Compositions and methods for treating alpha-synucleinopathies

Abstract

The present invention provides a method for treating or preventing alpha-synucleinopathies in a subject in need thereof, the method comprising administering to the subject a fixed dose of an anti-alpha-synuclein antibody or antigen-binding fragment thereof from 50 mg to 5,000 mg. Corresponding compositions and kits are also provided. As an example, the antibody may be aslo0452 ngl-3.

Description

1 Background Technology

[0001] The present invention relates to α-synuclein antibodies and their use in preventing or treating diseases, particularly α-synuclein diseases, more particularly Parkinson's disease (PD) and multiple system atrophy (MSA).

[0002] Alpha-synucleinopathies (alpha-synucleinopathies or α-synucleinopathies), also known as Lewy body diseases (LBD), are a family of neurodegenerative diseases that all have α-synuclein as a key pathological hallmark at their core (Jellinger (2003) Mov. Disord. 18 Suppl 6: S2-12; and Spillantini and Goedert (2000) Ann. NY Acad. Sci. 920: 16-27; both of which are incorporated herein by reference). α-Synucleinopathies include Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA).

[0003] PD is a slowly progressive, age-related movement disorder that affects more than 1% of people over the age of 65. The hallmark motor symptoms of PD include resting tremor, postural instability, bradykinesia, and rigidity. However, PD also involves non-motor symptoms, including loss of smell, cognitive dysfunction, sleep-wake disorders, autonomic dysfunction, depression, and constipation. PD is defined by the selective loss of dopaminergic neurons and the widespread accumulation of aggregates containing α-synuclein (called Lewy bodies) in surviving neurons. PD is the second most common neurodegenerative disorder after Alzheimer's disease.

[0004] MSA is a rare but universally fatal progressive neurodegenerative disorder clinically defined by parkinsonian, cerebellar, and autonomic dysfunction features. Isolated autonomic dysfunction, characterized by genitourinary dysfunction and orthostatic hypotension, and REM sleep behavior disorder are common features in the early stages of MSA. Like Parkinson's disease (PD), MSA is characterized by the widespread accumulation of α-synuclein aggregates. Currently, there are no disease-modifying therapies to halt or slow the progression of MSA.

[0005] The defining hallmark pathology of alpha-synuclein disease is Lewy bodies and Lewy neurites, which are insoluble inclusion bodies of aggregated proteins found in the interior of brain neurons during postmortem histopathological examination. Lewy pathology and neuronal loss may cause cognitive deficits and dementia, hyposmia, sleep disorders (including rapid eye movement sleep behavior disorder (RBD)), mood disorders (including depression and anxiety), autonomic dysfunction (including cardiovascular and gastrointestinal problems, such as constipation) and fatigue and sleepiness in non-motor brain areas (such as basal forebrain, midbrain pons system, amygdala, neocortex, vagus nerve dorsal motor nucleus, olfactory bulb, locus coeruleus and brainstem). Some symptoms in these non-motor symptoms seem to characterize the motor pre-stage or prodromal stage of Parkinson's disease (Kalia et al. Lancet (2015), 386 (9996): 896-912; The document is incorporated herein by reference).

[0006] Lewy pathology and neuronal loss in motor brain areas, including most notably the death of dopaminergic neurons in the substantia nigra, lead to the resting tremor, rigidity, bradykinesia, and postural instability that are hallmarks of Parkinson's disease (Spillantini and Goedert, Ann NY Acad Sci (2000), 920:16-27; incorporated herein by reference).

[0007] Alpha-synapse nucleoprotein (also referred to as "alpha-synapse nucleoprotein" or "alpha-syn") protein is the main structural component of Lewy bodies and Lewy neurites. Alpha-synapse nucleoprotein is a small acidic protein consisting of 140 amino acids (14kDa). People's natural wild-type alpha-synapse nucleoprotein has the amino acid sequence SEQ ID NO:1 as described according to UniProtKB accession number P37840. Unless otherwise apparent from the context, reference to alpha-synapse nucleoprotein or its fragment includes the natural human wild-type amino acid sequence indicated above, and its human allelic variant, particularly those related to Lewy body disease (e.g., E46K, A30P, H50Q, G51D and A53T, wherein the first letter indicates the amino acid in SEQ ID NO:1, the number is the codon position in SEQ ID NO:1, and the second letter is the amino acid in the allelic variant). Such variants may optionally exist alone or in any combination. The inducing mutations E83Q, A90V, and A76T that enhance α-synuclein aggregation can also be present alone or in combination with each other and / or in combination with the human allelic variants E46K, A30P, H50Q, G51D, and A53T. At the structural level, α-synuclein contains three distinct regions: an amphipathic N-terminal α-helical domain (residues 1-60) with lipid and membrane binding properties, a central hydrophobic amyloid-binding domain (residues 61-95) encoding the non-amyloid-β component (NAC) of the plaque, and an acidic proline-rich C-terminal tail (residues 96-140). Residues 71-82 of the NAC domain enable α-synuclein to transition from a random coil structure to a β-sheet structure and are therefore considered to be key to the aggregation / fibrillation properties of the protein (Bisaglia et al. (2009) FASEB J. 23(2):329-40; incorporated herein by reference). Although the C-terminal domain has no obvious secondary structure, it contains a key phosphorylation site at residue Ser129 and many tyrosine residues that are nitrated in cytoplasmic α-synuclein inclusion bodies. N-terminal and C-terminal truncated forms of α-synuclein also exist. Post-translational modifications of this protein can affect α-synuclein aggregation and toxicity (Oueslati et al. (2010) Prog. Brain. Res. 183: 115-45; which is incorporated herein by reference).

[0008] Under pathological conditions, abnormal α-synuclein aggregation may be key to the pathological changes seen in α-synucleinopathies (Lashuel et al. (2002) Nature 418:291; and Tsigelny et al. (2007) FEBS J. 274:1862-1877; both of which are incorporated herein by reference). In vitro studies have shown that α-synuclein monomers can form the starting point of the aggregation process. The monomers can aggregate into a variety of small oligomeric species, which are then stabilized by β-sheet interactions and go on to form fibrils, which can polymerize into insoluble fibrillar structures reminiscent of those identified in Lewy bodies (Cremades et al. (2012) Cell (2012), 149(5):1048-59; incorporated herein by reference). In vitro and in vivo studies have shown that the neurotoxic effects of α-synuclein appear to be caused by small, soluble oligomeric conformers or fibrils (Winner et al. (2011) Proc. Natl. Acad. Sci. USA 108(10):4194-9; and Danzer et al. (2007) J. Neurosci. 27(34):9220-32; both incorporated herein by reference). While fibrillar aggregates of α-synuclein are characteristic of PD, oligomeric forms of α-synuclein are toxic (Danzer et al. (2007) J. Neurosci. 27(34):9220-32; Lashuel et al. (2002) Nature 418:291; and Winner et al. (2011) Proc. Natl. Acad. Sci. USA 108:4194-4199; each of which is incorporated herein by reference).

[0009] α-Synuclein oligomers can be released into the extracellular environment and taken up by neighboring cells in a "spreading" mechanism (Angot and Brundin, (2009) Parkinsonism Relat. Disord. 15 Suppl 3:S143-147; Desplats et al. (2009) Proc. Natl. Acad. Sci. USA 106:13010-13015; and Lee et al. (2010) J. Biol. Chem. 285:9262-9272; each of which is incorporated herein by reference). Aggregates of α-synuclein can propagate misfolding through a prion-like spreading mechanism (Lee et al. (2010) Nat. Rev. Neurol. 6:702-706; Luk et al. (2012) J. Exp. Med. 209(5):975-86; and Luk et al. (2012) Science 338(6109):949-53; each of which is incorporated herein by reference). Therefore, α-synuclein can induce neurodegenerative diseases through oligomeric toxicity or propagation and prion-like spread.

[0010] It is now well established and recognized that under conditions of cellular stress, pathologically transmissible forms of α-synuclein can spread between neurons by releasing α-synuclein into the extracellular environment (Recasens and Dehay, Front Neuroanat (2014), 8:159; incorporated herein by reference).

[0011] The method for treating alpha-synuclein disease relies on passive immunotherapy. Passive immunotherapy methods (Lawand et al. (2015) Expert Opin. Ther. Targets 19: 1-10) using antibodies targeting alpha-synuclein have been tested in many preclinical alpha-synuclein disease mouse models; The document is incorporated herein by reference. Specifically, the research using the monoclonal antibody (9E4) for alpha-synuclein has shown that alpha-synuclein aggregates and pathological body clearance, behavioral movement improvement and neuroprotective effect (WO 2014 / 058924; The document is incorporated herein by reference).

[0012] Further studies using passive immunization of α-synuclein transgenic mice developed as an experimental model of PD / DLB have shown that the 9E4 monoclonal antibody clears α-synuclein pathology, reduces synaptic and axonal defects, eliminates the loss of striatal tyrosine hydroxylase fibers, and significantly reduces memory deficits and motor dysfunction (Games et al. (2014) J. Neurosci. 34(28):9441-54; Bae et al. (2012) J. Neurosci. 32(39):13454-69; and Masliah et al. (2011) PLoS One 6(4):e19338; each of which is incorporated herein by reference). Furthermore, passive administration of anti-α-synuclein monoclonal antibodies in wild-type mice injected intrastriatically with synthetic α-synuclein preformed fibrils (PFFs) resulted in a robust reduction in Lewy pathology, prevented dopamine neuron loss in the substantia nigra, and significantly improved movement disorders manifested in a mouse model after PFFs treatment (Tran et al. (2014) Cell Rep. 7(6):2054-65; incorporated herein by reference).

[0013] However, no passive immunotherapy approach has been approved for PD and other α-synucleinopathies, and therefore, current treatment approaches for these diseases primarily focus on treating the motor-related symptoms of the disease.

[0014] Therefore, there is a need in the art for a therapy for treating α-synucleinopathies, particularly in humans. 2 Summary of the invention

[0015] The present invention relates to an anti-α-synuclein antibody or an antigen-binding fragment thereof. The present invention relates to the use of such an antibody or antibody fragment to treat or prevent α-synuclein diseases. Specifically, the present invention relates to a specific dosage regimen that can be used when administering an anti-α-synuclein antibody or an antigen-binding fragment thereof to treat or prevent α-synuclein diseases. More specifically, the present invention relates to a treatment for α-synuclein diseases comprising administering to a subject a fixed dose of an anti-α-synuclein antibody or an antigen-binding fragment thereof, e.g., a dose that is independent of the subject's weight.

[0016] Thus, the present invention provides a method for treating or preventing alpha-synuclein disease in a subject in need thereof, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of an anti-alpha-synuclein antibody or antigen-binding fragment thereof. The present invention also provides an anti-alpha-synuclein antibody or antigen-binding fragment thereof for use in a method for treating or preventing alpha-synuclein disease in a subject, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of an anti-alpha-synuclein antibody or antigen-binding fragment thereof. The present invention also provides a use of an anti-alpha-synuclein antibody or antigen-binding fragment thereof in the manufacture of a medicament for treating or preventing alpha-synuclein disease in a subject, wherein the medicament comprises a fixed dose of 50 mg to 5,000 mg of the antibody or antigen-binding fragment thereof, or wherein the treating or preventing comprises administering a fixed dose of 50 mg to 5,000 mg of the antibody or antigen-binding fragment thereof. The present invention also provides a composition for use in a method of treating or preventing α-synucleinopathy, comprising a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof. The present invention also provides a kit comprising an anti-α-synuclein antibody or antigen-binding fragment thereof and instructions for using the same at a fixed dose of 50 mg to 5,000 mg to treat α-synucleinopathy.

[0017] The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 70 mg to 4,500 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,000 mg to 4,500 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,800 mg to 2,600 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 2,000 mg to 2,400 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be up to 2,600 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be up to 2,400 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be up to 2,200 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be up to 2,000 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,400 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,600 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,700 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 1,800 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 2,000 mg or 2,400 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 2,000 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 2,400 mg. The fixed dose of an anti-α-synuclein antibody or antigen-binding fragment thereof may be 4,500 mg.

[0018] The method may comprise administering the anti-α-synuclein antibody or antigen-binding fragment thereof intravenously, subcutaneously, intradermally, or intramuscularly. Preferably, the method may comprise administering the anti-α-synuclein antibody or antigen-binding fragment thereof intravenously.

[0019] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject once every 3 to 5 weeks. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject multiple times over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject once every four weeks over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years.

[0020] The α-synuclein disease may be selected from Parkinson's disease (PD), dementia with Lewy bodies (DLB), multiple system atrophy (MSA), Alzheimer's disease, pure autonomic failure, REM behavior disorder, prodromal synuclein disease, and neuroaxonal dystrophy. The α-synuclein disease may be Parkinson's disease (PD). The α-synuclein disease may be multiple system atrophy (MSA). The α-synuclein disease may be dementia with Lewy bodies (DLB). The subject may have a diagnosis of possible or probable MSA. The subject to be treated may have a Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score of 21 or less. As measured using the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score, the subject to be treated may have a severity score of 2 or less on the swallowing item; a severity score of 2 or less on the walking item; and / or a severity score of 2 or less on the falls item. The subject to be treated may have a UMSARS Part IV disability score of 3 or less.

[0021] As measured by the subject's score in a modified version of UMSARS Part I, the method can slow down the progression of alpha-synuclein disease. As measured by the subject's UMSARS total score, the method can slow down the progression of alpha-synuclein disease. As measured by the subject's UMSARS Part I score, the method can slow down the progression of alpha-synuclein disease. As measured by the subject's UMSARS Part II score, the method can slow down the progression of alpha-synuclein disease. As measured by the subject's 11 UMSARS scores, the method can slow down the progression of alpha-synuclein disease. As measured by the subject's Clinical Global Impression-Severity (CGI-S) scale score, the method can slow down the progression of alpha-synuclein disease.

[0022] The method can slow the progression of an alpha-synuclein disease as measured by the diffusion of alpha-synuclein in the subject. The method can slow the progression of an alpha-synuclein disease as measured by the level of free, unbound alpha-synuclein in the subject's cerebrospinal fluid (CSF).

[0023] The method can reduce a subject's score on a modified version of the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I. The method can reduce a subject's score on the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I. The method can reduce a subject's score on the Unified Multiple System Atrophy Rating Scale (UMSARS) Part II. The method can reduce a subject's score on the 11-item Unified Multiple System Atrophy Rating Scale (UMSARS). The method can reduce a subject's score on the Clinical Global Impression - Severity (CGI-S) scale.

[0024] The method can reduce the spread of alpha-synuclein in a subject. The method can reduce the level of free, unbound alpha-synuclein in the cerebrospinal fluid (CSF) of a subject.

[0025] The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to free, unbound α-synuclein in the CSF of a subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric and / or oligomeric α-synuclein in the CSF of a subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to α-synuclein in the CSF of a subject in vivo.

[0026] The anti-α-synuclein antibody or antigen-binding fragment thereof can specifically bind to a region between about amino acid 102 and about amino acid 130 within the C-terminal region of human α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric and aggregated forms of α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric and aggregated forms of α-synuclein in vivo. The anti-α-synuclein antibody or antigen-binding fragment thereof can sequester both monomeric and aggregated forms of α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can sequester both monomeric and aggregated forms of α-synuclein in vivo.

[0027] Anti-α-synuclein antibodies or antigen-binding fragments thereof with a K of less than 500 pM D Binds monomeric human α-synuclein.

[0028] Anti-α-synuclein antibodies or antigen-binding fragments thereof can reduce α-synuclein spread in vivo.

[0029] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise three heavy chain CDRs having the sequence H-CDR1 of SEQ ID NO: 5, the sequence H-CDR2 of SEQ ID NO: 15, and the sequence H-CDR3 of SEQ ID NO: 16; and three light chain CDRs having the sequence L-CDR1 of SEQ ID NO: 20, the sequence L-CDR2 of SEQ ID NO: 10, and the sequence L-CDR3 of SEQ ID NO: 21.

[0030] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable light chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 19. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 14 and a variable light chain region having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having the amino acid sequence of SEQ ID NO: 14 and a variable light chain region having the amino acid sequence of SEQ ID NO: 19.

[0031] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain having the amino acid sequence of SEQ ID NO: 17.

[0032] The anti-α-synuclein antibody or antigen-binding fragment thereof can be an antibody.

[0033] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a triple mutation in the Fc region corresponding to L234F / L235E / P331S numbered based on Kabat numbering. 3. Description of the accompanying drawings

[0034] These and other features of the present invention will become more apparent in the following detailed description taken in conjunction with the accompanying drawings, in which:

[0035] Figure 1 Shown is a simplified layout of a double-blind, placebo-controlled study of single ascending doses (SAD) of up to 4,500 mg aslo0452 ngl-3 in healthy subjects aged 18 to 65 years.

[0036] Figure 2 Shown are aslo0452 ngl-3 pharmacokinetic (PK) concentrations observed in the single ascending dose trial, stratified by dose level.

[0037] Figure 3 Shown is a simplified layout of a double-blind, placebo-controlled study of multiple ascending doses (MAD) of up to 4,200 mg aslo0452 ngl-3 in subjects 40 to 85 years of age with Parkinson's disease.

[0038] Figure 4 Shown are aslo0452 ngl-3 pharmacokinetic (PK) concentrations observed in the multiple ascending dose trial, stratified by dose level.

[0039] Figure 5 Shown are the dose-normalized (DN) values ​​of aslo0452 ngl-3 pharmacokinetic (PK) concentrations observed in single and multiple ascending dose trials.

[0040] Legend for SEQ ID NO:

[0041]

[0042] 4 Specific Implementation Methods

[0043] 4.1 Treatment and Prevention of α-Synucleinopathy

[0044] Unless otherwise defined herein, the scientific and technical terms used in conjunction with this application should have the meanings commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that the present invention is not limited to the particular methods, protocols, and reagents described herein, and therefore may vary. The terms used herein are only used to describe the purpose of specific examples and are not intended to limit the scope of the present invention, which is limited only by the claims.

[0045] The present invention provides a method for treating or preventing α-synucleinopathy in a subject in need thereof, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof. The present invention also provides an anti-α-synuclein antibody or antigen-binding fragment thereof for use in a method for treating or preventing α-synucleinopathy in a subject, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof. The present invention also provides use of an anti-α-synuclein antibody or antigen-binding fragment thereof in the manufacture of a medicament for treating or preventing α-synucleinopathy in a subject, wherein the medicament comprises a fixed dose of 50 mg to 5,000 mg of the antibody or antigen-binding fragment thereof, or wherein the treating or preventing comprises administering a fixed dose of 50 mg to 5,000 mg of the antibody or antigen-binding fragment thereof. References herein to the methods of the present invention are also intended to encompass equivalent uses of the present invention, or antibodies or antigen-binding fragments for use in the present invention, or compositions comprising such antibodies or antigen-binding fragments for use in the present invention. Therefore, when the present invention relates to an anti-α-synuclein antibody or antigen-binding fragment thereof for use in a method for treating or preventing α-synuclein disease, or the use of an anti-α-synuclein antibody or antigen-binding fragment thereof in the manufacture of a medicament for treating or preventing α-synuclein disease, the method for treating or preventing α-synuclein disease can be any method for treating or preventing α-synuclein disease as described herein. A kit is also provided, comprising: (a) an anti-α-synuclein antibody or antigen-binding fragment thereof; and (b) instructions for using it to treat α-synuclein disease at a fixed dose of 50 mg to 5,000 mg. The antibody or antigen-binding fragment thereof in the method of the present invention, the antibody or antigen-binding fragment thereof used in the present invention, and the use, composition or kit of the present invention can be any antibody or antigen-binding fragment thereof as described herein.

[0046] Preferably, the dosage as disclosed herein is a fixed dose. As used herein, the term "fixed dose" refers to a dose that is used for all subjects, for example, an amount that does not vary based on the subject's weight. A fixed dose can be a specific, unchanging amount, where the same dose is used for all subjects or all adult subjects, such as a unit dosage form as disclosed herein. A fixed dose is typically expressed in terms of the amount of active agent (such as an antibody or antigen-binding fragment thereof), rather than an amount expressed relative to the weight or mass of the subject to be treated.

