JP2021502955A - Administration regimen for the treatment of synuclein disease - Google Patents

Abstract

The present invention provides a dosage regimen for the treatment of synuclein disease. In one regimen, subjects receive 3000-5000 mg of antibody intravenously every 3-5 weeks. In another regimen, subjects receive 1300 to 1700 mg of antibody intravenously every 3-5 weeks.

Description

Sequence Listing A sequence listing containing SEQ ID NOs: 1-151 is attached and incorporated in its entirety by reference. 503725 SEQ. The list named txt was created in ASCII format on September 27, 2017 and is 127,151 bytes in size.

Lewy body disease (LBD) is characterized by degeneration of the dopaminergic system, motor changes, cognitive impairment, and the formation of Lewy bodies (LB) and / or Lewy neurites. [McKeith et al., Neurology (1996) 47: 1113-24]. Lewy body diseases include Parkinson's disease (including idiopathic Parkinson's disease) and dementia (DLB) with Lewy bodies. Lewy body disease is a common cause of movement disorders and cognitive decline in the aging population [Galasko et al., Arch. Neurol. (1994) 51: 888-95]. Constipation is another common symptom in subjects with Lewy body disease [Ondo et al., Neurology 2012; 78; 1650-1654; Ashraf et al., Movement Disorders 12, 946-951 (1997)]. ..

Alpha-synuclein is a protein normally associated with synapses and is thought to play a role in neuroplasticity, learning and memory. Alpha-synuclein is also associated with a central role in the pathogenesis of Lewy body dementias. The protein can aggregate in pathological conditions to form insoluble fibrils. For example, alpha-synuclein accumulates in LB [Spillantini et al., Nature (1997) 388: 839-40; Takeda et al., J. Pathol. (1998) 152: 367-72; Wakabayashi et al., Neurosci. Lett. (1997) 239: 45-8]. Mutations in the alpha-synuclein gene co-separate with the rare familial form of Parkinsonism [Kruger et al., Nature Gen. (1998) 18: 106-8; Polymeropoulos, et al., Science (1997) 276 : 2045-7]. Overexpression of alpha-synuclein in transgenic mice [Masliah et al., Science (2000) 287: 1265-9] and Drosophila [Feany et al., Nature (2000) 404: 394-8] is a Lewy body disease. It mimics a number of pathological aspects of. Soluble oligomers of alpha-synuclein can be neurotoxic [Conway et al., Proc Natl Acad Sci USA (2000) 97: 571-576]. Accumulation of alpha-synuclein with similar morphological and neurological changes in diverse species and animal models such as human, mouse, and fly suggests that this molecule contributes to the development of Lewy body disease. To do.

Immunotherapy for alpha-synuclein has been reported to reduce alpha-synuclein deposition and behavioral symptoms in a mouse model of Lewy body disease [Masliah et al., PLoS ONE 6 (4): e19338]. . doi: 10.1371 / journal.pone.0019338].

In one aspect, the invention is a method of performing treatment or prevention of a subject having or at risk of synuclein disease, wherein the subject is provided with 3-5 antibodies against alpha-synuclein at a dose of 3000-5000 mg. Methods are provided that include intravenous administration at weekly intervals, or administration of another regimen that delivers the antibody to the subject in substantially the same subcurve area. In some such methods, the dose is 3,000-4500 mg.

In some such methods, the antibody is administered to multiple subjects who have or are at risk for synuclein disease, and the subjects receive one or two fixed doses. In some such methods, subjects weighing less than 65 kg receive a dose of 3500 mg of antibody, and subjects weighing more than 65 kg receive a dose of 4500 mg.

For some such methods, the dose is 45-75 mg / kg. In some such methods, the dose is 50-70 mg / kg.

In some such methods, the interval is 28 days. In some such methods, the subject receives at least 6 doses of antibody at intervals of 3-5 weeks. In some such methods, the subject receives at least 12 doses of antibody at intervals of 3-5 weeks. In some such methods, the subject receives at least 18 doses of antibody at intervals of 3-5 weeks. In some such methods, the interval is 4 weeks. In some such methods, subjects receive antibodies at least every 52 weeks and 4 weeks.

Some such methods further include monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms.

In some such ways, monitoring is
(A) A mobile device programmed to receive and transmit data acquired from internal and / or external sensors of the mobile device regarding movement disorders of subjects with or suspected of having synuclein disease. Providing to the subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device acquire data on the movement,
It includes (b) collecting data transmitted from mobile devices and (c) comparing the data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject.

In some such methods, the mobile device is programmed to receive and transmit data from at least two external sensors attached to the subject's upper and lower limbs. In some such methods, the mobile device acquires data from sensors on the subject's upper and lower limbs. In some such methods, the mobile device is carried by the subject and acquires data from internal sensors. In some such methods, the sequence of exercises involves tapping, sitting and standing on the device. In some such methods, monitoring indicates reduction of movement disorders or cognitive impairment in response to administration of antibodies.

In some such methods, the antibody binds within residues 115-130 of alpha-synuclein.

In some such methods, the antibody binds within residues 118-126 of alpha-synuclein.

In some such methods, the antibody binds within residues 117-123 of alpha-synuclein. In some such methods, the antibody is NI-202.21D11. In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 139 to 141 and three light chain CDRs designated by SEQ ID NOs: 143 to 145, respectively. In some such methods, the antibody comprises a heavy chain variable region designated by SEQ ID NO: 138 and a light chain variable region designated by SEQ ID NO: 142.

In some such methods, the antibody comprises three light chain Kabat CDRs of SEQ ID NO: 5 and three heavy chain Kabat CDRs of SEQ ID NO: 10. In some such methods, the antibody comprises a heavy chain variable region of any of SEQ ID NOs: 8-11 and a light chain variable region of any of SEQ ID NOs: 3-5, preferably the heavy chain variable region. The heavy chain variable region of SEQ ID NO: 10 and the light chain variable region is the light chain variable region of SEQ ID NO: 5.

In some such methods, the antibody is a human IgG1 isotype antibody. In some such methods, the antibody comprises a heavy chain constant region of SEQ ID NO: 35 and a light chain constant region of SEQ ID NO: 30, where the C-terminal lysine of SEQ ID NO: 35 may not be present. In some such methods, the antibody comprises a heavy chain constant region of SEQ ID NO: 35 and a light chain constant region of SEQ ID NO: 13 in which the C-terminal lysine of SEQ ID NO: 35 may not be present.

In some such methods, the antibody is 9E4. In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 146 to 148 and three light chain CDRs designated by SEQ ID NOs: 149 to 151, respectively. In some such methods, the antibody comprises the heavy chain designated by SEQ ID NO: 37 and the light chain designated by SEQ ID NO: 32, and the C-terminal lysine of SEQ ID NO: 37 may be absent.

In some such methods, the antibody binds within residues 1-20 of alpha-synuclein. In some such methods, the antibody binds within residues 4-15 of alpha-synuclein.

In some such methods, the antibody is NI-202.12F4. In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 131-133 and three light chain CDRs designated by SEQ ID NOs: 135-137, respectively. In some such methods, the antibody comprises a heavy chain variable region designated by SEQ ID NO: 130 and a light chain variable region designated by SEQ ID NO: 134.

In some such methods, the subject weighs less than 65 kg and is given a dose of 3500 mg of antibody every 4 weeks.

In some such methods, prior to administration of antibodies in the range of 3000-5000 mg, a load dose of 2000 mg of antibody is administered, and until a dose of 3500 mg is reached, it is greater than or equal to 2000 mg but less than 3500 mg. Uptitration may be performed with one or more subsequent doses, all doses being separated at intervals of 3-5 weeks.

In some such methods, a dose of 3500 mg is administered at the next interval after the dose of 2000 mg. In some such methods, the dose is gradually increased over multiple intervals prior to administration of the 3500 mg dose. In some such methods, the subject weighs more than 65 kg and is given a dose of 4500 mg of antibody every 4 weeks.

In some such methods, prior to administering the antibody at 4500 mg, a loading dose of 2000 mg of the antibody is administered and one or more of which is greater than or equal to 2000 mg but less than 4500 mg until a dose of 4500 mg is reached. Subsequent doses may be used for incremental titration, with all doses separated at intervals of 3-5 weeks.

In some such methods, the 4500 mg dose is administered at the next interval after the 2000 mg dose. In some such methods, the dose is gradually increased over multiple intervals prior to administration of the 4500 mg dose.

Some such methods further include monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms.

In some such ways, monitoring is
(A) A mobile device programmed to receive and transmit data acquired from internal and / or external sensors of the mobile device regarding movement disorders of subjects with or suspected of having synuclein disease. Providing to the subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device acquire data on the movement,
It includes (b) collecting data transmitted from mobile devices and (c) comparing the data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject.

In some such methods, the mobile device is programmed to receive and transmit data from at least two external sensors attached to the subject's upper and lower limbs. In some such methods, the mobile device acquires data from sensors on the subject's upper and lower limbs. In some such methods, the mobile device is carried by the subject and acquires data from internal sensors. In some such methods, the sequence of exercises involves tapping, sitting and standing on the device. In some such methods, monitoring indicates reduction of movement disorders or cognitive impairment in response to administration of antibodies.

In some such methods, synuclein disease is Parkinson's disease. In some such methods, synuclein disease is dementia with Lewy bodies. In some such methods, synuclein disease is multiple system atrophy. In some such methods, synuclein disease is progressive supranuclear palsy. In some such methods, synuclein disease is REM sleep behavior disorder. In some such methods, synuclein disease is Alzheimer's disease with Lewy bodies in the amygdala. In some such methods, the subject has the disease.

In some such methods, the subject has not received symptomatological treatment for Parkinson's disease in combination with the antibody. In some such methods, the antibody is administered in combination with levodopa.

In another aspect, the invention is a method of performing treatment or prophylaxis of a subject with Lewy body disease, in which an antibody against alpha-synuclein at a dose of 1300 to 1700 mg is venous at 3-5 week intervals. A method is provided that comprises administering the antibody internally or administering to the subject another regimen that delivers the antibody in substantially the same area under the curve.

In some such methods, the dose is 1500 mg. In some such methods, the antibody is administered to multiple subjects with Lewy body dementias, who receive one or two fixed doses. In some such methods, the subject receives a dose of 18-25 mg / kg. In some such methods, the subject receives a dose of 20 mg / kg.

In some such methods, the interval is 28 days. In some such methods, the subject receives at least 6 doses of antibody at intervals of 3-5 weeks. In some such methods, the subject receives at least 12 doses of antibody at intervals of 3-5 weeks. In some such methods, the subject receives at least 18 doses of antibody at intervals of 3-5 weeks. In some such methods, the interval is 4 weeks. In some such methods, subjects receive antibodies at least every 52 weeks and 4 weeks.

Some such methods further include monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms.

In some such ways, monitoring is
(A) Programmed to receive and transmit data acquired from internal and / or external sensors of the device for movement disorders of subjects with or suspected of having Lewy body dementias. Providing a mobile device to a subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device acquire data about the movement.
It includes (b) collecting data transmitted from mobile devices and (c) comparing the data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject.

In some such methods, the mobile device is programmed to receive and transmit data from at least two external sensors attached to the subject's upper and lower limbs.

In some such methods, the mobile device acquires data from sensors on the subject's upper and lower limbs. In some such methods, the mobile device is carried by the subject and acquires data from internal sensors. In some such methods, the sequence of exercises involves tapping, sitting and standing on the device. In some such methods, monitoring indicates reduction of movement disorders or cognitive impairment in response to administration of antibodies.

In some such methods, the antibody binds within residues 115-130 of alpha-synuclein.

In some such methods, the antibody binds within residues 118-126 of alpha-synuclein.

In some such methods, the antibody binds within residues 117-123 of alpha-synuclein.

In some such methods, the antibody is NI-202.21D11. In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 139 to 141 and three light chain CDRs designated by SEQ ID NOs: 143 to 145, respectively. In some such methods, the antibody comprises a heavy chain variable region designated by SEQ ID NO: 138 and a light chain variable region designated by SEQ ID NO: 142.

In some such methods, the antibody comprises three light chain Kabat CDRs of SEQ ID NO: 5 and three heavy chain Kabat CDRs of SEQ ID NO: 10. In some such methods, the antibody comprises a heavy chain variable region of any of SEQ ID NOs: 8-11 and a light chain variable region of any of SEQ ID NOs: 3-5, preferably the heavy chain variable region. It is the heavy chain variable region of SEQ ID NO: 10, and the light chain variable region is the light chain variable region of SEQ ID NO: 5.

