.5 nmoles per dose, preferably 300 pg to 2400 pg per dose, corresponding to 89.1 nmoles to 712.8 nmols per dose, such as 300 pg, 900 pg, 1800 pg or 2400 pg per dose, corresponding to 89.1 nmoles, 267.3 nmoles, 534.6 nmoles or 712.8 nmoles per dose.
• the effective amount of liposomes comprises the toll-like receptor 4 agonist at an amount of 30 pg to 900 pg, preferably 100 pg to 585 pg, per dose. In certain embodiments, the effective amount of liposomes comprises the toll-like receptor agonist monophosphoryl hexa-acyl Lipid A, 3-deacyl at an amount of 30 pg to 900 pg, preferably 100 pg to 585 pg, per dose.
• the effective amount of liposomes comprises the helper T- cell epitope at an amount of 25 pg to 625 pg, preferably 75 pg to 550 pg, such as 75 pg to 450 pg, per dose.
• the effective amount of liposomes comprises a T50 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 13 at an amount of 25 pg to 625 pg, preferably 75pg to 450 pg, per dose.
• the effective amount of liposomes comprises the helper T-cell epitope at an amount of about 2 nmoles to about 110 nmoles per dose, such as about 4.02 nmoles to about 100.44 nmoles per dose, or about 4 nmoles to about 75 nmoles per dose, such as about 4.02 nmoles to about 72.32 nmoles per dose, or about 10 nmoles to about 105 nmoles per dose, such as about 12.06 nmoles to about 100.44 nmoles per dose, or about 70 to about 105 nmoles per dose, such as about 72.32 nmoles to about 100.44 nmoles per dose.
• the effective amount of liposomes comprises a T50 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 13 at an amount of about 3 nmoles to about 105 nmoles per dose, preferably about 10 nmoles to about 105 nmoles per dose, such as about 12.06 nmoles to about 100.44 nmoles per dose.
• the effective amount of liposomes comprises the helper T-cell epitope at an amount of 2 to 5 nmoles per dose, e.g., 2, 3, 4 or 5 nmoles per dose or any value in between, such as about 3.82, 3.92, 4.02 or 4.12 nmoles per dose.
• the effective amount of liposomes comprises the helper T-cell epitope at an amount of 10 to 15 nmoles per dose, such as 10, 11, 12, 13, 14 or 15 nmoles per dose, or any value in between, such as 11.86, 11.96, 12.06, 12.16 nmoles per dose.
• the effective amount of liposomes comprises the helper T-cell epitope at an amount of 70 to 75 nmoles per dose, such as 70, 71, 72, 73, 74 or 75 nmoles per dose, or any value in between, such as 72.02, 72.12, 72.22, 72.32, 72.42 nmoles per dose.
• the effective amount of liposomes comprises the helper T-cell epitope at an amount of 98 to 103 nmoles per dose, such as 98, 99, 100, 101, 102 or 103 nmoles per dose, or any value in between, such as 100.24, 100.34, 100.44, 100.54 or 100.64 nmoles per dose.
• the effective amount of liposomes comprises the lipidated CpG oligonucleotide at an amount of 50 pg to 1250 pg, preferably 100 pg to 1000 pg, such as 150 pg to 800 pg, per dose.
• the effective amount of liposomes comprises a CpG oligonucleotide consisting of the nucleotide sequence of SEQ ID NO: 18 at an amount of 50 pg to 1250 pg, preferably 150 pg to 800 pg, per dose.
• the liposomes are administered subcutaneously.
• the liposomes are administered intramuscularly.
• the liposome further comprises one or more lipids selected from the group consisting of l,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2- dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol (DMPG), and cholesterol.
• DMPC l,2-dimyristoyl-sn-glycero-3-phosphocholine
• DMPG 1,2- dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol
• the liposome comprises:
• the toll-like receptor 4 agonist comprising monophosphoryl hexa-acyl Lipid A, 3- deacyl;
• helper T-cell epitope comprising the amino acid sequence of SEQ ID NO: 39;
• lipid selected from the group consisting of 1,2-dimyristoyl-sn-glycero- 3 -phosphocholine (DMPC), l,2-dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol (DMPG), and cholesterol.
• DMPC 1,2-dimyristoyl-sn-glycero- 3 -phosphocholine
• DMPG l,2-dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol
• the antibody response comprises a specific IgG antibody response directed against the pTau.
• the specific IgG antibody response has an anti-pTau IgG titer at least 50, 60, 70, 80, 90, 100, or more times higher than that of a placebo control.
• the antibody response comprises a specific IgM antibody response directed against the pTau and a class switch of the specific IgM antibody response to a specific IgG antibody response directed against the pTau.
• the antibody response comprises an IgG antibody response that preferentially recognizes the pTau over non- phosphorylated Tau protein.
• the ratio of the anti-pTau IgG titer to the anti-Tau IgG titer is at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70, or more.
• the antibody response comprises an IgG antibody response against an enriched Paired Helical Filament (ePHF).
• the IgG antibody response has an anti-ePHF IgG titer at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more times higher than that of a placebo control.
• the anti- ePHF IgG has an increased binding avidity to the pathological ePHF Tan for at least 6 weeks after the initial administration of the effective amount of the liposome or after a boosting administration as measured at least 2 weeks after the boosting administration, preferably the anti- ePHF IgG has an avidity index of at least 0.3, 0.4, 0.5, 0.6, or 0.7.
• the antibody response can be boosted by a booster administration.
• a method of the application further comprises administering to the subject a second dose of the effective amount of liposome 4 to 12 weeks, such as 8 weeks, after the initial administration of the effective amount of liposome.
• the antibody response is boosted as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• the antibody response is boosted at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• a method of the application further comprises administering to the subject a third dose of the effective amount of liposome 20 to 28 weeks, such as 24 weeks, after the initial administration of the effective amount of liposome.
• the antibody response is boosted as measured at least 2 weeks after the administration of the third dose of the effective amount of liposome, preferably the antibody response is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more as measured at least 2 weeks after the administration of the third dose of the effective amount of liposome.
• a method of the application further comprises administering to the subject a fourth dose of the effective amount of liposome 44 to 52 weeks, such as 48 weeks, after the initial administration of the effective amount of liposome.
• the antibody response is boosted at least 2 weeks after the administration of the fourth dose of the effective amount of liposome, preferably the antibody response is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more by the administration of the fourth dose of the effective amount of liposome.
• the human subject is in need of clearance of aggregates of Tau.
• the subject is in need of a prevention or treatment of Alzheimer’s Disease, such as preclinical Alzheimer’s Disease, early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’s Disease, mild Alzheimer’s Disease, or mild to moderate Alzheimer’s Disease.
• the subject is amyloid positive in the brain but does not yet show significant cognitive impairment.
• the invention also relates to a vaccine combination for use in inducing an immune response, such as an antibody response against a phospho-Tau protein (pTau), in a human subject in need thereof, wherein the vaccine combination comprises a primer vaccine and a booster vaccine according to embodiments of the invention, and the immune response lasts at least 10 weeks, such as at least 10, 15, 20, 25, 30, 35, 40, 45, 50 weeks or more after the administration of the primer vaccine to the human subject.
• an immune response such as an antibody response against a phospho-Tau protein (pTau)
• pTau phospho-Tau protein
• FIG. 1 is an overview of the study design for cohort 1 (ACI-35.030 or placebo) in a Phase lb/2a study (NCT04445831).
• FIG. 2 shows the geometric mean of anti-pTau IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg or 900 pg doses at weeks 0, 8, and 24.
• FIG. 3 shows the geometric mean of anti-Tau (non-phosphorylated) IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg or 900 pg doses at weeks 0, 8, and 24.
• FIG. 4 shows the geometric mean of anti-ePHF IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg or 900 pg doses at weeks 0, 8, and 24.
• FIG. 5 shows the geometric mean of anti-Tau (non-phosphorylated) IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg, 900 pg or 1800 pg doses.
• FIG. 6 shows the geometric mean of anti-pTau IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg, 900 pg or 1800 pg doses.
• FIG. 7 shows the geometric mean of anti-ePHF IgG titers in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at 300 pg, 900 pg or 1800 pg doses.
• FIG. 8A and 8B show the epitope recognition profile of antibodies induced in patients with early Alzheimer’s Disease who were treated with placebo or ACI-35.030 at a 900 pg dose at weeks 0, 8, and 24.
• any numerical values such as a concentration or a concentration range described herein, are to be understood as being modified in all instances by the term “about.”
• a numerical value typically includes ⁇ 10% of the recited value.
• a concentration of 1 mg/mL includes 0.9 mg/mL to 1.1 mg/mL.
• a concentration range of 1% to 10% (w/v) includes 0.9% (w/v) to 11% (w/v).
• the use of a numerical range expressly includes all possible subranges, all individual numerical values within that range, including integers within such ranges and fractions of the values unless the context clearly indicates otherwise.
• compositions, a mixture, a process, a method, an article, or an apparatus that comprises a list of elements is not necessarily limited to only those elements but can include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus.
• “or” refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one 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).
• the invention provides a method of inducing anti-phosphorylated Tau antibodies without inducing a severe adverse event considered either possibly or probably related to the study vaccine, such as encephalitis, in a human subject in need thereof.
• the method comprises administering to the subject an effective amount of liposomes comprising a Tau phosphopeptide presented on the surface of the liposome and a toll-like receptor 4 agonist.
• anti-phosphorylated Tau antibody refers to an antibody that binds to Tau that has been phosphorylated on an amino acid residue at one or more locations of the amino acid sequence of Tau.
• the phosphorylated amino acid residues can be, e.g., serine (Ser), threonine (Thr) or tyrosine (Tyr).
• the site on phosphorylated Tau to which the anti -phosphorylated Tau antibody binds is preferably a site that is specifically phosphorylated in neurodegenerative diseases such as Alzheimer’ s Disease.
• sites of phosphorylated Tau to which the anti-phosphorylated Tau antibody binds include, for example, Tyrl8, Serl99, Ser202, Thr205, Thr212, Ser214, Ser396, Ser404, Ser409, Ser422, Thr427.
• amino acid positions are given in reference to the sequence of human microtubule-associated protein tau isoform 2 having the amino acid sequence represented in GenBank Accession No. NP_005901.2.
• the ability to induce anti-phosphorylated Tau antibodies upon administration can be determined by testing a biological sample (e.g., blood, plasma, serum, PBMCs, urine, saliva, feces, CSF or lymph fluid) from the subject for the presence of antibodies, e.g., IgG or IgM antibodies, directed to the immunogenic Tau peptide(s) administered in the pharmaceutical composition (see for example Harlow, 1989, Antibodies, Cold Spring Harbor Press).
• a biological sample e.g., blood, plasma, serum, PBMCs, urine, saliva, feces, CSF or lymph fluid
• antibodies e.g., IgG or IgM antibodies
• titers of antibodies produced in response to administration of a composition providing an immunogen can be measured by enzyme-linked immunosorbent assay (ELISA), other ELISA-based assays (e.g., MSD-Meso Scale Discovery), dot blots, SDS-PAGE gels, ELISPOT or Antibody-Dependent Cellular Phagocytosis (ADCP) Assay.
• ELISA enzyme-linked immunosorbent assay
• other ELISA-based assays e.g., MSD-Meso Scale Discovery
• dot blots e.g., SDS-PAGE gels
• ELISPOT Antibody-Dependent Cellular Phagocytosis
• AE adverse event
• mild referring to an AE that is easily tolerated by the subject, which causes minimal discomfort and does not interfere with everyday activities
• moderate referring to an AE that is sufficiently discomforting to interfere with normal everyday activities and intervention may be needed
• severe referring to an AE that prevents normal everyday activities, and treatment or other intervention is usually needed.