[0047] As used herein, the terms "treating" or "treatment" refer to an improvement in a disease or disorder or at least one discernible symptom thereof. "Treatment" or "treating" may refer to an improvement in at least one measurable physical parameter, but not necessarily discernible to the patient. "Treatment" or "treating" may refer to inhibiting, slowing, or delaying the progression of a disease or disorder, either physically, such as stabilization of an discernible symptom, or physiologically, such as stabilization of a physical parameter, or both. For example, "treating" or "treatment" may refer to slowing the progression of an alpha-synuclein disease in a subject as compared to the expected progression for a subject with an alpha-synuclein disease, or as compared to the expected progression for a subject at that stage of the alpha-synuclein disease. "Treatment" or "treating" may refer to delaying the onset of a disease or disorder.

[0048] As used herein, the term "preventing" or "prevention" refers to reducing the risk of developing a given disease or disorder. "Preventing" or "prevention" can refer to preventing or delaying the onset of one or more symptoms of a disease or disorder, for example, as compared to an untreated control or placebo.

[0049] "Subject" refers to an animal, such as a mammal, that has been or will be the subject of treatment, observation, or experimentation. The methods described herein can be used for both human therapy and veterinary applications. Preferably, the subject is a human. The subject to be treated can be an adult. The subject can be a human and can be at least 30 years old, at least 40 years old, at least 50 years old, at least 60 years old, at least 70 years old, or at least 75 years old. The subject can be at least 40 years old. The subject can be between 50 and 60 years old.

[0050] The subject to be treated may have a BMI of at least 15 kg / m at baseline. 2 The subject can be a person with a BMI of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 kg / m at baseline. 2 Subjects can be people with a BMI less than or equal to 50 kg / m at baseline. 2 The subjects may have a BMI less than or equal to 50, 49, 48, 47, 46, 45, 44, 43, 42, 41 or 40 kg / m at baseline. 2 Subjects can be people with a BMI of 15 to 40 kg / m at baseline. 2 Preferably, the subject has a BMI of 18 to 35 kg / m at baseline. 2 people.

[0051] The methods, uses, compositions and kits of the present invention utilize an anti-α-synuclein antibody or antigen-binding fragment thereof at a dose of 50 mg to 5,000 mg (such as a fixed dose), for example, comprising administering such a (fixed) dose to a subject. The 50 mg to 5,000 mg (fixed) dose can be at least 50 mg, such as at least 70 mg, at least 100 mg, at least 250 mg, at least 500 mg, at least 750 mg, at least 1,000 mg, at least 1,250 mg, at least 1,500 mg, at least 1,750 mg, at least 2,000 mg, at least 2,250 mg, at least 2,500 mg, at least 2,750 mg, at least 3,000 mg, at least 3,250 mg, at least 3,500 mg, at least 3,750 mg, at least 4,000 mg, at least 4,250 mg, at least 4,500 mg or at least 4,750 mg. The 50 mg to 5,000 mg (fixed) dose can be up to 5,000 mg, such as up to 4,750 mg, up to 4,500 mg, up to 4,250 mg, up to 4,000 mg, up to 3,750 mg, up to 3,500 mg, up to 3,250 mg, up to 3,000 mg, up to 2,750 mg, up to 2,500 mg, up to 2,250 mg, up to 2,000 mg, up to 1,750 mg, up to 1,500 mg, up to 1,250 mg, up to 1,000 mg, up to 750 mg, up to 500 mg, up to 250 mg, up to 100 mg to up to 70 mg. Any of these upper and lower endpoints can be combined within the 50 mg to 5,000 mg range. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose (such as a fixed dose) of 70 mg to 4,500 mg, 1,000 mg to 4,500 mg, 1,500 to 4,500 mg, 1,800 mg to 2,600 mg, 2,000 mg to 2,600 mg, or 2,000 mg to 2,400 mg.

[0052] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of up to 2,600 mg, such as a fixed dose. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of 70 mg to 2,600 mg, 1,000 mg to 2,600 mg, 1,500 to 2,600 mg, 1,800 mg to 2,600 mg, or 2,000 mg to 2,600 mg, such as a fixed dose.

[0053] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of up to 2,400 mg, such as a fixed dose. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of 70 mg to 2,400 mg, 1,000 mg to 2,400 mg, 1,500 to 2,400 mg, 1,800 mg to 2,400 mg, or 2,000 mg to 2,400 mg, such as a fixed dose.

[0054] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of up to 2,200 mg, such as a fixed dose. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of 70 mg to 2,200 mg, 1,000 mg to 2,200 mg, 1,500 to 2,200 mg, 1,800 mg to 2,200 mg, or 2,000 mg to 2,200 mg, such as a fixed dose.

[0055] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of up to 2,000 mg, such as a fixed dose. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose of 70 mg to 2,000 mg, 1,000 mg to 2,000 mg, 1,500 to 2,000 mg, or 1,800 mg to 2,000 mg, such as a fixed dose.

[0056] 4,000, 4,250, 4,500, 4,750, or 5,000 mg. The anti-alpha-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a dose, such as a fixed dose, of about 50, about 100, about 150, about 200, about 250, about 300, about 400, about 500, about 600, about 700, about 800, about 900, about 1,000, about 1,200, about 1,400, about 1,600, about 1,800, about 2,000, about 2,200, about 2,400, about 2,600, about 2,800, about 3,000, about 3,200, about 3,400, about 3,600, about 3,800, about 4,000, about 4,250, about 4,500, about 4,750, or about 5,000 mg. Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can be administered to a subject at a dose of about 70, 210, 400, 1,200, 2,000, 2,400, 4,200 or 4,500 mg (such as a fixed dose). Preferably, anti-alpha-synuclein antibodies or antigen-binding fragments thereof are administered to a subject at a dose of about 1,200 mg (such as a fixed dose), such as a dose of 1,200 mg. Preferably, anti-alpha-synuclein antibodies or antigen-binding fragments thereof are administered to a subject at a dose of about 2,000 mg (such as a fixed dose), such as a dose of 2,000 mg. Preferably, anti-alpha-synuclein antibodies or antigen-binding fragments thereof are administered to a subject at a dose of about 2,400 mg (such as a fixed dose), such as a dose of 2,400 mg. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 4,200 mg (such as a fixed dose), such as 4,200 mg. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject at a dose of about 4,500 mg (such as a fixed dose), such as 4,500 mg.

[0057] The anti-α-synuclein antibody or its antigen-binding fragment can be administered to the subject at a dose of about 1,400 mg (such as a fixed dose), such as a dose of 1,400 mg. The anti-α-synuclein antibody or its antigen-binding fragment can be administered to the subject at a dose of about 1,600 mg (such as a fixed dose), such as a dose of 1,600 mg. The anti-α-synuclein antibody or its antigen-binding fragment can be administered to the subject at a dose of about 1,700 mg (such as a fixed dose), such as a dose of 1,700 mg. The anti-α-synuclein antibody or its antigen-binding fragment can be administered to the subject at a dose of about 1,800 mg (such as a fixed dose), such as a dose of 1,800 mg.

[0058] Treatment or prevention can be achieved by a single administration or multiple administrations of an anti-α-synuclein antibody or antigen-binding fragment thereof. For example, treatment or prevention can include administering two, three, four, five, six, seven, eight, nine, ten, or more than ten doses of an anti-α-synuclein antibody or antigen-binding fragment thereof to a subject. Each dose can be a dose as disclosed herein, such as a fixed dose. Treatment or prevention can include multiple administrations of an anti-α-synuclein antibody or antigen-binding fragment thereof as described herein, such as repeated administrations, during the course of treatment.

[0059] By a course of treatment is meant a treatment plan comprising several rounds of administration as part of a method for treating or preventing α-synucleinopathy according to the present invention. A course of treatment may last for one or more weeks, one or more months, such as two, three, four, five, six, seven, eight, nine, ten, or eleven months, or one or more years, such as one, two, three, four, five, or more years. A course of treatment may be ongoing without a fixed endpoint, for example, a course of treatment may continue for the lifetime of the subject being treated. Multiple administrations, such as repeated administrations, may be continued at regular or irregular intervals during the course of treatment, as determined necessary by the practitioner.

[0060] Anti-α-synuclein antibodies or antigen-binding fragments thereof may be administered to a subject daily or less frequently (e.g., weekly or monthly). Anti-α-synuclein antibodies or antigen-binding fragments thereof may be administered to a subject once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or seven times a week. Anti-α-synuclein antibodies or antigen-binding fragments thereof may be administered to a subject once every two weeks, once every three weeks, once every four weeks, once a month, twice a month, three times a month, or four times a month. Anti-α-synuclein antibodies or antigen-binding fragments thereof may be administered to a subject once a year, twice a year, three times a year, four times a year, five times a year, six times a year, seven times a year, eight times a year, nine times a year, ten times a year, eleven times a year, or twelve times a year.

[0061] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject multiple times at intervals of one week, two weeks, three weeks, four weeks, one month, two months, three months, six months, or one year. Treatment or prevention can include administering the anti-α-synuclein antibody or antigen-binding fragment thereof to the subject weekly, every two weeks, every three weeks, every three to five weeks, every four weeks, monthly, every two months, every three months, every six months, or annually.

[0062] When an anti-α-synuclein antibody or its antigen-binding fragment is administered in a manner of multiple administrations (such as repeated administrations), each of the administrations can be performed at a fixed dose as described herein. For example, each administration in the methods or uses of the present invention may comprise a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or its antigen-binding fragment as disclosed herein. In such examples, the number of administrations or the frequency of administration can be any of the options disclosed herein. Multiple administrations of an anti-α-synuclein antibody or its antigen-binding fragment (such as repeated administrations) can each be performed at the same dose. Two or more administrations of an anti-α-synuclein antibody or its antigen-binding fragment can each be performed at the same dose. Multiple administrations of an anti-α-synuclein antibody or its antigen-binding fragment (such as repeated administrations) can each be performed at a dose within the dosage range disclosed herein. For example, each administration of an anti-α-synuclein antibody or its antigen-binding fragment in the methods or uses of the present invention can be performed at a fixed dose in the range of 70 mg to 4,500 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 1,000 mg to 4,500 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 1,800 mg to 2,600 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 2,000 mg to 2,400 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose of about 2,000 mg. Each administration in the methods or uses of the present invention may be performed at a fixed dose of about 2,400 mg.

[0063] Each administration in the methods or uses of the present invention may comprise a fixed dose of up to 2,600 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof as disclosed herein. For example, each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 70 mg to 2,600 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,000 mg to 2,600 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,500 to 2,600 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 2,000 mg to 2,600 mg.

[0064] Each administration in the methods or uses of the present invention may comprise a fixed dose of up to 2,400 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof as disclosed herein. For example, each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 70 mg to 2,400 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,000 mg to 2,400 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,500 to 2,400 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 2,000 mg to 2,400 mg.

[0065] Each administration in the methods or uses of the present invention may comprise a fixed dose of up to 2,200 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof as disclosed herein. For example, each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 70 mg to 2,200 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,000 mg to 2,200 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 1,500 to 2,200 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose in the range of 2,000 mg to 2,200 mg.

[0066] Each administration in the methods or uses of the present invention may comprise a fixed dose of up to 2,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof as disclosed herein. For example, each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 70 mg to 2,000 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 1,000 mg to 2,000 mg. Each administration of an anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention may be performed at a fixed dose ranging from 1,500 to 2,000 mg.

[0067] Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention can be performed at a fixed dose of about 1,400 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention can be performed at a fixed dose of about 1,600 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention can be performed at a fixed dose of about 1,700 mg. Each administration of the anti-α-synuclein antibody or antigen-binding fragment thereof in the methods or uses of the present invention can be performed at a fixed dose of about 1,800 mg.

[0068] The anti-α-synuclein antibody or antigen-binding fragment thereof may be administered to the subject every four weeks. Thus, the methods or uses of the present invention may comprise administering to the subject a fixed dose of 2,000, 2,400, or 4,500 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 2,000 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 2,400 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 4,500 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks.

[0069] The method may comprise administering to the subject a fixed dose of 1,400 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 1,600 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 1,700 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks. The method may comprise administering to the subject a fixed dose of 1,800 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof every four weeks.

[0070] According to the present invention, the anti-α-synuclein antibody or antigen-binding fragment thereof may be administered to the subject multiple times over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years. The anti-α-synuclein antibody or antigen-binding fragment thereof may be administered to the subject twice a month over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years. The anti-α-synuclein antibody or antigen-binding fragment thereof may be administered to the subject once a month over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject once every four weeks over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years. Particularly preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject once every four weeks over a period of at least 1 year. Thus, the method may comprise administering to the subject a fixed dose of 1,200, 2,000, 2,400, 4,200, or 4,500 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 1,200 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 2,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 2,400 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 4,200 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method can include administering to the subject a fixed dose of 4,500 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year.

[0071] The method may comprise administering to the subject a fixed dose of 1,400 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 1,600 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 1,700 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year. The method may comprise administering to the subject a fixed dose of 1,800 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof once every four weeks for a period of at least 1 year.

[0072] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered by any suitable route. For example, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered intravenously, intraperitoneally, transdermally, intracavitary, subcutaneously, intradermally, or intramuscularly. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered intravenously.

[0073] As used herein, and in accordance with the present invention, the term "α-synucleinopathy" ("α-synucleinopathy" or "alpha-synucleinopathy") refers to a neurodegenerative disease characterized by abnormal accumulation of insoluble α-synuclein in neurons and glial cells of a subject. The term "α-synucleinopathy" is further discussed in Mendoza-Velásquez et al. (2019) Front. Neurol. 10: 363, which is hereby incorporated by reference in its entirety. α-synucleinopathy can be selected from Parkinson's disease (PD), dementia with Lewy bodies (DLB), multiple system atrophy (MSA). α-synucleinopathy can be selected from Alzheimer's disease, pure autonomic failure, and REM behavioral disorder. α-synucleinopathy can be a prodromal synucleinopathy, for example, characterized by early signs or symptoms of α-synucleinopathy, such as REM behavioral disorder or autonomic symptoms, such as orthostatic hypotension. Preferably, the α-synucleinopathy is PD or MSA.

[0074] Alpha-synuclein disease can be Parkinson's disease (PD). The subject to be treated according to the present invention may suffer from PD. The subject may have been diagnosed with PD. The subject may have one or more symptoms of PD, such as one or more of tremor, bradykinesia, postural instability and muscle rigidity. The subject may have a diagnosis of PD based on the Movement Disorder Society (MDS) Parkinson's Disease Clinical Diagnostic Criteria (see, for example, Postuma et al. (2015) Move.Disord.30:1591-1599). The subject may be over 50 years old, such as over 55 years old, over 60 years old, over 65 years old, over 70 years old or over 75 years old. The subject may have early-onset PD. The subject may be less than 50 years old. The subject may be at risk of PD. The subject may be susceptible to PD, for example, having a family history of PD, having a genetic susceptibility to PD, and / or having been exposed to environmental factors associated with PD.

[0075] Preferably, the subject may have early PD. The subject may have been diagnosed with early PD. The subject may have a diagnosis of early PD. The subject may have one, two, three, four, five, six, seven, eight or more symptoms of early PD. Preferably, the subject has two or more symptoms of early PD. More preferably, the subject has two or more symptoms of early PD selected from tremor, slow movement and muscle stiffness. Symptoms of early PD may include one or more of the following: a) smaller handwriting; b) tremor; c) muscle stiffness; d) slow movement; e) hunched posture; f) lack of facial expression; g) reduced arm swing; and h) soft or deep voice.

[0076] Alpha-synuclein disease can be pure autonomic failure. Pure autonomic failure is a neurodegenerative disorder of the autonomic nervous system, clinically characterized by orthostatic hypotension. Pure autonomic failure is described in more detail in Coon et al. (2019) Mayo Clin. Proc. 94(10): 2087-2098.

[0077] Alpha-synuclein disease can be REM behavior disorder. REM behavior disorder (RBD) involves muscle atonia during REM sleep, and repeated nighttime dream enactment behaviors, accompanied by vocalizations (such as yelling) and motor behaviors (such as punching and kicking). RBD is described in more detail in Barone and Henchcliffe (2018) Clin. Neurophysiol. 129 (8): 1551-1564.

[0078] The α-synucleinopathy may be MSA. The subject to be treated according to the present invention may have MSA. The subject may have been diagnosed with MSA. The subject may have one or more symptoms of MSA. The subject may be at risk for MSA. The subject may be susceptible to MSA. The subject may be at least 30 years old, at least 40 years old, at least 50 years old, at least 60 years old, or at least 70 years old. The subject may be at least 40 years old. The subject may be between 50 and 60 years old.

[0079] Preferably, the subject has early-stage MSA. The subject may have been diagnosed with early-stage MSA. The subject may have a diagnosis of early-stage MSA. The subject may have one, two, three, four, or five or more symptoms of early-stage MSA. Preferably, the subject has two or more symptoms of early-stage MSA. More preferably, the subject has two or more symptoms of early-stage PD selected from tremor, slow movement, and muscle stiffness. Symptoms of early-stage MSA may include one or more of the following: a) slow movement, tremor, or stiffness (rigidity); b) clumsiness or incoordination; c) speech disorders, hoarseness, and trembling of the voice; d) fainting or dizziness due to orthostatic hypotension; and e) bladder control problems, such as sudden urgency to urinate or difficulty emptying the bladder.

[0080] The subject may have a diagnosis of possible or probable MSA. As used herein, the term "possible or probable MSA" means possible or probable MSA defined using the modified Gilman et al., 2008 diagnostic criteria as described in detail in Gilman et al. (2008) Neurol. 71(9):670-676.

[0081] The diagnostic criteria for possible MSA are as follows:

[0082] (1) A sporadic, progressive, adult-onset disease (over 30 years of age) characterized by the following:

[0083] a) Parkinsonism (bradykinesia with rigidity, tremor, or postural instability); or cerebellar syndrome (gait ataxia with cerebellar dysarthria, limb ataxia, or cerebellar oculomotor dysfunction), and

[0084] b) at least one feature suggestive of autonomic dysfunction (otherwise unexplained urinary urgency, frequency, or incomplete bladder emptying, erectile dysfunction in male subjects, or a significant orthostatic drop in blood pressure that does not reach the level required for an impossible MSA), and

[0085] (2) At least one of the following additional characteristics:

[0086] For probable MSA with predominant parkinsonism (MSA-P) or MSA with predominant cerebellar ataxia (MSA-C):

[0087] a) Babinski sign with hyperreflexia; and / or

[0088] b) Stridor.

[0089] For probable MSA with predominant parkinsonism (MSA-P):

[0090] a) Rapidly progressive parkinsonism;

[0091] b) poor response to levodopa;

[0092] c) postural instability within 3 years of motor onset;

[0093] d) gait ataxia, cerebellar dysarthria, limb ataxia, or cerebellar oculomotor dysfunction;

[0094] e) dysphagia within 5 years of motor onset;

[0095] f) MRI showing atrophy of the putamen, middle cerebellar peduncle, pons, or cerebellum; and / or

[0096] g) Fluorodeoxyglucose (FDG)-positron emission tomography (PET) shows hypometabolism in the putamen, brainstem, or cerebellum.

[0097] For possible MSA with predominant cerebellar ataxia (MSA-C):

[0098] a) Parkinsonism (bradykinesia and rigidity):

[0099] b) MRI shows atrophy of the putamen, middle cerebellar peduncle, or pons;

[0100] c) FDG-PET shows putamen hypometabolism; and / or

[0101] d) Single-photon emission computed tomography or PET shows presynaptic nigrostriatal dopaminergic denervation.

[0102] The diagnostic criteria for probable MSA are as follows:

[0103] A sporadic, progressive, adult-onset (over 30 years of age) disorder characterized by the following:

[0104] a) Autonomic failure, including urinary incontinence (inability to control urine flow from the bladder, and erectile dysfunction in male subjects) or a decrease in orthostatic blood pressure within 3 minutes of standing, with a decrease in systolic blood pressure of at least 30 mmHg or a decrease in diastolic blood pressure of at least 15 mmHg; and any of the following conditions are met

[0105] b) Parkinsonism that is poorly responsive to levodopa (bradykinesia with rigidity, tremor, or postural instability), or

[0106] c) Cerebellar syndrome (gait ataxia with cerebellar dysarthria, limb ataxia, or cerebellar oculomotor dysfunction).