In some such methods, the antibody is a human IgG1 isotype antibody.

In some such methods, the antibody comprises a heavy chain constant region of SEQ ID NO: 35 in which the C-terminal lysine may not be present, and a light chain constant region of SEQ ID NO: 30. In some such methods, the antibody comprises a heavy chain constant region of SEQ ID NO: 35 in which the C-terminal lysine may not be present, and a light chain constant region of SEQ ID NO: 13.

In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 146 to 148 and three light chain CDRs designated by SEQ ID NOs: 149 to 151, respectively. In some such methods, the antibody comprises the heavy chain designated by SEQ ID NO: 37 and the light chain designated by SEQ ID NO: 32, and the C-terminal lysine of SEQ ID NO: 37 may be absent.

In some such methods, the antibody binds within residues 1-20 of alpha-synuclein. In some such methods, the antibody binds within residues 4-15 of alpha-synuclein.

In some such methods, the antibody is NI-202.12F4. In some such methods, the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 131-133 and three light chain CDRs designated by SEQ ID NOs: 135-137, respectively. In some such methods, the antibody comprises a heavy chain variable region designated by SEQ ID NO: 130 and a light chain variable region designated by SEQ ID NO: 134.

In some such methods, synuclein disease is Parkinson's disease. In some such methods, synuclein disease is dementia with Lewy bodies. In some such methods, synuclein disease is plural atrophy. In some such methods, synuclein disease is progressive supranuclear palsy. In some such methods, synuclein disease is REM sleep behavior disorder. In some such methods, synuclein disease is Alzheimer's disease with Lewy bodies in the amygdala. In some such methods, the subject has the disease.

In some such methods, the subject has not received symptomatological treatment for Parkinson's disease in combination with the antibody. In some such methods, the antibody is administered in combination with levodopa.

Definitions The term "antibody" includes intact antibodies and their binding fragments. Typically, the fragments compete for specific binding to the target with the intact antibody from which they are derived. Fragments include separated heavy chains, separated light chains, Fab, Fab', F (ab') 2, F (ab) c, Fv, single chain antibodies, and single domain antibodies. The term "antibody" also includes bispecific antibodies. Bispecific or bifunctional antibodies are artificial hybrid antibodies with two different heavy / light chain pairs and two different binding sites [eg, Songsivilai and Lachmann, Clin. Exp. Immunol., 79: 315. -321 (1990); see Kostelny et al., J. Immunol., 148: 1547-53 (1992)].

The basic antibody structural unit is the subunit tetramer. Each tetramer comprises two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain contains variable regions of about 100-110 or more amino acids that are primarily involved in antigen recognition. When first expressed, this variable region is typically linked to a cleavable signal peptide. Variable regions that do not have a signal peptide are sometimes referred to as mature variable regions. Thus, for example, the light chain maturation variable region means a light chain variable region that does not have a light chain signal peptide. The carboxy-terminal portion of each chain defines a constant region primarily involved in effector function. The constant region can include any or all of the CH1 region, the hinge region, the CH2 region, and the CH3 region.

Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, defining antibody isotypes as IgG, IgM, IgA, IgD, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are linked by the "J" region of about 12 or more amino acids, and the heavy chain also contains the "D" region of about 10 or more amino acids. [In general, see Fundamental Immunology (Paul, W., ed., 2nd ed. Raven Press, N.Y., 1989), Ch. 7] (By reference, the whole is incorporated for all purposes).

The mature variable region of each light / heavy chain pair forms an antibody binding site. Therefore, intact antibodies have two binding sites. The two binding sites are the same, except for bifunctional or bispecific antibodies. All chains exhibit the same general structure of relatively conserved framework regions (FRs) linked by three hypervariable regions, also called complementarity determining regions (CDRs). The CDRs from the two strands of each pair are aligned by the framework region to allow binding to a particular epitope. From the N-terminus to the C-terminus, both the light and heavy chains contain regions FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each region is Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991), or Chothia & Lesk, J. Mol. Biol. 196: 901-917 (1987). ); According to the definition of Chothia et al., Nature 342: 878-883 (1989). Kabat also provides a widely used numbering rule (Kabat numbering) in which the same number of corresponding residues between different heavy chains or between different light chains is assigned.

The percentage of sequence identity is determined using antibody sequences that are maximally aligned according to the Kabat numbering rules. After alignment, if the target antibody region (eg, the entire maturation variable region of the heavy or light chain) is compared to the same region of the reference antibody, the sequence identity percentage between the target antibody region and the reference antibody region is the subject. The number of positions occupied by the same amino acid in both the antibody region and the reference antibody region divided by the total number of aligned positions in the two regions (gap is not counted) multiplied by 100 and converted to a percentage. Is.

For the purpose of classifying amino acid substitutions as conservative or non-conservative, amino acids are classified as follows: Group I (hydrophobic side chains): Norleucine, Met, Ala, Val, Leu, Ile; Group II (Neutral hydrophilic side chain): Cys, Ser, Thr; Group III (acidic side chain): Asp, Glu; Group IV (basic side chain): Asn, Gln, His, Lys, Arg; Group V (chain) Residues Affecting Orientation): Gly, Pro; and Group VI (Aromatic Side Chains): Trp, Tyr, Phe. Conservative substitutions involve substitutions between amino acids of the same class. Non-conservative replacement constitutes exchanging a member of one of these classes with a member of another class.

Antibodies of the invention typically bind to their designated targets with an affinity constant of at least 10 ⁶ , 10 ⁷ , 10 ⁸ , 10 ⁹ or 10 ¹⁰ M- ¹ . Such binding is specific in that it is more detectable and distinguishable from non-specific binding that occurs to at least one unrelated target. Specific bonds can be the result of the formation of bonds between specific functional groups or specific spatial fits (eg, lock and key types), while non-specific bonds are usually the result of van der Waals forces. Is. However, specific binding does not necessarily imply that the monoclonal antibody binds to one and only one target.

The term "symptom" refers to subjective evidence of the disease, such as gait changes, as perceived by the subject. "Signs" refer to objective evidence of the disease as observed by a physician.

Individuals at risk of developing a disease that is statistically significantly higher than individuals who have at least one known risk factor (eg, genetic, biochemical, family history, environmental exposure) and who do not have the risk factor. If placed, the individual is at increased risk of disease. Statistical significance means p ≦ 0.05.

Unless it is clear from the context that this is not the case, the term "about" includes the value that falls within the greater of the standard deviation of the mean of the values listed or +/- 5% of the values listed. ..

The term "9E4 antibody" refers to any antibody in which each CDR is substantially 9E4, and thus includes mouse, chimeric, veneered, and humanized 9E4. References to other antibodies such as 5C1 and 1H7 have corresponding implications.

The antibodies of the invention can be administered in combination with another treatment for the same indication as the antibody, meaning that the other treatments are administered at least once during the period in which the antibody is administered. However, such a period begins one month before the first dose of antibody and ends one month after the last dose. Other treatments can be administered at repetitive intervals during this period, which may or may not be the same as the interval at which the antibody is administered. Other treatments may be symptomatological treatments.

Treatment is symptomatic when it affects only one or more of the causes of the disease, i.e. not its etiology, but one or more of its symptoms.

As used herein, the term "mobile device" refers to any portable device, including sensors and data recording devices suitable for obtaining a dataset of activity measurements. Typically, mobile devices include sensors for measuring activity. It may also require a display for electronically simulating activity tests on mobile devices, in addition to data processors and storage devices. In addition, data from the activity of interest is recorded and edited into a dataset, which is evaluated by the methods of the invention on either the mobile device itself or a second device. Depending on the particular configuration envisioned, it may be necessary for the mobile device to include a data transmitting device in order to transmit the acquired dataset from the mobile device to one or more additional devices. Particularly well suited as mobile devices according to the invention are smartphones, smart watches, wearable sensors, portable multimedia devices or tablet computers. Alternatively, a data recording device and, as appropriate, a portable sensor having a processing device may be used. In addition, depending on the type of activity test being performed, the mobile device is configured to direct the subject regarding the activity to be performed for the test.

References to ranges include any integer within the range, unless it is clear from the context that this is not the case.

A treatment regimen refers to a combination of parameters that characterize the administration of an immunotherapeutic agent, including dose, frequency of administration, route of administration, and any or all of the total duration of administration.

Brief Description of Sequence SEQ ID NO: 1 is the m9E4VL variable region.

SEQ ID NO: 2 is the 63102889Hu9E4VLFr variable region.

SEQ ID NO: 3 is the Hu9E4VLv1 variable region.

SEQ ID NO: 4 is the Hu9E4VLv2 (no return mutation) variable region.

SEQ ID NO: 5 is the Hu9E4VLv3 variable region.

SEQ ID NO: 6 is the m9E4VH variable region.

SEQ ID NO: 7 is the 1791009Hu9E4VHFr variable region.

SEQ ID NO: 8 is the Hu9E4VHv1 variable region.

SEQ ID NO: 9 is a Hu9E4VHv2 variable region.

SEQ ID NO: 10 is the Hu9E4VHv3 variable region.

SEQ ID NO: 11 is a Hu9E4VHv4 (no return mutation) variable region.

SEQ ID NO: 12 is the amino acid sequence of natural human wild-type alpha-synuclein.

SEQ ID NO: 13 is the humanized 9E4 light chain constant region (with R) (common for v1, v2, v3).

SEQ ID NO: 14 is a humanized 9E4 light chain constant region (with R) nucleotide sequence (common for v1, v2, v3).

SEQ ID NO: 15 is an exemplary human IgG1 constant region amino acid sequence.

SEQ ID NO: 16 is a nucleic acid sequence encoding an exemplary human IgG1 constant region.

SEQ ID NO: 17 is the Hu9E4VLv1 nucleotide sequence variable region.

SEQ ID NO: 18 is the Hu9E4VLv2 nucleotide sequence (without reversion mutation) variable region.

SEQ ID NO: 19 is the Hu9E4VLv3 nucleotide sequence variable region.

SEQ ID NO: 20 is the Hu9E4VHv1 nucleotide sequence variable region.

SEQ ID NO: 21 is the Hu9E4VHv2 nucleotide sequence variable region.

SEQ ID NO: 22 is the Hu9E4VHv3 nucleotide sequence variable region.

SEQ ID NO: 23 is the Hu9E4VHv4 nucleotide sequence (without reversion mutation) variable region.

SEQ ID NO: 24 is the Hu9E4VL signal peptide (common for v1, v2, v3).

SEQ ID NO: 25 is the Hu9E4VL signal peptide nucleotide sequence (common for v1, v2, v3).

SEQ ID NO: 26 is the Hu9E4VH signal peptide (common for v1, v2, v3).

SEQ ID NO: 27 is the Hu9E4VH signal peptide nucleotide sequence (common for v1, v2, v3, v4).

SEQ ID NO: 28 is a Hu9E4VL alternative.

SEQ ID NO: 29 is a Hu9E4VH alternative.

SEQ ID NO: 30 is an exemplary human kappa light chain constant region amino acid sequence.

SEQ ID NO: 31 is a humanized 9E4 light chain constant region (without R) nucleotide.

Sequence (common for v1, v2, v3).

SEQ ID NO: 32 is humanized 9E4 light chain type 3 (variable region + constant region, with arginine).

SEQ ID NO: 33 is humanized 9E4 light chain type 3 (variable region + constant region, no arginine).

SEQ ID NO: 34 is humanized 9E4 heavy chain type 3 (variable region + constant region).

SEQ ID NO: 35 is a humanized 9E4 heavy chain constant region (G1m3 allotype; BIP type).

SEQ ID NO: 36 is humanized 9E4 heavy chain type 3 (variable region + constant region).

SEQ ID NO: 37 is humanized 9E4 heavy chain type 3 (variable region + alternative constant region G1m3 allotype).

SEQ ID NO: 38 is the m5C1 antibody mature heavy chain variable region nucleotide sequence.

SEQ ID NO: 39 is the m5C1 antibody heavy chain variable region amino acid sequence.

SEQ ID NO: 40 is the m5C1 antibody heavy chain signal peptide nucleotide sequence.

SEQ ID NO: 41 is the m5C1 antibody heavy chain signal peptide amino acid sequence.

SEQ ID NO: 42 is the m5C1 antibody mature light chain variable region nucleotide sequence.

SEQ ID NO: 43 is the m5C1 antibody light chain variable region amino acid sequence.

SEQ ID NO: 44 is the m5C1 antibody light chain signal peptide nucleotide sequence.

SEQ ID NO: 45 is the m5C1 antibody light chain signal peptide amino acid sequence.