• a serious AE can be any AE occurring at any dose that results in any of the following outcomes: death, where death is an outcome, not an event; life-threatening, referring to an event in which the patient is at risk of death at the time of the event; it does not refer to an event which could hypothetically have caused death had it been more severe; in patient hospitalization, i.e., an unplanned, overnight hospitalization, or prolongation of an existing hospitalization; persistent or significant incapacity or substantial disruption of the ability to conduct normal life functions; congenital anomaly/birth defect; important medical event (as deemed by the investigator) that may jeopardize the patients or may require medical or surgical intervention to prevent one of the other outcomes listed above (e.g., intensive treatment in an emergency room or at home for allergic bronchospasm or blood dyscrasias or convulsions that do not result in hospitalization).
• death where death is an outcome, not an event
• life-threatening referring to an event in which the patient is at risk of death at the time of the event; it
• Hospitalization is official admission to a hospital. Hospitalization or prolongation of a hospitalization constitutes criteria for an AE to be serious; however, it is not in itself considered an SAE. In the absence of an AE, hospitalization or prolongation of hospitalization should not be reported as a SAE by the participating investigator. This can be the case, in the following situations: the hospitalization or prolongation of hospitalization is needed for a procedure required by the protocol; or the hospitalization or prolongation of hospitalization is a part of a routine procedure followed by the center (e.g., stent removal after surgery). This should be recorded in the study file. Hospitalization for elective treatment of a pre-existing condition that did not worsen during the study is not considered an AE.
• Complications that occur during hospitalization are AEs. If a complication prolongs hospitalization, or meets any of the other SAE criteria, then the event is an SAE.
• encephalitis refers to an inflammation of the brain which can result from infectious and non-infectious causes.
• meningoencephalitis refers to a condition characterized by infection or inflammation of the brain meninges and of the brain. The diagnosis of encephalitis or meningoencephalitis can be determined by techniques known to those skilled in the art in view of the present disclosure, for example, by clinical, neurological and psychiatric examinations, biological sampling including blood and CSF samplings, MRI scanning and electroencephalography (EEG).
• EEG electroencephalography
• liposome refers generally to a lipid vesicle that is made of materials having high lipid content, e.g., phospholipids, cholesterol.
• the lipids of these vesicles are generally organized in the form of lipid bilayers.
• the lipid bilayers generally encapsulate a volume which is either interspersed between multiple onion-like shells of lipid bilayers, forming multilamellar lipid vesicles (ML Vs) or contained within an amorphous central cavity.
• ML Vs multilamellar lipid vesicles
• Lipid vesicles having an amorphous central cavity are unilamellar lipid vesicles, i.e., those with a single peripheral bilayer surrounding the cavity.
• LUVs Large unilamellar vesicles
• SUVs small unilamellar lipid vesicles
• Tau or “Tau protein”, also known as microtubule- associated protein Tau, MAPT, neurofibrillary tangle protein, paired helical filament-Tau, PHF-Tau, MAPTL, MTBT1 refers to an abundant central and peripheral nervous system protein having multiple isoforms.
• CNS human central nervous system
• Tau examples include, but are not limited to, Tau isoforms in the CNS, such as the 441-amino acid longest Tau isoform (4R2N), also named microtubule-associated protein tau isoform 2, that has four repeats and two inserts, such as the human Tau isoform 2 having the amino acid sequence represented in GenBank Accession No. NP_005901.2.
• Other examples of Tau include the 352-amino acid long shortest (fetal) isoform (3R0N), also named microtubule- associated protein tau isoform 4, that has three repeats and no inserts, such as the human Tau isoform 4 having the amino acid sequence represented in GenBank Accession No. NP_058525.1.
• Examples of Tau also include the “big Tau” isoform expressed in peripheral nerves that contains 300 additional residues (exon 4a). Friedhoff et al., Biochimica et Biophysica Acta 1502 (2000) 122-132.
• Examples of Tau include a human big Tau that is a 758 amino acid-long protein encoded by an mRNA transcript 6762 nucleotides long (NM_016835.4), or isoforms thereof.
• the amino acid sequence of the exemplified human big Tau is represented in GenBank Accession No. NP_058519.3.
• the term “Tau” includes homologs of Tau from species other than human, such as Macaca Fascicularis (cynomolgus monkey), rhesus monkeys or Pan troglodytes (chimpanzee).
• the term “Tau” includes proteins comprising mutations, e.g., point mutations, fragments, insertions, deletions and splice variants of full- length wild type 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.
• peptide or “polypeptide” refers to a polymer composed of amino acid residues, related naturally occurring structural variants, and synthetic non- naturally occurring analogs thereof linked via peptide bonds.
• the term refers to a peptide of any size, structure, or function. Typically, a peptide is at least three amino acids long.
• a peptide can be naturally occurring, recombinant, or synthetic, or any combination thereof. Synthetic peptides can be synthesized, for example, using an automated polypeptide synthesizer.
• Examples of Tau peptides include any peptide of Tau protein of about 5 to about 30 amino acids in length, preferably of about 10 to about 25 amino acids in length, more preferably of about 16 to about 21 amino acids in length. In the present disclosure, peptides are listed from N to C terminus using the standard three 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 SEQ ID NOs: 1-12, or Tau peptides having an amino acid sequence that is at least 75%, 80%, 85%, 90% or 95% identical to the amino acid sequence of any of SEQ ID NOs: 1- 12.
• phosphopeptide or “phospho-epitope” refers to a peptide that is phosphorylated at one or more amino acid residues.
• Tau phosphopeptides include any Tau peptide comprising one or more phosphorylated amino acid residues.
• the Tau peptides of the present invention can be synthesized by solid phase peptide synthesis or by recombinant expression systems.
• Automatic peptide synthesizers are commercially available from numerous suppliers, such as Applied Biosystems (Foster City, Calif.).
• 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 (C.S.H.P. Press, NY 2d ed., 1989).
• the liposome comprises one or more Tau peptides.
• the Tau peptides in the liposome can be the same or different. Any suitable Tau peptide known to those skilled in the art can be used in the invention in view of the present disclosure.
• one or more of the Tau peptides comprise the amino acid sequence of one of SEQ ID NOs: 1-12.
• 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 SEQ ID NOs: 1-12, wherein none of the amino acid residues are phosphorylated, or one or more amino acid residues are phosphorylated.
• one or more of the Tau peptides are Tau phosphopeptides.
• the one or more Tau phosphopeptides comprise the amino acid sequence of one of 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 SEQ ID NOs: 1-3 or 5-12, wherein one or more of the indicated amino acid residues are phosphorylated.
• the Tau phosphopeptide comprises the amino acid sequence of one of SEQ ID Nos: 1-3.
• the Tau peptide can have the C-terminus amidated.
• a Tau peptide is presented on the surface of the liposome.
• a Tau peptide preferably a Tau phosphopeptide
• the one or more Tau peptides, including phosphopeptides further comprise one or more modifications, such as palmitoylation or dodecyl modification to allow the Tau peptides to be presented on the surface of the liposome.
• Additional amino acid residues can be added to the Tau peptide to facilitate the modification. It was reported that the position of lipid anchors induces different conformations of the peptide sequence (Hickman et al., J. Biol. Chem. vol. 286, No. 16, pp. 13966-13976, April 22, 2011). While not wishing to be bound by theory, it is believed that adding hydrophobic moieties at both termini may increase the pathological beta-sheet conformation of the Tau peptide. Thus, the one or more Tau peptides further comprise hydrophobic moieties at both termini.
• the modified Tau peptide can have the C-terminus amidated.
• a Tau peptide presented on the surface of the liposome consists of the amino acid sequence of one of SEQ ID NO:27 to SEQ ID NO:29 and SEQ ID NO:31 to SEQ ID NO:38.
• tau liposomes useful for the present invention include, but are not limited, tau liposomes described in U.S. Patent Nos. 8,647,631 and 9,687,447, and International Patent Application No. PCT/US 18/57286, the disclose of each is herein incorporated by reference in its entirety.
• the term “effective amount” refers to an amount of an active ingredient or component that elicits the desired biological or medicinal response in a subject. Selection of a particular effective dose can be determined (e.g., via clinical trials) by those skilled in the art based upon the 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 by the skilled artisan. The precise dose to be employed in the formulation will also depend on the mode of administration, route of administration, target site, physiological state of the patient, other medications administered and the severity of disease, and should be decided according to the judgment of the practitioner and each patient’s circumstances.
• the effective amount of tau phosphopeptide also depends on whether adjuvant is also administered, with higher dosages being required in the absence of adjuvant. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.
• an effective amount of liposomes comprises an amount of Tau phosphopeptide that is sufficient to increase a level of antiphosphorylated Tau antibodies, without inducing a severe adverse event, such as encephalitis.
• an effective amount of liposomes comprises a Tau phosphopeptide at an amount of about 25 nmoles to about 750 nmoles per dose, such as about 29.7 nmoles to about 742.5 nmoles per dose, preferably about 90 nmoles to about 715 nmoles per dose, such as about 89.1 nmoles to about 712.8 nmoles per dose, or about 90 nmoles to about 535 nmoles per dose, such as about 89.1 nmoles to about 534.6 nmoles per dose, or about 90 nmoles to about 275 nmoles per dose, such as about 89.1 nmoles to about 26
• the amount of Tau phosphopeptide administered can also be expressed by weight.
• the tetrapalmitoylated Tau phosphopeptide has four lipidic chains that allow the presentation of the Tau phosphopeptide on the surface of the liposomes.
• the doses of 300, 900, 1800 pg of tetrapalmitoylated Tau phosphopeptide consisting of the amino acid sequence of SEQ ID NO: 28 correspond to 169, 508, 1016 pg, respectively of the corresponding “naked” peptide without any of the lipidic chains.
• an effective amount of liposomes comprises a Tau phosphopeptide at an amount of about 25 nmoles to about 750 nmoles per dose, such as about 25 nmoles, about 30 nmoles, about 35 nmoles, about 40 nmoles, about 45 nmoles, about 50 nmoles, about 55 nmoles, about 60 nmoles, about 65 nmoles, about 70 nmoles, about 75 nmoles, about 80 nmoles, about 85 nmoles, about 90 nmoles, about 95 nmoles, about 100 nmoles, about 125 nmoles, about 150 nmoles, about 175 nmoles, about 200 nmoles, about 225 nmoles, about 250 nmoles, about 275 nmoles, about 300 nmoles, about 325 nmol
• the Tau phosphopeptide consists of the amino acid sequence of one of SEQ ID NO:27 to SEQ ID NO:29 and SEQ ID NO:31 to SEQ ID NO:38. More preferably, the Tau phosphopeptide consists of the amino acid sequence of SEQ ID NO:28.
• an effective amount of liposomes comprises a tetrapalmitoylated Tau phosphopeptide at an amount of 100 pg to 2500 pg, 300 pg to 2400 j-ig, 300 j-ig to 1800 pg, or 300 pg to 900 pg per dose, such as 100 pg, 150 pg, 200 pg, 250 pg, 300 pg, 400 pg, 500 pg, 600 pg, 700 pg, 800 pg, 900 pg, 1000 pg, 1100 pg, 1200 pg, 1300 pg, 1400 pg, 1500 pg, 1600 pg, 1700 pg, 1800 pg, 1900 pg, 2000 pg, 2100 pg, 2200 pg, 2300 pg, 2400 pg, or 2500 pg per dose.