[0107] A subject may have a diagnosis of "clinically definite MSA," "clinically probable MSA," or "probable MSA." As used herein, the terms "clinically definite MSA," "clinically probable MSA," and "probable MSA" refer to clinically definite MSA, clinically probable MSA, and probable MSA, respectively, as defined in Wenning et al. (2022) Mov. Disor. 37(6): 1131-1148.

[0108] The diagnostic criteria for clinically confirmed MSA are as follows:

[0109] (1) A sporadic, progressive, adult-onset disease (over 30 years of age) characterized by the following:

[0110] At least one of the following:

[0111] (a) Unexplained dysuria with residual urine volume ≥100 mL after urination;

[0112] (b) unexplained urge incontinence; and

[0113] (c) neurogenic OH (≥20 / 10 mmHg blood pressure drop) within 3 minutes of standing or head-up tilt test;

[0114] AND at least one of the following:

[0115] (a) Parkinsonism that is poorly responsive to L-dopa; and

[0116] (b) cerebellar syndrome (at least two of gait ataxia, limb ataxia, cerebellar dysarthria, or oculomotor features; and

[0117] (2) At least two supportive clinical (motor or non-motor) features selected from the following:

[0118] (a) Rapid progression of motor seizures within 3 years;

[0119] (b) moderate to severe postural instability within 3 years of motor onset;

[0120] (c) L-dopa-induced or exacerbated craniocervical dystonia in the absence of limb movement disorders;

[0121] (d) severe speech impairment within 3 years of motor onset;

[0122] (e) severe dysphagia within 3 years of motor onset;

[0123] (f) Babinski sign of unknown cause;

[0124] (g) convulsive myoclonic postural or kinetic tremor;

[0125] (h) postural deformity;

[0126] (i) stridor;

[0127] (j) Inhale and sigh;

[0128] (k) cold and discolored hands and feet; and

[0129] (l) pathological laughing or crying; and

[0130] (3) At least one MRI marker selected from the following (each affected brain region as evidenced by atrophy or increased diffuseness counts as one MRI marker):

[0131] For MSA-P:

[0132] (a) Atrophy of the following parts:

[0133] (i) putamen (and reduced signal on iron-sensitive sequences);

[0134] (ii) middle cerebellar peduncle;

[0135] (iii) pons; or

[0136] (iv) cerebellum;

[0137] (b) the “hot cross bun” sign; and

[0138] (c) Increased diffuseness in the following areas:

[0139] (i) putamen; or

[0140] (ii) middle cerebellar peduncle;

[0141] For MSA-C:

[0142] (a) Atrophy of the following parts:

[0143] (i) putamen (and reduced signal at the iron-sensitive sequence); or

[0144] (ii) infratentorial structures (pons and middle cerebellar peduncles);

[0145] (b) the “hot cross bun” sign; and

[0146] (c) increased putaminal diffusion; and

[0147] (4) None of the following characteristics exist:

[0148] (a) substantial and lasting beneficial response to dopaminergic drugs;

[0149] (b) unexplained loss of smell during olfactory testing;

[0150] (c) Fluctuating cognitive impairment, with marked changes in attention and alertness and early decline in visual perception;

[0151] (d) recurrent non-drug-induced visual hallucinations within 3 years of disease onset;

[0152] (e) dementia consistent with DSM-V within 3 years of disease onset;

[0153] (f) downward gaze supranuclear palsy or slowed vertical saccades;

[0154] (g) Brain MRI findings suggest an alternative diagnosis (e.g., PSP, multiple sclerosis, vascular parkinsonism, symptomatic cerebellar disease, etc.); and

[0155] (h) Documentation of alternative conditions (similar to MSA, including hereditary

[0156] hereditary or symptomatic ataxia and parkinsonism).

[0157] The diagnostic criteria for clinically probable MSA are as follows:

[0158] (1) A sporadic, progressive, adult-onset disease (over 30 years of age) characterized by the following:

[0159] At least two of the following:

[0160] (a) Autonomic dysfunction, which is defined as:

[0161] (i) Unexplained dysuria with residual urine volume after urination;

[0162] (ii) unexplained urge incontinence; and

[0163] (iii) neurogenic OH (≥20 / 10 mmHg blood pressure drop) within 10 minutes of standing or head-up tilt test;

[0164] (b) Parkinson's disease; and

[0165] (c) cerebellar syndrome (defined as at least one of gait ataxia, limb ataxia, cerebellar dysarthria, and oculomotor features);

[0166] (2) At least one supportive clinical (motor or non-motor) feature selected from the following:

[0167] (a) Rapid progression of motor seizures within 3 years;

[0168] (b) moderate to severe postural instability within 3 years of motor onset;

[0169] (c) L-dopa-induced or exacerbated craniocervical dystonia in the absence of limb movement disorders;

[0170] (d) severe speech impairment within 3 years of motor onset;

[0171] (e) severe dysphagia within 3 years of motor onset;

[0172] (f) Babinski sign of unknown cause;

[0173] (g) convulsive myoclonic postural or kinetic tremor;

[0174] (h) postural deformity;

[0175] (i) stridor;

[0176] (j) Inhale and sigh;

[0177] (k) cold and discolored hands and feet; and

[0178] (l) pathological laughing or crying; and

[0179] (3) None of the following characteristics exist:

[0180] (a) substantial and lasting beneficial response to dopaminergic drugs;

[0181] (b) unexplained loss of smell during olfactory testing;

[0182] (c) Fluctuating cognitive impairment, with marked changes in attention and alertness and early decline in visual perception;

[0183] (d) recurrent non-drug-induced visual hallucinations within 3 years of disease onset;

[0184] (e) dementia consistent with DSM-V within 3 years of disease onset;

[0185] (f) downward gaze supranuclear palsy or slowed vertical saccades;

[0186] (g) Brain MRI findings suggest an alternative diagnosis (e.g., PSP, multiple sclerosis, vascular parkinsonism, symptomatic cerebellar disease, etc.); and

[0187] (h) Alternative conditions known to cause autonomic failure, ataxia, or parkinsonism that appear to be related to the patient's symptoms are documented (similar to MSA, including hereditary or symptomatic ataxia and parkinsonism).

[0188] The diagnostic criteria for possible prodromal MSA are as follows:

[0189] (1) A sporadic, progressive, adult-onset disease (over 30 years of age) characterized by the following:

[0190] At least one of the following:

[0191] (a) RBD (confirmed by polysomnography);

[0192] (b) neurogenic OH (≥20 / 10 mmHg drop in blood pressure) within 10 minutes of standing or head-up tilt test; and

[0193] (c) genitourinary failure (erectile dysfunction in men under 60 years of age, accompanied by at least one unexplained dysuria, post-void residual volume >100 mL, and unexplained urge incontinence); and

[0194] (2) At least one of the following:

[0195] (a) Mild parkinsonism; and

[0196] (b) mild cerebellar signs; and

[0197] (3) None of the following exist:

[0198] (a) Unexplained anosmia or abnormal cardiac sympathetic nerve imaging during olfactory testing ( 123 at least one of the following: I-MIBG-scintigraphy);

[0199] (b) Fluctuating cognitive impairment, with significant changes in attention and alertness and early decline in visual perception;

[0200] (c) recurrent non-drug-induced visual hallucinations within 3 years of disease onset;

[0201] (d) dementia consistent with DSM-V within 3 years of disease onset;

[0202] (e) downward gaze supranuclear gaze palsy or slowed vertical saccades;

[0203] (f) Brain MRI findings suggest an alternative diagnosis (e.g., PSP, multiple sclerosis, vascular parkinsonism, symptomatic cerebellar disease, etc.); and

[0204] (g) Alternative conditions known to cause autonomic failure, ataxia, or parkinsonism that appear to be related to the patient's symptoms are documented (similar to MSA, including hereditary or symptomatic ataxia and parkinsonism).

[0205] The subject to be treated according to the present invention can be defined or selected based on a score on the Unified Multiple System Atrophy Rating Scale. The subject may have a score of 40 or less on the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I. The subject may have a score of 30 or less on the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I. Preferably, the subject has a score of 24 or less on the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I. The UMSARS score (described in more detail in Wenning et al. (2004) Mov. Disord. 19(12): 1391-1402) is a clinical rating scale designed to assess the severity of a subject's MSA, with higher scores indicating a more severe condition. UMSARS includes four subscales: UMSARS Part I (12 questions) assesses patient-reported functional disability; UMSARS Part II (14 questions) assesses clinician-assessed movement disorders; UMSARS Part III records blood pressure and heart rate measurements in the supine and standing positions; and UMSARS Part IV (1 question) assesses disability based on housework. Answers to the questions or items in UMSARS Part I and Part II are scored on a scale of 0 to 4, with a score of 0 indicating normal function and a score of 1 indicating mild disability. A subject may have a severity score of 2 or less on the swallowing item. A subject may have a severity score of 2 or less on the walking item. A subject may have a severity score of 2 or less on the falls item. A subject may have a UMSARS Part IV disability score of 3 or less. A subject may have a UMSARS Part I score of 21 or less. Preferably, when these scores are used herein to define a subject to be treated according to the invention, they refer to the subject's score at baseline, i.e., the subject's score immediately before starting treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof.

[0206] The subject to be treated (e.g., at baseline) may have a Montreal Cognitive Assessment (MoCA) score of ≥18. The MoCA test was developed to detect mild cognitive impairment, but is now often used as a screening tool for dementia. The MoCA has a maximum score of 30 points and is a brief test of cognitive function that takes 10 minutes. It assesses short-term memory, visual-spatial function, executive function, attention, concentration and working memory, language, and orientation. The MoCA test is described in more detail in Nasreddine et al. (2005) J. Amer. Geriatrics Soc. 53(4): 659-9.

[0207] 4.1.1 Clinical Outcomes

[0208] The present invention relates to treating or preventing α-synuclein diseases. Treatment or prevention can be observed as a reduction in one or more markers and / or symptoms of α-synuclein diseases, and / or a reduction in scores from one or more relevant scoring systems for α-synuclein diseases. For example, in a subject treated according to the present invention, the subject's condition can improve. In a subject treated according to the present invention, one or more symptoms and / or markers and / or scores of α-synuclein diseases can improve. Treatment or prevention can be observed as a slowing of the progression of α-synuclein diseases in the subject or a delay in the progression of α-synuclein diseases in the subject. For example, in a subject treated according to the present invention, the subject's condition can stabilize or remain stable. In a subject treated according to the present invention, one or more symptoms and / or markers and / or scores of α-synuclein diseases can remain stable, can be stable, or can not worsen. In a subject treated according to the present invention, α-synuclein diseases can continue to progress, but at a slower rate than would be expected for a subject with that type of α-synuclein disease or a subject at that stage of the disease. The progression of alpha-synuclein disease can be measured by assessing one or more markers and / or symptoms and / or scores of alpha-synuclein disease in a subject at two or more time points (e.g., before treatment according to the invention (e.g., at baseline) and during and / or after treatment), and determining whether there are any changes during the time period. Slowing or delaying progression can be measured by comparing the progression of a subject treated with an antibody as described herein, or an antigen-binding fragment thereof, with changes in the same markers and / or symptoms and / or scores over the same time period in a subject not treated with such an antibody or antigen-binding fragment thereof (e.g., a control or placebo-treated subject or a control or placebo-treated subject group). Preferably, the comparison is to a placebo-treated subject group, such as the typical or average change in the scores obtained from the placebo-treated subject group. Preferably, the control or placebo-treated subject is a subject with the same disease or disorder as the treated subject. Preferably, the control or placebo-treated subject has the same or similar level of disease progression as the treated subject at the start of treatment. Preferably, the control or placebo treated subject has the same range or similar score as the treated subject at the start of treatment. Slowing or delaying of progression can be measured by comparing the progress of a subject treated with an antibody as described herein or its antigen-binding fragment with changes in the same markers and / or symptoms and / or scores that would be expected or predicted (e.g., based on a disease progression model, e.g., a reference subject with the same disease or disorder or the same α-synuclein disease as the subject or a reference subject with the same degree of disease progression as the treated subject) over the same time period for a subject not treated with such an antibody or its antigen-binding fragment.

[0209] The treatment or prevention of the present invention can be observed as such a reduction, stabilization or delay in progression or slowing of: (i) the spread of alpha-synuclein in the subject; (ii) the level of alpha-synuclein in the cerebrospinal fluid (CSF) of the subject; and / or (iii) the level of alpha-synuclein in the interstitial fluid (ISF) of the subject. Thus, the method can slow the progression of alpha-synuclein disease as measured by: the spread of alpha-synuclein in the subject; the level of free unbound alpha-synuclein in the CSF of the subject; and / or the level of free unbound alpha-synuclein in the ISF of the subject. The method can slow the progression of alpha-synuclein disease as measured by the spread of alpha-synuclein in the subject and the level of free unbound alpha-synuclein in the CSF of the subject. The method can slow the progression of alpha-synuclein disease as measured by the spread of alpha-synuclein in the subject. Preferably, the method slows the progression of alpha-synucleinopathy as measured by the level of free, unbound alpha-synuclein in the subject's CSF. The method can slow the progression of alpha-synucleinopathy as measured by the level of free, unbound alpha-synuclein in the subject's brain interstitial fluid. Advantageously, these effects can be achieved by intravenous administration of an anti-alpha-synuclein antibody or antigen-binding fragment thereof.

[0210] The methods disclosed herein can provide advantages over other therapeutic agents by reducing the spread of α-synuclein in a subject; reducing the level of α-synuclein in the CSF of a subject; and / or reducing the level of α-synuclein in the interstitial fluid of the brain of a subject. Thus, the method can reduce the spread of α-synuclein in a subject; reduce the level of free, unbound α-synuclein in the CSF of a subject; and / or reduce the level of free, unbound α-synuclein in the ISF of a subject. The method can reduce the spread of α-synuclein in a subject and reduce the level of free, unbound α-synuclein in the CSF of a subject. The method can reduce the spread of α-synuclein in a subject. Preferably, the method reduces the level of free, unbound α-synuclein in the cerebrospinal fluid (CSF) of a subject. The method can reduce the level of free, unbound α-synuclein in the interstitial fluid (ISF) of a subject. Advantageously, when administered intravenously, the anti-α-synuclein antibody or antigen-binding fragment thereof can reduce α-synuclein diffusion, reduce the level of free unbound α-synuclein in cerebrospinal fluid, and / or reduce the level of free unbound α-synuclein in brain interstitial fluid.

[0211] The α-synuclein spread of a subject can be measured by any suitable means. α-Synuclein is abundant in the central nervous system (CNS) and brain, being present both intracellularly in neurons and glia, and extracellularly in cerebrospinal fluid (CSF) (Mollenhauer et al. (2012) J. Neural. Transm. (2012), 119(7):739-46; incorporated herein by reference) and in the interstitial fluid (ISF) that bathes and surrounds brain cells (Emmanouilidou et al. (2011) PLoS One (2011), 6(7):e22225; incorporated herein by reference). α-Synuclein is a synaptic protein expressed primarily in neurons of the neocortex, hippocampus, substantia nigra, thalamus, and cerebellum (Iwai et al. (1995) Neuron (1995), 14:467-475; incorporated herein by reference). Under physiological conditions, it is localized to neuronal synaptic terminals and is specifically upregulated at presynaptic terminals during acquisition-related synaptic rearrangements (Fortin et al. (2005) J. Neurosci. (2005), 25: 10913-10921; incorporated herein by reference). A variety of experimental models have demonstrated cell-to-cell transmission of α-synuclein in cultured cells or the spread and propagation of α-synuclein pathology in vivo. For example, preformed recombinant α-synuclein fibrils and α-synuclein oligomers can be internalized by cultured cells and neurons, and direct transfer of α-synuclein from donor to recipient cells and the formation of α-synuclein inclusions resembling Lewy pathology have been demonstrated (Danzer et al. (2007) J. Neurosci. (2007), 27(34):9220-32; Volpicelli-Daley et al. (2011) Neuron (2011), 72(1):57-71; and Luk et al. (2009) Proc. Natl. Acad. Sci. USA (2009), 106(47):20051-6; each of which is incorporated herein by reference). Furthermore, the translocation and delivery of monomeric and oligomeric α-synuclein from the olfactory bulb to interconnected brain structures has been demonstrated in mice (Rey et al. (2013) Acta Neuropathol. (2013), 126(4):555-73; incorporated herein by reference).

[0212] As used herein, the term "α-synuclein spread" refers to the expansion of an area in which α-synuclein is found. This may involve the expansion of an area in which misfolded α-synuclein or aggregated α-synuclein (such as Lewy bodies or Lewy body-like aggregates) is found. α-synuclein spread may involve the spread of α-synuclein to a wider brain region or additional brain region. α-synuclein spread may involve the spread of α-synuclein to a wider peripheral region or additional peripheral region. α-synuclein spread may involve the movement of α-synuclein by cell-to-cell transmission. For example, α-synuclein oligomers can be released into the extracellular environment and taken up by neighboring cells in a "spreading" mechanism (Angot and Brundin (2009) Parkinsonism Relat. Disord. 15 Suppl 3: S143-147; Desplats et al. (2009) Proc. Natl. Acad. Sci. USA 106: 13010-13015; and Lee et al. (2010) J. Biol. Chem. 285: 9262-9272). α-synuclein spread can involve the spread of misfolded α-synuclein (such as α-synuclein oligomers or multimers) or other forms of aggregated α-synuclein (such as the formation or spread of Lewy bodies or Lewy body-like aggregates) to a wider brain region or additional brain regions. For example, this can occur by propagating α-synuclein misfolding through a prion-like spreading mechanism (Lee et al. (2010) Nat. Rev. Neurol. 6:702-706; Luk et al. (2012) J. Exp. Med. 209(5):975-86; and Luk et al. (2012) Science 338(6109):949-54). α-synuclein spread can occur within a single brain region and / or can involve spreading to different brain regions, such as to interconnected or adjacent brain regions. Preferably, the present invention causes a reduction in α-synuclein spread in a subject. The reduction in α-synuclein spread can include a reduction in the extent of spread in the subject compared to that observed in a control subject to whom an anti-α-synuclein antibody or antigen-binding fragment thereof according to the present invention was not administered. The reduction in α-synuclein spread can include a reduction in the extent of spread compared to a reference level of spread. The reference level of α-synuclein spread can be based on a model of disease progression, for example, based on normal or average levels of α-synuclein spread in reference subjects having the same disease or disorder or the same α-synucleinopathy as the subject, or at the same degree of disease progression as the subject being treated.The reference level of alpha-synuclein diffusion can be based on the amount of diffusion observed in one or more control subjects over a similar time period, the one or more control subjects, such as a control subject population that is not administered an anti-alpha-synuclein antibody or antigen-binding fragment thereof according to the present invention, such as one or more placebo-treated subjects. A decrease in alpha-synuclein diffusion can be a decrease in the amount of alpha-synuclein in the diffusion region compared to a reference level, such as a lower amount of alpha-synuclein in the diffusion region. A decrease in alpha-synuclein diffusion can be a decrease in the extent of diffusion compared to a reference level, such as a decrease in the number of regions to which alpha-synuclein diffuses or a decrease in the area or volume of diffusion. A decrease in alpha-synuclein diffusion can be a slowing or delay in the progression of alpha-synuclein diffusion compared to a reference level, such as a decrease or delay in the diffusion of alpha-synuclein within a region or to a different region.