SEQ ID NO: 46 is a 5C1 immunogen.

SEQ ID NO: 47 is m5C1 heavy chain CDR1.

SEQ ID NO: 48 is m5C1 heavy chain CDR2.

SEQ ID NO: 49 is m5C1 heavy chain CDR3.

SEQ ID NO: 50 is m5C1 light chain CDR1.

SEQ ID NO: 51 is m5C1 light chain CDR2.

SEQ ID NO: 52 is m5C1 light chain CDR3.

SEQ ID NO: 53 is a mouse 5C1 heavy chain variable region nucleotide sequence having a sequence encoding a signal peptide.

SEQ ID NO: 54 is a mouse 5C1 heavy chain variable region sequence having a signal peptide.

SEQ ID NO: 55 is a mouse 5C1 light chain variable region nucleotide sequence having a sequence encoding a signal peptide.

SEQ ID NO: 56 is a mouse 5C1 light chain variable region sequence having a signal peptide.

SEQ ID NO: 57 is human VH acceptor FR (accession number AAY42876.1).

SEQ ID NO: 58 is humanized 5C1H1.

SEQ ID NO: 59 is humanized 5C1H2.

SEQ ID NO: 60 is humanized 5C1H3.

SEQ ID NO: 61 is humanized 5C1H4.

SEQ ID NO: 62 is humanized 5C1H5.

SEQ ID NO: 63 is a nucleic acid sequence encoding humanized 5C1 H1.

SEQ ID NO: 64 is a nucleic acid sequence encoding humanized 5C1 H2.

SEQ ID NO: 65 is a nucleic acid sequence encoding humanized 5C1H3.

SEQ ID NO: 66 is a nucleic acid sequence encoding humanized 5C1 H4.

SEQ ID NO: 67 is a nucleic acid sequence encoding humanized 5C1H5.

SEQ ID NO: 68 is human VL acceptor FR (accession number CAB51293.1).

SEQ ID NO: 69 is humanized 5C1L1.

SEQ ID NO: 70 is humanized 5C1L2.

SEQ ID NO: 71 is humanized 5C1L3.

SEQ ID NO: 72 is humanized 5C1L4.

SEQ ID NO: 73 is a nucleic acid sequence encoding humanized 5C1 L1.

SEQ ID NO: 74 is a nucleic acid sequence encoding humanized 5C1 L2.

SEQ ID NO: 75 is a nucleic acid sequence encoding humanized 5C1L3.

SEQ ID NO: 76 is a nucleic acid sequence encoding humanized 5C1 L4.

SEQ ID NO: 77 is a nucleic acid sequence encoding an exemplary human kappa light chain constant region.

SEQ ID NO: 78 can be found in Jensen et al. The non-amyloid component (NAC) domain of alpha-synuclein as reported by.

SEQ ID NO: 79 can be found in Ueda et al. The non-amyloid component (NAC) domain of alpha-synuclein as reported by.

SEQ ID NO: 80 is the m1H7 antibody heavy chain variable nucleotide sequence.

SEQ ID NO: 81 is the m1H7 antibody heavy chain variable amino acid sequence.

SEQ ID NO: 82 is the m1H7 antibody light chain variable nucleotide sequence.

SEQ ID NO: 83 is the m1H7 antibody light chain variable amino acid sequence.

SEQ ID NO: 84 is a mature m1H7 antibody heavy chain variable nucleotide sequence.

SEQ ID NO: 85 is a mature m1H7 antibody heavy chain variable amino acid sequence.

SEQ ID NO: 86 is a mature m1H7 antibody light chain variable nucleotide sequence.

SEQ ID NO: 87 is a mature m1H7 antibody light chain variable amino acid sequence.

SEQ ID NO: 88 is m1H7 antibody heavy chain CDR1 (definition of Kabat).

SEQ ID NO: 89 is m1H7 antibody heavy chain CDR2 (definition of Kabat).

SEQ ID NO: 90 is m1H7 antibody heavy chain CDR3 (definition of Kabat).

SEQ ID NO: 91 is m1H7 antibody light chain CDR1 (definition of Kabat).

SEQ ID NO: 92 is m1H7 antibody light chain CDR2 (definition of Kabat).

SEQ ID NO: 93 is the m1H7 antibody light chain CDR3 (definition of Kabat).

SEQ ID NO: 94 is the Hu1H7VHv1 nucleotide sequence.

SEQ ID NO: 95 is the Hu1H7VHv1 amino acid sequence.

SEQ ID NO: 96 is the Hu1H7VHv2 nucleotide sequence.

SEQ ID NO: 97 is the Hu1H7VHv2 amino acid sequence.

SEQ ID NO: 98 is the Hu1H7VHv3 nucleotide sequence.

SEQ ID NO: 99 is the Hu1H7VHv3 amino acid sequence.

SEQ ID NO: 100 is the Hu1H7VHv4 nucleotide sequence.

SEQ ID NO: 101 is the Hu1H7VHv4 amino acid sequence.

SEQ ID NO: 102 is the Hu1H7VHv5 nucleotide sequence.

SEQ ID NO: 103 is the Hu1H7VHv5 amino acid sequence.

SEQ ID NO: 104 is the Hu1H7VH signal peptide nucleotide sequence.

SEQ ID NO: 105 is the Hu1H7VH signal peptide amino acid sequence.

SEQ ID NO: 106 is the Hu1H7VH signal peptide nucleotide sequence.

SEQ ID NO: 107 is the Hu1H7VH signal peptide amino acid sequence.

SEQ ID NO: 108 is the Hu1H7VLv1 nucleotide sequence.

SEQ ID NO: 109 is the Hu1H7VLv1 amino acid sequence.

SEQ ID NO: 110 is the Hu1H7VLv2 nucleotide sequence.

SEQ ID NO: 111 is the Hu1H7VLv2 amino acid sequence.

SEQ ID NO: 112 is the Hu1H7VLv3 nucleotide sequence.

SEQ ID NO: 113 is the Hu1H7VLv3 amino acid sequence.

SEQ ID NO: 114 is the Hu1H7VLv4 nucleotide sequence.

SEQ ID NO: 115 is the Hu1H7VLv4 amino acid sequence.

SEQ ID NO: 116 is the Hu1H7VL signal peptide nucleotide sequence.

SEQ ID NO: 117 is the amino acid sequence of the BAC02037 (GI-21670055) human acceptor used for the heavy chain framework.

SEQ ID NO: 118 is the amino acid sequence of AAY33358 GI-63102905) human acceptor used for the light chain framework.

SEQ ID NO: 119 is Hu1H7VH without a reversion or CDR mutation.

SEQ ID NO: 120 is Hu1H7VL without a reversion or CDR mutation.

SEQ ID NO: 121 is a Hu1H7VH alternative.

SEQ ID NO: 122 is a Hu1H7VL alternative.

SEQ ID NO: 123 is a Hu1H7VH CDR3 alternative.

SEQ ID NO: 124 is humanized 1H7 light chain type 3 (variable region + constant region, with arginine).

SEQ ID NO: 125 is humanized 1H7 light chain type 3 (variable region + constant region, no arginine).

SEQ ID NO: 126 is humanized 1H7 heavy chain type 3 (variable region + constant region).

SEQ ID NO: 127 is humanized 1H7 heavy chain type 3 (variable region + constant region G1m3 allotype).

SEQ ID NO: 128 is a humanized 1H7 heavy chain constant region (IgG2).

SEQ ID NO: 129 is a humanized 1H7 heavy chain constant region (G1m1 allotype).

SEQ ID NO: 130 is the NI-20.12.F4-VHA1b variable heavy chain (VH) sequence.

SEQ ID NO: 131 is the NI-202.12F4-VHA1b CDR1 sequence.

SEQ ID NO: 132 is the NI-202.12F4-VHA1b CDR2 sequence.

SEQ ID NO: 133 is the NI-20.12.F4-VHA1b CDR3 sequence.

SEQ ID NO: 134 is the NI-202.12F4-VLa1 variable light chain (VL) sequence.

SEQ ID NO: 135 is the NI-20.12.F4-VLa1 CDR1 sequence.

SEQ ID NO: 136 is the NI-20.12.F4-VLa1 CDR2 sequence.

SEQ ID NO: 137 is the NI-202.12F4-VLa1 CDR3 sequence.

SEQ ID NO: 138 is the NI-202.21D11-VH sequence.

SEQ ID NO: 139 is the NI-202.21D11-VH CDR1 sequence.

SEQ ID NO: 140 is the NI-202.21D11-VH CDR2 sequence.

SEQ ID NO: 141 is the NI-202.21D11-VH CDR3 sequence.

SEQ ID NO: 142 is the NI-202.21D11-VK sequence.

SEQ ID NO: 143 is the NI-202.21D11 VK CDR1 sequence.

SEQ ID NO: 144 is the NI-202.21D11 VK CDR2 sequence.

SEQ ID NO: 145 is the NI-202.21D11 VK CDR3 sequence.

SEQ ID NO: 146 is the 9E4 VH CDR1 sequence.

SEQ ID NO: 147 is the 9E4 VH CDR2 sequence.

SEQ ID NO: 148 is the 9E4 VH CDR3 sequence.

SEQ ID NO: 149 is the 9E4 VL CDR1 sequence.

SEQ ID NO: 150 is the 9E4 VL CDR2 sequence.

SEQ ID NO: 151 is the 9E4 VL CDR3 sequence.

I. Abstract The present invention provides a dosing regimen for immunotherapy of synuclein disease. Subjects receive antibodies to alpha-synuclein intravenously every 3-5 weeks at doses of 3000-5000 mg or 1300-1700 mg. One or more lower loading doses may be given prior to such doses.

II. Target molecule Natural human wild-type alpha-synuclein has the following amino acid sequence:
MDVFMKGLSK AKEGVVAAAE KTKQGVAEAA GKTKEGVLYV GSKTKEGVVH GVATVAEKTK EQVTNVGGAV VTGVTAVAQK TVEGAGSIAA ATGFVKKDQL GKNEEGAPQE GILEDMPVDP DNEAYEMPSE EGYQDYEPEA
It is a 140 amino acid peptide having [Ueda et al., Proc. Natl. Acad. Sci. USA (1993) 90: 11282-6]; GenBank Accession No .: P37840. The protein has three recognized domains: an N-terminal KTKE repeat domain covering amino acids 1-41; a NAC (non-amyloid component) domain covering approximately amino acids 60-95; and a C-terminal acidic domain covering approximately amino acids 98-140. Have.

Unless it is clear from the context that this is not the case, references to alpha-synuclein or fragments thereof refer to the natural human wild-type amino acid sequence pointed to above, and human allelic variants thereof, especially human allelic variants associated with Levy body disease. The body (eg, mutants E46K, A30P and A53T; the first letter points to the amino acid in SEQ ID NO: 12, the number points to the codon position in SEQ ID NO: 12, and the second letter points to the amino acid in the allelic variant. Includes). Such variants can be present individually or in any combination of any of the aspects of the invention described below, as appropriate.

III. Synuclein Disease Synuclein disease is characterized by an excessive accumulation of alpha-synuclein, which may or may not be associated with the formation of Lewy bodies (LBs). For example, Lewy body disease (LBD) is characterized by degeneration of the dopaminergic system, motor changes, cognitive impairment, and formation of Lewy bodies. [McKeith et al., Neurology (1996) 47: 1113-24]. Lewy bodies are globular protein deposits found in nerve cells. Their presence in the brain disrupts the normal functioning of the brain and interferes with the action of chemical messengers, including acetylcholine and dopamine. Parkinson's disease (including idiopathic Parkinson's disease), dementia with Levy's body (DLB) [also known as diffuse Levy's body disease (DLBD)], and Levy's body variant of Alzheimer's disease (Synucrane's disease) LBV), Alzheimer's and Parkinson's disease, progressive supranuclear palsy (PSP), and multiple system atrophy (MSA; eg, Olive Bridge cerebral atrophy, striatal melanosis) , And Shy-Drager's syndrome). DLB shares the symptoms of both Alzheimer's disease and Parkinson's disease. DLB differs from Parkinson's disease primarily in the location of Lewy bodies. In DLB Lewy bodies are formed primarily in the cortex. In Parkinson's disease, they are formed primarily in the substantia nigra. Other synuclein diseases include pure autonomic failure, Lewy body dementias, accidental LBD, and hereditary LBD (eg, mutations in the alpha-synuclein gene, PARK3 and PARK4).

IV. Immunotherapeutic agents Passive immunotherapy involves treatment with antibodies that specifically bind to alpha-synuclein.