• the Tau phosphopeptide is presented on the surface of the liposomes.
• the Tau phosphopeptide comprises the amino acid sequence of one of SEQ ID NOs: 1-3 or 5-12.
• the Tau phosphopeptide consists of the amino acid sequence of one of SEQ ID NO:27 to SEQ ID O:29 and SEQ ID NO:31 to SEQ ID NO:38. More preferably, the Tau phosphopeptide consists of the amino acid sequence of SEQ ID NO:28.
• an effective amount of liposomes further comprises a toll-like receptor 4 agonist at an amount of 30 pg to 900 pg, preferably 100 pg to 585 pg, per dose.
• the effective amount of liposomes can comprise a toll-like receptor 4 agonist at an amount of 30 pg, 50 pg, 100 pg, 150 pg, 200 pg, 250 pg, 300 pg, 330 pg, 360 pg, 390 pg, 420 pg, 450 pg, 480 pg, 500 pg, 520 pg, 540 pg, 560 pg, 580 pg, 600 pg, 700 pg, 800 pg or 900 pg per dose.
• the toll-like receptor 4 comprises 3D- (6-acyl) PHAD®.
• an effective amount of liposomes further comprises a helper T-cell epitope at an amount of 25 pg to 625 pg, preferably 75 pg to 550 pg, such as 75 pg to 450 pg, 80 pg to 540 pg, 82.5 pg to 535 pg, 85 pg to 530 pg, 87.5 pg to 525 pg, or 90 pg to 520 pg, per dose.
• a helper T-cell epitope at an amount of 25 pg to 625 pg, preferably 75 pg to 550 pg, such as 75 pg to 450 pg, 80 pg to 540 pg, 82.5 pg to 535 pg, 85 pg to 530 pg, 87.5 pg to 525 pg, or 90 pg to 520 pg, per dose.
• the effective amount of liposomes can comprise a helper T-cell epitope at an amount of 25 pg, 50 pg, 70 pg, 72.5 pg, 75 pg, 77.5 pg, 80 pg, 82.5 pg, 85 pg, 87.5 pg, 90 pg, 100 pg, 125 pg, 150 pg, 175 pg, 200 pg, 225 pg, 250 pg, 275 pg, 300 pg, 325 pg, 350 pg, 375 pg, 400 pg, 425 pg, 450 pg, 475 pg, 500 pg, 525 pg, 550 pg, 575 pg, 600 pg, or 625 pg per dose.
• an effective amount of liposomes further comprises a helper T-cell epitope at an amount of about 3 nmoles to about 105 nmoles per dose, such as about 4 nmoles, about 5 nmoles, about 6 nmoles, about 7 nmoles, about 8 nmoles, about 9 nmoles, about 10 nmoles, about 15 nmoles, about 20 nmoles, about 25 nmoles, about 30 nmoles, about 35 nmoles, about 40 nmoles, about 45 nmoles, about 50 nmoles, about 55 nmoles, about 60 nmoles, about 65 nmoles, about 70 nmoles, about 75 nmoles, about 80 nmoles, about 85 nmoles, about 90 nmoles, about 95 nmoles, about 100 nmol
• the helper T-cell epitope is a T50 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 13, a T46 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 14, a T48 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 15, a T51 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 16, or a T52 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 17, preferably the helper T-cell epitope is a T50 helper T-cell epitope consisting of the amino acid sequence of SEQ ID NO: 13.
• an effective amount of liposomes further comprises a lipidated CpG oligonucleotide at an amount of 50 pg to 1250 pg, preferably 100 pg to 1000 pg, such as 150 pg to 800 pg, 150-900 pg, 125 pg to 950 pg or 150 pg to 850 pg per dose.
• the effective amount of liposomes can comprise a lipidated CpG oligonucleotide at an amount of 50 pg, 100 pg, 150 pg, 200 pg, 250 pg, 300 pg, 350 pg, 400 pg, 450 pg, 500 pg, 550 pg, 600 pg, 650 pg, 700 pg, 750 pg, 800 pg, 850 pg, 900 pg, 950 pg, 1000 pg, 1050 pg, 1100 pg, 1200 pg, or 1250 pg per dose.
• the lipidated CpG oligonucleotide is a CpG oligonucleotide comprising a nucleotide sequence of one of SEQ ID NOs: 18-22, preferably the lipidated CpG oligonucleotide is a CpG oligonucleotide comprising a nucleotide sequence of SEQ ID NO: 18.
• the lipidated CpG oligonucleotide is a CpG oligonucleotide comprising a nucleotide sequence of SEQ ID NO: 18 which has one or more phosphorothioate intemucleotide linkages and is covalently linked to cholesterol via a linker comprising polyethylene glycol (PEG).
• PEG polyethylene glycol
• the effective amount of liposomes comprise 50 pg, 100 pg, 150 pg, 200 pg, 250 pg, 300 pg, 350 pg, 400 pg, 450 pg, 500 pg, 550 pg, 600 pg, 650 pg, 700 pg, 750 pg, 800 pg, 850 pg, 900 pg, 950 pg, 1000 pg, 1050 pg, 1100 pg, 1200 pg, or 1250 pg per dose of the CpG oligonucleotide covalently linked to cholesterol via the PEG linker.
• a “sustained immune response” or a “sustainable immune response” refers to an immune response that lasts at least six weeks after the initial administration of an effective amount of a liposome.
• a “sustained immune response” is a sustained antibody response that lasts at least six weeks, at least 12 week, at least 24 weeks, at least 36 weeks, at least 48 weeks, at least 60 weeks, at least 72 weeks or longer, and the antibody response can be characterized by the presence of antiphosphorylated Tau IgG, anti-phosphorylated Tau IgM, or anti-ePHF.
• Anti-phosphorylated Tau IgG, anti -phosphorylated Tau IgM and anti-ePHF can be detected and measured by any method known to one of skill in the art, including those described herein.
• an “antibody response that lasts” refers to an antibody response that is maintained at a level equal to or higher than a defined threshold level during a specified period of time after the initial administration of an effective amount of a liposome, and the defined threshold level is higher than a baseline level measured before the initial administration of the effective amount of the liposome.
• the baseline level is determined based on the average measured level of antibody titers before the initial administration, preferably two measurements are performed.
• the antibody response comprises a specific IgG antibody response directed against the pTau, and the defined threshold level is at least 1.5 or more times of the baseline level, such as at least 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 or more times of the baseline level.
• the antibody response comprises an IgG immune response against ePHF
• the defined threshold level is at least 2.0 or more times of the baseline level, such as at least 2.0, 2.1, 2.2, 2.3, 2.4, 2.5 or more times of the baseline level.
• the human subject is in need of treatment of a neurodegenerative disease, disorder, or condition.
• neurodegenerative disease, disorder, or condition includes any neurodegenerative disease, disorder, or condition known to those skilled in the art in view of the present disclosure.
• Examples of neurodegenerative diseases, disorders, or conditions include neurodegenerative diseases or disorders caused by or associated with the formation of neurofibrillary lesions, such as Tau-associated diseases, disorders or conditions, referred to as Tauopathies.
• the neurodegenerative disease, disorder, or condition includes any of the diseases or disorders which show co-existence of Tau and amyloid pathologies including, but not is limited to, Alzheimer’s Disease, Parkinson’s Disease, Creutzfeldt- Jacob disease, Dementia pugilistica, Down Syndrome, Gerstmann- Straussler-Scheinker disease, inclusion body myositis, prion protein cerebral amyloid angiopathy, traumatic brain injury, amyotrophic lateral sclerosis, parkinsonism-dementia complex of Guam, Non-Guamanian motor neuron disease with neurofibrillary tangles, argyrophilic grain dementia, corticobasal degeneration, Dementia Lewy Amyotrophic Lateral sclerosis, diffuse neurofibrillary tangles with calcification, frontotemporal dementia, preferably frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar dementia, Hallervor
• the neurodegenerative disease, disorder, or condition is Alzheimer’s Disease or another Tauopathy.
• the neurodegenerative disease, disorder, or condition is Alzheimer’s Disease.
• the clinical course of Alzheimer's Disease can be divided into stages, with progressive patterns of cognitive and functional impairments. The stages can be defined using grading scales known in the art including, e.g., NIA-AA Research Framework.
• the neurodegenerative disease, disorder, or condition is early Alzheimer’ s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease, mild Alzheimer’s Disease, or mild to moderate Alzheimer’s Disease.
• MCI mild cognitive impairment
• the subject in need of a treatment is amyloid positive in the brain but does not yet show significant cognitive impairment.
• the amyloid deposition in the brain can be detected using methods known in the art, such as PET scan, immunoprecipitation mass spectrometry or other methods.
• TLR refers to a class of pattern recognition receptor (PRR) proteins that play a key role in the innate immune response.
• PRR pattern recognition receptor
• TLRs recognize pathogen-associated molecular patterns (PAMPs) from microbial pathogens, such as bacteria, fungi, parasites and viruses, which can be distinguished from host molecules.
• PAMPs pathogen-associated molecular patterns
• 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.
• TLR4 recognizes lipopolysaccharides (LPS), a component present in many Gram- negative bacteria, as well as viral proteins, polysaccharide, and endogenous proteins such as low-density lipoprotein, beta-defensins and heat shock protein
• TLR9 is a nucleotide-sensing TLR which is activated by unmethylated cytosine-phosphate-guanine (CpG) single- stranded or double-stranded dinucleotides, which are abundant in prokaryotic genomes but rare in vertebrate genomes.
• CpG cytosine-phosphate-guanine
• TLRs Activation of TLRs leads to a series of signaling events 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 then results in the clearance of the invading pathogens and the infected cells.
• IFNs type I interferons
• cytokines inflammatory cytokines
• chemokines chemokines
• agonist refers to a molecule that binds to one or more TLRs and induces a receptor mediated response.
• an agonist can induce, stimulate, increase, activate, facilitate, enhance, or up regulate the activity of the receptor.
• activities are referred to as “agonistic activities.”
• a TER4 or TER9 agonist can activate or increase 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 or interact with receptors. Agonists can mimic the activity of a natural receptor ligand.
• Agonists can be homologous to these natural receptor ligands with respect to sequence, conformation, charge or other characteristics such that they can be recognized by the receptors. This recognition can result in physiologic and/or biochemical changes within the cell, such that the cell reacts to the presence of the agonist in the same manner as if the natural receptor ligand were present.
• the toll-like receptor agonist is at least one of a toll-like receptor 4 agonist and a toll-like receptor 9 agonist.
• induction of an immune response can include, for example, activation, proliferation, or maturation of a population of immune cells, increasing the production of a cytokine, and/or another indicator of increased immune function.
• induction of an immune response can include increasing the proliferation of B cells, producing antigen-specific antibodies, increasing the proliferation of antigen- specific T cells, improving dendritic cell antigen presentation and/or an increasing expression of certain cytokines, chemokines and co- stimulatory markers.
• toll-like receptor 4 agonist refers to any compound that acts as an agonist of TER4. Any suitable toll-like receptor 4 agonist known to those skilled in the art in view of the present disclosure can be used in the invention.
• Examples of toll-like receptor 4 ligand useful for the invention include TER4 agonist, including, but not limited to, monophosphoryl lipid A (MPEA).
• MPEA monophosphoryl lipid A
• MPLA refers to a modified form of lipid A, which is the biologically active part of Gramnegative bacterial lipopolysaccharide (EPS) endotoxin. MPLA is less toxic than LPS while maintaining the immunostimulatory activity.
• MPLA stimulates both cellular and humoral responses to the vaccine antigen.