[0213] The level of free unbound α-synuclein in the CSF of a subject can be measured by any suitable means. As used herein, the term "free unbound α-synuclein" refers to α-synuclein that is not bound to an anti-α-synuclein antibody or its antigen-binding fragment. The free unbound α-synuclein can be applied to α-synuclein in its monomeric or oligomeric form or aggregated form. These terms are generally applicable to any pathological form of α-synuclein. Therefore, the level of free unbound α-synuclein in a sample can be assessed by a method comprising separating α-synuclein bound to an antibody or fragment from α-synuclein that is not bound to the antibody or fragment. The level of free unbound α-synuclein in the CSF of a subject can be measured by a method comprising removing α-synuclein bound to an antibody or its antigen-binding fragment from a CSF sample and then assessing the amount of α-synuclein retained in the sample. For example, the CSF sample can be immunoprecipitated (IP) to remove α-synuclein bound to the antibody or its antigen-binding fragment, so that unbound "free" α-synuclein remains in the supernatant. The free level of α-synuclein retained in the CSF sample (e.g., supernatant) can then be determined by any suitable method, such as by ELISA. For example, a commercial ELISA kit (e.g., Mesoscale Discovery ELISA kit (see Example 1) or Sensolyte TMThe level of free α-synuclein in the CSF supernatant can be assessed using a quantitative ELISA kit, human / mouse / rat, AnaSpec, US, AS-55550). As used herein, a "sample" may refer to an in vitro or ex vivo sample that has been obtained from a suitable subject. A CSF sample can be collected from a subject by lumbar puncture. A change or decrease in the level of free unbound α-synuclein in the CSF can be determined by comparing the level of free unbound α-synuclein in CSF samples collected from the subject at different time points (e.g., before treatment according to the present invention (e.g., at baseline) and during and / or after treatment), and determining whether there is any change during that time period. A CSF sample can be obtained from a subject before the start of the method of the present invention or before administration that constitutes a part of the method of the present invention, and the CSF sample can be compared to one or more CSF samples obtained from the subject during and / or after the method of the present invention (such as a CSF sample collected after one or more administrations of an antibody or antigen-binding fragment thereof according to the present invention). The free unbound α-synuclein reduction in CSF can include a decrease in the α-synuclein level in the CSF of a subject during the methods of the invention, for example, compared to the level in the CSF of a subject before the methods of the invention (for example, at baseline), after the methods of the invention, the level is lower. The free unbound α-synuclein reduction in CSF can be a decrease in the progression of the free unbound α-synuclein level in CSF compared to that observed in a control subject not administered with an anti-α-synuclein antibody or its antigen-binding fragment according to the invention. The α-synuclein diffusion reduction can include a decrease in the progression of the free unbound α-synuclein level in CSF compared to a reference level of free unbound α-synuclein. The reference level of free unbound α-synuclein can be based on a model of disease progression, for example, based on a normal or average level of free unbound α-synuclein in a reference subject with the same disease or disorder or the same α-synuclein disease as the subject or a reference subject with the same degree of disease progression as the subject being treated. The reference level of free unbound α-synuclein can be based on the amount of free unbound α-synuclein observed in the CSF of one or more control subjects, such as a population of control subjects to which an anti-α-synuclein antibody or antigen-binding fragment thereof according to the present invention is not administered, such as one or more placebo-treated subjects. One or more CSF samples obtained from a subject during and / or after the methods of the present invention (such as CSF samples collected after one or more administrations of an antibody or antigen-binding fragment thereof according to the present invention) can be compared with one or more CSF samples obtained from one or more placebo-treated subjects.The level of free unbound α-synuclein can be assessed in a CSF sample, or the relative level of free unbound α-synuclein can be compared between CSF samples. The amount (or relative amount) of free unbound α-synuclein can then be compared between CSF samples collected at different time points associated with the methods of the present invention to determine whether the methods of the present invention or the administration of one or more doses of an antibody or antigen-binding fragment thereof has reduced the amount of free unbound α-synuclein in the CSF. A lower level of free unbound α-synuclein in the CSF during or after the methods of the present invention compared to the level of free unbound α-synuclein in the subject's CSF before the methods of the present invention may indicate that the methods of the present invention cause a reduction in free unbound α-synuclein in the CSF. A lower level of free unbound α-synuclein in the subject's CSF during or after the methods of the invention, compared to the level of free unbound α-synuclein in a control or placebo-treated subject, or compared to a reference level of free unbound α-synuclein, can indicate that the methods of the invention result in a decrease in free unbound α-synuclein in the CSF. A delay or slowing of the increase in the level of free unbound α-synuclein in the subject's CSF, when compared to the change during or after the methods of the invention, compared to the level of free unbound α-synuclein in the CSF of a control or placebo-treated subject over the same time period, or compared to a reference change in the level of free unbound α-synuclein, (based on a comparison between the level before the methods of the invention (e.g., at baseline) and the level during or after the methods of the invention), indicates that the methods of the invention result in a decrease in free unbound α-synuclein in the CSF.

[0214] The level of free, unbound α-synuclein in the brain ISF of a subject can be measured by any suitable means. For example, an ISF sample can be collected by microdialysis, for example as described in Herukka et al. (2015) J. Alzheimers Dis. 46(1): 261-9. The level of free, unbound α-synuclein in the ISF sample can be assessed by methods as discussed above with respect to the measurement of free, unbound α-synuclein levels in CSF (such as by methods comprising separating α-synuclein bound to the antibody or fragment from α-synuclein not bound to the antibody or fragment). For example, the ISF sample can be immunoprecipitated to remove α-synuclein bound to the antibody or its antigen-binding fragment. The free level of α-synuclein in the remaining supernatant can be determined, for example, by ELISA. A change in the level of free, unbound α-synuclein in the ISF can be determined by comparing the levels of α-synuclein in ISF samples collected from the subject at different time points in the same manner as discussed above with respect to assessing reduction in CSF samples. A change or decrease in the level of free, unbound α-synuclein in the ISF can be determined by comparing the levels of free, unbound α-synuclein in ISF samples collected from the subject at different time points (e.g., before treatment according to the invention (e.g., at baseline) and during and / or after treatment), and determining whether there is any change over that time period. An ISF sample obtained from the subject before the start of the method of the invention or before administration that forms part of the method of the invention can be compared to one or more ISF samples obtained from the subject during and / or after the method of the invention (such as an ISF sample collected after one or more administrations of an antibody or antigen-binding fragment thereof according to the invention). Alternatively, one or more ISF samples obtained from a subject during and / or after the methods of the invention (such as ISF samples collected after one or more administrations of an antibody or antigen-binding fragment thereof according to the invention) can be compared to one or more ISF samples obtained from a placebo-treated subject. The level of free, unbound α-synuclein can be assessed in the ISF samples, or the relative levels of free, unbound α-synuclein can be compared between ISF samples. A reduction in free, unbound α-synuclein in the ISF can include a decrease in the level of α-synuclein in the subject's ISF during the methods of the invention, e.g., a lower level after the methods of the invention compared to the level in the subject's ISF before the methods of the invention (e.g., at baseline). A reduction in free, unbound α-synuclein in the ISF can be a slowing or delay in the progression of free, unbound α-synuclein levels in the ISF compared to that observed in a control subject not administered an anti-α-synuclein antibody or antigen-binding fragment thereof according to the invention.Reduction in α-synuclein diffusion can include a slowed or delayed progression of free, unbound α-synuclein levels in the ISF compared to a reference level of free, unbound α-synuclein. The reference level of free, unbound α-synuclein can be based on a model of disease progression, for example, based on normal or average levels of free, unbound α-synuclein in the ISF of a reference subject having the same disease or disorder or the same α-synuclein disease as the subject, or a reference subject having the same degree of disease progression as the subject being treated. The reference level of free, unbound α-synuclein can be based on the amount of free, unbound α-synuclein observed in the ISF of one or more control subjects, such as a control population of subjects who have not been administered an anti-α-synuclein antibody or antigen-binding fragment thereof according to the invention, such as one or more placebo-treated subjects. One or more ISF samples obtained from a subject during and / or after the methods of the invention (such as ISF samples collected after one or more administrations of an antibody or antigen-binding fragment thereof according to the invention) can be compared to one or more ISF samples obtained from one or more placebo-treated subjects. The level of free, unbound α-synuclein can be assessed in the ISF samples, or the relative levels of free, unbound α-synuclein can be compared between ISF samples. The amount (or relative amount) of free, unbound α-synuclein can then be compared between ISF samples collected at different time points associated with the methods of the invention to determine whether the methods of the invention or the administration of one or more doses of an antibody or antigen-binding fragment thereof has reduced the amount of free, unbound α-synuclein in the ISF. A lower level of free, unbound α-synuclein in the ISF during or after the methods of the invention compared to the level of free, unbound α-synuclein in the subject's ISF before the methods of the invention can indicate that the methods of the invention result in a reduction in free, unbound α-synuclein in the ISF. A lower level of free, unbound α-synuclein in the subject's ISF during or after the methods of the invention, compared to the level of free, unbound α-synuclein in a control or placebo-treated subject, or compared to a reference level of free, unbound α-synuclein, can indicate that the methods of the invention result in a decrease in free, unbound α-synuclein in the ISF. A delay or slowing of the increase in the level of free, unbound α-synuclein in the subject's ISF (based on a comparison of the level before the methods of the invention (e.g., at baseline) to the level during or after the methods of the invention), when compared to the change during or after the methods of the invention, compared to the level of free, unbound α-synuclein in the ISF of a control or placebo-treated subject over the same time period, or compared to a reference change in the level of free, unbound α-synuclein, can indicate that the methods of the invention result in a decrease in free, unbound α-synuclein in the ISF.

[0215] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof according to the present invention can alter (e.g., improve, inhibit, reduce, stabilize, delay, or slow the increase in) one or more symptoms or features of an α-synuclein disease in a subject, based on a score or measurement of one or more clinical indicators, outcomes, or symptoms of the subject (e.g., based on a clinical scoring system). There are a variety of well-known clinical scoring systems for evaluating subjects with α-synuclein diseases. Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof according to the present invention can alter (e.g., improve, inhibit, reduce, stabilize, delay, or slow the increase in) the score of a subject in any such scoring system, such as the UMSARS, CGI-S, SCOPA AUT, or MDS-UPDRS score of a subject as described herein. Preferably, the reduction in any of the scores or levels described herein is a reduction compared to the score of a subject not treated with such an anti-α-synuclein antibody or antigen-binding fragment thereof (e.g., a control or placebo-treated subject or a control or placebo-treated subject group). Thus, compared to such a control, a reduction in the score or level described herein indicates a slowing of progression of alpha-synuclein disease. For example, based on a comparison between the score of a subject before (e.g., at baseline) and the score of a subject during or after the method of the present invention, the method of the present invention may reduce the score of the subject. The method of the present invention may delay or slow the increase in the score of the subject. This may reflect a delay or slowing of progression of the subject's alpha-synuclein disease. For example, based on a comparison between the score of a subject before (e.g., at baseline) and the score of a subject during or after the method of the present invention, the score may be stable, e.g., the score may not increase. The change in score (e.g., based on a comparison between the score of a subject before (e.g., at baseline) and the score of a subject during or after the method of the present invention) may be less than the change expected based on a control (such as a control subject to which an anti-alpha-synuclein antibody or antigen-binding fragment thereof according to the present invention is not administered). For example, compared to a reference value representing the expected score change of such a subject during the time period, the change in the score of the subject may be slowed or delayed. This may reflect a delay or slowing of progression of the subject's alpha-synuclein disease. The reference value can be based on a model of disease progression, for example, based on normal levels or average scores or changes in scores observed in reference subjects with the same disease or disorder or the same α-synuclein disease as the subject or with the same degree of disease progression as the subject being treated. The reference value can be based on the scores of one or more control subjects, such as a control subject population that is not administered an anti-α-synuclein antibody or antigen-binding fragment thereof according to the invention, such as one or more placebo-treated subjects.The reference value can be based on the change in score over the same time period in one or more control subjects, such as a population of control subjects who are not administered an anti-α-synuclein antibody or antigen-binding fragment thereof according to the invention, such as one or more placebo-treated subjects.

[0216] The score can be a UMSARS score. The score can be a modified version of UMSARS Part I. The method can slow the progression of α-synucleinopathy as measured by the modified version of UMSARS Part I. Thus, treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the subject's score in the modified version of UMSARS Part I. Preferably, the method slows the progression of α-synucleinopathy as measured by the modified version of UMSARS Part I. UMSARS Part I can be modified to include fewer than 12 questions. UMSARS Part I can be modified so that the answers to the questions are scored on a scale of 0 to 3 or 1 to 4.

[0217] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the subject's UMSARS total score. As measured by the UMSARS total score, the method can slow the progression of α-synuclein disease. The UMSARS total score can include all items from UMSARS Part I, Part II, Part III, and Part IV. The UMSARS total score can include all items from the modified UMSARS Part I and all items from UMSARS Part II, Part III, and Part IV.

[0218] Treatment with the anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the subject's UMSARS Part I score. The method can slow the progression of α-synucleinopathy as measured by the UMSARS Part I score.

[0219] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the subject's UMSARS Part II score. The method can slow the progression of α-synucleinopathy as measured by the UMSARS Part II score.

[0220] Treatment with anti-α-synuclein antibodies or their antigen-binding fragments can reduce the subject's 11 UMSARS scores. As measured by the 11 UMSARS scores, the method can slow the progression of α-synuclein disease. The 11 UMSARS scores are described in Palma et al. (2021) Clinical Auton. Res. 31: 157-164 and consist of UMSARS Part I questions 2, 3, 6, 7, and 11 and UMSARS Part II questions 1, 2, 9, 11, 12, and 14. These 11 items showed the highest cumulative standardized effect in a longitudinal natural history analysis from a natural history study of synuclein disease.

[0221] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can reduce a subject's Clinical Global Impression-Severity (CGI-S) scale score. This approach can slow the progression of α-synuclein disease, as measured by the CGI-S scale score. The CGI-S (described in more detail in Busner and Targum (2007) Psychiatry (Edgmont) 4(7):28-37) is used to assess a clinician's impression of a subject's clinical condition. Clinicians rate the severity of a subject's illness on a 7-point scale, ranging from 1 for normal, completely healthy, to 7 for the most severely ill patient. The rating is based on symptoms, behaviors, and function observed and reported over the past 7 days and should reflect the average severity over the 7 days.

[0222] Treatment with anti-α-synuclein antibodies or their antigen-binding fragments can reduce the subject's score in the SCOPA-AUT test. As measured by the SCOPA-AUT test, this method can slow the progression of α-synuclein disease. The SCOPA-AUT (described in more detail in Visser et al. (2004) Mov. Disord. 19(11): 1306-12) is used to evaluate autonomic symptoms in patients with Parkinson's disease. The scale is completed by the patient themselves and consists of 25 items that assess gastrointestinal, urinary, cardiovascular, thermoregulatory, pupillary movement, and sexual symptoms.

[0223] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof (eg, a fixed dose every 4 weeks for at least 1 year) improves survival compared with placebo.

[0224] Treatment with an anti-α-synuclein antibody or its antigen-binding fragment can reduce the subject's score on the Movement Disorder Society-sponsored revised Unified Parkinson's Disease Rating Scale (MDS-UPDRS). As measured by the MDS-UPDRS, this method can slow the progression of α-synuclein disease. For example, as assessed by an increase of at least 5 points from baseline in the MDS-UPDRS Part III score, this method can increase the time to meaningful progression of disease motor signs. The MDS-UPDRS (described in more detail in Goetz et al. (2008) Mov. Disord. 23 (15): 21129-2170) is used to evaluate various aspects of Parkinson's disease, including non-motor and motor experiences of daily life and motor complications.

[0225] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can improve a subject's cognition. For example, treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can increase a subject's score on the MoCA. This method can slow the progression of α-synucleinopathy, as measured by the subject's score on the MoCA.

[0226] Treatment with an anti-alpha-synuclein antibody or antigen-binding fragment thereof (e.g., a fixed dose once every 4 weeks for at least 1 year) can improve gait, balance, and / or parkinsonism compared to a control or reference value, for example, compared to the progression of a subject treated with a placebo. The method can slow the progression of alpha-synuclein disease as measured by gait, balance, and / or parkinsonism.

[0227] For example, as measured using the Orthostatic Hypotension Questionnaire (OHQ), treatment with an anti-α-synuclein antibody or its antigen-binding fragment (e.g., a fixed dose once every 4 weeks for at least 1 year) can improve blood pressure and / or heart rate. This approach can slow the progression of α-synuclein disease, as measured using the OHQ. The OHQ (described in more detail in Kaufmann et al. (2011) Clin. Auton. Res. 22(2): 79-90) is used to assess the symptom burden associated with orthostatic hypotension and includes two parts: a six-item symptom assessment scale; and a four-item daily activities scale.

[0228] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof (e.g., a fixed dose once every 4 weeks for at least 1 year) can improve one or more symptoms of REM behavior disorder, for example, as measured using: polysomnography with or without video; the Innsbruck REM Sleep Behavior Disorder Questionnaire (described in more detail in Frauscher et al. (2012) Mov. Disord. 27(13):1673-1678); the Hong Kong REM Behavior Disorder Questionnaire (described in more detail in Shen et al. (2014) Sleep Med. 15(8):952-958); the CGI (e.g., CGI-C); the PGI (e.g., PGI-I or PGI-C; described in Guy (1976) ECDEU assessment manual for psychopharmacology, US Department of Health, Education, and The method can slow the progression of one or more symptoms of REM behavior disorder, for example, as measured using polysomnography with or without video; the Innsbruck REM Sleep Behavior Questionnaire; the Hong Kong REM Behavior Disorder Questionnaire; the CGI-C; the PGI-I / C; and / or the CIRUS-RBD questionnaire.

[0229] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof (e.g., a fixed dose every 4 weeks for at least 1 year) can reduce plasma neurofilament light chain levels, reduce total α-synuclein levels, reduce free unbound CSF α-synuclein levels, and / or reduce CSF neurofilament light chain levels. As measured by plasma neurofilament light chain levels, total α-synuclein levels, free unbound CSF α-synuclein levels, and / or CSF neurofilament light chain levels, this method can slow the progression of α-synuclein disease.

[0230] A reduction in any of the scores or levels described herein can be a reduction compared to a control measurement of the subject (e.g., the subject's baseline score immediately before starting treatment). For example, treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the score or level by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% compared to baseline.

[0231] Preferably, the reduction in any score or level described herein can be a reduction compared to the score or level of a subject or subject group (e.g., a control subject, a placebo-treated subject, a control subject group, or a placebo-treated subject group) to which an anti-alpha-synuclein antibody or its antigen-binding fragment is not administered. For example, treatment with an anti-alpha-synuclein antibody or its antigen-binding fragment can provide a score or level that is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% lower than the corresponding score or level of a control subject, a placebo-treated subject, a control subject group, or a placebo-treated subject group. Preferably, the comparison is made with a placebo-treated subject group, such as a typical or average change in the score obtained from a placebo-treated subject group. Preferably, the control or placebo-treated subject is a subject who has the same disease or disorder as the subject being treated. Preferably, the control or placebo-treated subject has the same or similar level of disease progression as the subject being treated at the start of treatment. Preferably, control or placebo-treated subjects have scores in the same range or similar to those of treated subjects at the start of treatment.

[0232] Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject, for example, at a fixed dose once every 4 weeks for at least 1 year, and the method reduces the level of free unbound α-synuclein in the subject's cerebrospinal fluid (CSF). The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free unbound α-synuclein in the subject's CSF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's CSF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's CSF by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's CSF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, e.g., compared to the level in a placebo-treated subject. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject at a fixed dose, e.g., once every 4 weeks for at least 1 year, and the method reduces the level of free, unbound α-synuclein in the subject's CSF by at least 30%, compared to a control or reference value, e.g., compared to the level in a placebo-treated subject.

[0233] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's interstitial fluid (ISF). The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's ISF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's ISF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's ISF by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's ISF by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, e.g., compared to the level in a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks for at least 1 year, and the method can reduce the level of free, unbound α-synuclein in the subject's ISF by at least 30%, compared to a control or reference value, e.g., compared to the level in a placebo-treated subject.

[0234] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the spread of α-synuclein in a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the spread of α-synuclein in the subject by at least 30%, compared to a control or reference value, for example, compared to the spread of α-synuclein in a placebo-treated subject.

[0235] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's score on a modified version of UMSARS, Part I. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's score on a modified version of UMSARS, Part I by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every four weeks for at least one year, and the method can reduce the subject's score on a modified version of UMSARS Part I by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The methods or dosage regimens disclosed herein can reduce the subject's modified UMSARS Part I score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every four weeks for at least one year, and the method can reduce the subject's score on a modified version of UMSARS Part I by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's score by at least 30% compared to a control or reference value, e.g., compared to the score of a placebo-treated subject.

[0236] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score from baseline by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS total score by at least 30%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0237] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, e.g., compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part I score by at least 30%, compared to a control or reference value, e.g., compared to the score of a placebo-treated subject.

