Pharmaceuticals and pharmaceutical compositions

Abstract

Compositions and methods for treating Parkinson's disease are provided. The present specification provides compounds, compositions, uses, and methods for increasing cell viability of dopaminergic neurons or for preventing or treating the death of dopaminergic neurons. In certain embodiments, methods are provided for alleviating symptoms and / or preventing or treating Parkinson's disease in an individual in need thereof, the methods comprising increasing GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP The method includes treating with one or more expressible nucleic acids encoding the compound of formula (I), or a combination thereof.

Description

[Technical Field]

[0001] The present invention relates generally to the treatment of Parkinson's disease. More particularly, the present invention relates to compositions and methods for enhancing microautophagy to treat various forms of Parkinson's disease and related diseases or conditions. [Background technology]

[0002] In eukaryotes, there are three known cellular processes by which cytoplasmic contents and / or subcellular organelles are delivered to or engulfed by lysosomes for degradation: macroautophagy (commonly referred to as autophagy), microautophagy, and chaperone-mediated autophagy. Unlike macroautophagy or chaperone-mediated autophagy, microautophagy is a stepwise, mediated autophagic process in which lysosomes (in mammals) or vacuoles (in plants and fungi) directly engulf cytoplasmic targets or cargo (e.g., proteins, lipids, glycogen, or pathogens).

[0003] Microautophagy can promote the degradation of cytosolic proteins by late endosomes (MVBs). Microautophagy can also support the direct delivery of glycogen to lysosomes for degradation. In this regard, dysfunction or insufficiency of microautophagy may be associated with the development of, for example, a variety of metabolic and / or neurological diseases.

[0004] Clearly, the accumulation of specific proteins, lipids and / or glycogen can be associated with a variety of important diseases, conditions and pathologies, and methods for targeting these proteins, lipids and / or glycogen are needed.

[0005] Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzheimer's disease (Zheng et al., 2018). PD is a progressive neurodegenerative disorder characterized by severe motor symptoms, including resting tremor, postural instability, bradykinesia, and muscle rigidity. Although genetic mutations that cause familial Parkinson's disease occur in some individuals, the majority of cases are sporadic (Schapira, 2008). Unfortunately, the underlying causes of PD remain poorly understood. Studies of mutations that cause familial PD and postmortem brain studies of idiopathic and sporadic PD have shown that mitochondrial dysfunction, oxidative stress, and protein metabolism via the ubiquitin-proteasome and autophagy pathways are central components of the pathogenic mechanism (Schapira and Tolosa, 2010).

[0006] Alpha-synuclein, encoded by the SNCA gene, is a neuronal protein that regulates synaptic vesicle trafficking and subsequent neurotransmitter release. One of the hallmarks of Parkinson's disease is the presence of Lewy bodies, characterized by the aggregation of alpha-synuclein to form insoluble fibrils within presynaptic dopaminergic neurons in the midbrain, particularly the substantia nigra (Zeng et al., 2018). Therefore, strategies to treat synucleinopathy by developing compounds that inhibit alpha-synuclein aggregation have been explored, but their effectiveness has been limited. Indeed, since the introduction of levodopa (L-Dopa) in the 1960s, relatively little progress has been made in PD treatment, and no dopamine agonist-based disease-modifying therapy or disease management exists (Stoker et al., 2018). Unfortunately, long-term use of L-Dopa causes significant side effects, including movement disorders, hallucinations, and impulse control disorders.

[0007] Alternatively, there is a need for additional and / or improved anti-Parkinson's agents, compositions and / or methods for the treatment of Parkinson's disease and related diseases or conditions. Summary of the Invention

[0008] As detailed herein, GDP-bound Rab1a, e.g., Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N or another dominant negative (DN) GDP-bound form of Rab1a, or these Rab1a GDP It has been recognized that treatment with a microautophagy-enhancing agent, comprising one or more expressible nucleic acids encoding:

[0009] In one specific embodiment, the present disclosure provides a method for alleviating, or preventing or treating, a symptom of Parkinson's disease in an individual in need thereof, the method comprising: GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof; and This allows the Rab1a GDP thereby reducing the symptoms of Parkinson's disease in the individual; and Includes:

[0010] In one specific embodiment, the symptoms of Parkinson's disease include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior (punding), movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0011] In another specific embodiment, the present disclosure provides a method for increasing cell viability of dopaminergic neurons in vitro or in vivo, the method comprising: GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP treating the cells with one or more expressible nucleic acids encoding: This results in Rab1a expression in dopaminergic neurons. GDP increasing the cell content of the dopaminergic neurons, thereby increasing cell viability of the dopaminergic neurons; Includes:

[0012] In another specific embodiment of any of the one or more methods described above, Rab1a GDP Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N or is or comprises another dominant-negative (DN) GDP-bound form of Rab1a.

[0013] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0014] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0015] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or comprises or consists of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0016] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP is in the form of a fusion protein, and Rab1a GDP optionally, via a linker, to a signaling or targeting peptide, a fluorescent peptide or other marker or tracer, or a peptide or peptides for targeted delivery, enhanced cellular uptake, increased stability or in vivo half-life, or Rab1a GDP or are fused or otherwise linked, directly or indirectly, to other peptide or non-peptide moieties for the improvement of other therapeutic, diagnostic or in vivo properties of the compound.

[0017] In yet another specific embodiment of any of the one or more methods preceding, the fusion protein has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0018] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP is in the form of a fusion protein, having the following amino acid sequence: MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDR DKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLD TGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKLMSSMNPEYDYLFKLLL IGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 21) or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0019] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are selected from the group consisting of Rab1a, Rab2a, Rab3a, Rab4a, Rab5a, Rab6a, Rab7a, Rab8a, Rab9a, Rab10a, Rab11a, Rab12a, Rab13a, Rab14a, Rab15a, Rab16a, Rab17a, Rab18a, Rab19a, Rab19b, Rab19c, Rab19d, Rab19e, Rab19f, Rab19g, Rab19i, Rab19i, Rab19i, Rab20a, Rab21a, Rab22a, Rab23a, Rab24a, Rab25b, Rab26a, Rab27a, Rab28a, Rab29b, Rab30a GDP The gene encoding the nucleotide sequence encoding one or more of the following:

[0020] In yet another specific embodiment of any of the preceding one or more methods, the one or more expressible nucleic acids are DNA-based or RNA-based.

[0021] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are transiently expressed in the individual as Rab1a GDP or one or more expressible nucleic acids are integrated into the genome of the individual and Rab1a is expressed in the individual. GDP is expressed.

[0022] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids transiently express Rab1a in the dopaminergic neuron. GDP or one or more expressible nucleic acids are integrated into the cellular genome and express Rab1a in dopaminergic neurons. GDP is expressed.

[0023] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids express Rab1a in the cell. GDP The expression vectors include expression vectors, plasmids, or mRNA capable of expressing the

[0024] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are Rab1a GDP and Rab1a GDP The present invention includes one or more expression vectors, plasmids or mRNAs capable of expressing the above.

[0025] In yet another specific embodiment of any of the one or more methods preceding, the one or more expressible nucleic acids comprise the following nucleic acid sequence: ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a S25N ORF codon sequence, SEQ ID NO: 5), ATGGGGGACTACAAGGACGACGATGACAAGGGGGGTAGCGGTGGATCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGATTCTGGGGTTGGAAAGTCCTGCCTTCTCCTTAGGTTTGCAGATGATACGTATACGGAAAGCTACATC AGCACAATTGGTGTGGATTTCAAGATACGAACTATAGAGTTAGATGGGAAAACAATCAAGCTACAGATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACTTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAG GAGTCCTTCAATAACGTTAAACAGTGGCTGCAGGAGATAGATCGCTACGCCAGTGAAAATGTCAACAAGTTGTTGGTAGGGATCAAATGTGACCTGACCACAAAGAAAGTAGTAGACTACACAACAGCAAAGGAATTTGCAGATTCCCTTGGAATTCCATTTTTG GAAACCAGTGCTAAGAACGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAAGCGAATGGGTCCTGGAGCTACAGCTGGTGGTGCCGAGAAGTCCAATGTTAAAATCCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGCTGCTGCTAA (Mouse Rab1a N124I ORF codon sequence, SEQ ID NO: 8), or ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGTCTTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGAACTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a D47N ORF codon sequence, SEQ ID NO: 17), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0026] In yet another specific embodiment of any of the one or more methods preceding, the one or more expressible nucleic acids comprise the following nucleic acid sequence: (SEQ ID NO: 19, MG-008 ORF DNA sequence with 5' luciferase tag), or (SEQ ID NO: 20, MG-008 ORF mRNA sequence with 5' luciferase tag), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0027] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof, for alleviating the symptoms of Parkinson's disease in an individual, or for preventing or treating Parkinson's disease in an individual in need thereof.