1. 1. Antibody A. Binding Specificity and Functional Properties The methods of the invention are epitopes within residues 1-15, 4-15, 1-20, 91-99, 115-130, 117-123 or 118-126 of human alpha-synuclein. Can include antibodies that specifically bind to. The antibody can be a monoclonal antibody or a polyclonal antibody. Polyclonal sera can be specific for an epitope or range in alpha-synuclein in that they lack antibody binding outside the epitope or range of amino acids. The antibody can be a non-human, chimeric, veneered, humanized, or human antibody, among other possibilities.

Some antibodies are bispecific, one arm is an antibody against alpha-synthulein and the other arm is an insulin receptor, insulin-like growth factor (IGF) receptor, leptin. An antibody that binds to a receptor, or a receptor expressed at the blood-brain barrier, such as a lipoprotein receptor, or preferably a transferrin receptor (Friden et al., PNAS 88: 4771-4775, 1991; Friden et al. ., Science 259: 373-377, 1993). Such bispecific antibodies can be translocated across the blood-brain barrier by receptor-mediated transcytosis. Brain uptake of bispecific antibodies can be further enhanced by manipulating the bispecific antibodies to reduce their affinity for blood-brain barrier receptors. The reduced affinity for the receptor resulted in a broader distribution in the brain (eg Athal. Et al. Sci. Trans. Med. 3, 84ra43, 2011; Yu et al. Sci. Trans). . Med. 3, 84ra44, 2011).

An exemplary bispecific antibody can also be: (1) a bivariable domain antibody (DVD-Ig) containing two variable domains in tandem through a short peptide linkage of each light and heavy chain: Wu et al., Generation and characterization of a Dual Variable Domain Immunoglobulin (DVD-Ig ^TM ) Molecule, Antibody Engineering, Springer Berlin Heidelberg (2010)]; (2) Tetravalent having two binding sites for each target antigen Tandab, which is a fusion of two single-stranded diabodies resulting in a bispecific antibody; (3) flexibody, which is a combination of scFv with a diabody resulting in a polyvalent molecule; (4). So-called "dimerization and docking domains" in protein kinase A that, when applied to a Fab, can result in a trivalent bispecific binding protein consisting of two identical Fab fragments linked to different Fab fragments. A "dock and lock"molecule; (5), for example, a so-called scorpion molecule comprising two scFv fused to both ends of the human Fc region. Examples of useful platforms for preparing bispecific antibodies are BiTE (Micromet), DART (MacroGenics), Fcab and Mab2 (F-star), Fc-engineered IgGl (Xencor) or DuoBody (Fab arm exchange). Based on Genmab).

Another exemplary antibody is mAb 9E4, which binds to an epitope within residues 118-126 of human alpha-synuclein. The cell line, called JH17.9E4.3.337.1.14.2, which produces antibody 9E4, has ATCC Accession No. PTA-8221 and is American on February 26, 2007 under the provisions of the Budapest Treaty. It was deposited with the Type Culture Collection (ATCC, PO Box 1549, Manassas, Va. 20108). The mouse light chain and heavy chain variable sequences are SEQ ID NO: 1 and SEQ ID NO: 6, respectively. Mice mAb 9E4 were of VH-CDR1 of SEQ ID NO: 146, VH-CDR2 of SEQ ID NO: 147, VH-CDR3 of SEQ ID NO: 148, VL-CDR1 of SEQ ID NO: 149, VL-CDR2 of SEQ ID NO: 150, and SEQ ID NO: 151. Includes VL-CDR3.

Another exemplary antibody is mAb 5C1. The mouse light chain and heavy chain variable sequences are SEQ ID NO: 43 and SEQ ID NO: 39, respectively.

Another exemplary antibody is mAb 1H7. The cell line, called JH17.1H7.4.24.34, which produces antibody 1H7, has ATCC Accession No. PTA-8220 and is subject to the American Type Culture Collection (American Type Culture Collection) on February 26, 2007 under the provisions of the Budapest Treaty. It was deposited with ATCC, PO Box 1549, Manassas, Va. 20108). The mouse light chain and heavy chain variable sequences are SEQ ID NO: 87 and SEQ ID NO: 85, respectively.

Such antibodies include WO06 / 02581 and US Applications 61 / 591,835, 61 / 711,207, 13 / 750,983, 61 / 711,204, 61. / 719,281, 61 / 840,432, 61 / 872,366, 14 / 049,169, 61 / 533, 131, 61 / 711,204 , 61 / 843,011 and 13 / 662,261 (all of which are incorporated herein by reference for all purposes).

Another exemplary antibody is NI-202.12F4, which binds to an epitope within residues 4-15 of alpha-synuclein, the major contributor to the epitope being residue 10. It is a human antibody that has been reported (see US8,940,276; WO2012177792, US9,580,493). NI-202.12F4 comprises the variable heavy chain (VHA1b) sequence of SEQ ID NO: 130 and the variable light chain (VLa1) sequence of SEQ ID NO: 134. NI-202.12F4 contains VHA1b-CDR1 of SEQ ID NO: 131, VHA1b-CDR2 of SEQ ID NO: 132, VHA1b-CDR3 sequence of SEQ ID NO: 133 (Ala-His), VLa1-CDR1 sequence of SEQ ID NO: 135, and SEQ ID NO: 136. Includes the VLa1-CDR2 sequence and the VLa1-CDR3 sequence of SEQ ID NO: 137. Another exemplary antibody is NI-202.21D11, which binds to epitopes within residues 117-123 of alpha-synuclein, with residues 121 and 122 making the most contribution to the epitopes. (See US9,580,493). NI-202.21D11 comprises the variable heavy chain (VH) sequence of SEQ ID NO: 138 and the variable light chain (VK) sequence of SEQ ID NO: 142. NI-202.21D11 includes VH-CDR1 of SEQ ID NO: 139, VH-CDR2 of SEQ ID NO: 140, VH-CDR3 sequence of SEQ ID NO: 141, VK-CDR1 sequence of SEQ ID NO: 143, VK-CDR2 sequence of SEQ ID NO: 144, And the VK-CDR3 sequence of SEQ ID NO: 145.

B. Humanized Antibodies Humanized antibodies are genetically engineered antibodies in which CDRs from non-human "donor" antibodies are grafted onto human "acceptor" antibody sequences (eg, Queen et al., US 5,530,101 and the same. 5,585,089; see Winter et al., US5,225,539, Carter, US6,407,213, Adair, US5,859,205, 6,881,557, Foote, US6,881,557) .. The acceptor antibody sequence may be, for example, a mature human antibody variable region sequence, a complex of such sequences, a consensus sequence of human antibody sequences (eg, Kabat, 1991, light chain and heavy chain variable region consensus sequences listed above), or It can be a germline variable region sequence. Thus, the humanized antibodies of the invention are three light chain CDRs and three heavy chain CDRs, as defined by Kabat from mouse antibodies (donor antibodies) such as 9E4, 5C1 or 1H7, either globally or substantially. Also includes antibodies with mature variable region framework sequences and constant regions, if present, from whole or substantially human antibody sequences. Similarly, humanized heavy chains are three heavy chain CDRs as defined by Kabat from the heavy chain of the donor antibody, as well as mature weights from the overall or substantially human antibody heavy chain sequence, if present. Includes heavy chains with variable chain sequences and heavy chain constant region sequences. Similarly, the humanized light chain matures from the three light chain CDRs as defined by Kabat from the light chain of the donor antibody, as well as the overall or substantially human antibody light chain sequence, if present. Includes light chains with light chain variable sequences and light chain constant region sequences. The maturation variable region framework sequence of the antibody chain or the constant region sequence of the antibody chain is at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or of the corresponding residues defined by Kabat. When 100% identical, they are substantially from the human mature variable region framework sequence or the human constant region sequence, respectively. A CDR is substantially from the corresponding donor CDR if it exhibits at least 85% identity to the corresponding donor CDR, except for CDRH2, which requires at least 65% identity. belongs to.

Certain amino acids from human mature variable region framework residues may be selected for substitution based on their possible effects on CDR conformation and / or binding to antigen. Investigation of such possible effects is by modeling, examining the characteristics of amino acids at specific positions, or empirically observing the effects of substitution or mutagenesis of specific amino acids.

For example, if an amino acid differs between a mouse maturation variable region framework residue and a selected human maturation variable region framework residue, then the amino acid is:
(1) Directly binds to an antigen by non-covalent bond,
(2) Adjacent to the CDR regions,
(3) Other interactions with the CDR regions (eg, within about 6 Å of the CDR regions).
(4) Human framework amino acids can be replaced by equivalent framework amino acids from mouse antibodies if reasonably predicted to mediate interactions between heavy and light chains.

The exemplary humanized mouse 9E4 antibody includes three exemplified humanized light chain maturation variable regions (Hu9E4VLv1 to v3; SEQ ID NOs: 3-5) and four exemplified humanized heavy chain maturation variable regions (Hu9E4VHv1). ~ V4; SEQ ID NOs: 8-11) are included.

The exemplary light and heavy chain maturation variable regions can be paired in any combination. An exemplary combination is Hu9E4VLv3 (SEQ ID NO: 5) and Hu9E4VHv3 (SEQ ID NO: 10). An exemplary heavy chain constant region is SEQ ID NO: 35 with or without C-terminal lysine. This constant region can be linked to any of the heavy chain variable regions. An exemplary light chain constant region is SEQ ID NO: 30. This constant region can be linked to any of the light chain variable regions. An exemplary full-length heavy chain has the amino acid sequence of SEQ ID NO: 37 with or without C-terminal lysine. An exemplary full-length light chain has the amino acid sequence of SEQ ID NO: 32. An exemplary combination is an antibody comprising SEQ ID NO: 37 (with or without C-terminal lysine) and SEQ ID NO: 32. Nucleic acids encoding the various heavy and light chains described immediately above are provided in SEQ ID NOs: 17-23. The humanized antibodies exemplified have heavy and light chains characterized by SEQ ID NOs: 37 (with or without C-terminal lysine) and 32.

The exemplary humanized mouse 5C1 antibody includes four exemplified humanized light chain maturation variable regions (5C1L1-L4; SEQ ID NOs: 69-72) and five exemplified humanized heavy chain maturation variable regions (5C1L1-L4; SEQ ID NOs: 69-72). 5C1H1-H4; SEQ ID NOs: 58-62) are included. The exemplary light and heavy chain maturation variable regions can be paired in any combination. An exemplary heavy chain constant region is SEQ ID NO: 35 with or without C-terminal lysine. This constant region can be linked to any of the heavy chain variable regions. An exemplary light chain constant region is SEQ ID NO: 30. This constant region can be linked to any of the light chain variable regions. Nucleic acids encoding the various heavy and light chains described immediately above are provided in SEQ ID NOs: 73-76 and 63-67.

The exemplary humanized mouse 1H7 antibody includes four exemplified humanized light chain maturation variable regions (Hu1H7VLv1-v4; SEQ ID NOs: 109, 111, 113, and 115, respectively) and five exemplified humanizations. Includes heavy chain maturation variable regions (Hu1H7VHv1 to v5; SEQ ID NOs: 95, 97, 99, 101, and 103, respectively). The exemplary light and heavy chain maturation variable regions can be paired in any combination. Illustrative combinations are Hu1H7VHv3 (SEQ ID NO: 99) and Hu1H7VLv3 (SEQ ID NO: 113). An exemplary heavy chain constant region is SEQ ID NO: 35 with or without C-terminal lysine. This constant region can be linked to any of the heavy chain variable regions. An exemplary light chain constant region is SEQ ID NO: 30. This constant region can be linked to any of the light chain variable regions. The illustrated full-length heavy chain has the amino acid sequence of SEQ ID NO: 127 (with G1m3). An exemplary full-length light chain has the amino acid sequence of SEQ ID NO: 125 (without arginine). An exemplary combination is an antibody comprising SEQ ID NO: 127 and SEQ ID NO: 125. Nucleic acids encoding the various heavy and light chains described immediately above are provided in SEQ ID NOs: 108, 110, 112, 114, 94, 96, 98, 100, 102.

Such humanized antibodies are described in WO06 / 02581 and US13 / 750,983, 14 / 049,169, 13 / 662,261, and 61 / 843,011.

In these or other antibodies described herein, one or more terminal residues may be cleaved during post-translational processing, especially for the terminal lysine of the heavy chain constant region.

C. Chimeric and Veneered Antibodies The present invention further provides chimeric and veneered non-human antibodies, in particular 9E4, 5C1 or 1H7.