• MPLA include, but are not limited to, 3-O-desacyl-4 ' -monophosphoryl lipid A, Monophosphoryl Hexa-acyl Lipid A, 3-Deacyl (Synthetic) (also referred to as 3D-(6-acyl) PHAD®), monophosphoryl 3-deacyl 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®, PHADO-504, 3D-PHAD® from Avanti Polar Lipids (Alabaster, Alabama, USA) or MPLTM from various commercial sources.
• the toll-like receptor 4 agonist is MPLA.
• the liposome comprising a Tau phosphopeptide and a toll-like receptor 4 agonist also comprises a helper T- cell epitope that is capable of binding most or all HLA DR (Human Leukocyte Antigen - antigen D Related) molecules.
• the helper T-cell epitope is then able to activate CD4 + T-cells and provides essential maturation and survival signals to the Tau-specific B-cells.
• the Tau liposomes can be used to generate high-quality antibodies against the pTau antigen in homologous or heterologous immunization schemes, with the liposome used in the prime and/or in the boost.
• helper T-cell epitope refers to a polypeptide comprising an epitope that is capable of recognition by a helper T-cell.
• helper T-cell epitopes include, but are not limited to, tetanus toxoid (e.g., the P2 and P30 epitopes, also named, respectively as T2 and T30), Hepatitis B surface antigen, cholera toxin B, diphtheria toxoid, measles virus F protein, Chlamydia trachomatis major outer membrane protein, Plasmodium falciparum circumsporozite T, P.
• HA hemagglutinin
• 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.
• the helper T-cell epitope comprises two or more of the amino acid sequences of SEQ ID NO:23 to SEQ ID NO:26 fused together via a linker, such as a peptide linker comprising one or more amino acids, e.g., Vai (V), Ala (A), Arg (R), Gly (G), Ser (S), Lys (K).
• the length of the linker can vary, preferably 1-5 amino acids.
• the helper T-cell epitope comprises three or more of the amino acid sequences of SEQ ID NO:23 to SEQ ID NO:26 fused together via one or more linkers selected from the group consisting of VVR, GS, RR, RK.
• the helper T-cell epitope can have its C-terminus amidated.
• the helper T-cell epitopes can be incorporated on the liposomal surface, e.g. anchored by a covalently bound hydrophobic moiety wherein said hydrophobic moiety is an alkyl group, a fatty acid, a triglyceride, diglyceride, steroid, sphingolipid, glycolipid or a 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.
• a covalently bound hydrophobic moiety is an alkyl group, a fatty acid, a triglyceride, diglyceride, steroid, sphingolipid, glycolipid or a phospholipid, particularly an alkyl group or a fatty acid, particularly with a carbon backbone of
• the hydrophobic moiety is palmitic acid.
• the helper T-cell epitopes can be encapsulated in the liposomes. According to particular embodiments, the helper T-cell epitope is encapsulated in the liposome.
• helper T-cell epitope can be modified for its desired location in the liposomes using methods known in the art in view of the present disclosure.
• the helper T-cell epitope useful for the invention comprises an amino acid sequence of one of SEQ ID NO:39 to SEQ ID NO:44.
• 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.
• the liposome comprising a Tau phosphopeptide and a toll-like receptor 4 agonist also comprises a toll-like receptor 9 agonist.
• the term “toll-like receptor 9 agonist” refers to any compound that acts as an agonist of TLR9. Any suitable toll-like receptor 9 agonist known to those skilled in the art in view of the present disclosure can be used in the invention. Examples of toll-like receptor 9 ligand useful for the invention include TLR9 agonist including, but not limited to, CpG oligonucleotides.
• CpG oligonucleotide refers to an oligonucleotide comprising at least one CpG motif.
• oligonucleotide refers to a polynucleotide formed from a plurality of linked nucleotide units. Such oligonucleotides can be obtained from existing nucleic acid sources or can be produced by synthetic methods.
• CpG motif refers to a nucleotide sequence which contains 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 intemucleotide linkages.
• CpG oligonucleotide is lipidated, i.e., conjugated (covalently linked) to a lipid moiety.
• 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 a phospholipid, particularly a sterol such as cholesterol, or fatty acids, when attached to highly hydrophilic molecules, such as nucleic acids, can substantially enhance plasma protein binding and consequently circulation half-life of the hydrophilic molecules.
• lipid moiety conjugated to the phosphopeptides and/or CpG oligonucleotide allows anchoring the said peptides and/or oligonucleotides into the membrane of a liposome via a hydrophobic moiety.
• the CpG oligonucleotide can comprise any suitable internucleotide linkages.
• nucleotide linkage refers to a chemical linkage to join two nucleotides through their sugars consisting of a phosphorous atom and a charged or neutral group between adjacent nucleosides.
• intemucleotide linkage 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.
• Oligonucleotide phosphorothioates are typically synthesized as a random racemic mixture of Rp and Sp phosphorothioate linkages.
• CpG oligonucleotide Any suitable CpG oligonucleotide known to those skilled in the art can be used in the invention in view of the present disclosure.
• CpG oligonucleotides include, but are not limited to CpG2006 (also known as CpG 7909) (SEQ ID NO: 18), CpG 1018 (SEQ ID NO: 19), CpG2395 (SEQ ID NO: 20), CpG2216 (SEQ ID NO: 21) or CpG2336 (SEQ ID NO: 22).
• a CpG oligonucleotide can be lipidated using methods known in the art in view of the present disclosure.
• the CpG oligonucleotide is covalently linked to a cholesterol molecule directly.
• the 3’ terminus of a CpG oligonucleotide is covalently linked to a cholesterol molecule through a phosphate bond, optionally via a PEG linker.
• the 5’ terminus of a CpG oligonucleotide is covalently linked to a cholesterol molecule through a phosphate bond, optionally via a PEG linker.
• a CpG oligonucleotide can be covalently linked to a lipid anchor of the same length as the phospholipids from liposome: one palmitic acid chain (using Pal-OH or similar, activated for coupling) or two palmitic acids (e.g., using 1 ,2-dipalmitoyl- sn-glycero-3-phosphoethanolamine-N-(succinyl) or similar, activated for coupling), optionally via a PEG linker.
• the length of PEG can vary, from example, from 1 to 5 PEG units.
• linkers can also be used to covalently connect a CpG oligonucleotide to a lipophilic moiety (such as a cholesterol molecule), examples of which include, but are not limited to an alkyl spacer having 3 to 12 carbons.
• a short linker compatible with oligonucleotide chemistry is needed as aminodiol.
• no linker is used for the covalent bonding. See e.g., Ries et al., “Convenient synthesis and application of versatile nucleic acid lipid membrane anchors in the assembly and fusion of liposomes,” Org. Biomol. Chem., 2015, 13, 9673, the relevant disclosure of which is incorporated herein by reference.
• lipidated 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 intemucleotide linkages, and the nucleotide sequence is covalently linked to at least one cholesterol via a linker.
• the lipidated CpG oligonucleotide comprises a nucleotide sequence of SEQ ID NO: 18, has one or more phosphorothioate intemucleotide linkages, and is covalently linked to cholesterol.
• Any suitable linkers can be used to covalently link a CpG oligonucleotide to a cholesterol molecule.
• the linker comprises polyethylene glycol (PEG).
• the liposome further comprises one or more lipids selected from the group consisting of l,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), l,2-dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol (DMPG), and cholesterol.
• the liposome further comprises a buffer. Any suitable buffer known to those skilled in the art in view of the present disclosure can be used in the invention.
• the liposome comprises a phosphate-buffered saline.
• the buffer comprises histidine and sucrose.
• An exemplary liposome used in the present invention comprises a Tau tetrapalmitoylated phosphopeptide (pTau Peptide T3, SEQ ID NO: 28) that is presented on the surface of the liposome via two palmitic acids at each terminus of the Tau peptide; A TLR-9 ligand comprising lipidated CpG (Adjuvant CpG7909 (CpG2006); SEQ ID NO: 18) incorporated into the liposome membrane via a cholesterol molecule that is covalently linked to the CpG via a PEG linker; a TLR-4 ligand (Monophosphoryl lipid A (e.g., 3D-(6-acyl) PHAD®)) incorporated into the membrane; an encapsulated helper T-cell epitope (PAN-DR binder T50; SEQ ID NO: 13); and l,2-dimyristoyl-sn-glycero-3 -phospho-choline (DMPC), l,2-dimy
• Liposomes of the invention can be made using methods known in the art in view of the present disclosure.
• the optimal ratios of each component of the liposomes can be determined by techniques known to those skilled in the art in view of the present disclosure.
• the liposomes can be administered by suitable means for prophylactic and/or therapeutic treatment.
• the liposomes are administered by subcutaneous or intramuscular injection. Intramuscular injection is most typically performed in the arm or leg muscles.
• the invention relates to pharmaceutical compositions comprising a therapeutically effective amount of liposome, together with a pharmaceutically acceptable excipient and/or carrier.
• Pharmaceutically acceptable excipients and/or carriers 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 can include pharmaceutically-acceptable, non-toxic carriers or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration. The diluent is selected so as not to affect the biological activity of the combination.
• diluents examples include distilled water, physiological phosphate- buffered saline, Ringer’s solutions, dextrose solution, and Hank’s solution.
• the pharmaceutical composition or formulation may also include other carriers, adjuvants, or non-toxic, non-therapeutic, non-immunogenic stabilizers, and the like. It will be understood that the characteristics of the carrier, excipient or diluent will depend on the route of administration for a particular application.
• the target antigen for the vaccine is located in the brain, and the brain is separated from the circulation by a specialized cellular structure called the blood-brain barrier (BBB).
• BBB blood-brain barrier
• the BBB restricts passage of substances from the circulation into the brain. This prevents the entry of toxins, microbes, etc. into 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 that surrounds the brain.
• the avidity of an antibody can be measured by avidity index using methods known in the art in view of the present disclosure.
• the titers of antibodies against a particular antigen are measured at two different concentrations of the coated antigen: one is the saturated concentration, where all antibodies can bind to the antigen and another one is at a low concentration, where only antibodies with the highest binding capacity can bind to the antigen.
• “avidity index” refers to the ratio of the levels of antibody titers measured at the low- and the high-density coating of the antigen.
• avidity of antibodies against an antigen can be measured at different time points after an immunization or following different immunizations, to evaluate whether the avidity (as measured by the avidity index) increases over time.
• antibodies with an “increased avidity” or “increased binding avidity” to an antigen refers to antibodies with an increased avidity index to the antigen over time during the course of a treatment or immunization.
• An increased avidity suggests a potential affinity maturation of the antibodies.
• the pharmaceutical compositions of the present invention therefore further comprise one or more suitable adjuvants to achieve the desired immune response in the subject.
• Suitable adjuvants can be administered before, after, or concurrent with administration of the liposome.
• Preferred adjuvants augment the intrinsic response to an immunogen without causing conformational changes in the immunogen that affect the qualitative form of the response.
• examples of adjuvants are the aluminum salts (alum), such as aluminum hydroxide, aluminum phosphate, and aluminum sulfate.
• Such adjuvants can be used with or without other specific immunostimulating agents, such as MPLA Class (3 De-O-acylated monophosphoryl lipid A (MPLTM), monophosphoryl hexaacyl Lipid A 3-deacyl synthetic (3D-(6-acyl) PHAD®, PHADTM, PHAD®-504, 3D-PHAD®) lipid A), polymeric or monomeric amino acids, such as polyglutamic acid or polylysine.
• MPLTM De-O-acylated monophosphoryl lipid A
• PHADTM monophosphoryl hexaacyl Lipid A 3-deacyl synthetic (3D-(6-acyl) PHAD®, PHADTM, PHAD®-504, 3D-PHAD®) lipid A
• polymeric or monomeric amino acids such as polyglutamic acid or polylysine.