[0238] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS Part II score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS Part II score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's UMSARS Part II score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part II score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part II score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, e.g., compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, e.g., once every 4 weeks, for at least 1 year, and the method can reduce the subject's UMSARS Part II score by at least 30%, compared to a control or reference value, e.g., compared to the score of a placebo-treated subject.

[0239] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score from baseline by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score by at least 10%, at least 20%, or at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's 11-item UMSARS score by at least 30% compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0240] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's CGI-S score by at least 30%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0241] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score from baseline by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's SCOPA-AUT score by at least 30%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0242] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can improve the subject's cognitive, motor, and / or speech function. Thus, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can improve the subject's cognitive, motor, and / or speech function. The method can slow the progression of α-synuclein disease as measured by the subject's cognitive, motor, and / or speech function.

[0243] Treatment with an anti-α-synuclein antibody or antigen-binding fragment thereof can improve autonomic function, such as orthostatic blood pressure and / or orthostatic heart rate. Thus, an anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to a subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can improve the subject's orthostatic blood pressure and / or orthostatic heart rate. The method can slow the progression of α-synuclein disease as measured by orthostatic blood pressure and / or orthostatic heart rate.

[0244] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score from baseline by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can reduce the subject's MDS-UPDRS score by at least 30%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0245] The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score by at least 30% from baseline. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject. The anti-α-synuclein antibody or antigen-binding fragment thereof can be administered to the subject at a fixed dose, for example, once every 4 weeks for at least 1 year, and the method can increase the subject's MoCA score by at least 30%, compared to a control or reference value, for example, compared to the score of a placebo-treated subject.

[0246] Prevention or treatment as defined herein or according to the present invention may be applied as a separate therapy, or may involve the administration of other agents or other therapies in addition to anti-α-synuclein antibodies or antigen-binding fragments. Anti-α-synuclein antibodies or antigen-binding fragments may be administered in combination with another agent or therapy for treating or preventing α-synuclein disease. Anti-α-synuclein antibodies or antigen-binding fragments may be administered in combination with established agents or therapies typically used to treat α-synuclein disease, such as L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine (receptor) agonists, catechol-O-methyltransferase (COMT) inhibitors, and / or type B monoamine oxidase (MAO-B) inhibitors. The administration of other agents or therapies may be combined with the antibodies or antigen-binding fragments used in the present invention, or as an adjuvant thereof, or in combination therewith, and may be performed by administering the various components of the treatment simultaneously, sequentially, or separately. Antibodies or antigen-binding fragments may be provided in the same composition with additional therapeutic agents. The antibody or antigen-binding fragment can be provided in a different composition from any other therapeutic agent administered as part of the same treatment. The anti-α-synuclein antibody or antigen-binding fragment can be administered in combination with one or more other anti-α-synuclein antibodies or antigen-binding fragments thereof (such as one or more other anti-α-synuclein antibodies or antigen-binding fragments thereof as described herein). Each anti-α-synuclein antibody as defined herein can be administered in a fixed dose of 50 mg to 5,000 mg or a fixed dose as defined herein. When more than one anti-α-synuclein antibody as defined herein is administered in the methods of the present invention, each administration can be performed in a fixed dose of 50 mg to 5,000 mg or a fixed dose as defined herein.

[0247] Combination therapy may be performed in any manner deemed necessary or convenient by one skilled in the art, and for the purposes of this specification, no limitation is considered as to the order, amount, repetition or relative amounts of the compounds to be used in combination.

[0248] 4.2 Anti-α-synuclein Antibodies or Antigen-Binding Fragments thereof

[0249] The present invention uses anti-α-synuclein antibodies or antigen-binding fragments thereof. For example, in the methods of the present invention, anti-α-synuclein antibodies or antigen-binding fragments thereof are administered to treat or prevent α-synuclein diseases. Similarly, the present invention provides an anti-α-synuclein antibody or antigen-binding fragment thereof for use in a method of treating or preventing α-synuclein diseases. The kits and compositions of the present invention comprise anti-α-synuclein antibodies or antigen-binding fragments thereof. In any aspect of the present invention, the anti-α-synuclein antibody or antigen-binding fragment thereof can be an anti-α-synuclein antibody or antigen-binding fragment thereof as described below.

[0250] As used herein, the term "anti-α-synuclein antibody" means an antibody that binds to α-synuclein. An anti-α-synuclein antibody or an antigen-binding fragment thereof can specifically bind to α-synuclein, for example, it can bind to α-synuclein without binding to other related molecules. For example, an anti-α-synuclein antibody or an antigen-binding fragment thereof can bind to α-synuclein, but not to β-synuclein or γ-synuclein. An anti-α-synuclein antibody or an antigen-binding fragment thereof can bind to human α-synuclein, but not to human β-synuclein or human γ-synuclein. An anti-α-synuclein antibody or an antigen-binding fragment thereof can bind to human, rat and / or cynomolgus monkey α-synuclein. An anti-α-synuclein antibody or an antigen-binding fragment thereof can bind to human, rat and cynomolgus monkey α-synuclein.

[0251] 4.2.1 Amino Acid Sequences of aslo0452ngl-3 and Its Antigen-Binding Fragments

[0252] The anti-α-synuclein antibody or antigen-binding fragment thereof can be the antibody aslo0452 ngl-3 or an antigen-binding fragment thereof. Aslo0452 ngl-3 is a human α-synuclein monoclonal antibody (mAb) derived from the parent clone asyn0087 and is directed against the C-terminus of human α-synuclein. Aslo0452 ngl-3 comprises a variable heavy chain region of SEQ ID NO: 14 and a variable light chain region of SEQ ID NO: 19. Aslo0452 ngl-3 comprises a light chain having an amino acid sequence of SEQ ID NO: 17. Due to lysine cleavage during the manufacturing process, the final lysine of the aslo0452 ngl-3 heavy chain may not be present. Therefore, aslo0452 ngl-3 may comprise a heavy chain having an amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62. Preferably, aslo0452 ngl-3 comprises a heavy chain having an amino acid sequence of SEQ ID NO: 12. Aslo0452 ngl-3 may comprise a heavy chain having the amino acid sequence of SEQ ID NO: 12 and a light chain having the amino acid sequence of SEQ ID NO: 17. Aslo0452 ngl-3 may comprise a heavy chain having the amino acid sequence of SEQ ID NO: 62 and a light chain having the amino acid sequence of SEQ ID NO: 17. Aslo0452 ngl-3 binds to both monomeric and aggregated forms of human α-synuclein. Aslo0452 ngl-3 has been shown to reduce α-synuclein spread in vivo. In addition, aslo0452 ngl-3 has been shown to bind to reduce levels of free, unbound α-synuclein in interstitial fluid (ISF) and cerebrospinal fluid (CSF) in animal models of the disease. Aslo0452 ngl-3 and its production are described in more detail in WO2017 / 207739, which is incorporated herein by reference in its entirety. As shown herein, aslo0452 ngl-3 is the first anti-α-synuclein antibody demonstrated to reduce the levels of free, unbound α-synuclein in the CSF of human subjects. This allows for measurement of aslo0452 ngl-3 target engagement during treatment.

[0253] The anti-α-synuclein antibody or antigen-binding fragment thereof used in the present invention may have the six CDRs of antibody aslo0452 ng1-3. Therefore, preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises:

[0254] a) three heavy chain CDRs having the following sequences:

[0255] (i) H-CDR1 of SEQ ID NO: 5,

[0256] (ii) H-CDR2 of SEQ ID NO: 15, and

[0257] (iii) H-CDR3 of SEQ ID NO: 16, and

[0258] b) three light chain CDRs having the following sequences:

[0259] (i) L-CDR1 of SEQ ID NO: 20,

[0260] (ii) L-CDR2 of SEQ ID NO: 10, and

[0261] (iii) L-CDR3 of SEQ ID NO: 21.

[0262] As used herein, "H-CDR" refers to the complementarity determining region (CDR) on the heavy chain region of an antibody or antigen-binding fragment thereof, and "L-CDR" refers to the complementarity determining region (CDR) on the light chain region of an antibody or antigen-binding fragment thereof.

[0263] The anti-α-synuclein antibody or antigen-binding fragment thereof can comprise a variable heavy chain region encoded by a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence defined by SEQ ID NO: 13, and / or a variable light chain region encoded by a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence defined by SEQ ID NO: 18.

[0264] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region encoded by the nucleotide sequence defined by SEQ ID NO: 13 and / or a variable light chain region encoded by the nucleotide sequence defined by SEQ ID NO: 18. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region encoded by the nucleotide sequence defined by SEQ ID NO: 13 and a variable light chain region encoded by the nucleotide sequence defined by SEQ ID NO: 18.

[0265] The anti-α-synuclein antibody or antigen-binding fragment thereof can comprise a variable heavy chain region having an amino acid sequence as defined by SEQ ID NO: 14, or at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence defined by SEQ ID NO: 14, and / or a variable light chain region having an amino acid sequence as defined by SEQ ID NO: 19, or at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence defined by SEQ ID NO: 19.

[0266] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having the amino acid sequence of SEQ ID NO: 14 and / or a variable light chain region having the amino acid sequence of SEQ ID NO: 19. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having the amino acid sequence of SEQ ID NO: 14 and a variable light chain region having the amino acid sequence of SEQ ID NO: 19. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region consisting of the amino acid sequence defined by SEQ ID NO: 14 and a variable light chain region consisting of the amino acid sequence defined by SEQ ID NO: 19.

[0267] Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence defined by SEQ ID NO: 14 and a variable light chain region that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence defined by SEQ ID NO: 19, and may further comprise:

[0268] a) three heavy chain CDRs having the following sequences:

[0269] (i) H-CDR1 of SEQ ID NO: 5,

[0270] (ii) H-CDR2 of SEQ ID NO: 15, and

[0271] (iii) H-CDR3 of SEQ ID NO: 16, and

[0272] b) three light chain CDRs having the following sequences:

[0273] (i) L-CDR1 of SEQ ID NO: 20,

[0274] (ii) L-CDR2 of SEQ ID NO: 10, and

[0275] (iii) L-CDR3 of SEQ ID NO: 21.

[0276] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having the amino acid sequence defined by SEQ ID NO: 12 or SEQ ID NO: 62. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence defined by SEQ ID NO: 12. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a light chain having the amino acid sequence defined by SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having the amino acid sequence defined by SEQ ID NO: 12 or SEQ ID NO: 62 and / or a light chain having the amino acid sequence defined by SEQ ID NO: 17. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence defined by SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain having the amino acid sequence defined by SEQ ID NO: 17. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence defined by SEQ ID NO: 12 and a light chain having the amino acid sequence defined by SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain consisting of the amino acid sequence defined by SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain consisting of the amino acid sequence defined by SEQ ID NO: 17. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain consisting of the amino acid sequence defined by SEQ ID NO: 12 and a light chain consisting of the amino acid sequence defined by SEQ ID NO: 17.

[0277] 4.2.2 Amino Acid Sequences of aslo0543 and Its Antigen-Binding Fragments

[0278] Anti-alpha-synuclein antibody or its antigen-binding fragment can be antibody aslo0543 or its antigen-binding fragment. Aslo0543 is a human alpha-synuclein monoclonal antibody (mAb) derived from parent clone asyn0087 and is directed against the C-terminus of human alpha-synuclein. Aslo0543 comprises a variable heavy chain region of SEQ ID NO:24 and a variable light chain region of SEQ ID NO:30. Aslo0543 comprises a heavy chain (SEQ ID NO:22) and a light chain (SEQ ID NO:28). Aslo0543 binds to both monomers and aggregated forms of human alpha-synuclein. Aslo0543 and its production are described in more detail in WO2017 / 207739, which is incorporated herein by reference in its entirety.

[0279] The anti-α-synuclein antibody or antigen-binding fragment thereof may have the six CDRs of antibody aslo0543. Thus, the anti-α-synuclein antibody or antigen-binding fragment thereof may comprise:

[0280] a) three heavy chain CDRs having the following sequences:

[0281] (i) H-CDR1 of SEQ ID NO: 25,

[0282] (ii) H-CDR2 of SEQ ID NO: 26, and

[0283] (iii) H-CDR3 of SEQ ID NO: 27, and

[0284] b) three light chain CDRs having the following sequences:

[0285] (i) L-CDR1 of SEQ ID NO: 31,

[0286] (ii) L-CDR2 of SEQ ID NO: 32, and

[0287] (iii) L-CDR3 of SEQ ID NO: 33.

[0288] The anti-α-synuclein antibody or antigen-binding fragment thereof can comprise a variable heavy chain region encoded by a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence defined by SEQ ID NO: 23, and / or a variable light chain region encoded by a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleotide sequence defined by SEQ ID NO: 29.

[0289] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region encoded by the nucleotide sequence defined by SEQ ID NO: 23 and / or a variable light chain region encoded by the nucleotide sequence defined by SEQ ID NO: 29. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region encoded by the nucleotide sequence defined by SEQ ID NO: 23 and a variable light chain region encoded by the nucleotide sequence defined by SEQ ID NO: 29.

[0290] The anti-α-synuclein antibody or antigen-binding fragment thereof can comprise a variable heavy chain region having an amino acid sequence as defined by SEQ ID NO: 24, or at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence defined by SEQ ID NO: 24, and / or a variable light chain region having an amino acid sequence as defined by SEQ ID NO: 30, or at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence defined by SEQ ID NO: 30.

[0291] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region having the amino acid sequence of SEQ ID NO: 24 and / or a variable light chain region having the amino acid sequence of SEQ ID NO: 30. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having the amino acid sequence of SEQ ID NO: 24 and a variable light chain region having the amino acid sequence of SEQ ID NO: 30. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region consisting of the amino acid sequence defined by SEQ ID NO: 24 and a variable light chain region consisting of the amino acid sequence defined by SEQ ID NO: 30.

[0292] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence defined by SEQ ID NO: 24 and a variable light chain region that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence defined by SEQ ID NO: 30, and may further comprise:

[0293] a) three heavy chain CDRs having the following sequences:

[0294] (i) H-CDR1 of SEQ ID NO: 25,

[0295] (ii) H-CDR2 of SEQ ID NO: 26, and

[0296] (iii) H-CDR3 of SEQ ID NO: 27, and

[0297] b) three light chain CDRs having the following sequences:

[0298] (i) L-CDR1 of SEQ ID NO: 31,

[0299] (ii) L-CDR2 of SEQ ID NO: 32, and

[0300] (iii) L-CDR3 of SEQ ID NO: 33.

[0301] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region that is at least 80%, 85%, 90%, or 95% identical to the sequence defined by SEQ ID NO: 14 and a variable light chain region that is at least 80%, 85%, 90%, or 92% identical to the sequence defined by SEQ ID NO: 19, and may further comprise:

[0302] a) three heavy chain CDRs having the following sequences:

[0303] (i) H-CDR1 of SEQ ID NO: 25,

[0304] (ii) H-CDR2 of SEQ ID NO: 26, and

[0305] (iii) H-CDR3 of SEQ ID NO: 27, and

[0306] b) three light chain CDRs having the following sequences:

[0307] (i) L-CDR1 of SEQ ID NO: 31,

[0308] (ii) L-CDR2 of SEQ ID NO: 32, and

[0309] (iii) L-CDR3 of SEQ ID NO: 33.

[0310] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence defined by SEQ ID NO: 24 and a variable light chain region that is at least 80%, 85%, 90%, or 92% identical to the sequence defined by SEQ ID NO: 30, and may further comprise:

[0311] a) three heavy chain CDRs having the following sequences:

[0312] (i) H-CDR1 of SEQ ID NO: 5,

[0313] (ii) H-CDR2 of SEQ ID NO: 15, and

[0314] (iii) H-CDR3 of SEQ ID NO: 16, and

[0315] b) three light chain CDRs having the following sequences:

[0316] (i) L-CDR1 of SEQ ID NO: 20,

[0317] (ii) L-CDR2 of SEQ ID NO: 10, and

[0318] (iii) L-CDR3 of SEQ ID NO: 21.

[0319] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having the amino acid sequence defined by SEQ ID NO: 22 and / or a light chain having the amino acid sequence defined by SEQ ID NO: 28. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain having the amino acid sequence defined by SEQ ID NO: 22 and a light chain having the amino acid sequence defined by SEQ ID NO: 28. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a heavy chain consisting of the amino acid sequence defined by SEQ ID NO: 22 and a light chain consisting of the amino acid sequence defined by SEQ ID NO: 28.

[0320] 4.2.3 Functional Properties of Antibodies and Antigen-Binding Fragments thereof Used in the Present Invention

[0321] Any of the anti-α-synuclein antibodies or antigen-binding fragments thereof disclosed herein, such as any anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention, can have any one or more of the functional properties of aslo0452 ngl-3, such as any of the functional properties of aslo0452 ngl-3 listed herein. Any of the anti-α-synuclein antibodies or antigen-binding fragments thereof disclosed herein, such as any anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention, can have any one or more of the functional properties of aslo0543, such as any of the functional properties of aslo0543 listed herein.

[0322] Like the parent clone asyn0087, aslo0452 ng1-3 and aslo0543 antibodies bind to the C-terminal region of human α-synuclein (residues 96 to 140). More specifically, aslo0452 ng1-3 and aslo0543 antibodies bind to a region comprising about amino acid 102 and about amino acid 130 of human α-synuclein (e.g., SEQ ID NO: 1). Thus, the present invention can use anti-α-synuclein antibodies or antigen-binding fragments thereof that specifically bind to the C-terminal region of human α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can specifically bind to a region comprising about amino acid 102 to about amino acid 130 of human α-synuclein (e.g., SEQ ID NO: 1), for example, it can specifically bind to a stretch of amino acids 102 to 130 of human α-synuclein, or it can specifically bind to one or more of amino acids 102 to 130 of human α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can specifically bind to a region comprising about amino acid 120 to about amino acid 130 of human α-synuclein (SEQ ID NO: 1). The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acids in a region comprising about amino acid 102 to about amino acid 130 of human α-synuclein (SEQ ID NO: 1), such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acids in a region comprising about amino acid 102 to about amino acid 130 of human α-synuclein (SEQ ID NO: 1). The epitope of the anti-α-synuclein antibody or antigen-binding fragment thereof can comprise amino acids 102 to 130 of human α-synuclein (SEQ ID NO: 1), or can comprise one or more amino acids from a region comprising amino acids 102 to 130 of human α-synuclein. For example, the epitope of the anti-α-synuclein antibody or antigen-binding fragment thereof can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acids from amino acids 102 to 130 of human α-synuclein (SEQ ID NO: 1). The anti-α-synuclein antibody or antigen-binding fragment thereof binds to a different epitope than the antibody that binds to the 9E4 antibody.

[0323] Aslo0452 ngl-3 and aslo0543 antibodies are selective for alpha-synuclein. Therefore, preferably, the present invention can use anti-alpha-synuclein antibodies or their antigen-binding fragments that bind to human alpha-synuclein but not to human beta-synuclein or human gamma-synuclein. The specificity of anti-alpha-synuclein antibodies or their antigen-binding fragments can be determined using any suitable means, for example, HTRF epitope competition assay, which measures the binding of biotinylated human alpha-synuclein to the antibody or its antigen-binding fragment in solution. In the HTRF assay, relevant synuclein (e.g., alpha-synuclein, beta-synuclein, and gamma-synuclein) can be titrated into the assay, and the selectivity of the antibody or its antigen-binding fragment can be measured by assessing the degree of inhibition of biotinylated human alpha-synuclein binding to the antibody or its antigen-binding fragment. IC can be determined by fitting the data curve to a four-parameter logistic equation using PRISM 6 software (Graphpad). 50 value.