A chimeric antibody is an antibody in which the light and heavy chain maturation variable regions of a non-human antibody (eg, mouse) are combined with the light and heavy chain constant regions from antibodies of different species. Typically, the light and heavy chain constant regions are of human origin, whereas constant regions are optionally (eg, to facilitate testing of non-human antibodies in suitable animal models) in rats. Can originate from different non-human species such as. Such antibodies are approximately two-thirds of the human (or different non-human species) sequences that substantially or totally retain the binding specificity of the non-human (eg, mouse) antibody that supplies the variable region. ..

Veneerized antibodies retain some and usually all of the CDRs of non-human antibodies as well as some of the non-human variable region framework residues, but other variable region framework residues that can contribute to B or T cell epitopes. A type of humanized antibody that replaces a group, eg, an exposed residue (Padlan, Mol. Immunol. 28: 489, 1991), with a residue from the corresponding position in the human antibody sequence. The result is an antibody in which the CDR is entirely or substantially from a non-human antibody, and the variable region framework of the non-human antibody has been made more human-like by substitution. The veneer type 9E4 is included in the present invention.

D. Selection of constant regions The heavy and light chain variable regions of chimeric, veneered or humanized antibodies can be linked to at least a portion of the human constant region. The choice of constant region depends in part on whether antibody-dependent cell-mediated cytotoxicity, antibody-dependent cellular cytotoxicity and / or complement-dependent cytotoxicity is desired. For example, human isotopes IgG1 and IgG3 have complement-dependent cytotoxicity and human isotopes IgG2 and IgG4 do not. Human IgG1 and IgG3 also induce stronger cell-mediated effector function than human IgG2 and IgG4. The light chain constant region can be a lambda or kappa. An exemplary human light chain kappa constant region has the amino acid sequence of SEQ ID NO: 13. Some such light chain kappa constant regions can be encoded by the nucleic acid sequence. The N-terminal arginine of SEQ ID NO: 13 may be omitted, in which case the light chain kappa constant region has the amino acid sequence of SEQ ID NO: 30. Some such light chain kappa constant regions can be encoded by the nucleic acid sequence. An exemplary human IgG1 heavy chain constant region has the amino acid sequence of the heavy chain constant region component of SEQ ID NO: 15 (with or without C-terminal lysine) or SEQ ID NO: 37 (with or without C-terminal lysine). Some such heavy chain constant regions can be encoded by nucleic acid sequences. Antibodies are as tetramers containing two light chains and two heavy chains, as separate heavy chains, as light chains, as Fab, Fab', F (ab') 2, and Fv, or as heavy and light chains. The chain maturation variable domain can be expressed as a single chain antibody linked through a spacer.

The human constant region exhibits allotype and isoarotype variations between different individuals, i.e., the constant region can differ in one or more polymorphic positions in different individuals. Isotypes differ from allotypes in that serum that recognizes isotypes binds to non-polymorphic regions of one or more other isotypes. Thus, for example, another heavy chain constant region is an IgG1 G1 m3 allotype heavy chain constant region, which has an amino acid sequence encoding the constant region of SEQ ID NO: 37. Yet another heavy chain constant region has an amino acid sequence encoding the constant region of SEQ ID NO: 37, except that it lacks a C-terminal lysine.

One or more amino acids at the amino or carboxy terminus of the light and / or heavy chain, such as the C-terminal lysine of the heavy chain, may be deleted or derivatized in some or all of the molecule. Substitution is performed in the constant region to reduce or increase effector function such as complement-mediated cytotoxicity or ADCC (eg, Winter et al., US5,624,821; Tso et al., US5,834,597; And Lazar et al., Proc. Natl. Acad. Sci. USA 103: 4005, 2006) or prolong the half-life in humans (eg Hinton et al., J. Biol. Chem. 279: 6213, See 2004). Illustrative substitutions include Gln at position 250 and / or Leu at position 428 to increase the half-life of the antibody (EU numbering is used for the constant region in this paragraph). Substitution at any or all of positions 234, 235, 236 and / or 237 reduces affinity for Fcγ receptors, especially FcγRI receptors (see, eg, US6,624,821). Some antibodies have alanine substitutions at positions 234, 235 and 237 of human IgG1 to reduce effector function. Positions 234, 236 and / or 237 in human IgG2 may be replaced by alanine and position 235 by glutamine (see, eg, US 5,624,821).

E. Expression of recombinant antibodies Antibodies can be produced by recombinant expression. The nucleic acid encoding the antibody can be codon-optimized for expression in the desired cell type (eg, CHO or Sp2 / 0). The recombinant nucleic acid construct typically comprises an expression control sequence operably linked to the coding sequence of the antibody strand, which comprises a naturally associated or heterologous promoter region. The expression control sequence can be a eukaryotic promoter system in a vector capable of transforming or transfecting a eukaryotic host cell. Once the vector is incorporated into a suitable host, the host is maintained under suitable conditions for expression of high levels of nucleotide sequences, as well as for recovery and purification of cross-reactive antibodies. One or more vectors encoding the antibody strand may also contain selectable genes such as dihydrofolate reductase to allow amplification of copy number of the nucleic acid encoding the antibody strand.

E. coli is a prokaryotic host that is particularly useful for the expression of antibodies, especially antibody fragments. Microorganisms such as yeast are also useful for expression. Saccharomyces is an exemplary yeast host, and suitable vectors have expression control sequences, origins of replication, termination sequences and the like, if desired. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include, in particular, promoters from alcohol dehydrogenase, isocytochrome C, and enzymes involved in maltose and galactose utilization.

Mammalian cells can be used to express a nucleotide segment that encodes an immunoglobulin or fragment thereof. See Winnacker, From Genes to Clones, (VCH Publishers, NY, 1987). Several suitable host cell lines capable of secreting intact heterologous proteins have been developed in the art, and the host cell lines include CHO cell lines, various COS cell lines, HeLa cells, and HEK293 cells. , L cells, and non-antibody-producing myeloma containing Sp2 / 0 and NS0. It may be advantageous to use non-human cells. Expression vectors for these cells include expression control sequences such as origin of replication, promoters, enhancers [Queen et al., Immunol. Rev. 89:49 (1986)], as well as ribosome binding sites, RNA splice sites, and polyadenylation. It may include sites and required processing information sites such as transcription terminator sequences. A suitable expression control sequence is a promoter derived from an endogenous gene, cytomegalovirus, SV40, adenovirus, bovine papillomavirus and the like. See Co et al., J. Immunol. 148: 1149 (1992).

Once the vectors encoding the antibody heavy and light chains have been introduced into the cell culture, the cell pool can be screened for growth productivity and product quality in serum-free medium. The top-producing cell pool can then be subjected to FACS-based single cell cloning to generate monoclonal strains. Specific productivity greater than 50 pg or 100 pg per cell per day, corresponding to a product titer greater than 7.5 g per liter of culture, may be advantageous. Antibodies produced by single cell clones can also be tested for turbidity, filtration properties, PAGE, IEF, UV scans, HP-SEC, carbohydrate-oligosaccharide mapping, mass spectrometry, and binding assays such as ELISA or Biacore. .. The selected clones can then be stored in multiple vials and cryostored for subsequent use.

Once the antibody is expressed, the antibody can be purified according to standard procedures in the art, including protein A capture, column chromatography (eg, hydrophobic interaction or ion exchange), virus-free. Examples include low pH for activation [generally see Scopes, Protein Purification (Springer-Verlag, NY, 1982)].

Methods for commercial production of antibodies include codon optimization, selection of promoters, transcription elements, and terminators, serum-free single cell cloning, cell banking, and copy count amplification. Use of selective markers, CHO terminators, serum free single cell cloning, improved protein titers (eg, US5,786,464, US6,114,148, US6,063,598). , US7,569,339, WO2004 / 050884, WO2008 / 021424, WO2008 / 021442, WO2005 / 019442, WO2008 / 107388, and WO2009 / 027471, and US5,888,809).

V. Formulations The formulations of the present invention (also known as pharmaceutical compositions) are antibodies (eg, chimeric, veneered or humanized mouse 9E4 (ATCC Accession No. PTA-8221)) or antigen-binding fragments thereof, buffered. The agent comprises one or more sugars and / or polyols and a surfactant and has a pH in the range of about 5 to about 7.5. The formulations may be prepared for storage in liquid or lyophilized form. When stored in lyophilized form, the formulation can be reconstituted with liquids (eg, sterile water) to the concentrations and properties described herein. If it is stated that the lyophilized composition can be reconstituted by adding water to produce a formulation of the specified component concentration and pH, the lyophilized formulation simply adds water (ie, additional). It means that it can be so reconstituted by supplying an amount of the component or adding an acid or base to change the pH). The concentration and properties of the pre-lyophilized liquid formulation may also be as described below if the lyophilized formulation is reconstituted to the same volume as the pre-lyophilized formulation. If the volumes are different, the concentration of the formulation should be adjusted proportionally. For example, if the reconstituted volume is half the pre-lyophilized volume, the concentration of the component in the pre-lyophilized formulation should be half the concentration in the reconstituted formulation.

Some formulations include a filler, which may or may not be the same as the sugar / polyol component. Typically, the formulation is sterile, which is achieved, for example, by sterile filtration using a 0.2 μm or 0.22 μm filter. Some formulations have ≦ about 3 CFU / 30 mL bioburden. Some formulations contain ≦ about 0.1 EU / mg of bacterial endotoxin. The formulations of the present invention are also generally stable upon freezing and thawing by fragmentation and / or aggregation from low to undetectable levels as further defined below. Other formulations are stable at 40 ° C. for at least 3 months after reconstitution of the lyophilized cake. In some formulations, less than about 10% of the antibody is present as aggregates in the formulation. In some formulations, about 5% or less of the antibody is present as aggregates in the formulation.

In some formulations, the antibody is present at a concentration in the range of about 5 mg / mL to about 100 mg / mL. In some formulations, the antibody is present at a concentration in the range of about 5 mg / mL to about 50 mg / mL. In some formulations, the antibody is present at a concentration in the range of about 25 mg / mL to about 50 mg / mL. For example, the antibody may be present at a concentration of about 35-45 mg / ml or about 40 mg / mL. The antibody may be present in about 50 mg / vial to about 500 mg / vial, or more sterile liquid dosage forms. The antibody may be present in the lyophilized form of about 40 mg / vial to about 500 mg / vial. For example, the antibody may be present in a sterile liquid or lyophilized form of about 250-350 mg / vial or about 200 mg / vial.

Antibodies used in the disclosed formulations promote the activity of cytotoxic agents, radiotherapeutic agents, immunomodulators, second antibodies (eg, to form antibody heteroconjugates), or antibodies to be formulated. Alternatively, it can be coupled with a therapeutic portion such as any other bioactive agent that enhances. However, antibodies are typically used in naked form (ie, not conjugated to any of these moieties).

The antibody to be blended may be any of the above antibodies, including any of the above chimeric, veneered or humanized antibodies 9E4.

Buffers are used in formulations disclosed to achieve suitable pH for antibodies, such as histidine, succinic acid, and citric acid buffers. Some formulations have a pH in the range of about 5.5 to about 7, such as 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, It has a pH of 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0. Some formulations have a pH between about 5.5 and about 6.5. Some formulations have a pH of about 6.0 and others have a pH of about 6.5. In some formulations, the citrate or succinic acid buffer is present at a concentration in the range of about 10 mM to about 30 mM, eg, a concentration of about 15 to 25 mM or about 20 mM. Some citrate buffers contain sodium citrate dehydrated and citrate monohydrate at concentrations in the range of about 15 mM to about 20 mM and in the range of about 2 mM to about 6 mM, respectively.

Suitable sugars and / or polyols for the formulation include trehalose, sucrose, mannitol, or a combination thereof. The sugar / polyol acts as a filler, cryoprotectant, and / or tonicity adjuster. For example, some formulations have trehalose present at concentrations in the range of about 220 mM to about 260 mM, sucrose present at concentrations in the range of about 220 mM to about 260 mM, or concentrations in the range of about 20 mM to about 40 mM. Contains a mixture of sucrose present in and mannitol present at concentrations in the range of about 200 mM to about 220 mM. Some formulations contain trehalose, which is present at a concentration of about 230 mM or 240 mM. Other formulations include sucrose present at a concentration of about 230 mM or 240 mM. Other formulations include a mixture of sucrose present at a concentration of about 50 mM and mannitol present at a concentration of about 200 mM. Another formulation comprises a mixture of sucrose present at a concentration of about 28 mM and mannitol present at a concentration of about 212 mM. Some such formulations are characterized by a weight osmolal concentration in the range of about 250-400, 300-400, or 300-350 mOsm / kg, eg, 335 mOsm / kg.