• Such 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-acetyl- normuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-alanyl-D- isoglutaminyl-L-alanine-2-(l ' -2' dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)- ethylamine (MTP-PE), N-acetylglucsaminyl-N-acetylmuramyl-L-Al-D-isoglu-L-Ala- dipalmitoxy propylamide (DTP-DPP) TheramideTM), or other bacterial cell wall components.
• muramyl peptides
• Oil-in-water emulsions include MF59 (see WO 90/14837), containing 5% Squalene, 0.5% Tween 80, and 0.5% Span 85 (optionally containing various 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 thr-MDP, either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion; and the RibiTM adjuvant system (RAS) (Ribi ImmunoChem, Hamilton, Mont.) 0.2% Tween 80, and one or more bacterial cell wall components selected from the group consisting of monophosphoryl lipid A (MPLTM), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPLTM+CWS (DetoxTM).
• MPLTM monophosphoryl lipid A
• CFA Complete Freund’s Adjuvant
• cytokines such as interleukins (IL-1, IL-2, and IL-12), macrophage colony stimulating factor (M-CSF), and tumor necrosis factor (TNF).
• the term “in combination,” in the context of the administration of two or more therapies to a subject, refers to the use of more than one therapy.
• the use of the term “in combination” does not restrict the order in which therapies are administered to a subject.
• a first therapy e.g., a composition described herein
• a first therapy can be administered prior to (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 subsequent to (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 subject.
• a pharmaceutical composition according to the invention can be used in combination with a biologically active substance such as, for example, known compounds used in the medication of tauopathies and/or of amyloidosis, a group of diseases and disorders associated with amyloid or amyloid-like protein such as the amyloid protein involved in Alzheimer's Disease.
• a biologically active substance such as, for example, known compounds used in the medication of tauopathies and/or of amyloidosis, a group of diseases and disorders associated with amyloid or amyloid-like protein such as the amyloid protein involved in Alzheimer's Disease.
• the other biologically active compound may include neutron-transmission enhancers, psychotherapeutic drugs, acetylcholine esterase inhibitors, calcium channel blockers, biogenic amines, benzodiazepine tranquillizers, acetylcholine synthesis, storage or release enhancers, acetylcholine postsynaptic receptor agonists, monoamine oxidase-A or -B inhibitors, N-methyl-O-aspartate glutamate receptor antagonists, non-steroidal anti-inflammatory drugs, antioxidants, and serotonergic receptor antagonists.
• neutron-transmission enhancers may include neutron-transmission enhancers, psychotherapeutic drugs, acetylcholine esterase inhibitors, calcium channel blockers, biogenic amines, benzodiazepine tranquillizers, acetylcholine synthesis, storage or release enhancers, acetylcholine postsynaptic receptor agonists, monoamine oxidase-A or -B inhibitors, N-
• the other biologically active compound may be selected from the group consisting of compounds against oxidative stress, anti- apop to tic compounds, metal chelators, inhibitors of DNA repair such as pirenzepin and metabolites, 3- amino-1 -propanesulfonic acid (3APS), 1,3-propanedisulfonate (1,3PDS), secretase activators, and v -secretase inhibitors, tau proteins, neurotransmitter, P-sheet breakers, anti-inflammatory molecules, or cholinesterase inhibitors (ChEls) such as tacrine, rivastigmine, donepezil, and/or galantamine and other drugs and nutritive supplements, together with an therapeutic vaccine according to the invention and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.
• a pharmaceutically acceptable carrier and/or a diluent and/or an excipient such as tacrine, rivastigmine, donepezil, and/or gal
• the other biologically active compound may comprise niacin or memantine together with a liposome according to the invention and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.
• other compounds comprises "atypical antipsychotics" such as, for example clozapine, ziprasidone, risperidone, aripiprazole or olanzapine for the treatment of positive and negative psychotic symptoms including hallucinations, delusions, thought disorders (manifested by marked incoherence, derailment, tangentiality), and playful or disorganized behavior, as well as anhedonia, flattened affect, apathy, and social withdrawal, together with a liposome of the invention.
• the timing of administrations can vary significantly from once a day, to once a year, to once a decade.
• a typical regimen consists of an immunization followed by booster injections at time intervals, such as 1 to 24-week intervals.
• Another regimen consists of an immunization followed by booster injections 1, 2, 4, 6, 8, 10 and 12 months later.
• Another regimen entails an injection every two months for life.
• booster injections can be on an irregular basis as indicated by monitoring of immune response.
• the regimen for the priming and boosting administrations can be adjusted based on the measured immune responses after the administrations.
• the boosting compositions are generally administered weeks or months after administration of the priming composition, for example, about 1 week, or 2 weeks, or 3 weeks, or 4 weeks, or 8 weeks, or 16 weeks, or 20 weeks, or 24 weeks, or 28 weeks, or 32 weeks, or 36 weeks, or 40 weeks, or 44 weeks, or 48 weeks, or 52 weeks, or 56 weeks, or 60 weeks, or 64 weeks, or 68 weeks, or 72 weeks, or 76 weeks, or one to two years after administration of the priming composition.
• one or more boosting immunizations can be administered.
• the antigens in the respective priming and boosting compositions need not be identical, but should share antigenic determinants or be substantially similar to each other.
• the administration of an effective amount of a liposome described herein is able to induce a potent antibody response against pTau in a patient in need thereof, such as a patient in need of treating an Alzheimer’s Disease (e.g., mild to moderate Alzheimer’s Disease or early Alzheimer’ s Disease) or mild cognitive impairment (MCI) due to Alzheimer’s Disease.
• the antibody response is sustainable, e.g., lasting at least 6 weeks after the initial administration of the liposome.
• the antibody response is also boosted by one or more subsequent boosting administrations.
• boosted in the context of an antibody response refers to the antibody response that is maintained or enhanced after a subsequent administration as measured at least two weeks after the administration of the subsequent administration.
• an antibody response is “boosted” by a subsequent administration, if there is an increase of the antibody titer when measured 2 weeks after the subsequent administration as compared with the antibody titer before the subsequent administration.
• Pharmaceutical compositions of the present invention can be formulated according to methods known in the art in view of the present disclosure. The optimal ratios of each component in the compositions can be determined by techniques known to those skilled in the art in view of the present disclosure.
• a Tau peptide via administration of a pharmaceutical composition according to an embodiment of the invention, induces an active immune response in the subject, such as an antibody response to the Tau peptide and to the pathological form of Tau, thereby facilitating the clearance of related Tau aggregates, slowing the progression of Tau-pathology related behavior and/or treating the underlying Tauopathy.
• an active immune response such as an antibody response to the Tau peptide and to the pathological form of Tau, thereby facilitating the clearance of related Tau aggregates, slowing the progression of Tau-pathology related behavior and/or treating the underlying Tauopathy.
• Tau is a human “self’ protein. This means that, in principle, all lymphocytes bearing a receptor specific for tau should have been deleted during development (central tolerance) or rendered unresponsive by a peripheral tolerance mechanism. This problem has proved to be a significant roadblock to the development of vaccines against self or “altered self’ proteins (e.g., tumor antigens). Generating high-quality antibodies against an antigen (self or infectious) requires the action of not only B lymphocytes, which produce the antibody, but also of CD4 + T “helper” lymphocytes. CD4 + T-cells provide critical survival and maturation signals to B lymphocytes, and CD4 + T-cell deficient animals are profoundly immunosuppressed.
• CD4 + T-cells are also subject to tolerance mechanisms, and an additional roadblock to generating strong anti-self (e.g., anti-tau) antibody responses is that tau-reactive CD4 + T-cells are also likely to be rare to non-existent in the human/animal repertoire.
• an immune response involves the development of a beneficial humoral (antibody mediated) response directed against the Tau peptide and a cellular (mediated by antigen-specific T cells or their secretion products) response directed against the T-cell epitope or the immunogenic carrier.
• a Tau-pathology related behavioral phenotype includes, without limitation, cognitive impairments, early personality change and disinhibition, apathy, abulia, mutism, apraxia, perseveration, stereotyped movements/behaviors, hyperorality, disorganization, inability to plan or organize sequential tasks, selfishness/callousness, antisocial traits, a lack of empathy, halting, agrammatic speech with frequent paraphasic errors but relatively preserved comprehension, impaired comprehension and word-finding deficits, slowly progressive gait instability, retropulsions, freezing, frequent falls, non- levodopa responsive axial rigidity, supranuclear gaze palsy, square wave jerks, slow vertical saccades, pseudobulbar palsy, limb apraxia, dystonia, cortical sensory loss, and tremor.
• a subject having or at risk of having Alzheimer’s Disease or other Tauopathy a subject having Tau aggregates in the brain, or a subject exhibiting a tangle related behavioral phenotype prior to administering the immunogenic peptides or antibodies of the present invention.
• Subjects amenable to treatment include individuals at risk of disease but not showing symptoms, as well as patients presently showing symptoms. In the case of Alzheimer’s Disease, virtually anyone is at risk of suffering from Alzheimer’s Disease. Therefore, the present methods can be administered prophylactically to the general population without the need for any assessment of the risk of the subject patient.
• the present methods are especially useful for individuals who have a known genetic risk of Alzheimer’s disease for the prevention or treatment of the disease.
• Such individuals include those having relatives who have experienced the disease, and those whose risk is determined by analysis of genetic or biochemical markers.
• the subject is in need of a treatment of Alzheimer’s Disease, preferably early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’s Disease, mild Alzheimer’ s Disease, or mild to moderate Alzheimer’ s Disease.
• the subject is in need of a prevention of Alzheimer’s Disease, preferably preclinical Alzheimer’ s Disease, early Alzheimer’ s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease, mild Alzheimer’ s Disease, or mild to moderate Alzheimer’s Disease.
• the preclinical Alzheimer’s Disease is a stage before early Alzheimer’ s Disease.
• treatment can begin at any age (e.g., 10, 20, 30 years of age). Usually, however, it is not necessary to begin treatment until a patient reaches 40, 50, 60, or 70 years of age. Treatment typically entails multiple dosages over a period of time. Treatment can be monitored by assaying antibody, or activated T-cell or B-cell responses to the therapeutic agent over time. If the response decreases, a booster dosage is indicated.
• age e.g. 10, 20, 30 years of age.
• Treatment typically entails multiple dosages over a period of time. Treatment can be monitored by assaying antibody, or activated T-cell or B-cell responses to the therapeutic agent over time. If the response decreases, a booster dosage is indicated.
• compositions containing the Tau peptides are administered to a patient susceptible to, or otherwise at risk of, Alzheimer’s Disease or other Tauopathy in an amount sufficient to eliminate or reduce the risk, lessen the severity, or delay the outset of the disease, including biochemical, histologic and/or behavioral symptoms of the disease, its complications and intermediate pathological phenotypes presented during development of the disease.
• pharmaceutical compositions containing a Tau peptide are administered to a patient suspected of, or already suffering from, such a disease in an amount sufficient to cure, or at least partially arrest, the symptoms of the disease (biochemical, histologic and/or behavioral), including its complications and intermediate pathological phenotypes in development of the disease.
• the composition can, if desired, be presented in a kit, pack or dispenser, which can contain one or more unit dosage forms containing the active ingredient.
• the kit for example, can comprise metal or plastic foil, such as a blister pack.
• the kit, pack, or dispenser can be accompanied by instructions for administration.