[0324] The aslo0452 ngl-3 and aslo0543 antibodies bind to each of human, rat, and cynomolgus monkey α-synuclein proteins. The ability of aslo0452 ngl-3 and aslo0543 to bind to human, cynomolgus monkey, and rat α-synuclein proteins, compared to antibodies that do not bind to human, cynomolgus monkey, and rat α-synuclein proteins, indicates binding to distinct epitopes on human α-synuclein. Therefore, the aslo0452 ngl-3 and aslo0543 antibodies can be used for in vivo safety evaluation and studies in cynomolgus monkey and rat disease models. Therefore, preferably, the present invention can utilize anti-α-synuclein antibodies or antigen-binding fragments thereof that bind to human, rat, and / or cynomolgus monkey α-synuclein proteins. Preferably, the anti-α-synuclein antibodies or antigen-binding fragments thereof bind to human, rat, and cynomolgus monkey α-synuclein proteins. Binding to human, rat, or cynomolgus monkey α-synuclein proteins can be determined using any suitable method, for example, an HTRF epitope competition assay. The HTRF epitope assay measures the binding of biotinylated human α-synuclein to an antibody or antigen-binding fragment thereof. Human, cynomolgus monkey, and rat α-synuclein can be titrated into the assay, and the selectivity of the antibody or antigen-binding fragment thereof can be assessed by measuring the degree of inhibition of binding of biotinylated human α-synuclein to the antibody or antigen-binding fragment thereof. The IC is determined, for example, by fitting the data curve to a four-parameter logistic equation using PRISM 6 software (Graphpad). 50 The species cross-reactivity of an antibody or antigen-binding fragment thereof can be confirmed using a direct binding HTRF assay format. Using the HTRF assay, the antibody or antigen-binding fragment thereof is titrated into the assay to compete for binding of human, cynomolgus monkey, or rat α-synuclein to the antibody or antigen-binding fragment thereof.

[0325] Aslo0452 ngl-3 and aslo0543 antibodies bind to human α-synuclein with high affinity. WO 2017 / 207739 has reported that aslo0452 ngl-3 and aslo0543 antibodies bind to human α-synuclein with K values ​​of 106 pM (95% CI 10 to 292 pM) and 113 pM (95% CI 5 to 333 pM), respectively. D Binds to α-synuclein as measured using an Octet assay. Thus, the present invention can use anti-α-synuclein antibodies or antigen-binding fragments thereof that bind to human α-synuclein with high affinity. The anti-α-synuclein antibodies or antigen-binding fragments thereof can bind to α-synuclein with a K of less than 500 picomolar (pM), less than 400 pM, less than 300 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, or 106 pM or less. D Binding to α-synuclein, as measured, for example, using Octet analysis. The affinity of the antibody or its antigen binding fragment can be assessed using an avi-tag α-synuclein-Flag-His tag molecule. The antibody or antigen binding fragment can be premixed with different concentrations of ligand until equilibrium is reached. The amount of free antibody or its antigen binding fragment can then be measured using Octet by capturing the free antibody or its antigen binding fragment using biotinylated α-synuclein fixed to a streptavidin-coated sensor. The amount of free antibody or its antigen binding fragment detected at each α-synuclein concentration can then be plotted relative to the concentration of the ligand, and the equilibrium dissociation constant (K) can be calculated using software. D ). The anti-α-synuclein antibody or antigen-binding fragment thereof can be expressed at a K of less than 500 picomolar (pM), less than 400 pM, less than 300 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, or 106 pM or less. D Binds to monomeric α-synuclein as measured, for example, using an Octet assay. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric α-synuclein with a K of less than 500 pM. D Binds to monomeric α-synuclein as measured, for example, using an Octet assay. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric α-synuclein with a K of less than 500 picomolar (pM), less than 400 pM, less than 300 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, or 10<6 > pM or less. D Binds to monomeric human α-synuclein as measured, for example, using Octet analysis. Particularly preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric human α-synuclein with a K of less than 500 pM. DBinding to monomeric human α-synuclein as measured, for example, using Octet analysis.

[0326] WO 2017 / 207739 has reported that aslo0452 ngl-3 and aslo0543 antibodies have K values ​​of 74 pM (95% CI 15 to 177 pM) and 108 pM (95% CI 34 to 223 pM), respectively. D Binding to α-synuclein as measured using the KinExA assay. WO2017 / 207739 reported that the aslo0452 ngl-3 Fab fragment binds to α-synuclein with a K of 174 pM (95% CI 15 to 177 pM). D Binding to α-synuclein as measured using a KinExA assay. Thus, the anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can have a K of less than 400 picomolar (pM), less than 300 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, less than 100 pM, less than 80 pM, or 74 pM or less. D Binding to α-synuclein, as measured, for example, using the KinExA assay. The solution phase affinity (K) of antibodies or antigen-binding fragments thereof can be assessed using a KinExA instrument (Sapidyne Instruments) using monomeric human biotinylated α-synuclein. D ). The antibody or antigen-binding fragment thereof can be premixed with different concentrations of each ligand until equilibrium is reached. The amount of free antibody can then be measured using KinExA by capturing the free antibody or antigen-binding fragment thereof using an α-synuclein-coated plate, washing away unbound material, and detecting bound antibody using a fluorescently labeled species-specific antibody. The amount of free antibody detected at each α-synuclein concentration can then be plotted against the concentration of the ligand, and the equilibrium dissociation constant (K) can be calculated using KinExA software. D The anti-α-synuclein antibody or antigen-binding fragment thereof can be expressed at a K of less than 400 picomolar (pM), less than 300 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, less than 100 pM, less than 80 pM, or 74 pM or less. D Binds to monomeric α-synuclein as measured, for example, using a KinExA assay. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric α-synuclein with a K of less than 400 pM. DBinding to monomeric α-synuclein as measured, for example, using a KinExA assay. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric α-synuclein at a K of less than 400 picomolar (pM), less than 300 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 120 pM, less than 110 pM, less than 100 pM, less than 80 pM, or 74 pM or less. D Binds to monomeric human α-synuclein as measured, for example, using a KinExA assay. Particularly preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric human α-synuclein with a K of less than 400 pM. D Binding to monomeric human α-synuclein as measured, for example, using the KinExA assay.

[0327] Aslo0452 ngl-3 and aslo0543 antibodies bind to natural endogenous human α-synuclein. As used herein, "natural endogenous α-synuclein" means the α-synuclein present in the tissue of the subject to be treated. Therefore, the present invention can use anti-α-synuclein antibodies or antigen-binding fragments thereof that are bound to natural endogenous human α-synuclein. The combination of antibody or its antigen-binding fragment with natural endogenous human α-synuclein can be determined using any suitable means, for example, using flow cytometry of α-synuclein positive and negative cell lines. For example, α-synuclein positive cells (for example, SHSY5Y neuroblastoma cells) and α-synuclein negative cells (for example, BT-20 breast cancer cells) can be fixed in formaldehyde and permeabilized, and then incubated with anti-α-synuclein antibodies or its antigen-binding fragment, positive control or isotype control antibodies. Subsequently, the bound antibody can be detected by incubation with an appropriate second antibody. The cells can then be analyzed using a FACS machine (e.g., a FACS Canto II instrument (Becton Dickinson, Franklin Lakes, NJ)) and appropriate software (e.g., FlowJo software (Tree Star, Ashland, OR) for data analysis. Binding to native endogenous human α-synuclein can be confirmed by a shift in the fluorescence signal in the presence of the antibody or its antigen-binding fragment in α-synuclein-positive cells compared to an isotype control and / or a second antibody alone, and a lack of a shift in the fluorescence signal in the presence of the antibody or its antigen-binding fragment in α-synuclein-negative cells compared to an isotype control and / or a second antibody alone. Preferably, the anti-α-synuclein antibody or its antigen-binding fragment binds to native endogenous human α-synuclein in vivo. The ability of the anti-α-synuclein antibody or its antigen-binding fragment to bind to native endogenous human α-synuclein in vivo can be determined by administering the antibody or its antigen-binding fragment to a subject and then determining binding of the antibody or its antigen-binding fragment to native endogenous α-synuclein in a sample obtained from the subject. The anti-α-synuclein antibody or its antigen-binding fragment can bind to native endogenous human α-synuclein in the brain and / or CSF of the subject to be treated.

[0328] Aslo0452 ngl-3 and aslo0543 antibodies bind to aggregates of human α-synuclein. Therefore, the anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can bind to aggregates of human α-synuclein, such as oligomers and / or multimers and / or aggregates of human α-synuclein. The anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can bind to aggregates of human α-synuclein, such as Lewy bodies or Lewy body-like aggregates. Antibody capture assay determines the binding of anti-α-synuclein antibodies or their antigen-binding fragments to human α-synuclein aggregates. For example, a mouse IgG1 form of an antibody or its antigen-binding fragment can be fixed to the wells of a 96-well microtiter plate (Nunc). After blocking, aggregated or monomeric human α-synuclein can be incubated in the wells. After washing, the captured human α-synuclein can be detected by adding the human IgG1TM form of the same antibody or its antigen-binding fragment, followed by adding an anti-human IgG-Europium conjugate (PerkinElmer) or an anti-human IgG-HRP conjugate. After incubation and washing, a detection substrate (e.g., TMB or DELFIA enhancer solution) can be added. The plate can then be read on a microtiter plate reader. This assay cannot detect monomeric α-synuclein.

[0329] Aslo0452 ngl-3 and Aslo0543 antibodies bind to monomeric human α-synuclein. The anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can bind to monomeric human α-synuclein. The anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can bind to monomeric and / or oligomeric human α-synuclein. Binding of the anti-α-synuclein antibodies or antigen-binding fragments thereof to monomeric forms of human α-synuclein can be determined using any suitable method, for example, a KinExA assay. The anti-α-synuclein antibodies or antigen-binding fragments thereof can bind to monomeric and / or oligomeric α-synuclein in the CSF of a subject. The anti-α-synuclein antibodies or antigen-binding fragments thereof can bind to monomeric α-synuclein in the CSF of a subject. The anti-α-synuclein antibodies or antigen-binding fragments thereof can bind to oligomeric α-synuclein in the CSF of a subject. The binding of an antibody or antigen-binding fragment thereof to monomeric α-synuclein in a subject's CSF can be determined using any suitable means, for example, using an ELISA that specifically detects the presence of monomeric α-synuclein to measure the binding of the antibody or antigen-binding fragment thereof to monomeric α-synuclein from a subject's CSF. The binding of an antibody or antigen-binding fragment thereof to oligomeric α-synuclein in a subject's CSF can be determined using any suitable means, for example, using an ELISA that specifically detects the presence of oligomeric α-synuclein to measure the binding of the antibody or antigen-binding fragment thereof to oligomeric α-synuclein from a subject's CSF. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric and / or oligomeric α-synuclein in a subject's CSF or ISF in vivo. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to monomeric α-synuclein in a subject's CSF in vivo.

[0330] Aslo0452 ngl-3 and aslo0543 antibodies can bind to and chelate both monomeric and aggregated forms of alpha-synuclein. The anti-alpha-synuclein antibody or its antigen-binding fragment used in the present invention can chelate alpha-synuclein. The term "chelate" means binding a molecule in a certain way so as to prevent or inhibit the molecule from forming aggregates, or to prevent or inhibit the molecule from forming further aggregates. The antibody or its binding fragment that chelates alpha-synuclein can be bound to alpha-synuclein outside the cell (such as in CSF or ISF) and prevent it from entering the cell. The antibody or its binding fragment that chelates alpha-synuclein can be bound to monomeric alpha-synuclein and prevent it from forming aggregates. The antibody or its binding fragment that chelates alpha-synuclein can be bound to alpha-synuclein and prevent it from forming aggregates or prevent it from forming Lewy bodies.

[0331] The anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention can bind to both monomeric and aggregated forms of α-synuclein. The anti-α-synuclein antibodies or antigen-binding fragments thereof can chelate both monomeric and aggregated forms of α-synuclein. The anti-α-synuclein antibodies or antigen-binding fragments thereof can bind to both monomeric and aggregated forms of α-synuclein in vivo. The anti-α-synuclein antibodies or antigen-binding fragments thereof can chelate both monomeric and aggregated forms of α-synuclein in vivo. The ability of an antibody or antigen-binding fragment thereof to bind to monomeric and aggregated forms of α-synuclein can be determined using any suitable means, for example, using any of the methods described herein, such as using the KinExA assay and Antibody capture assay.

[0332] Aslo0452 ngl-3 and aslo0543 antibodies bind to the disease-associated pathological forms of the alpha-synuclein in Parkinson's disease brain tissue, for example, Lewy bodies, Lewy neurites, and Lewy dots. Therefore, preferably, anti-alpha-synuclein antibodies or their antigen-binding fragments bind to the disease-associated pathological forms of alpha-synuclein. This can be assessed by immunohistochemical staining using antibodies or their antigen-binding fragments in diseased tissue. For example, immunohistochemical staining of Parkinson's disease brain tissue can show the presence of Lewy bodies, Lewy neurites, neuronal aggregates, Lewy dots, and background brain tissue. Minimal staining is observed in normal (non-diseased) brain or in tissues that do not contain alpha-synuclein. The combination with brain tissue can be assessed in vitro or in vivo. Preferably, anti-alpha-synuclein antibodies or their antigen-binding fragments bind to the disease-associated pathological forms of alpha-synuclein in vivo (for example, in a suitable animal model of alpha-synuclein disease).

[0333] The aslo0452 ngl-3 antibody reduces the level of alpha-synuclein in the interstitial fluid (ISF) of the brain. Specifically, the aslo0452 ngl-3 antibody reduces the level of free, unbound alpha-synuclein in the interstitial fluid of the brain (e.g., in an animal model of alpha-synuclein disease). Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of free, unbound alpha-synuclein in the interstitial fluid of the brain. The reduction in the level of free, unbound alpha-synuclein in the interstitial fluid of the brain can be assessed using any suitable means, for example, as described above. Therefore, anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of alpha-synuclein in the interstitial fluid of the brain. Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of free, unbound alpha-synuclein in the interstitial fluid of the brain of an animal (e.g., a rat). The anti-α-synuclein antibody or antigen-binding fragment thereof can reduce the level of free, unbound α-synuclein in the brain interstitial fluid (e.g., rat brain interstitial fluid) 1 hour after administration (e.g., intravenous administration). The level of free, unbound α-synuclein can be reduced for at least 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, or preferably 10 hours after administration.

[0334] The aslo0452 ngl-3 antibody reduces the level of alpha-synuclein in cerebrospinal fluid (CSF). Specifically, the aslo0452 ngl-3 antibody reduces the level of free, unbound alpha-synuclein in the cerebrospinal fluid. Therefore, anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of alpha-synuclein in the cerebrospinal fluid. Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of free, unbound alpha-synuclein in the cerebrospinal fluid. The reduction in the level of free, unbound alpha-synuclein in the cerebrospinal fluid can be assessed using any suitable means, for example, as described above. Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of free, unbound alpha-synuclein in the CSF of animals (e.g., rats). Anti-alpha-synuclein antibodies or antigen-binding fragments thereof can reduce the level of free, unbound alpha-synuclein in the CSF (e.g., rats) 6 hours after administration (e.g., intravenous administration). The level of free unbound alpha-synuclein may be reduced over at least 12 hours, 18 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 ​​hours, 54 hours, 60 hours, 66 hours, or preferably 72 hours.

[0335] Aslo0452 ngl-3 antibody reduces alpha-synapse nucleoprotein diffusion in vivo. Compared with the antibody that does not inhibit diffusion, this effect of inhibiting alpha-synapse nucleoprotein diffusion indicates that it is bound to different epitopes on human alpha-synapse nucleoprotein. Therefore, preferably, anti-alpha-synapse nucleoprotein antibody or its antigen-binding fragment reduces alpha-synapse nucleoprotein diffusion in vivo. The reduction of alpha-synapse nucleoprotein diffusion in vivo can be determined using any suitable means, for example, as described above. For example, a lentiviral vector (LV-α-syn) expressing alpha-synapse nucleoprotein is injected into the right hippocampus of the transgenic mice (α-syn tg) overexpressing alpha-synapse nucleoprotein, and then the transgenic mice are immunized with anti-alpha-synapse nucleoprotein antibody or its antigen-binding fragment or control. At the end of the immune period, mice are euthanized and their brains are fixed. The fixed brain is sliced, and the alpha-synapse nucleoprotein immunoreactivity level of the ipsilateral and contralateral LV-α-syn injection site is analyzed by immunocytochemistry. A reduction in α-synuclein diffusion was observed by comparing the level of α-synuclein diffusion from the ipsilateral to the contralateral side of the hippocampus in mice treated with anti-α-synuclein antibodies or antigen-binding fragments thereof with that in control mice.

[0336] 4.2.4 Additional anti-α-synuclein antibodies or antigen-binding fragments thereof

[0337] The anti-α-synuclein antibody or antigen-binding fragment thereof can compete with the antibody aslo0452 ngl-3 and / or the antibody aslo0543 for binding to human α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can bind to the same epitope on human α-synuclein as the antibody aslo0452 ngl-3 and / or the antibody aslo0543, or to an overlapping epitope on human α-synuclein. The anti-α-synuclein antibody or antigen-binding fragment thereof can compete with the antibody aslo0452 ngl-3 for binding to human α-synuclein. Preferably, the anti-α-synuclein antibody or antigen-binding fragment thereof binds to the same epitope on human α-synuclein as the antibody aslo0452 ngl-3 or to an overlapping epitope on human α-synuclein.

[0338] Can easily determine whether antibody or its Fab is bound to identical or overlapping epi-position with reference antibody or Fab.Such method is the routine procedure in this area.For example, the comparison of antibody and another antibody can be carried out by biochemical competition assay, wherein two antibodies (a kind of is labeled for detection purpose, a kind of is not labeled) are incubated with given antigen simultaneously.If the binding signal of labeled antibody is obtained, then it is believed that these two antibodies recognize different, non-overlapping epi-positions on the protein of interest.If binding signal is not obtained, then on the contrary, they will be characterized as having overlapping epi-positions on protein sequence, because the combination of a kind of antibody hinders the combination of the second antibody spatially.In addition, also can use modified albumen (such as truncation), derive from the linear peptide sequence of the primary amino acid sequence of antigen, species ortholog and identify the amino acid position of given epi-position by proteolytic digestion and mass spectrometry analysis of the antibody combined with given protein.These methods are used to generate the interaction zone between antibody and antigen.

[0339] Additional routine experiments (such as peptide mutations and binding analysis) can be performed to confirm whether any observed lack of binding is actually due to binding to an epitope of the invention or is due to some other phenomenon (such as steric hindrance). Such experiments can be performed using ELISA, RIA, Biacore, flow cytometry or other known antibody binding assays.

[0340] For example, for the fine mapping of a specific epitope, a mathematical model of the epitope:paratope interface can be derived from data generated by analyzing the structure of the antigen:antibody complex using high-resolution imaging methods (such as co-crystallization and X-ray diffraction). In order to confirm the relevance of the derived mathematical model in identifying the key contact residues that define the epitope, it is necessary to subsequently perform point mutagenesis of the antigen and analyze the effect of such mutations on the binding strength between the antigen and the antibody. Using this combination of methods, a precise map of the key contact residues that make up the epitope can be established.

[0341] An antibody or antigen-binding fragment thereof of interest, such as an antibody or antigen-binding fragment thereof that binds to the same epitope as a reference antibody or antigen-binding fragment thereof, can be generated by generating variants of the reference antibody or antigen-binding fragment thereof. Thus, the antibody or antigen-binding fragment thereof used in the present invention can be a variant of a known anti-α-synuclein antibody or antigen-binding fragment thereof, such as a variant of aslo0452ngl-3 or an antigen-binding fragment thereof, a variant of aslo0543 or an antigen-binding fragment thereof, or a variant of asyn0087 or an antigen-binding fragment thereof. Thus, with respect to the variant antibodies or antigen-binding fragments thereof disclosed herein, the reference antibody can be aslo0452ngl-3 or an antigen-binding fragment thereof, aslo0543 or an antigen-binding fragment thereof, or asyn0087 or an antigen-binding fragment thereof.

[0342] Such variant antibodies or antigen-binding fragments thereof may retain one or more CDRs of the reference antibody or antigen-binding fragment thereof, or may have CDRs that share a high level of identity with the CDRs of the reference antibody or antigen-binding fragment thereof. For example, an antibody or antigen-binding fragment for use in the present invention may have one or more CDRs that differ from any one or more of the specific CDR sequences mentioned herein (e.g., the CDRs of aslo0452 ng1-3, aslo0543, or asyn0087, e.g., any one or more CDRs of the CDRs of SEQ ID NOs: 5, 15, 16, 20, 10, and 21, any one or more CDRs of the CDRs of SEQ ID NOs: 25, 26, 27, 31, 32, and 33) by 1 or 2 amino acid residues (e.g., 1 or 2 conservative amino acid substitutions). The antibodies or antigen-binding fragments used in the present invention may have a CDR set that differs from any of the specific CDR sequence sets mentioned herein (e.g., the CDR set of aslo0452 ng1-3, aslo0543, or asyn0087, e.g., a CDR set having SEQ ID NOs: 5, 15, 16, 20, 10, and 21, or a CDR set having SEQ ID NOs: 25, 26, 27, 31, 32, and 33) by 1 or 2 amino acid residues (e.g., 1 or 2 conservative amino acid substitutions). The antibodies or antigen-binding fragments used in the present invention may have a CDR set that includes one or more of the CDRs listed in SEQ ID NOs: 5, 6, 7, 9, 10, and 11. The antibodies or antigen-binding fragments used in the present invention may have a CDR set that includes one or more of the CDRs listed in SEQ ID NOs: 5, 6, 7, 9, 10, and 11.