The formulation may contain a surfactant to reduce antibody aggregation and absorption on the surface. Suitable surfactants include polysorbate 20 (PS20), which is present at a concentration in the range of about 0.005% by weight to about 0.05% by weight. PS20 protects against significant increases in agglutination or turbidity that would otherwise occur in the formulation of 9E4 antibody. The polysorbate 20 may be present at a concentration in the range of about 0.01% to about 0.05%. For example, the concentrations are 0.005%, 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.035%, 0.04%, 0.045%, Or it can be 0.05%. Alternatively, in some formulations, the polysorbate 20 is approximately 0.05 g / L, 0.1 g / L, 0.15 g / L, 0.2 g / L, 0.25 g / L, 0.3 g / L. It is present at a concentration in the range of L, 0.35 g / L, 0.4 g / L, 0.45 g / L, or 0.5 g / L. Some formulations contain polysorbate 20 at a concentration of 0.2 g / L (ie, 0.163 mmol / L).

An exemplary formulation (liquid, pre-sucrose-dried or post-sucrose reconstruction) is characterized by a pH in the range of about 5.5 to about 7, and (a) about 10 mg / ml to about 50 mg / ml. Chimeric, veneered, or humanized antibodies 9E4 at concentrations within the range of, or fragments thereof that specifically compete with 9E4 for binding to the antigen; (b) at concentrations within the range of about 10 mM to about 30 mM. Citrate or succinate buffers present; (c) trehalose present at concentrations in the range of about 220 mM to about 260 mM, sucrose present at concentrations in the range of about 220 mM to about 260 mM, and sucrose present at concentrations in the range of about 220 mM to about 260 mM, and about 20 mM to about. One or more sugars and polyols (“sugar / polyol”); and ("sugar / polyol") selected from a mixture of sucrose present at a concentration in the range of 40 mM and mannitol present at a concentration in the range of about 200 mM to about 220 mM. d) Contains polysolvate 20 present at a concentration in the range of about 0.005% by weight to about 0.05% by weight. For example, the formulation comprises (a) a light chain having the amino acid sequence set forth in SEQ ID NO: 32 and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 37 with or without C-terminal lysine at a concentration of approximately 40 mg / mL. Amino acids present in (b) citrate buffer at a concentration of about 20 mM; (c) trehalose at a concentration of about 230 mM; (d) polysolvate 20 at a concentration of about 0.02%; and a pH of about 6.0. Can include.

Some lyophilized formulations include (a) humanized antibody 9E4 or antigen-binding fragments thereof; (b) citrate; (c) trehalose; and polysorbate 20. The lyophilized formulation may contain about 200 mg of antibody. Some lyophilized formulations can be reconstituted with sterile water. Some lyophilized formulations are 100-300 or 150-250 mg of 9E4 antibody, 15-35 or 20-25 mg of sodium citrate dehydrate, 1.65-2.75 or 2-3 mg of citric acid. It contains 360-500 or 400-470 mg of trehalose dehydrated product, and 0.5-1.5 mg or 0.75-1.25 mg of polysorbate 20. An exemplary lyophilized formulation is 200 mg 9E4 antibody (eg, humanized 9E4 antibody), 25 mg sodium citrate dehydration, 2.15 mg citrate monohydrate, 435 mg trehalose dehydration, and 1 mg. Includes polysorbate 20. Another exemplary lyophilized formulation is 200 mg 9E4 antibody (eg, humanized 9E4 antibody), 25 mg sodium citrate dehydration, 3.15 mg citrate monohydrate, 435 mg trehalose dehydration, and. Contains 1 mg of polysorbate 20. Such formulations can be reconstituted to a volume of about 5 ml. Other lyophilized formulations have the same proportions as any of those disclosed in this paragraph, but in different amounts (eg, 400 mg antibody, 50 mg sodium citrate, 4.3 mg citrate monohydrate, 870 mg trehalose). Contains dehydrated product, and the same components of 2 mg of polysorbate 20).

The lyophilized formulation has an antibody concentration of about 30-50 or 35-45 mg / mL, such as about 40 mg / mL; (b) citrate buffer present at a concentration of about 10-30 or 15-25 mM, preferably about 20 mM. Agents; (c) trehalose present at a concentration of about 160-330 or 200-260 mM, eg about 230 mM; (d) about 0.1-0.3 or 0.15-0.25 g / L, eg about 0. Polysorbate 20 present at a concentration of 2 g / L; and (e) can be reconstituted to a pH of about 5.5-6.5, for example about 6.0.

The liquid or reconstituted lyophilized formulation can be substantially isotonic, implying a weight osmolal concentration of about 250-350 mOsm / kg water. Some formulations have a weight osmolal concentration of about 335 mOsm / kg. Some formulations have a weight osmolal concentration of 270-300 mOsm / kg. The liquid or reconstituted lyophilized formulation can also be hypertonic or hypotonic (<250 mOsm / kg water) of> 350 mOsm / kg water.

Any of the formulations described can be made without the use of pharmaceutical excipients or carriers other than those described as ingredients herein. Such formulations may consist of or essentially consist of the listed ingredients, in the presence of small amounts of other ingredients that do not affect the properties of the formulation. Formulations can be made under FDA-approved or approved Good Manufacturing Practices (GMP) for the preparation of drugs for administration to humans.

VI. Diagnostic Criteria for Synuclein Disease The methods of the invention are such that they have been diagnosed with Lewy body disease by a qualified healthcare professional, or are substantiated by the disease's genetic or biochemical markers, family history or prodrome. It is commonly performed on subjects at increased risk compared to the general population. Such individuals include anyone who has been given a previous prescription for the treatment or prevention of Lewy body disease. Diagnosis of synuclein disease is recognized in the art for the likelihood or high likelihood of Lewy body disease such as DSM-V or DSM IV-TR, the Lewy Body dementia association, and the Parkinson's disease societies. Obtained based on criteria. However, the diagnosis can also be based on the presence of any sign or symptom of Lewy body disease that leads the therapist to conclude that the subject is likely to have Lewy body disease. Illustrative criteria for diagnosing the likelihood or high likelihood of PD are shown below.
Group A: Occurrence of resting tremor, bradykinesia, rigidity and asymmetry Group B characteristics: Suggestions for selective diagnosis Significant postural instability in the first 3 years after symptom onset Freezing phenomenon in the first 3 years Drug-unrelated hallucinations in the first 3 years Prior to motor symptoms or dementia in the first year Nuclear gaze palsy (other than upward gaze restrictions) or slowing of vertical sacking Severe drug-unrelated symptoms Sexual autonomic imbalance

A record of conditions known to cause Parkinson's disease and plausibly associated with the subject's condition (eg, well-positioned local brain injury or use of neuroleptic agents within the last 6 months).

Criteria for diagnosing the possibility of Parkinson's disease:
There are at least two of the four features in group A; at least one of these is tremor or slow movement, none of the features in group B is present, or symptoms are present for less than 3 years. , None of the features in Group B to date; and a substantive and sustained response to levodopa or dopamine agonists has been recorded, or subjects have been thoroughly tested for both levodopa and dopamine agonists. Not done.

Diagnostic criteria for high likelihood of Parkinson's disease:
At least three or four features in group A are present, none of the features in group B are present, and a substantive and sustained response to levodopa or dopamine agonists has been recorded.

Illustrative criteria for the diagnosis of Lewy body dementias are:
Presence of dementia At least two of the three core features:
Upset of attention and concentration,
Recurrent, well-formed hallucinations, as well as spontaneous Parkinson's disease-like motor signs.
Suggestive clinical features include:
Rapid eye movement (REM) sleep behavior disorder Severe neuroleptic hypersensitivity Low dopamine transporter uptake in the basal ganglia demonstrated by SPECT or PET imaging

In the absence of two core features, a diagnosis of the high likelihood of DLB can also be made if at least one suggestive feature is present with one core feature in addition to dementia.

The potential for DLB can be diagnosed with the presence of one core or suggestive feature in addition to dementia.

Early warning signs of REM behavior disorder include, for example, EEG slowing, neuropsychiatric manifestations (depression, dementia, illusion, anxiety, indifference, insomnia), autonomic changes (hypersomnia). Orthostatic hypotension, bladder disorder, constipation, fecal incontinence, salivation, swallowing disorder, sexual dysfunction, changes in cerebral blood flow), sensory changes (smell, pain, abnormal sensation of color discrimination), sleep disorders [REM sleep behavior Disorder (RBD), Restless Leg Syndrome / Regular Extreme Exercise, Hypersomnia, Insomnia] and various other signs and symptoms (fatigue, compound vision, blurred vision, seborrhea, weight loss / gain) ). Genetic markers of risk to PD include mutations in the alpha-synuclein or Parkin, UCHLI, and CYP2D6 genes, in particular mutations at positions 30 and 53 of the alpha-synuclein gene. Neither of these genetic markers or early warning signs are themselves a diagnosis of Lewy body disease, but they may contribute to the diagnosis of Lewy body disease individually or in combination.

VII. Therapeutic regimens In therapeutic applications, regimens known or presumed to be effective in ameliorating at least one sign or symptom of a disease or at least inhibiting further exacerbations (dose, frequency and route of administration). The antibody is administered to a subject diagnosed with synuclein disease in. In prophylactic applications, there is still an increased risk of synuclein disease in regimens known or presumed to be effective in inhibiting or delaying the development of at least one sign or symptom of the disease. Antibodies are administered to subjects who do not have sufficient symptoms to be diagnosed with the disease.

P <0.2, 0.1, 0.05 or 0.01 or even 0.001 in controlled animal models or clinical trials (eg, phase II, phase II / III or phase III trials) The regimen is considered to be therapeutically or prophylactically effective if a more favorable outcome is demonstrated in the treated subject compared to the control subject at the level of. However, due to changes in genetics, subject characteristics and environment, and disease subtypes between individual subjects, a regimen that is effective in one subject may or may not be effective in another. It can be effective.

An exemplary dosage range of the antibody is 3000-5000 mg of antibody to alpha-synuclein administered intravenously at intervals of 3-5 weeks, such as every 4 weeks. For some subjects, the dose is 3500-4500 mg every 3-5 weeks, such as every 4 weeks. Subjects may receive the same or different doses from each other (eg, depending on the subject's weight). In some methods, the subject receives one of two fixed doses. For example, a subject weighing less than 65 kg can receive 3500 mg and a subject weighing 65 kg or more can receive 4500 mg. In some methods, the dosage range for at least some subjects is in the range of 45-75 mg / kg, eg, 50-70 mg / kg, 45 mg / kg, 60 mg / kg or 65 mg / kg. Dosages are usually administered at intervals of 3-5 weeks, eg, every 28 days or every 4 weeks, or every calendar month on multiple occasions. The subject may receive at least 6, 9, 12 or 18 doses at such intervals, or may be administered while the symptoms of the condition persist or for the rest of the subject's life. In some regimens, after the initial loading dose of 2000 mg is administered, it is administered in a range of 2000 mg or more but less than the intended target dose until the intended target dose is reached. For example, a subject may be titrated to a dose of 3500 mg or a dose of 4500 mg after receiving an initial loading dose of 2000 mg. Increasing titration can be performed in a single subsequent dose or in a gradual increase over multiple doses until a target dose or dose within the target range is reached. For example, a subject can receive a starting dose of 2000 mg followed by a subsequent dose of 3500 mg. Alternatively, the subject may receive a starting dose of 2000 mg followed by one or more subsequent doses of 2000 mg or more but less than 3500 mg, and a subsequent dose of 3500 mg. Similarly, a subject can receive a starting dose of 2000 mg followed by a subsequent dose of 4500 mg. Alternatively, the subject may receive a starting dose of 2000 mg followed by a subsequent dose of 2000 mg or more but less than 4500 mg and a subsequent dose of 4500 mg. In some regimens, subjects receive a dose of 3000-5000 mg of antibody intravenously every 4 weeks for at least 52 weeks. For subjects receiving multiple dose regimens at doses within a specified range, such as 3500-5000 mg, the subject may receive the same or different doses within the specified range at each dose. For some regimens, the subject receives the same dose within the specified range for each dose.

In another exemplary regimen, a dose of 1300 to 1700 mg of antibody is administered intravenously to the subject at intervals of 3-5 weeks. An exemplary dose is 1500 mg. The subject may receive a single fixed dose or two or more different doses within this range, eg, based on the subject's body weight. Some subjects administered within this range receive 18-25 mg / kg, eg 20 mg / kg of antibody. As in other methods, the intervals can be 3-5 weeks, for example every 4 weeks or every calendar month. The subject may receive at least 6, at least 9, at least 12, or at least 18 doses, or may be administered at such intervals during the residual symptoms or for the rest of the subject's life.