• Embodiment 1 A method of inducing an immune response, such as an antibody response against a phosphorylated Tau protein (pTau), in a human subject in need thereof, comprising administering to the subject an effective amount of a liposome comprising:
• a Tau phosphopeptide consisting of the amino acid sequence of SEQ ID NO:27 to SEQ ID NO:29 and SEQ ID NO:31 to SEQ ID NO:38 at an amount of 300 pg to 1800 pg per dose;
• a toll-like receptor 4 agonist comprising monophosphoryl lipid A
• helper T-cell epitope having an amino acid sequence selected from the group consisting of SEQ ID NO: 13 to SEQ ID NO: 17, SEQ ID NO:23 to SEQ ID NO:26, and SEQ ID NO:39 to SEQ ID NO:44;
• a CpG oligonucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO: 18 to SEQ ID NO:22 , wherein: the Tau phosphopeptide is presented on the surface of the liposome, and the antibody response lasts at least 6 weeks after the initial administration of the effective amount of the liposome to the human subject.
• Embodiment 2 The method of Embodiment 1, wherein the effective amount of the liposome comprises:
• Tau phosphopeptide consisting of the amino acid sequence of SEQ ID NO: 28 at the amount of 300 pg to 1800 pg per dose;
• Embodiment 3 The method of Embodiment 1 or 2, wherein the effective amount of the liposome comprises 300 pg, 900 pg or 1800 pg per dose of the Tau phosphopeptide.
• Embodiment 3a The method of Embodiment 1 or 2, wherein the effective amount of the liposome comprises 300 pg per dose of the Tau phosphopeptide.
• Embodiment 3b The method of Embodiment 1 or 2, wherein the effective amount of the liposome comprises 900 pg per dose of the Tau phosphopeptide.
• Embodiment 3c The method of Embodiment 1 or 2, wherein the effective amount of the liposome comprises 1800 pg per dose of the Tau phosphopeptide.
• Embodiment 4 The method of any one of Embodiments l-3c, wherein the effective amount of the liposome is administered subcutaneously.
• Embodiment 5 The method of any one of Embodiments l-3c, wherein the effective amount of the liposome is administered intramuscularly.
• Embodiment 6 The method of any one of Embodiments 1-5, wherein the CpG oligonucleotide has one or more phosphorothioate intemucleotide linkages, and the CpG oligonucleotide is covalently linked to at least one lipophilic group, optionally via a PEG linker.
• Embodiment 7 The method of any one of Embodiments 1-6, wherein the Tau phosphopeptide consists of the amino acid sequence of SEQ ID NO:28, the toll-like receptor 4 agonist comprises monophosphoryl hexa-acyl Lipid A, 3-deacyl, the helper T-cell epitope comprises the amino acid sequence of SEQ ID NO: 39, the CpG oligonucleotide comprises the nucleotide sequence of SEQ ID NO: 18, and the liposome further comprises at least one lipid selected from the group consisting of 1,2-dimyristoyl-sn-glycero- 3 -phosphocholine (DMPC), l,2-dimyristoyl-sn-glycero-3-phosphoryl-3’-rac-glycerol (DMPG), and cholesterol.
• DMPC 1,2-dimyristoyl-sn-glycero- 3 -phosphocholine
• DMPG 1,2-dimyristoyl-sn-glycero-3-
• Embodiment 8 The method of any one of Embodiments 1-7, wherein the antibody response has conformation specificity against pathological Tau protein, which increases over time after the initial administration of the effective amount of the liposome to the human subject, preferably wherein the antibody response comprises a specific IgG antibody response directed against the pTau, preferably the specific IgG antibody response has an anti-pTau IgG titer at least 50, 60, 70, 80, 90, 100 or more times higher than that of a placebo control.
• Embodiment 9 The method of any one of Embodiments 1-8, wherein the antibody response induces a class switch of a specific IgM antibody response to a specific IgG antibody response directed against the pTau, with indication for memory building.
• Embodiment 10 The method of any one of Embodiments 1-9, wherein the antibody response comprises an IgG immune response that preferentially recognizes the pTau over non- phosphorylated Tan protein, preferably the ratio of the anti-pTau IgG titer to the anti- Tau IgG titer is at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70.
• Embodiment 10a The method of any one of Embodiments 1-10, wherein the IgG immune response against pTau is maintained over time.
• Embodiment 10b The method of any one of Embodiments l-10a, wherein the IgG immune response against non- phosphorylated Tau protein becomes lower over time.
• Embodiment 11 The method of any one of Embodiments l-10b, wherein the antibody response comprises an IgG immune response against an enriched Paired Helical Filament (ePHF).
• ePHF Paired Helical Filament
• Embodiment 12 The method of Embodiment 11, wherein the IgG immune response has an anti-ePHF IgG titer at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more times higher than that of a placebo control.
• Embodiment 12a The method of Embodiments 11 or 12, wherein the IgG immune response matures toward a stronger preference for binding to ePHF while concomitantly lowering antibody titers towards the non-phosphorylated Tau.
• Embodiment 12b The method of Embodiments 11 or 12, wherein the IgG immune response has a higher IgG titer towards ePHF than an IgG titer towards non-phosphorylated Tau.
• Embodiment 13 The method of Embodiments 11, 12, 12a or 12b, wherein the anti- ePHF IgG has an increased binding avidity to the pathological ePHF Tau for at least 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, 22 weeks, 24 weeks or longer after the initial administration of the effective amount of the liposome, preferably the anti-ePHF IgG has an avidity index of at least 0.3, 0.4, 0.5, 0.6, or 0.7.
• Embodiment 14 The method of any one of Embodiments 1-13, further comprising administering to the subject a second dose of the effective amount of liposome 4 to 12 weeks, such as 8 weeks, after the initial administration of the effective amount of liposome.
• Embodiment 14a The method of Embodiment 14, wherein the effective amount of liposome comprises 300 pg per dose of the Tau phosphopeptide for each of the initial administration and the second dose, and the second dose is administered to the subject 8 weeks after the initial administration.
• Embodiment 14b The method of Embodiment 14, wherein the effective amount of liposome comprises 900 pg per dose of the Tau phosphopeptide for each of the initial administration and the second dose, and the second dose is administered to the subject 8 weeks after the initial administration.
• Embodiment 14c The method of Embodiment 14, wherein the effective amount of liposome comprises 1800 pg per dose of the Tau phosphopeptide for each of the initial administration and the second dose, and the second dose is administered to the subject 8 weeks after the initial administration.
• Embodiment 15 The method of any one of Embodiments 14- 14c, wherein the antibody response comprising the IgG immune response against pTau is boosted after the administration of the second dose of the effective amount of liposome, preferably the antibody response is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• Embodiment 15a The method of Embodiment 15, wherein the anti-ePHF IgG response is boosted after the administration of the second dose of the effective amount of liposome as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• Embodiment 16 The method of any one of Embodiments 14- 15a, further comprising administering to the subject a third dose of the effective amount of liposome 20 to 28 weeks, such as 24 weeks, after the initial administration of the effective amount of liposome.
• Embodiment 16a The method of Embodiment 16, wherein the effective amount of liposome comprises 300 pg per dose of the Tau phosphopeptide for each of the initial administration, the second dose and the third dose, and the second dose and the third dose are respectively administered to the subject 8 weeks and 24 weeks after the initial administration.
• Embodiment 16b The method of Embodiment 16, wherein the effective amount of liposome comprises 900 pg per dose of the Tau phosphopeptide for each of the initial administration, the second dose and the third dose, and the second dose and the third dose are respectively administered to the subject 8 weeks and 24 weeks after the initial administration.
• Embodiment 16c Embodiment 16c.
• Embodiment 16 wherein the effective amount of liposome comprises 1800 pg per dose of the Tau phosphopeptide for each of the initial administration, the second dose and the third dose, and the second dose and the third dose are respectively administered to the subject 8 weeks and 24 weeks after the initial administration.
• Embodiment 17 The method of any one of Embodiments 16- 16c, wherein the antibody response comprising the IgG immune response against pTau is boosted after the administration of the third dose of the effective amount of the liposome, preferably the antibody response is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more as measured at least 2 weeks after the administration of the third dose of the effective amount of liposome.
• Embodiment 17a The method of Embodiment 17, wherein the anti-ePHF IgG response is boosted after the administration of the third dose of the effective amount of liposome as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• Embodiment 18 The method of any one of Embodiments 16- 17a, further comprising administering to the subject a fourth dose of the effective amount of liposome 44 to 52 weeks, such as 48 weeks, after the initial administration of the effective amount of liposome.
• Embodiment 18a The method of Embodiment 18, wherein the effective amount of liposome comprises 300 pg per dose of the Tau phosphopeptide for each of the initial administration, the second, third and fourth doses, and the second, third and fourth doses are respectively administered to the subject 8 weeks, 24 weeks and 48 weeks after the initial administration.
• Embodiment 18b The method of Embodiment 18, wherein the effective amount of liposome comprises 900 pg per dose of the Tau phosphopeptide for each of the initial administration, the second, third and fourth doses, and the second, third and fourth doses are respectively administered to the subject 8 weeks, 24 weeks and 48 weeks after the initial administration.
• Embodiment 18c The method of Embodiment 18, wherein the effective amount of liposome comprises 1800 pg per dose of the Tau phosphopeptide for each of the initial administration, the second, third and fourth doses, and the second, third and fourth doses are respectively administered to the subject 8 weeks, 24 weeks and 48 weeks after the initial administration.
• Embodiment 19 The method of Embodiment 18, wherein the antibody response comprising the IgG immune response against pTau is boosted after the administration of the fourth dose of the effective amount of liposome, preferably the antibody response is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more as measured at least 2 weeks by the administration of the fourth dose of the effective amount of liposome.
• Embodiment 19a The method of Embodiment 19, wherein the anti-ePHF IgG response is boosted after the administration of the fourth dose of the effective amount of liposome as measured at least 2 weeks after the administration of the second dose of the effective amount of liposome.
• Embodiment 20 A method of inducing a sustained immune response against a phosphorylated Tau protein (pTau) in a human subject in need thereof, comprising: i. intramuscularly administering to the subject a primer vaccine comprising an effective amount of a liposome; and ii. intramuscularly administering to the subject a first booster vaccine comprising the effective amount of the liposome 6-10 weeks after the administration of the primer vaccine, wherein: the sustained immune response lasts at least about 20 weeks after the administration of the primer vaccine; the liposome comprises:
• a toll-like receptor 4 agonist comprising monophosphoryl lipid A
• helper T-cell epitope having an amino acid sequence selected from the group consisting of SEQ ID NOs:23, 24, 25. and 26;
• helper T-cell epitope at an amount of 75 pg to 550 pg per dose, such as 75 pg to 450 pg, 80 pg to 540 pg, 82.5 pg to 535 pg, 85 pg to 530 pg, 87.5 pg to 525 pg, or 90 pg to 520 pg per dose; and
• the CpG oligonucleotide at an amount of 100 pg to 1000 pg, such as 150-800 pg, 125 pg to 950 pg, 150 pg to 900 pg, or 150 pg to 850 pg per dose.
• Embodiment 20a The method of Embodiment 20, wherein the helper T-cell epitope has an amino acid sequence selected from the group consisting of SEQ ID NOs:39, 40, 41 , 42, and 43.
• Embodiment 20b The method of Embodiment 20, wherein the helper T-cell epitope has an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 14, 15, 16, 17, and 44.
• Embodiment 20c The method of any one of Embodiments 20 - 20b, wherein the lipidaied CpG oligonucleotide has the nucleotide sequence of SEQ ID NO:18 and the CpG oligonucleotide is covalently linked to at least one cholesterol via a linker.