[0343] The anti-α-synuclein antibody or antigen-binding fragment thereof can have one or more variations (e.g., one or more conservative amino acid substitutions) in the CDR amino acid sequence such that at least 80%, at least 85%, at least 90%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, and up to 99% sequence identity is maintained with any one or more of the CDRs of antibody aslo0452ngl-3 (e.g., any one or more of the CDRs having SEQ ID NOs: 5, 15, 16, 20, 10, and 21). The anti-α-synuclein antibody or antigen-binding fragment thereof can have one or more variations in the CDR amino acid sequence such that at least 80%, at least 85%, at least 90%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, and up to 99% sequence identity is maintained with any one or more of the CDRs of antibody aslo0452 ngl-3 (e.g., any one or more of the CDRs having SEQ ID NOs: 5, 15, 16, 20, 10, and 21) and binding to α-synuclein is maintained.

[0344] The anti-α-synuclein antibody or antigen-binding fragment thereof may have at least one CDR of a reference antibody. The anti-α-synuclein antibody or antigen-binding fragment thereof may have at least one CDR selected from the CDRs of antibody aslo0452 ngl-3, i.e., at least one CDR selected from any one of SEQ ID NO: 5, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 10, and SEQ ID NO: 21.

[0345] The anti-α-synuclein antibody or antigen-binding fragment thereof can have at least one, at least two, at least three, at least four, at least five, or all six of the CDRs of the reference antibody. The anti-α-synuclein antibody or antigen-binding fragment thereof can have at least one, at least two, at least three, at least four, at least five, or all six of the CDRs selected from the CDRs of antibody aslo0452 ngl-3 (i.e., SEQ ID NO: 5, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 10, and SEQ ID NO: 21).

[0346] The anti-α-synuclein antibody or antigen-binding fragment thereof may have all six CDRs of the reference antibody. The anti-α-synuclein antibody or antigen-binding fragment thereof may have all six CDRs of the antibody aslo0452 ng1-3. The anti-α-synuclein antibody or antigen-binding fragment thereof may have a CDR set in which CDRH1 has the amino acid sequence of SEQ ID NO: 5, CDRH2 has the amino acid sequence of SEQ ID NO: 15, CDRH3 has the amino acid sequence of SEQ ID NO: 16, CDRL1 has the amino acid sequence of SEQ ID NO: 20, CDRL2 has the amino acid sequence of SEQ ID NO: 10, and CDRL3 has the amino acid sequence of SEQ ID NO: 21.

[0347] The CDR3 of the heavy chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the heavy chain of a reference antibody, and / or the CDR3 of the light chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the light chain of a reference antibody. The CDR3 of the heavy chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the heavy chain of the antibody aslo0452 ngl-3; and / or the CDR3 of the light chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the light chain of the antibody aslo0452 ngl-3. Thus, the CDR3 of the heavy chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the heavy chain of the antibody aslo0452 ngl-3 according to SEQ ID NO: 16; and / or the CDR3 of the light chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the light chain of the antibody aslo0452 ngl-3 according to SEQ ID NO: 21. The CDR3 of the heavy chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the heavy chain of the antibody aslo0452 ngl-3. The CDR3 of the light chain of the anti-α-synuclein antibody or antigen-binding fragment thereof can be the CDR3 of the light chain of the antibody aslo0452 ngl-3.

[0348] A variant antibody or antigen-binding fragment thereof may have one or more variations (eg, conservative amino acid substitutions) outside of the CDRs, such as in one or more of the framework regions.

[0349] Variant antibodies or antigen-binding fragments thereof for use in the present invention may have a VH region and / or VL region that shares a high level of identity with the variable heavy chain region (VH) and variable light chain region (VL) of a reference antibody or antigen-binding fragment thereof. For example, an antibody or antigen-binding fragment for use in the present invention may have a VH region and / or VL region that differs from the VH and / or VL of a reference antibody or antigen-binding fragment thereof (e.g., the VH and / or VL of aslo0452 ngl-3, aslo0543, or asyn0087, e.g., the VH and / or VL of SEQ ID NOs 14 and 19, respectively, or the VH and / or VL of SEQ ID NOs 24 and 30, respectively, or the VH and / or VL of SEQ ID NOs 2 and 3, respectively, by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acid residues (e.g., conservative amino acid substitutions) compared to the VH and / or VL of a reference antibody or antigen-binding fragment thereof. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a VH and a VL, wherein one or both of the VH and the VL have at least 80%, at least 85%, at least 90%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, and up to 99% sequence identity with the corresponding VH and / or VL of a reference antibody (e.g., the VH and / or VL of aslo0452 ngl-3, aslo0543, or asyn0087, e.g., the VH and / or VL of SEQ ID NOs 14 and 19, respectively, or the VH and / or VL of SEQ ID NOs 24 and 30, respectively, or the VH and / or VL of SEQ ID NOs 2 and 3, respectively).

[0350] The differences from the reference VH and / or VL sequences may be located outside of the CDRs, i.e., the variant may comprise the CDRs of the VH and / or VL from a reference antibody, and the differences from the reference VH and / or VL sequences may be located in the framework sequences of the variable domains. The differences from the relevant reference sequences may be located anywhere in the VH and / or VL subsequences, e.g., the variant antibody or antigen-binding fragment thereof may comprise variations in the CDR sequences, as discussed above.

[0351] Variant antibodies or antigen-binding fragments thereof can be used in the present invention. Such variant antibodies or antigen-binding fragments may retain one or more functional properties of the reference antibody from which they are derived, such as any one or more functional properties or characteristics of the antibody aslo0542ngl-3 described herein. The variant antibody or antigen-binding fragment may specifically bind to α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may specifically bind to human α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may bind to monomeric α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may bind to human monomeric α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may bind to aggregated α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may bind to aggregated human α-synuclein, for example, as defined herein. The variant antibody or antigen-binding fragment may bind to monomeric α-synuclein and may bind to aggregated α-synuclein, for example, as defined herein. Variant antibodies or antigen-binding fragments can bind to monomeric human α-synuclein and can bind to aggregated human α-synuclein, for example, as defined herein. Variant antibodies or antigen-binding fragments used in the present invention can bind to human α-synuclein, rat α-synuclein and cynomolgus monkey α-synuclein, for example, as defined herein. Variant antibodies or antigen-binding fragments can bind to the same epitope as a reference antibody or its antigen-binding fragment, for example, as described herein. Variant antibodies or antigen-binding fragments can compete with reference antibodies (e.g., aslo0542ngl-3) for binding to human α-synuclein, for example, as described herein. Variant antibodies or antigen-binding fragments can bind to the C-terminal region (residues 96 to 140) of human α-synuclein, for example, as defined herein. Variant antibodies or antigen-binding fragments can bind to the region between about amino acid 102 and about amino acid 130 comprising human α-synuclein, for example, as defined herein.

[0352] Specifically, conservative amino acid substitutions are contemplated. Conservative substitutions are substitutions that occur within a family of amino acids with related side chains. Genetically encoded amino acids are generally divided into the following families: (1) acidic: aspartic acid, glutamic acid; (2) basic: lysine, arginine, histidine; (3) nonpolar: alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar: glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. These families can be further classified as follows: serine and threonine are the aliphatic-hydroxyl family; asparagine and glutamine are the amide-containing family; alanine, valine, leucine, and isoleucine are the aliphatic family; and phenylalanine, tryptophan, and tyrosine are the aromatic family. Thus, in general, it is expected that replacing a leucine with isoleucine or valine alone, replacing an aspartic acid with glutamic acid alone, replacing a threonine with serine alone, or similarly replacing an amino acid with a structurally related amino acid will not significantly affect the binding function or properties of the resulting antibody, particularly when the replacement does not involve an amino acid within a CDR position.

[0353] The anti-α-synuclein antibody or antigen-binding fragment thereof can be a variant antibody or antigen-binding fragment thereof, wherein the reference antibody is asyn0087 or an antigen-binding fragment thereof comprising the VH of SEQ ID NO: 2 and the VL of SEQ ID NO: 3. Specifically, the anti-α-synuclein antibody or antigen-binding fragment thereof can be a variant antibody or antigen-binding fragment thereof, wherein the reference antibody is asyn0087 or an antigen-binding fragment thereof comprising the VH of SEQ ID NO: 2 and the VL of SEQ ID NO: 3, wherein the variant antibody or antigen-binding fragment has a K of less than 500 nM. D , and binds to the same epitope as any of the antibodies asyn0087, aslo0452ngl-3, and aslo0543 described herein.

[0354] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain region (VH) having an amino acid sequence of SEQ ID NO: 2 and a variable light chain region (VL) having an amino acid sequence of SEQ ID NO: 3.

[0355] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain having a sequence defined by SEQ ID NO: 4 and a variable light chain having a sequence defined by SEQ ID NO: 8. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a variable heavy chain having a sequence defined by SEQ ID NO: 4 and a variable light chain having a sequence defined by SEQ ID NO: 8, and has a K of less than 500 pM. DBinds to human α-synuclein and binds to the same epitope as asyn0087, aslo0452ngl-3, or aslo0543.

[0356] The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise at least one CDR selected from:

[0357] (i) H-CDR1 of SEQ ID NO: 5,

[0358] (ii) H-CDR2 of SEQ ID NO: 6,

[0359] (iii) H-CDR3 of SEQ ID NO: 7,

[0360] (iv) L-CDR1 of SEQ ID NO: 9,

[0361] (v) L-CDR2 of SEQ ID NO: 10,

[0362] (vi) L-CDR3 of SEQ ID NO:11.

[0363] For example, the anti-α-synuclein antibody or antigen-binding fragment thereof may comprise 1, 2, 3, 4, 5, or all 6 of the CDR sequences of SEQ ID NOs 5, 6, 7, 9, 10, and 11. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise H-CDR1 of SEQ ID NO: 5, H-CDR2 of SEQ ID NO: 6, H-CDR3 of SEQ ID NO: 7, L-CDR1 of SEQ ID NO: 9, L-CDR2 of SEQ ID NO: 10, and L-CDR3 of SEQ ID NO: 11. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise H-CDR3 of SEQ ID NO: 7.

[0364] The framework regions and CDRs of an antibody can be precisely defined (see Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., US Department of Health and Human Services, pp. 91-3242; and Chothia et al. (1987) J. MoI. Biol. 196:901-917, both of which are incorporated herein by reference).

[0365] Minor variations in the amino acid sequence of an anti-α-synuclein antibody or antigen-binding fragment thereof are considered to be encompassed by the present invention, provided that the variation in the amino acid sequence retains at least 75%, more preferably at least 80%, at least 90%, at least 95%, and most preferably at least 99% sequence identity with the equivalent sequence of a reference anti-α-synuclein antibody or antigen-binding fragment thereof as defined anywhere herein. In particular, conservative amino acid substitutions are contemplated.

[0366] The present invention also relates to the use of a single-chain amino acid sequence comprising a light chain of an anti-α-synuclein antibody or antigen-binding fragment thereof as defined anywhere herein. The present invention also relates to the use of a single-chain amino acid sequence comprising a heavy chain of an anti-α-synuclein antibody or antigen-binding fragment thereof as defined anywhere herein.

[0367] Optimal alignment of sequences for comparison can be achieved, for example, by the local homology alignment algorithm of Smith and Waterman (Smith and Waterman (1981) Adv. Appl. Math. 2:484; incorporated herein by reference), the algorithm of Needleman and Wunsch (Needleman and Wunsch (1970) J. Mol. Biol. 48:443; incorporated herein by reference), the search for similarity method of Pearson and Lipman (1988; Proc. Natl. Acad. Sci. USA 85:2444; incorporated herein by reference), by computer implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA - sequence analysis software packages of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705), or by visual inspection (see Current Protocols in Molecular Biology, 1996). Biology, FM Aubel et al., eds., Current Protocols, a joint venture of Greene Publishing Associates, Inc. and John Wiley & Sons, Inc. (1995 supplement); incorporated herein by reference).

[0368] Examples of suitable algorithms for determining percent sequence similarity or identity are BLAST and BLAST 2.0 algorithms (see Altschul et al. (1990) J. Mol. Biol. 215(3):403-410; and the National Center for Biotechnology Information at "http: / / www.ncbi.nlm.nih.gov / "; both of which are incorporated herein by reference).

[0369] The anti-α-synuclein antibody or antigen-binding fragment thereof as defined anywhere above may be an IgA, IgD, IgE, IgM, IgG, such as an IgG1, IgG2, IgG3 or IgG4 antibody or antigen-binding fragment thereof.

[0370] The anti-α-synuclein antibody or its antigen-binding fragment may not have an Fc region. Preferably, the anti-α-synuclein antibody or its antigen-binding fragment has an Fc region. The anti-α-synuclein antibody or its antigen-binding fragment may have a modified Fc region. Suitable modifications are well known to those skilled in the art and may include, among others, modifications that increase or decrease half-life, ablate, reduce or enhance effector function, and provide substituted cysteine ​​with free thiols for conjugation. Examples of such modifications are YTE to increase half-life and / or TM to reduce effector function. The anti-α-synuclein antibody or its antigen-binding fragment may have reduced binding affinity for IgG Fc receptors. Thus, the anti-α-synuclein antibody or its antigen-binding fragment may have a low immunogenic effect. Any of the antibodies or antigen-binding fragments disclosed herein may comprise the mutations M252Y / S254T / T256E (YTE) in the Fc region of the antibody (Dall'Acqua et al. (2006) J. Biol. Chem. 281: 23514-23524). Any of the antibodies or antigen-binding fragments disclosed herein may comprise a triple mutation (abbreviated herein as "TM") in the Fc region corresponding to the L234F / L235E / P331S mutation disclosed in Oganesyan et al. (2008) Acta Crystallogr. D. Biol. Crystallogr. 64: 700–704. An anti-α-synuclein antibody or antigen-binding fragment thereof may comprise a triple mutation in the Fc region corresponding to L234F / L235E / P331S numbered according to Kabat numbering. The anti-α-synuclein antibody or antigen-binding fragment thereof may be an IgG1 TM antibody or antigen-binding fragment thereof. IgG1 TM is an IgG1 triple mutant containing three point mutations (L234F / L235E / P331S) in the Fc domain that reduce the binding affinity of the antibody or antigen-binding fragment thereof for Fc-γ receptors (FcγRs) (Oganesyan et al. (2008) Acta Crystallogr. D. Biol. Crystallogr. 64:700–704; incorporated herein by reference). Antibody-mediated prevention of α-synuclein spread may not require Fc-associated effector functions as a key mechanism of action. The anti-α-synuclein antibody or antigen-binding fragment thereof may comprise an Fc region with a YTE mutation. The Fc region may be mutated or exchanged for alternative protein sequences, or the anti-α-synuclein antibody or antigen-binding fragment thereof may be chemically modified to increase its blood-brain barrier penetration.

[0371] The anti-α-synuclein antibody or antigen-binding fragment thereof can be linked to a half-life extender. As used herein, the term "half-life extender" means any molecule that increases the half-life of an anti-α-synuclein antibody or antigen-binding fragment thereof when linked to an anti-α-synuclein antibody or antigen-binding fragment thereof disclosed herein. Any half-life extender can be covalently or non-covalently linked to an anti-α-synuclein antibody or antigen-binding fragment thereof. The half-life extender can be polyethylene glycol, a peptide, or human serum albumin.

[0372] Anti-alpha-synuclein antibodies or their antigen-binding fragments can be associated with molecules that can penetrate the blood-brain barrier (BBB). The term "molecule that can penetrate the BBB" means any molecule that increases the brain penetration ability of an anti-alpha-synuclein antibody or its antigen-binding fragment when connected to an anti-alpha-synuclein antibody disclosed herein or its antigen-binding fragment. For example, a molecule that can penetrate the BBB can be a molecule that can specifically bind to brain microvascular endothelial cells (BMVEC) of one or more species and pass through BMVEC (for example, from a peripheral blood vessel into a CNS blood vessel) in vitro or in vivo. Whether a molecule is a molecule that can penetrate the BBB can be tested by a variety of in vitro or in vivo assays known to those of ordinary skill in the art. For example, a transporter molecule can be tested in an in vitro transcytosis assay, in an in vivo assay (such as a diuresis assay, as described in US 62 / 094,503). Other assays that can be used to measure the in vivo delivery of payloads across the BBB include, but are not limited to, chronic constrictive injury (CCI); retained nerve injury model (SNI) or spinal nerve ligation (SNL), all of which can be measured by paw flick or Hargreaves method (Hargreaves K et al., Pain; 1988; 32; 77-88). In some aspects, molecules that can penetrate the BBB as provided herein can be bound to BMVEC from one or more species, for example, humans, cynomolgus monkeys, mice, rats, or bovine BMVEC. Binding can be demonstrated in various ways known to those of ordinary skill in the art (for example, in FMAT assays as described in US 62 / 094,503). In some aspects, BMVEC is a brain capillary endothelial cell (BCEC). In some aspects, molecules that can penetrate the BBB as provided herein can pass through a monolayer of BCEC in an in vitro transcytosis assay. In some aspects, the activity of molecules that can penetrate the BBB can be demonstrated by visualization of molecules that can penetrate the BBB in the CNS. For example, tritium-labeled transporter molecules can be delivered to the subject, for example, and periphery (for example, intravenously) is delivered to mice, and then visualized in CNS by quantitative whole-body radiography.In some aspects, the BBB penetrable molecule is positioned in the specific area of ​​CNS, for example, cerebellar cortex, cerebral gray matter, spinal cord gray matter, pons or their combination.

[0373] Anti-α-synuclein antibodies or antigen-binding fragments thereof can be isolated. Anti-α-synuclein antibodies or antigen-binding fragments thereof can be purified.

[0374] The anti-α-synuclein antibody or antigen-binding fragment thereof may be a monoclonal antibody, a humanized antibody, or a human antibody.

[0375] Antigen-binding fragments for use in the present invention include Fab, Fv, scFv, dAb, Fd, Fab', F(ab')2, or isolated complementary determining regions (CDRs) with sufficient framework binding to α-synuclein. A Fab fragment can be a monovalent fragment consisting of VL, VH, CL, and CH1 domains. A F(ab')2 fragment can be a bivalent fragment comprising two Fab fragments connected by a disulfide bond at the hinge region. An Fc fragment can be composed of CH2 and CH3 domains. An Fv fragment can be composed of the VL and VH domains of a single antibody arm. A dAb fragment (Ward et al. (1989) Nature 341:544-546; incorporated herein by reference) can be composed of a VH domain. Isolated complementary determining regions (CDRs) with sufficient framework binding can be the antigen-binding portion of a variable region.

[0376] The antigen-binding portion of the light chain variable region and the antigen-binding portion of the heavy chain variable region (e.g., the two domains VL and VH of the Fv fragment) can be joined using recombinant methods through a synthetic linker that enables them to be made into a single protein chain in which the VL and VH regions pair to form a monovalent molecule (called single-chain Fv (scFv); see, e.g., Bird et al. (1988) Science 242(4877):423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; both of which are incorporated herein by reference). These antibody fragments are obtained using conventional techniques known to those skilled in the art and are screened for utility in the same manner as intact antibodies.

[0377] The anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention may have any or all of the advantageous properties as defined above, or a combination thereof. Specifically, the anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention may be selective for α-synuclein and be able to slow down or prevent the intercellular transmission and spread of α-synuclein in vivo.