For any of the above treatment regimens, the area under the curve can be calculated to indicate the amount of antibody delivered over time. Alternative treatment regimens can be devised to deliver substantially the same area under the curve (eg, within 25, 20 or 15%) (eg, using different routes of administration, frequency or dose). Preferably, such an alternative treatment regimen does not substantially exceed the Cmax of the designated regimen (eg, 25, 20 or 15% or less). Other routes of administration include topical, oral, subcutaneous, intraarterial, intracranial, intramedullary, intraperitoneal, intranasal or intramuscular.

Any treatment regimen may involve monitoring the subject being treated for changes in motor and / or cognitive impairment. Preferably, such monitoring involves at least one assessment before and after the start of treatment. Preferably, monitoring indicates a reduction in exercise and / or cognitive impairment in response to treatment compared to before initiation of treatment, or in a control patient who has not been given a previous rate of reduction or any immunotherapy in the subject. At least indicates a reduction in the rate of decline compared to the rate of decline. Subjects may also be monitored for autonomic dysfunction, gastrointestinal dysfunction, hallucinations or changes in one or more psychological symptoms, among other symptoms or symptoms.

Symptoms of the subject, such as tremor, stiffness and slowness of movement, may be monitored. A wearable system or an on-body sensor may be used to assess and quantify the motor symptoms of the subject. "On-body sensors" may be used in laboratory situations or in free living conditions [S. Del Din, et al., J. of NeuroEngineering and Rehabilitation, 2016 13:46].

Subjects may be monitored using mobile device-based monitoring. The mobile device may be a smartphone, smartwatch, wearable sensor, portable multimedia device or tablet computer. A built-in mobile device sensor may be used to record the subject's daily activities. The subject may carry a mobile device to record daily activities. Mobile device-based assessments and sensors may be used for remote passive monitoring of gait and mobility in a subject being treated, for example for Parkinson's disease. [For example, Lipsmeier, F., et al .. Mov Disord. 2017; 32 (suppl 2); WY Cheng et al., 2017 IEEE / ACM International Conference on Connected Health: Applications, Systems and Engineering Technologies (CHASE), Philadelphia , PA, 2017, pp. 249-250]. Sensor data may be analyzed by machine learning based activity profiling. Gait and mobility can correlate with MDS-UPDRS used in the clinic to assess the severity of Parkinson's disease.

Some mobile device-based monitoring (a) receives and receives data obtained from mobile device internal and / or external sensors regarding movement disorders in subjects with or suspected of having sinucrane disease. A mobile device programmed to transmit is provided to the subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device relate to the movement. Obtaining data, (b) collecting data transmitted from mobile devices, and (c) comparing the data obtained from the subject with control data to assess the presence or extent of movement disorders in the subject. May include that. In some mobile device-based monitoring, the mobile device is programmed to receive and transmit data from at least two external sensors attached to the subject's upper and lower limbs. In some mobile device-based monitoring, the mobile device acquires data from sensors on the subject's upper and lower limbs. In some mobile device-based monitoring, the mobile device is carried by the subject and acquires data from internal sensors. In some mobile device-based monitoring, the sequence of exercises involves tapping, sitting and standing on the device.

The regimen of the present invention may be administered in combination with another agent that is effective in treating or preventing the disease being treated. Other agents may be other immunotherapeutic agents described herein or other agents for treating Parkinson's disease, such as levodopa, benzaseride, carbidopa, dopamine agonists, non-wheat dopamine agonists, catechol. -O-Methyl (“COMT”) inhibitors such as monoamine oxidase (“MAO”) inhibitors such as entacopone or turcopone (“MAO”) inhibitors such as rasagaline, amantadine and the like can be mentioned or anti. Choliners can be used in combination with the regimens of the invention. Some such other agents reduce one or more symptoms of the disease without affecting the causative factors.

All cited publications (including GenBank accession numbers, UniProtKB / Swiss-Prot accession numbers, etc.), patents and patent applications are all by reference by each individual publication, patent and patent application. All of them are incorporated herein by reference in their entirety to the same extent as specifically and individually indicated to be incorporated for the purposes of. If there are any changes in the sequence or website, or tissue disease criteria associated with Genbank and UniProt KB / Swiss-Prot accession numbers, etc., this application refers to one valid on its effective filing date.

VIII. Example

Phase II clinical trial design for alpha-synuclein antibody: Phase II trials are heavy chain variable regions designated by SEQ ID NO: 10 and light chains designated by SEQ ID NO: 5 for subjects with Parkinson's disease. It is performed on alpha-synuclein antibodies with variable regions. The study has two treatment groups and one control group. Subjects are randomized to groups 1: 1: 1 (N = 300). The initial phase of the study is a 52-week double-blind treatment. During the early phase of the study, subjects do not receive other treatments for Parkinson's disease, including symptomatological treatments. Subjects in one treatment group receive a fixed dose of 1500 mg of antibody intravenously every 4 weeks. Subjects in other treatment groups receive 3500 mg or 4500 mg of antibody intravenously every 4 weeks, depending on body weight, subjects under 65 kg receive lower doses, subjects weighing 65 kg or more receive higher doses. Receive. Subjects in the second group receive a loading dose of 2000 mg and an additional titrated dose of 2000 mg or more until a target dose of 3500 mg or 4500 mg is reached as appropriate. Administration is continued for 1 year. The study then has an extension period, in which the subject in the placebo group first receives one of the two treatment regimens from the early phase and the subject from the early phase treatment group receives the same treatment as before. to continue. During the extended phase of the study, the subject may receive systematic treatment with the antibody subject of the study in addition to levodopa, but no other treatment for Parkinson's disease.

Purpose:
The primary objective is to use the Movement Disorder Society (MDS) -sponsored revision of the Unified Parkinson's Disease Racing Scale (MDS-UPDRS) and sequence number 5 variable to sequence number 10. To evaluate the efficacy of an alpha-synuclein antibody having a designated light chain variable region.
The secondary purpose is
-Evaluating the effect of the antibody on DaT-SPECT signals and signs and symptoms of Parkinson's disease, including changes in motor and cognitive impairment-Evaluating the safety and tolerability of the antibody for up to 104 weeks, and-Antibody Is to evaluate the pharmacokinetics of.

Passive monitoring of mobility in patients with early-stage Parkinson's disease in Phase I alpha-synuclein antibody clinical trials using smartphone sensors

1. 1. Abstract-Walking and mobility in patients with early stage Parkinson's disease (PD) were measured using smartphone-based passive monitoring. In a multiple escalating dose clinical trial of an alpha-synuclein antibody having a heavy chain variable region designated by SEQ ID NO: 10 and a light chain variable region designated by SEQ ID NO: 5, 44 PD patients and 35 age and sex matches Healthy individuals were subjected to smartphone-based assessments for up to 24 weeks and up to 6 weeks, respectively [Lipsmeier, F., et al .. Mov Disord. 2017; 32 (suppl 2); WY Cheng et al., 2017 IEEE / ACM International Conference on Connected Health: Applications, Systems and Engineering Technologies (CHASE), Philadelphia, PA, 2017, pp. 249-250].

For "passive monitoring", the subject carried a smartphone as part of his daily routine and continuously recorded exercise data with the smartphone's sensor. A total of over 30,000 hours of passive monitoring data was collected. To classify sensor signals into activity profiles, a Human Activity Recognition (HAR) model was constructed using a deep neural network (DNN) trained with previously published data. Participant activity profiles determined by the HAR model showed significant differences between PD patients and healthy controls in the percentage of walking time and the frequency with which controls changed posture (seating and standing and standing). ..

2. 2. Method

A. Data collection and analysis focused solely on investigating the differences between the HC cohort and the PD cohort, not antibody-related effects. In total, 24,104 hours of passive monitoring data were recorded for the PD cohort and 8,614 hours for the HC cohort. Accelerometer data were filtered out when the standard deviation of the Euclidean norm below 0.03 m / s2 was longer than 30 minutes, in line with the approach of Rai, A et al. (MobiCom'12, August 22-26, 2012]. However, this is because the smartphone appears to have not been carried by the subject during these spans. This step removed 14% of passive monitoring data.

B. HAR
A 9-layer neural network model structure was used. A similar structure has been previously used for HAR and has been shown to surpass traditional machine learning methods (FJ Ordonez and D. Roggen, Sensors 2016, 16, 115). HAR model with two public data Set (GM Weiss and JW Lockhart, Proceedings of the AAAI-12 Workshop on Activity Context Representation: Techniques and Languages, Toronto, CA. 2012; A. Stisen, et al., 13th ACM Conference on Embedded Networked Sensor Systems, Seoul, Korea , 2015) trained to classify 6 activities: walking, stairs, jogging, sitting, standing, and lying down. Continuous accelerometer data downsampled to 20 Hz with 75% overlap with adjacent ones 4 Segmented into a seconds window.

3. 3. result

A. Verification of human activity recognition results:
To ensure that the HAR model can accurately translate the sensor data into activity profiles, the model's performance was first analyzed in a holdout validation set. The HAR model was able to correctly distinguish between resting activity (sitting, standing, lying down) and walking activity (walking, stairs, jogging) with greater than 98% accuracy. Additional validation of labeled gait and rest data from test data also shows that the HAR model can successfully profile the gait segment with 96.9% accuracy and the rest segment with 99.5% accuracy. showed that.

B. Comparison of activity profiles The mobility of each subject was quantified by calculating the ratio of the time the subject engaged in walking activity (walking, stairs, jogging) to the total passive monitoring coverage time of the patient. The overall ratio of different gait activities to total coverage for the PD and HC cohorts was calculated. A median with a gait span of 9.7% of all coverage spans was detected in the PD cohort, as opposed to 15.1% of the HC cohort. The HC cohort had significantly higher levels of gait activity per subject than the PD cohort, with a Mann-Whitney test P-value of 2.43E-8.

C. Comparison of the Number of Sitting-to-Standing and Standing-to-Sitting Switches One manifestation of the functional effects of PD is sitting-to-standing and standing-to-sitting (sit-to-stand and). It has been observed to be in a stand-to-sit (STS) event (A. Zijlstra, et al., J. NeuroEngineering and Rehabilitation 2012, 9:75]. From the activity profile, each coverage normalized STS event Calculated for subjects. The number of STS medians per hour was observed in 1.44 PD patients, which was significantly lower than 1.74 in HC subjects. The P value of the Man-Whitney test between the two groups was It was 1.60E-8.

4. Conclusion

The results from this study show that it is feasible to measure gait and mobility in patients with early stage PD using smartphone-based passive monitoring. Sensor data collected during passive monitoring provides previously inaccessible, ecologically valid insights into the patient's daily behavior and function. Significant differences were observed between PD patients and healthy controls (HC).