• Embodiment 20d The method of any one of Embodiments 20 - 20c, wherein the first booster vaccine is administered 8 weeks after the administration of the primer vaccine, and the sustained immune response lasts at least about 24 weeks after the administration of the primer vaccine.
• Embodiment 20e The method of any one of Embodiments 20 - 20d, further comprising intramuscularly administering to the subject a second booster vaccine comprising the effective amount of the liposome 22-26 weeks after the administration of the primer vaccine, and the sustained immune response lasts at least about 36 weeks after the administration of the primer vaccine.
• Embodiment 20f The method of Embodiment 20e, wherein the second booster vaccine is administered 24 weeks after the administration of the primer vaccine, and the sustained immune response lasts at least about 48 weeks after the administration of the primer vaccine.
• Embodiment 20g The method of Embodiment 20e or 20f, further comprising intramuscularly administering to the subject a third booster vaccine comprising the effective amount of the liposome 45-50 weeks after the administration of the primer vaccine, and the sustained immune response lasts at least about 60 weeks after the administration of the primer vaccine.
• Embodiment 20h The method of Embodiment 20g, wherein the third booster vaccine is administered 48 weeks after the administration of the primer vaccine, and the sustained immune response lasts at least about 72 weeks after the administration of the primer vaccine.
• Embodiment 20i The method of any one of Embodiments 20 - 20h, wherein the effective amount of the liposome comprises 300 pg per dose of the Tau phosphopeptide.
• Embodiment 20j The method of any one of Embodiments 20 - 20h, wherein the effective amount of the liposome comprises 900 pg per dose of the Tau phosphopeptide.
• Embodiment 20k The method of any one of Embodiments 20 - 20h, wherein the effective amount of the liposome comprises 1800 pg per dose of the Tau phosphopeptide. [00178] Embodiment 20k 1.
• Embodiment 20k The method of any one of Embodiments 20i - 20k, wherein the effective amount of the liposome comprises 80 pg to 540 pg per dose of the helper T-cell epitope, and 125 pg to 950 pg per dose of the CpG oligonucleotide. [00179] Embodiment 20k2. The method of any one of Embodiments 20i - 20k, wherein the effective amount of the liposome comprises 82.5 pg to 535 pg per dose of the helper T-cell epitope, and 125 pg to 950 pg per dose of the CpG oligonucleotide.
• Embodiment 20k 3. The method of any one of Embodiments 20i - 20k, wherein the effective amount of the liposome comprises 87.5 pg to 525 pg per dose of the helper T-cell epitope, and 150 pg to 900 pg per dose of the CpG oligonucleotide.
• Embodiment 20k 4.
• Embodiment 201 The method of any one of Embodiments 20 - 20k4, wherein the sustained immune response comprises an IgG immune response that preferentially recognizes the pTau over non- phosphorylated Tau protein, preferably the ratio of the anti-pTau IgG titer to the anti-Tau IgG titer is at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70.
• Embodiment 20m The method of any one of Embodiments 20 - 201, wherein the sustained immune response comprises an IgG immune response against enriched Paired Helical Filament (ePHF) having an anti-ePHF IgG titer at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more times higher than that of a placebo control.
• ePHF Paired Helical Filament
• Embodiment 21 The method of any one of Embodiments l-20m, wherein the subject is in need of clearance of aggregates of Tau.
• Embodiment 22 The method of any one of Embodiments 1-21, wherein the subject is in need of a treatment of Alzheimer’s Disease, such as preclinical Alzheimer’s Disease, mild to moderate Alzheimer’s Disease or early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease.
• Alzheimer’s Disease such as preclinical Alzheimer’s Disease, mild to moderate Alzheimer’s Disease or early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease.
• Embodiment 23 The method of any one of Embodiments 1-21, wherein the subject is in need of a prevention of Alzheimer’s Disease, such as preclinical Alzheimer’s Disease, mild to moderate Alzheimer’s Disease or early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease.
• Alzheimer’s Disease such as preclinical Alzheimer’s Disease, mild to moderate Alzheimer’s Disease or early Alzheimer’s Disease, mild cognitive impairment (MCI) due to Alzheimer’ s Disease.
• Embodiment 24 The method of any one of Embodiments 1-23, wherein the immune response comprises an anti-phosphorylated Tau antibody that binds specifically to an epitope comprising phosphorylated Ser396.
• Embodiment 25 The method of Embodiment 24, wherein the epitope further comprises phosphorylated Ser404.
• ACI-35.030 is a liposome formulation according to embodiments of the invention that contains a phosphorylated Tau peptide having the amino acid sequence of SEQ ID NO: 28, MPLA (3D-(6-acyl) PHAD®), DMPC, DMPG, cholesterol, a helper T-cell epitope of SEQ ID NO: 13, a CpG2006 oligonucleotide covalently linked to a cholesterol group via a PEG linker, and a buffer
• ACI-35 is a liposome formulation according to embodiments of the invention that contains a phosphorylated Tau peptide having the amino acid sequence of SEQ ID NO: 28, MPLA, DMPC, DMPG, cholesterol, and a buffer.
• ACI-35.030 vaccine (ACI-35.030) is evaluated in a clinical Phase lb/2a multicenter, double blind, randomized, placebo-controlled study conducted in patients with early AD (mild cognitive impairment (MCI) due to AD or mild AD) in Europe (ACL35-1802 study). See also, study number NCT04445831 in clinicaltrials.gov. A summary of the design of the study is shown in FIG. 1.
• Each of 3 sub-cohorts consists of patients receiving placebo or different dosages of ACL35.030, referred to by the amount of pTau Peptide T3 in the composition (300 pg, 900 pg or 1800 pg of tetrapalmitoylated phosphopeptide pTau Peptide T3, SEQ ID NO: 28) spread over 48 weeks (dose administrations at weeks 0, 8, 24 and 48), followed by a 24-week follow-up period.
• a safety assessment was/is performed immediately after each dosing and 48 to 72 hours thereafter by telephone call for all study patients.
• the first dosing of the first 4 patients was/is performed once the safety assessment at 48 to 72 hours of the previous patient has been performed to confirm there is no clinically relevant safety issue related to study vaccine, according to the site principal investigator.
• All treated patients have a follow-up period of 24 weeks after the end of the treatment period. During this period, patients are asked to attend a first follow-up visit 19 weeks after the last administration and a last visit at the end of the follow-up period (26 weeks after the last administration). Patients’ safety is monitored throughout the study with regular review of safety data by an independent Data and Safety Monitoring Board (DSMB).
• DSMB Data and Safety Monitoring Board
• the first interim analyses were conducted in all sub-cohorts conducted in cohort 1 once all subjects in their respective sub-cohorts have completed visit 4 (Week 10), i.e., 2 weeks after the second injection.
• the objective was to review safety, tolerability, and immunogenicity data for ACL35.030 up to this time point.
• the second interim analyses were/are conducted in sub-cohorts 1.1, 1.2, and 1.3 once all subjects in their respective sub-cohort have completed visit 6 (Week 26), i.e., 2 weeks after the third injection.
• the objective is to decide to expand potentially cohort 1 in order to collect additional safety /tolerability data at the dose presenting the most favorable profile in terms of immunogenicity, safety and tolerability.
• the third interim analysis was performed at the end of the treatment period (i.e., 2 weeks after the 4th injection).
• the objective is to review the safety/tolerability and immunogenicity data up to this time point, including data from patients of sub-cohort expansion if applicable.
• Biomarker results can be included as supportive exploratory data. The results are compared with those subsequently obtained for other cohorts in order to select, among all study cohorts, the best strategy for further clinical development.
• the fourth interim analysis was/is performed at the end of the follow-up period, i.e., once all cohort 1 patients have completed visit 11 (Week 74).
• the objective is the same as in the fourth interim analysis and the results are subsequently compared across all cohorts.
• the study population is 50-75 years of age (male and female) with a diagnosis of mild AD or MCI due to AD according to NIA-AA criteria.
• MCI Mild Cognitive Impairment due to AD or mild AD according to NIA-AA criteria and a Clinical Dementia Rating scale (CDR) global score of 0.5 or 1.
• CDR Clinical Dementia Rating scale
• MMSE Mini mental state examination
• Immune suppression including but not limited to the use of immunosuppressant drugs or systemic steroids unless they have been prescribed transiently more than 3 months prior to screening.
• DSM-V Diagnostic and Statistical Manual of Mental Disorders-V
• Any clinically significant medical condition likely to interfere with the evaluation of safety and tolerability of the study treatment and/or the adherence to the full study visit schedule.
• Any clinically significant medical condition likely to impact on the immune system and/or expected to potentially impair the immunization potential of the study vaccine in patients (e.g., any history of acquired or innate immunodepressive disorder).
• Concomitant psychiatric or neurologic disorder other than those considered to be related to AD e.g., head injury with loss of consciousness, symptomatic stroke, Parkinson’s disease, severe carotid occlusive disease, TIAs.
• MRI examination cannot be done for any reason, including but not limited to metal implants contraindicated for MRI studies and/or severe claustrophobia.
• Safety and tolerability - adverse events e.g., immediate and delayed reactogenicity (e.g., anaphylaxis, local and systemic reactogenicity, including pain, redness, immune-complex disease, swelling, fever); global assessment of tolerability; suicidal ideation (C-SSRS); behavior (NPI); cognitive and functional assessments (RBANS, CDR-SB) to assess safety; vital signs; MRI imaging; electrocardiogram; routine hematology and biochemistry evaluation in blood and urine; evaluation of autoimmune antibodies including anti-DNA antibodies in blood; inflammatory markers in blood and CSF.
• immediate and delayed reactogenicity e.g., anaphylaxis, local and systemic reactogenicity, including pain, redness, immune-complex disease, swelling, fever
• C-SSRS suicidal ideation
• NPI behavior
• cognitive and functional assessments RBANS, CDR-SB
• Immune response - anti-pTau IgG titers in serum (geometric mean, change from baseline, responder rate, peak and area under the curve).
• Immune response - anti-Tau IgG, anti-pTau IgM, anti-ePHF IgG and anti-Tau IgM titers in serum (geometric mean, change from baseline, responder rate, peak and area under the curve), determination of IgG response profile by avidity testing.
• Three (3) sub-cohorts have received ACI-35.030 at the dosage level of 300 pg, 900 pg or 1800 pg of tetrapalmitoylated phosphopeptide pTau Peptide T3 (SEQ ID NO: 28), and placebo as in Table 2.
• This anti-pTau IgG response showed a preference for pTau over non-pTau peptide in all actively treated early AD subjects, and the antibody response was boosted by the additional administrations of ACI-35.030, as shown by increased anti- pTau-specific IgG titers and/or increased anti-ePHF IgG titers measured 2 weeks after the additional administrations of ACI-35.030.
• No antibody response was observed in subjects receiving the placebo, except one single limited elevation of IgM and IgG anti-pTau titers after the study treatment period at week 67 in one subject in sub-cohort 1.2 receiving placebo, for which the responses were close to the threshold set for defining a responder.
• Table 3 shows the anti-pTau IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 at the dosage level of 300 pg of tetrapalmitoylated phosphopeptide pTau Peptide T3 (ACI-35.030 300 pg) or placebo in subcohort 1.1.
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgG titers > 1.81x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• the anti-pTau IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 at the dosage level of 900 pg of tetrapalmitoylated phosphopeptide pTau Peptide T3 (ACI-35.030 900 pg) or placebo in sub-cohort 1.2 are summarized in Table 4.
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgG titers > 1.81x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior the date and time of the first injection.