[0378] The functionality of the anti-α-synuclein antibodies or antigen-binding fragments thereof used in the present invention, in particular (i) their ability to bind to an epitope of α-synuclein; and (ii) their ability to slow or prevent the intercellular transmission and spread of α-synuclein in vivo can be readily determined by measuring their specific activity using the techniques described in WO2017 / 207739, which is hereby incorporated by reference in its entirety.

[0379] 4.3 Pharmaceutical Compositions and Kits

[0380] The anti-α-synuclein antibody or antigen-binding fragment thereof can be provided in a pharmaceutical composition. According to the present invention, the anti-α-synuclein antibody or antigen-binding fragment thereof can be administered in the form of a pharmaceutical composition. Suitable pharmaceutical compositions can comprise an anti-α-synuclein antibody or antigen-binding fragment thereof as defined anywhere herein and a pharmaceutically acceptable excipient.

[0381] The phrase "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The composition may also contain other active compounds that provide supplemental, additional, or enhanced therapeutic function. The pharmaceutical composition may also be contained in a container, package, or dispenser together with instructions for administration.

[0382] The present invention also relates to a pharmaceutical composition comprising a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof (or any other fixed dose as described herein). The anti-α-synuclein antibody or antigen-binding fragment thereof can be any anti-α-synuclein antibody or antigen-binding fragment thereof as described herein. Therefore, the present invention provides a pharmaceutical composition comprising a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof (or any other fixed dose as described herein) and a pharmaceutically acceptable excipient. Suitable pharmaceutically acceptable excipients can facilitate processing of the active compound into a formulation suitable for pharmaceutical administration. The present invention provides a unit dosage form of a pharmaceutical composition comprising a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof (or any other fixed dose as described herein) and a pharmaceutically acceptable excipient. The present invention also provides a unit dosage form of an anti-α-synuclein antibody or antigen-binding fragment thereof of 50 mg to 5,000 mg (or any other fixed dose as described herein). As used herein, "unit dosage form" refers to the amount of a drug (e.g., an anti-α-synuclein antibody or antigen-binding fragment thereof) to be administered to a subject in a single dose.

[0383] The pharmaceutical compositions disclosed herein are formulated to be compatible with their intended route of administration. Administration can be, for example, intravenous, intraperitoneal, intramuscular, intracavitary, subcutaneous, intradermal or transdermal.

[0384] The present invention also relates to a kit comprising an anti-α-synuclein antibody or antigen-binding fragment thereof (which may be any anti-α-synuclein antibody or antigen-binding fragment thereof as described herein) and instructions for use thereof, optionally at a dose of 50 mg to 5,000 mg, to treat α-synuclein disease. Preferably, the instructions are instructions for use of the anti-α-synuclein antibody or antigen-binding fragment thereof at a fixed dose of 50 mg to 5,000 mg (or any other fixed dose as described herein) to treat α-synuclein disease.

[0385] The kit may comprise 50 mg to 5,000 mg (or any other fixed dose as described herein) of an anti-α-synuclein antibody or antigen-binding fragment thereof. The anti-α-synuclein antibody or antigen-binding fragment thereof may be aslo0452ngl-3.

[0386] The pharmaceutical compositions disclosed herein can be formulated for, but are not limited to, parenteral delivery, such as intramuscularly, subcutaneously, or intravenously. Compositions suitable for intramuscular, subcutaneous, or intravenous injection include sterile aqueous solutions.

[0387] The pharmaceutical composition may be in the form of an aqueous solution and may include a physiologically compatible buffer, such as Hank's solution, Ringer's solution, or physiologically buffered saline. The pharmaceutical composition may additionally or alternatively contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. The pharmaceutical composition may be prepared as an appropriate oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Optionally, the pharmaceutical composition may contain suitable stabilizers or agents that increase the solubility of the compound to allow the preparation of highly concentrated solutions.

[0388] 4.4 Informal Sequence Listing

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395] 5 Examples

[0396] 5.1 Example 1: Effects of different doses of aslo0452ngl-3 in humans

[0397] 5.1.1 Example 1.1: Effects of a Single Dose of aslo0452ngl-3 in Healthy Humans

[0398] aslo0452 ngl-3 was obtained using the methods described in WO2017 / 207739A1, which is incorporated herein by reference in its entirety.

[0399] A randomized, double-blind, placebo-controlled study of single ascending doses (SAD) of aslo0452 ngl-3, ranging from 70 mg to 4,500 mg, was designed for healthy human subjects aged 18 to 65. SAD Cohorts 1 to 6 each included 8 subjects, with six subjects in each cohort receiving aslo0452 ngl-3 and two subjects in each cohort receiving placebo. Figure 1 A simplified layout of the study design is provided in .

[0400] Subjects received a single intravenous (IV) infusion of aslo0452 ngl-3 (70, 210, 400, 1200, 2400, or 4500 mg) or placebo. C values ​​of aslo0452 ngl-3 were typically observed at the end of the 1-hour infusion. max- value; median t max It was approximately 1 hour after the start of the infusion at the 70, 210, and 2400 mg dose levels, 17 hours at the 400 mg dose level, 5 hours at the 4500 mg dose level, and 9 hours at the 1200 mg dose level. Following completion of the intravenous infusion, serum concentrations of aslo0452 ngl-3 declined in a generally multiphasic manner. Figure 2 As shown, aslo0452 ngl-3 exposure increased in a dose-dependent manner across the entire dose range. Increasing the aslo0452 ngl-3 dose from 1,200 mg to 2,400 mg and from 2,400 mg to 4,500 mg resulted in C max The increases in AUC∞ and AUC∞ were greater than dose-proportional (Table 1). Terminal elimination half-lives were comparable across single dose levels (16.6 to 24.3 days).

[0401] Table 1: Geometric Means (%CV) of Serum PK Parameters of aslo0452ngl-3 in a Phase 1 SAD Study of Healthy Subjects

[0402]

[0403]

[0404] %CV: Percent coefficient of variation; AUC∞ : area under the concentration-time curve from time 0 to infinity; C max : maximum observed concentration; PK: pharmacokinetics; t 1 / 2z : Terminal elimination phase half-life.

[0405] The pharmacodynamic (PD) effects of drugs were determined using free levels of α-synuclein (αSYN) in the cerebrospinal fluid (CSF). Following an immunoprecipitation procedure to remove αSYN bound to aslo0452 ngl-3, free αSYN protein in the CSF was quantified by Mesoscale Discovery enzyme-linked immunosorbent assay (ELISA). The percentage of CSF αSYN reduction at each dose is summarized in Table 2 below and is expressed as the mean change from the pre-dose baseline.

[0406] Table 2: Geometric reduction of CSF α-synuclein following aslo0452ngl-3 in a Phase I SAD study in healthy subjects Mean (%CV)

[0407]

[0408] %CV: percent coefficient of variation; CSF: cerebrospinal fluid; SAD: single ascending dose; values ​​in parentheses are standard deviations

[0409] 5.1.2 Example 1.2: Effects of Multiple Doses of aslo0452ngl-3 in Parkinson's Disease Patients

[0410] A multicenter, randomized, double-blind, placebo-controlled study of multiple ascending doses (MADs) of aslo0452 ngl-3 at 1,200 mg, 2,000 mg, or 4,200 mg was designed for subjects aged 40 to 85 years with Parkinson's disease (PD). MAD Cohorts 1 to 3 each included 12 subjects, with nine subjects in each cohort receiving aslo0452 ngl-3 and three subjects in each cohort receiving placebo. Figure 3 A simplified layout of the study design is provided in .

[0411] Subjects in MAD cohorts 1 and 2 received three intravenous infusions of aslo0452 ngl-3 or placebo, separated by 4 weeks (one infusion each on days 1, 29, and 57). Cohort 3 (4,200 mg) was not included. Following multiple intravenous infusions of 1200 mg aslo0452 ngl-3, C values ​​of aslo0452 ngl-3 were typically observed at the end of the 1-hour infusion. max values, and for the 1200 mg and 2000 mg multiple dose levels, the median t max The serum concentration of aslo0452 ngl-3 decreased in a generally heterogeneous manner after the end of the intravenous infusion. Figure 4As shown, aslo0452 ngl-3 exposure increased in a dose-dependent manner throughout the dose range investigated. Figure 5 As shown, all concentration time curves from healthy subjects in the above-mentioned SAD test are in the lower part of the observed distribution of concentration time curves from PD patients. The terminal elimination half-life is slightly shorter than the half-life in the SAD test (14.0 days). In the SAD and MAD tests, at all dose levels, the inter-subject variability as measured by the geometric mean percentage coefficient of variation (%CV) is low to moderate (11.3% to 29.8%) for AUC, and low to high (0.2% to 102.8%) for Cmax.

[0412] The free levels of αSYN in the cerebrospinal fluid (CSF) were used to determine the drug PD effect. After an immunoprecipitation procedure to remove αSYN bound to aslo0452ngl-3, free αSYN protein in the CSF was quantified by Mesoscale Discovery ELISA. Multiple intravenous doses of 1,200 mg or 2,000 mg of aslo0452 ngl-3 reduced CSF levels of αSYN. The mean changes in CSF free αSYN levels from baseline are shown in Table 3 below. In general, free α-synuclein in the CSF of the aslo0452ngl-3 treated groups was significantly reduced compared to placebo, and the percentage change in α-synuclein from baseline appeared to be dose-dependent.

[0413] Table 3: Reduction from Baseline in CSF Free α-Synuclein

[0414]

[0415] Aslo0452 ngl-3 appeared safe and well tolerated in both the SAD and MAD trials. There were no clinically significant abnormalities / changes in laboratory assessments, vital signs, and ECGs. Positive antidrug antibody (ADA) results were observed at each single dose level. None of the positive ADA results appeared to affect PK and PD parameters and were not expected to impact safety parameters.

[0416] 5.1.3 Example 1.3: Effects of Multiple Doses of aslo0452ngl-3 in Patients with Multiple System Atrophy

[0417] Given the surprisingly favorable PK profile, target engagement, and lack of safety concerns, a multicenter, randomized, double-blind, placebo-controlled, Phase 2 study was designed in subjects at least 40 years of age with possible or probable multiple system atrophy (MSA) to test the efficacy, safety, tolerability, PK, and PD of aslo0452 ngl-3 administered as multiple intravenous infusions every 4 weeks for 52 weeks.

[0418] The study includes a 42-day (6-week) screening period, a 52-week double-blind treatment period, and a follow-up safety visit approximately 90 days after the last infusion. During the treatment period, each subject will receive a total of 13 IV infusions of aslo0452 ngl-3 or placebo, each approximately 60 minutes, with an interval of approximately 4 weeks between infusions, i.e., Q4W dosing. The early PK cohort, consisting of the first approximately 15 subjects, will be randomly assigned in a 2:1 ratio to receive aslo0452 ngl-3 or placebo (approximately 10 subjects will receive aslo0452 ngl-3 and 5 subjects will receive placebo). Dosing for subjects in the early PK cohort will begin with 2400mg Q4W. After the early PK cohort is fully enrolled, additional subjects will be included in the main cohort. While the PK data from the early PK cohort are being analyzed, subjects in the main cohort will receive 2000mg aslo0452 ngl-3. Based on PK, safety, immunogenicity, and tolerability data from early PK cohorts, the aslo0452 ngl-3 dose level may be adjusted for subjects receiving active treatment.

[0419] The President's Multiple System Atrophy Rating Scale (UMSARS) will be determined for all subjects. The primary objective is to evaluate the change from baseline in the modified UMSARS Part I at Week 52.

[0420] Secondary Objectives :

[0421] Secondary objectives include:

[0422] a) evaluate the serum PK and CSF concentrations of aslo0452 ngl-3 in subjects with MSA;

[0423] b) Evaluate the efficacy of aslo0452 ngl-3 relative to placebo, as in Palma et al. (2021)

[0424] Changes from baseline to week 52 in 11 designated UMSARS measures;

[0425] c) evaluate the efficacy of aslo0452 ngl-3 relative to placebo as measured by the change from baseline to week 52 in total UMSARS;

[0426] d) Evaluate the efficacy of aslo0452 ngl-3 relative to placebo, as determined by Part II

[0427] as measured by the change in UMSARS from baseline to Week 52;

[0428] e) evaluate the efficacy of aslo0452 ngl-3 relative to placebo as measured by the change from baseline to week 52 on the Clinical Global Impression-Severity (CGI-S) scale;

[0429] f) evaluate the efficacy of aslo0452 ngl-3 compared to placebo on the Scale for Outcome Assessment of Parkinson's Disease-Autonomic Dysfunction (SCOPA-AUT); and

[0430] g) To evaluate the efficacy of aslo0452 ngl-3 relative to placebo as measured by overall survival at 52 weeks.

[0431] Exploratory goals :

[0432] Exploratory molecular and imaging biomarker targets include:

[0433] a) Evaluate the effect of aslo0452 ngl-3 compared to placebo on molecular biomarkers as measured by change from baseline to Week 52 in plasma and CSF biomarker levels, including but not limited to total αSYN levels and CSF free αSYN levels.

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Claims

1. A method for treating or preventing α-synucleinopathy in a subject in need thereof, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of an anti-α-synuclein antibody or antigen-binding fragment thereof. 2 . The method of claim 1 , wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 70 mg to 4,500 mg.

3. The method of claim 1 or claim 2, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 1,000 mg to 4,500 mg.

4. The method of any one of the preceding claims, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 1,800 mg to 2,600 mg.

5. The method of any one of the preceding claims, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 2,000 mg to 2,400 mg.

6. The method of any one of the preceding claims, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 2,000 mg or 2,400 mg.

7. The method of claim 6, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 2,000 mg.

8. The method of claim 6, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is 2,400 mg.

9. The method of claim 1, wherein the fixed dose of the anti-α-synuclein antibody or antigen-binding fragment thereof is: (a) 1,400 mg; (b) 1,600 mg; (c) 1,700 mg; (d) 1,800 mg; or (e) 4,500 mg.

10. The method of any one of the preceding claims, wherein the method comprises administering the anti-α-synuclein antibody or antigen-binding fragment thereof intravenously, subcutaneously, intradermally, or intramuscularly.

11. The method of claim 10, wherein the method comprises administering the anti-α-synuclein antibody or antigen-binding fragment thereof intravenously.

12. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject once every 3 to 5 weeks.

13. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject multiple times over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years.

14. The method of claim 13, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof is administered to the subject once every four weeks for a period of at least 3 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, or at least 5 years.

15. The method of any one of the preceding claims, wherein the alpha-synucleinopathy is selected from Parkinson's disease (PD), dementia with Lewy bodies (DLB), multiple system atrophy (MSA), Alzheimer's disease, pure autonomic failure, REM behavior disorder, prodromal synucleinopathy, and axonal dystrophy.

16. The method of claim 15, wherein the α-synuclein disease is Parkinson's disease.

17. The method of claim 15, wherein the α-synucleinopathy is multiple system atrophy (MSA).

18. The method of claim 15, wherein the α-synucleinopathy is dementia with Lewy bodies (DLB).

19. The method of claim 15, wherein the subject has a diagnosis of possible or probable MSA.

20. The method of any one of the preceding claims, wherein the subject to be treated has a Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score of 21 or less.

21. The method of any one of the preceding claims, wherein the subject to be treated, as measured using the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score: a) Severity score of 2 or less on the swallowing item; b) a severity score of 2 or less on the walking item; and / or c) A severity score of 2 or less on the falls item.

22. The method according to any one of the preceding claims, wherein the subject to be treated has a Unified Multiple System Atrophy Rating Scale (UMSARS) Part IV disability score of 3 or less.

23. The method of any one of the preceding claims, wherein the method slows progression of the alpha-synucleinopathy as measured by a modified version of the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I.

24. A method according to any one of the preceding claims, wherein: a) the method slows the progression of the α-synucleinopathy as measured by the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) total score; b) the method slows the progression of the alpha-synucleinopathy as measured by the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score; c) the method slows progression of the alpha-synucleinopathy as measured by the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) Part II score; d) the method slows the progression of the alpha-synucleinopathy as measured by the subject's 11-item Unified Multiple System Atrophy Rating Scale (UMSARS) score; and / or e) the method slows progression of the alpha-synucleinopathy as measured by the subject's Clinical Global Impression-Severity (CGI-S) scale score.

25. The method of any one of the preceding claims, wherein the method slows progression of the alpha-synuclein disease as measured by the spread of alpha-synuclein in the subject.

26. The method of any one of the preceding claims, wherein the method slows progression of the alpha-synuclein disease as measured by levels of free, unbound alpha-synuclein in the subject's cerebrospinal fluid (CSF).

27. The method of any of the preceding claims, wherein the method reduces the subject's score on a modified version of the Unified Multiple System Atrophy Rating Scale (UMSARS) Part I.

28. A method according to any one of the preceding claims, wherein: a) the method reduces the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) total score; b) the method reduces the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) Part I score; c) the method reduces the subject's Unified Multiple System Atrophy Rating Scale (UMSARS) Part II score; d) the method reduces the subject's 11-item Unified Multiple System Atrophy Rating Scale (UMSARS) score; and / or e) The method reduces the subject's Clinical Global Impression-Severity (CGI-S) scale score.

29. The method of any one of the preceding claims, wherein the method reduces alpha-synuclein spread in the subject.

30. The method of any one of the preceding claims, wherein the method reduces the level of free, unbound α-synuclein in the subject's cerebrospinal fluid (CSF).

31. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof binds to free, unbound α-synuclein in the subject's CSF.

32. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric and / or oligomeric α-synuclein in the subject's CSF.

33. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof binds to α-synuclein in the subject's CSF in vivo.

34. The method of any of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof specifically binds to a region comprising between about amino acid 102 and about amino acid 130 within the C-terminal region of human α-synuclein.

35. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric and aggregated forms of α-synuclein.

36. The method of claim 35, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof binds to monomeric and aggregated forms of α-synuclein in vivo.

37. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof sequesters both monomeric and aggregated forms of α-synuclein.

38. The method of claim 37, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof sequesters both monomeric and aggregated forms of α-synuclein in vivo.

39. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof has a K of less than 500 pM. D Binds monomeric human α-synuclein.

40. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof reduces α-synuclein spread in vivo.

41. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises: a) three heavy chain CDRs having the following sequences: (i) H-CDR1 of SEQ ID NO: 5; (ii) H-CDR2 of SEQ ID NO: 15; and (iii) H-CDR3 of SEQ ID NO: 16; and b) three light chain CDRs having the following sequences: (i) L-CDR1 of SEQ ID NO: 20; (ii) L-CDR2 of SEQ ID NO: 10; and (iii) L-CDR3 of SEQ ID NO:

21.

42. The method of claim 41, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

14.

43. The method of claim 41 or 42, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having the amino acid sequence of SEQ ID NO:

14.

44. The method of any one of claims 41 to 43, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable light chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

19.

45. The method of any one of claims 41 to 44, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable light chain region having the amino acid sequence of SEQ ID NO:

19.

46. ​​The method of claim 41, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 14 and a variable light chain region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:

19.

47. The method of claim 46, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a variable heavy chain region having the amino acid sequence of SEQ ID NO: 14 and a variable light chain region having the amino acid sequence of SEQ ID NO:

19.

48. The method of claim 41, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO:

62.

49. The method of claim 48, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO:

62.

50. The method of any one of claims 41, 48, or 49, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a light chain region having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

17.

51. The method of claim 50, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a light chain region having the amino acid sequence of SEQ ID NO:

17.

52. The method of claim 41, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:

17.

53. The method of claim 52, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence of SEQ ID NO: 12 or SEQ ID NO: 62 and a light chain having the amino acid sequence of SEQ ID NO:

17.

54. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof is an antibody.

55. The method of any one of the preceding claims, wherein the anti-α-synuclein antibody or antigen-binding fragment thereof comprises a triple mutation in the Fc region corresponding to L234F / L235E / P331S numbering based on Kabat numbering.

56. An anti-α-synuclein antibody or antigen-binding fragment thereof for use in a method of treating or preventing an α-synuclein disease in a subject, the method comprising administering to the subject a fixed dose of 50 mg to 5,000 mg of the anti-α-synuclein antibody or antigen-binding fragment thereof.

57. A kit comprising: a) anti-α-synuclein antibodies or antigen-binding fragments thereof; as well as b) instructions for use thereof in the treatment of alpha-synucleinopathies at a fixed dose of 50 mg to 5,000 mg.

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