Claims (107)

A method of performing treatment or prevention of a subject with or at risk of synuclein disease, the subject is given an antibody against alpha-synuclein at a dose of 3000-5000 mg intravenously at intervals of 3-5 weeks. That, or a method comprising administering to the subject another regimen that delivers the antibody in substantially the same subcurve area. The method of claim 1, wherein the dose is 3,000 to 4500 mg. The method of claim 1, wherein the method is administered to a plurality of subjects who have or are at risk of synuclein disease and the subjects receive one or two fixed doses. The method of claim 3, wherein a subject weighing less than 65 kg receives a dose of 3500 mg of the antibody and a subject weighing 65 kg or more receives a dose of 4500 mg. The method of claim 1, wherein the dose is 45-75 mg / kg. The method of claim 1, wherein the dose is 50-70 mg / kg. The method of claim 1, wherein the interval is 28 days. The method of claim 1, wherein the subject receives at least 6 doses of antibody at intervals of 3-5 weeks. The method of claim 1, wherein the subject receives at least 12 doses of antibody at intervals of 3-5 weeks. The method of claim 1, wherein the subject receives at least 18 doses of antibody at intervals of 3-5 weeks. The method according to any one of claims 8 to 10, wherein the interval is 4 weeks. The method of claim 1, wherein the subject receives the antibody at least every 52 weeks and 4 weeks. The method of claim 1, further comprising monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms. To monitor
(A) A mobile device programmed to receive and transmit data acquired from internal and / or external sensors of the mobile device regarding movement disorders of subjects with or suspected of having synuclein disease. Providing to the subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device acquire data on the movement,
Claims include (b) collecting data transmitted from mobile devices and (c) comparing data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject. 13. The method according to 13. 14. The method of claim 14, wherein the mobile device is programmed to receive and transmit data from at least two external sensors attached to the upper and lower limbs of the subject. The method of claim 14 or 15, wherein the mobile device acquires data from sensors on the upper and lower limbs of the subject. The method of claim 14 or 15, wherein the mobile device is carried by the subject and acquires data from an internal sensor. The method of claim 14 or 15, wherein the series of exercises comprises tapping, sitting and standing on the device. 13. The method of claims 13-18, wherein monitoring points to a reduction in movement or cognitive impairment in response to administration of the antibody. The method of claim 1, wherein the antibody binds within residues 115-130 of alpha-synuclein. The method of claim 1, wherein the antibody binds within residues 118-126 of alpha-synuclein. The method of claim 1, wherein the antibody binds within residues 117-123 of alpha-synuclein. The method according to any one of claims 1 to 19, wherein the antibody is NI-202.21D11. The method of any of claims 1-19, wherein the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 139 to 141 and three light chain CDRs designated by SEQ ID NOs: 143 to 145, respectively. The method of any of claims 1-19, wherein the antibody comprises a heavy chain variable region designated by SEQ ID NO: 138 and a light chain variable region designated by SEQ ID NO: 142. The method of claim 1, wherein the antibody comprises three light chain Kabat CDRs of SEQ ID NO: 5 and three heavy chain Kabat CDRs of SEQ ID NO: 10. The antibody comprises the heavy chain variable region of any of SEQ ID NOs: 8-11 and the light chain variable region of any of SEQ ID NOs: 3-5, preferably the heavy chain variable region is the heavy chain variable region of SEQ ID NO: 10. The method of claim 1, wherein the light chain variable region is the light chain variable region of SEQ ID NO: 5. The method according to any one of claims 1 to 27, wherein the antibody is an antibody of the human IgG1 isotype. 28. The method of claim 28, wherein the antibody comprises a heavy chain constant region of SEQ ID NO: 35 and a light chain constant region of SEQ ID NO: 30, where the C-terminal lysine of SEQ ID NO: 35 may not be present. 28. The method of claim 28, wherein the antibody comprises a heavy chain constant region of SEQ ID NO: 35 and a light chain constant region of SEQ ID NO: 13 in which the C-terminal lysine of SEQ ID NO: 35 may not be present. The method according to any one of claims 1 to 21, wherein the antibody is 9E4. The method of any of claims 1-21, wherein the antibody comprises three heavy chain CDRs, respectively designated by SEQ ID NOs: 146 to 148, and three light chain CDRs, respectively designated by SEQ ID NOs: 149 to 151. The invention according to any one of claims 1 to 21, wherein the antibody comprises a heavy chain designated by SEQ ID NO: 37 and a light chain designated by SEQ ID NO: 32, and the C-terminal lysine of SEQ ID NO: 37 may not be present. the method of. The method of claim 1, wherein the antibody binds within residues 1-20 of alpha-synuclein. 34. The method of claim 34, wherein the antibody binds within residues 4-15 of alpha-synuclein. The method according to any one of claims 1 to 19, wherein the antibody is NI-202.12F4. The method of any of claims 1-19, wherein the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 131-133 and three light chain CDRs designated by SEQ ID NOs: 135-137, respectively. The method of any of claims 1-19, wherein the antibody comprises a heavy chain variable region designated by SEQ ID NO: 130 and a light chain variable region designated by SEQ ID NO: 134. The method of any of claims 1-38, wherein the subject weighs less than 65 kg and is administered a dose of 3500 mg of antibody every 4 weeks. Prior to administration of the antibody in the range of 3000-5000 mg, administer the antibody at a loading dose of 2000 mg and in one or more subsequent doses of 2000 mg or more but less than 3500 mg until the dose of 3500 mg is reached. The method of any of claims 1-39, wherein incremental titration may be performed and all doses are separated at intervals of 3-5 weeks. 40. The method of claim 40, wherein a dose of 3500 mg is administered at the next interval after the dose of 2000 mg. 40. The method of claim 40, wherein the dose is gradually increased over multiple intervals prior to administration of a dose of 3500 mg. The method of any one of claims 1-38, wherein the subject weighs more than 65 kg and is administered a dose of 4500 mg of antibody every 4 weeks. Prior to administration of the antibody at 4500 mg, a loading dose of 2000 mg of antibody is administered and incremental titration is performed using one or more subsequent doses of 2000 mg or more but less than 4500 mg until a dose of 4500 mg is reached. The method of any of claims 1-38, wherein all doses are separated at intervals of 3-5 days. 44. The method of claim 44, wherein a dose of 4500 mg is administered at the next interval after the dose of 2000 mg. 44. The method of claim 44, wherein the dose is gradually increased over multiple intervals prior to administration of the 4500 mg dose. The method of any of claims 44-46, further comprising monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms. To monitor
(A) A mobile device programmed to receive and transmit data acquired from internal and / or external sensors of the mobile device regarding movement disorders of subjects with or suspected of having synuclein disease. Providing to the subject, after which the subject performs a series of exercises to reveal the movement disorder if there is a movement disorder, and sensors inside or outside the device acquire data on the movement,
Claims include (b) collecting data transmitted from mobile devices and (c) comparing data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject. 47. 48. The method of claim 48, wherein the mobile device is programmed to receive and transmit data from at least two external sensors attached to the upper and lower limbs of the subject. 48. The method of claim 48 or 49, wherein the mobile device acquires data from sensors on the upper and lower limbs of the subject. The method of claim 48 or 49, wherein the mobile device is carried by the subject and acquires data from an internal sensor. The method of claim 48 or 49, wherein the series of exercises comprises tapping, sitting and standing of the device. The method of any of claims 47-52, wherein monitoring points to a reduction in movement or cognitive impairment in response to administration of the antibody. The method according to any one of claims 1 to 53, wherein the synuclein disease is Parkinson's disease. The method according to any one of claims 1 to 53, wherein the synuclein disease is dementia with Lewy bodies. The method according to any one of claims 1 to 53, wherein the synuclein disease is plural atrophy. The method according to any one of claims 1 to 53, wherein the synuclein disease is progressive supranuclear palsy. The method according to any one of claims 1 to 53, wherein the synuclein disease is REM sleep behavior disorder. The method according to any one of claims 1 to 53, wherein the synuclein disease is Alzheimer's disease with Lewy bodies of the amygdala. The method of any of claims 1-59, wherein the subject has the disease. The method of any of claims 1-60, wherein the subject has not received symptomatological treatment for Parkinson's disease in combination with the antibody. The method according to any one of claims 1 to 60, wherein the antibody is administered in combination with levodopa. A method of performing treatment or prophylaxis for a subject with Lewy body disease, in which an antibody against alpha-synuclein at a dose of 1300 to 1700 mg is intravenously administered to the subject at intervals of 3 to 5 weeks, or substantially A method comprising administering another regimen that delivers the antibody in an area under the same curve. 63. The method of claim 63, wherein the dose is 1500 mg. 63. The method of claim 63, wherein the method is administered to a plurality of subjects with Lewy body disease and the subjects receive one or two fixed doses. 63. The method of claim 63, wherein the subject receives a dose of 18-25 mg / kg. The method of claim 66, wherein the subject receives a dose of 20 mg / kg. 63. The method of claim 63, wherein the interval is 28 days. 63. The method of claim 63, wherein the subject receives at least 6 doses of antibody at intervals of 3-5 weeks. 63. The method of claim 63, wherein the subject receives at least 12 doses of antibody at intervals of 3-5 weeks. 63. The method of claim 63, wherein the subject receives at least 18 doses of antibody at intervals of 3-5 weeks. The method according to any one of claims 68 to 70, wherein the interval is 4 weeks. The method of claim 63, wherein the subject receives the antibody at least every 52 weeks and 4 weeks. 63. The method of claim 63, further comprising monitoring the subject for motor changes, cognitive impairment, autonomic dysfunction, gastrointestinal dysfunction, hallucinations or psychological symptoms. To monitor
(A) Programmed to receive and transmit data acquired from internal and / or external sensors on mobile devices regarding movement disorders of subjects with or suspected of having Lewy body dementias. A mobile device is provided to the subject, after which the subject performs a series of exercises to identify the movement disorder in the event of a movement disorder, and sensors inside or outside the device acquire data on the movement.
Claims include (b) collecting data transmitted from mobile devices and (c) comparing data obtained from the subject with control data to assess the presence or degree of movement disorders in the subject. 74. 75. The method of claim 75, wherein the mobile device is programmed to receive and transmit data from at least two external sensors attached to the upper and lower limbs of the subject. The method of claim 75 or 76, wherein the mobile device acquires data from sensors on the upper and lower limbs of the subject. The method of claim 75 or 76, wherein the mobile device is carried by the subject and acquires data from an internal sensor. The method of claim 75 or 76, wherein the series of exercises comprises tapping, sitting and standing of the device. The method of any of claims 74-79, wherein monitoring points to a reduction in movement or cognitive impairment in response to administration of the antibody. The method of claim 63, wherein the antibody binds within residues 115-130 of alpha-synuclein. The method of claim 63, wherein the antibody binds within residues 118-126 of alpha-synuclein. The method of claim 63, wherein the antibody binds within residues 117-123 of alpha-synuclein. The method of any of claims 63-80, wherein the antibody is NI-202.21D11. The method of any of claims 63-80, wherein the antibody comprises three heavy chain CDRs, respectively designated in SEQ ID NOs: 139-141, and three light chain CDRs, respectively designated in SEQ ID NOs: 143 to 145. The method of any of claims 63-80, wherein the antibody comprises a heavy chain variable region designated by SEQ ID NO: 138 and a light chain variable region designated by SEQ ID NO: 142. 63. The method of claim 63, wherein the antibody comprises three light chain Kabat CDRs of SEQ ID NO: 5 and three heavy chain Kabat CDRs of SEQ ID NO: 10. The antibody comprises the heavy chain variable region of any of SEQ ID NOs: 8-11 and the light chain variable region of any of SEQ ID NOs: 3-5, preferably the heavy chain variable region is the heavy chain variable region of SEQ ID NO: 10. 63. The method of claim 63, wherein the light chain variable region is the light chain variable region of SEQ ID NO: 5. The method according to any one of claims 63 to 88, wherein the antibody is an antibody of the human IgG1 isotype. 89. The method of claim 89, wherein the antibody comprises a heavy chain constant region of SEQ ID NO: 35 in which the C-terminal lysine may not be present, and a light chain constant region of SEQ ID NO: 30. 89. The method of claim 89, wherein the antibody comprises a heavy chain constant region of SEQ ID NO: 35 in which the C-terminal lysine may not be present, and a light chain constant region of SEQ ID NO: 13. The method of any of claims 63-82, wherein the antibody comprises three heavy chain CDRs designated by SEQ ID NOs: 146 to 148 and three light chain CDRs designated by SEQ ID NOs: 149 to 151, respectively. 23. The invention of any of claims 63-82, wherein the antibody comprises the heavy chain designated by SEQ ID NO: 37 and the light chain designated by SEQ ID NO: 32 and the C-terminal lysine of SEQ ID NO: 37 may be absent. the method of. The method of claim 63, wherein the antibody binds within residues 1-20 of alpha-synuclein. The method of claim 94, wherein the antibody binds within residues 4-15 of alpha-synuclein. The method of any of claims 63-80, wherein the antibody is NI-202.12F4. The method of any of claims 63-80, wherein the antibody comprises three heavy chain CDRs, respectively designated by SEQ ID NOs: 131-133, and three light chain CDRs, respectively designated by SEQ ID NOs: 135-137. The method of any of claims 63-80, wherein the antibody comprises a heavy chain variable region designated by SEQ ID NO: 130 and a light chain variable region designated by SEQ ID NO: 134. The method of any of claims 63-98, wherein the synuclein disease is Parkinson's disease. The method of any of claims 63-98, wherein synuclein disease is dementia with Lewy bodies. The method of any of claims 63-98, wherein the synuclein disease is plural atrophy. The method of any of claims 63-98, wherein the synuclein disease is progressive supranuclear palsy. The method of any of claims 63-98, wherein synuclein disease is REM sleep behavior disorder. The method of any of claims 63-98, wherein the synuclein disease is Alzheimer's disease with Lewy bodies of the amygdala. The method of any of claims 63-104, wherein the subject has the disease. The method of any of claims 63-105, wherein the subject has not received symptomatological treatment for Parkinson's disease in combination with the antibody. The method of any of claims 63-105, wherein the antibody is administered in combination with levodopa.