• a responder is defined as the number of subjects with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgG titers > 1.81x baseline f
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior the date and time of the first injection.
• All subjects treated with ACI-35.030 at the 900 pg dose level were responders at all timepoints between week 2 and week 74, while 1 subject receiving placebo generated a limited anti-pTau IgG response at week 67 (after the treatment period) (1.9 x baseline) just slightly higher than the threshold (1.81 x baseline) set for defining a responderAll subjects treated with ACI-35.030 at the 1800 pg dose level were responders at all timepoints between week 2 and week 50, while no subjects receiving placebo generated an anti-pTau IgG response up to week 42.
• High responder rates were observed as early as 2 weeks post vaccination for subjects treated with either 300 pg or 900 pg of ACI-35.030. Overall, high responder rates were observed after the first vaccination and following all vaccinations.
• At the 900 pg dose of ACI-35.030 there was a 100% responder rate for phosphorylated Tau when analyzed at any study timepoint from week 2 to week 74.
• the responder rate for pathological ePHF at the 900 pg dose of ACI-35.030 ranged from 66.7% to 100% at any timepoint during the treatment period between week 2 and week 48 and from 50% to 100% at any timepoint during the posttreatment period from week 50 to week 74.
• Table 6 Responders were defined as subjects with an antibody response higher than a positivity threshold, i.e., a pretreatment value (baseline antibody titer), multiplied by a threshold factor (> ⁇ 2x). Any post-baseline result greater than or equal to this value defines a positive antibody response.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• a responder has anti-Tau IgG titers 3.38x baseline, anti-pTau IgG titers 1.81x baseline, and/or anti-ePHF IgG titers 2.21x baseline.
• NA data not yet available
• each additional immunization at weeks 8, 24 or 48 at both the 300 and 900 j-ig dose levels led to a boosting of the anti-pTau IgG response as shown by increased anti-pTau- specific IgG titers measured 2 weeks after the administration of the additional immunization, at weeks 10, 26 and 50, respectively (FIGs. 2 and 6).
• Additional immunization at weeks 8, 24, and 48 with the 1800 pg dose level also led to a boosting of the anti-pTau IgG response as shown by increased anti-pTau- specific IgG titers measured 2 weeks after the administration of the additional immunization at weeks 10, 26 and 50 (FIGs. 4 and 7).
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgM titers > 2.11 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• the anti-pTau IgM titers and responder rate (ITT population) following immunization with either ACI-35.030900 pg or placebo in sub-cohort 1.2 are shown in Table 8.
• Table 8 ⁇ A responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgM titers > 2.11 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• a responder is defined as the number of subjects with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-pTau IgM titers > 2.11 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• FIGs. 3 and 5 See FIGs. 3 and 5 for the IgG titers against nonphosphorylated Tau (anti-Tau IgG titers) induced by the various dosages of ACI-35.030 or placebo over time.
• the anti-Tau IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 300 pg or placebo in sub-cohort 1.1 are shown in
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-Tau IgG titers > 3.38 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-Tau IgG titers > 3.38 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior the date and time of the first injection.
• the anti-Tau IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 1800 pg or placebo in sub-cohort 1.3 are shown in Table 12.
• a responder is defined as the number of subjects with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-Tau IgG titers > 3.38 x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior the date and time of the first injection.
• Table 14 shows the ratio of the anti-pTau IgG I anti-Tau IgG titers (ITT population) following immunization with ACI-35.030 900 pg in sub-cohort 1.2.
• Table 15 shows the ratio of the anti-pTau IgG I anti-Tau IgG titers (ITT population) following immunization with ACI-35.030 1800 pg in sub-cohort 1.3.
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-ePHF IgG titers > 2.21x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• Table 17 shows the anti-ePHF IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 900 pg or placebo in sub-cohort 1.2.
• a responder is defined as a subject with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-ePHF IgG titers > 2.21x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection.
• Table 18 shows the anti-ePHF IgG titers and responder rate (ITT population) following immunization with either ACI-35.030 1800 pg or placebo in sub-cohort 1.3.
• a responder is defined as the number of subjects with an antibody response above the positivity threshold.
• a post-baseline value is considered positive if > an analytical threshold x baseline.
• the analytical threshold is defined from samples from human donors (obtained during the validation of each assay).
• a responder has anti-ePHF IgG titers > 2.21x baseline.
• Baseline antibody titer value is the mean value of the titers measured at screening and visit 1 (including unscheduled visits) provided that they occur prior to the first injection. # For sub-cohort 1.3, data at weeks 31, 48 and 50 data from subjects on active treatment are pooled to avoid potential unblinding
• Responder rates for IgG anti-ePHF in subjects treated with ACI-35.030 300 pg were respectively 66.7% at week 2, 83.3% at weeks 8 and 10 and 33.3% at week 36 whereas prior injection at week 24 was not performed in 7/8 subjects in the sub-cohort due to Co vid- 19 pandemic. In order to avoid potential study unblinding, the percentage of responders beyond week 36 in the sub-cohort are not currently reported.
• Responder rates for anti-ePHF IgG in subjects treated with ACI-35.030 900 pg were respectively 100% at weeks 2, 10 and 26, 66.7% at weeks 8 and 24, 83.3% at weeks 36 and 48, and 100% at week 50. It appears that the same 2 actively treated subjects were nonresponders at these 2 timepoints (weeks 8 and 24) whereas they were responders at weeks 2, 10 and 26 (e.g., 2 weeks after the injections performed at weeks 0, 8 and 24 respectively). Responder rates for IgG anti-ePHF IgG in subjects treated with ACI-35.030 1800 pg at weeks 2, 8 and 10 were 66.7%, 33.3% and 83.3% respectively, then 50% at week 24, 66.7% at week 26, and 80% at week 50.
• Table 20 shows the avidity index on ePHF following immunization with ACI-35.030 at 900 j-tg in sub-cohort 1.2.
• Table 21 shows the avidity index on ePHF following immunization with ACI-35.030 at 1800 j-ig in sub-cohort 1.3.
• ACI-35.030 induced a high, specific and sustained antibody response oriented toward Tau-pathological species (phospho-Tau and ePHF), with an apparent dose-response between the low- and mid-dose with evidence of immunoglobulin class switch from IgM to IgG. Individual responder rates were high and consistent, especially for anti-pTau and ePHF antibodies.
• the administration of ACI-35.030 did not appear to have caused any particular safety concerns related to the study vaccine as of the date of the data analyses providing support for ACI-35.030’ s favorable safety and tolerability profile, and ACI-35.030 was able to induce a lasting antibody response above the baseline values in immunized patients.
• the data demonstrates that the IgG response matures towards a stronger preference for binding ePHF, the more pathologic species, while concomitantly lowering antibody titers towards the non- pathological, non-phosphorylated Tau.
• the preliminary results support the further development of this vaccine as an effective AD disease-modifying treatment as well as an approach for a potential prevention of AD.
• Example 2 Vaccination with ACI-35.030 induces relatively homogeneous antibody response with a broad epitope coverage
• the epitope recognition profile of antibodies was determined by epitope mapping ELISA before the first immunization (VI, week 0) and after the third immunization (V6, week 26) using a library of N-terminally biotinylated 8-mer peptides, shifted by one amino acid and covering the entire sequence of phospho-Tau peptide T3.30 (SEQ ID NO: 45) as well as the corresponding sequence of non-phosphorylated Tau peptide T3.56 (SEQ ID NO: 46).
• Tables 22 and 23 show the epitope recognition profile of antibodies induced by vaccination with ACI-35.030, as determined by epitope mapping ELISA on short 8-mer overlapping peptides, covering phospho-peptides T3.30 and T3.85 and non-phospho-peptides T3.56 and T3.86. [00275] Table 22
• Table 23 [00277] Table 22 and Figure 8 A shows that two AD patients essentially did not produce any IgG antibodies after three immunizations at week 26 against the sequences of or within phospho-Tau peptides T3.30 and T3.85 (patients #1 and #2), whereas the other five AD patients generated IgG antibodies against the sequences of or within phospho-Tau peptides T3.30 and T3.85 with overall similar binding to the sequences of phospho-Tau peptides T3.30 and T3.85. O.D.
• Table 23 and Figure 8B shows that two AD patients essentially did not produce any IgG antibodies at week 26 against the sequence of non-phosphorylated Tau peptides T3.56 and T3.86 (patients #1 and #2).
• Four AD patients generated IgG antibodies with minor recognition of the sequences of or within the non-phosphorylated Tau peptides T3.56 and T3.86, and the binding appeared to be linked to binding of the C-terminal 8-mer peptide (tau401-408).
• One AD patient (patient #3) produced IgG antibodies against the sequences of or within the non-phosphorylated Tau peptides T3.56 and T3.86, and the binding appears to be mostly linked to the N-terminal part of the sequence.
• the IgG response of the subjects was relatively homogenous displaying a broad epitope coverage as binding occurred across the pTau sequences tested and importantly, without substantial specificity for terminal end of the peptide sequence or substantial binding to non-phosphorylated sequences.
• SEQ ID NO: 28 palmitoylated phospho-Tau peptide (T3, palmitoylated T3.5) K(pal)K(pal)VYK[pS]PVVSGDT[pS]PRHLK(pal)K(pal)
• SEQ ID NO: 46 biotinylated non-phosphorylated tau peptide (T3.56) Biotin-LC linker (Ahx)-GVYKSPVVSGDTSPRHL-NH2

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Epidemiology (AREA)
• Pharmacology & Pharmacy (AREA)
• Organic Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Neurology (AREA)
• Biomedical Technology (AREA)
• Immunology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Dispersion Chemistry (AREA)
• Neurosurgery (AREA)
• Microbiology (AREA)
• Mycology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Biochemistry (AREA)
• Biophysics (AREA)
• Genetics & Genomics (AREA)
• Molecular Biology (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Hospice & Palliative Care (AREA)
• Psychiatry (AREA)
• Toxicology (AREA)
• Zoology (AREA)
• Gastroenterology & Hepatology (AREA)
• Rheumatology (AREA)
• Medicinal Preparation (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Peptides Or Proteins (AREA)

Priority Applications (12)

Applications Claiming Priority (6)

Related Child Applications (1)

Publications (2)

Family

ID=85201019

Family Applications (1)

Country Status (11)

Family Cites Families (4)

• 2022
  • 2022-08-12 JP JP2024508568A patent/JP2024532787A/ja active Pending
  • 2022-08-12 KR KR1020247007914A patent/KR20240042508A/ko active Pending
  • 2022-08-12 AU AU2022328342A patent/AU2022328342A1/en active Pending
  • 2022-08-12 WO PCT/US2022/074902 patent/WO2023019241A2/en not_active Ceased
  • 2022-08-12 MX MX2024001906A patent/MX2024001906A/es unknown
  • 2022-08-12 US US18/683,001 patent/US20250134973A1/en active Pending
  • 2022-08-12 IL IL310761A patent/IL310761A/en unknown
  • 2022-08-12 EP EP22856833.3A patent/EP4384216A4/en active Pending
  • 2022-08-12 CA CA3228878A patent/CA3228878A1/en active Pending
• 2023
  • 2023-06-28 US US18/343,349 patent/US20230338535A1/en not_active Abandoned
• 2024
  • 2024-02-09 CL CL2024000408A patent/CL2024000408A1/es unknown
  • 2024-02-12 JO JOJO/P/2024/0028A patent/JOP20240028A1/ar unknown
• 2025
  • 2025-01-06 US US19/010,408 patent/US20250144212A1/en active Pending

Also Published As

Similar Documents

Legal Events