[0028] In a specific embodiment, the symptoms of Parkinson's disease include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0029] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof for increasing cell survival of dopaminergic neurons or preventing or treating death of dopaminergic neurons in an individual in need thereof.

[0030] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDPThe present invention provides the use of one or more expressible nucleic acids encoding the following, or combinations thereof, for increasing cell survival of dopaminergic neurons in vitro or in vivo.

[0031] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof in the manufacture of a medicament for alleviating the symptoms of, or preventing or treating Parkinson's disease in an individual in need thereof.

[0032] In a specific embodiment, the symptoms of Parkinson's disease include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0033] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof in the manufacture of a medicament for increasing cell survival of, or preventing or treating death of, dopaminergic neurons in an individual in need thereof.

[0034] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof in the manufacture of a medicament for increasing cell survival of dopaminergic neurons in vitro or in vivo.

[0035] In another specific embodiment of any of the one or more uses above, Rab1a GDP Rab1aS25N , Rab1a N124I , Rab1a D41N , Rab1a D47N or another dominant negative (DN) GDP-bound form of Rab1a.

[0036] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0037] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0038] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or comprises or consists of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0039] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP is in the form of a fusion protein, and Rab1a GDP optionally, via a linker, to a signaling or targeting peptide, a fluorescent peptide or other marker or tracer, or a peptide or peptides for targeted delivery, enhanced cellular uptake, increased stability or in vivo half-life, or Rab1a GDP or are fused or otherwise linked, directly or indirectly, to other peptide or non-peptide moieties for the improvement of other therapeutic, diagnostic, or in vivo properties of the compound.

[0040] In yet another specific embodiment of any of the one or more uses above, the fusion protein has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0041] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP is in the form of a fusion protein and has the following amino acid sequence: MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDR DKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLD TGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKLMSSMNPEYDYLFKLLL IGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 21), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0042] In yet another specific embodiment of any of the one or more uses described above, the one or more expressible nucleic acids are one or more Rab1a as described herein. GDP Code the following.

[0043] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids are DNA-based or RNA-based.

[0044] In yet another specific embodiment of any of the one or more uses described above, the one or more expressible nucleic acids are GDP or one or more expressible nucleic acids are integrated into the cellular genome and express Rab1a in dopaminergic neurons. GDP is expressed.

[0045] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids are Rab1a GDP and Rab1a GDP The present invention includes one or more expression vectors, plasmids or mRNAs capable of expressing the above.

[0046] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids comprise the following nucleic acid sequence: ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a S25N ORF codon sequence, SEQ ID NO: 5), ATGGGGGACTACAAGGACGACGATGACAAGGGGGGTAGCGGTGGATCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGATTCTGGGGTTGGAAAGTCCTGCCTTCTCCTTAGGTTTGCAGATGATACGTATACGGAAAGCTACATC AGCACAATTGGTGTGGATTTCAAGATACGAACTATAGAGTTAGATGGGAAAACAATCAAGCTACAGATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACTTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAG GAGTCCTTCAATAACGTTAAACAGTGGCTGCAGGAGATAGATCGCTACGCCAGTGAAAATGTCAACAAGTTGTTGGTAGGGATCAAATGTGACCTGACCACAAAGAAAGTAGTAGACTACACAACAGCAAAGGAATTTGCAGATTCCCTTGGAATTCCATTTTTG GAAACCAGTGCTAAGAACGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAAGCGAATGGGTCCTGGAGCTACAGCTGGTGGTGCCGAGAAGTCCAATGTTAAAATCCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGCTGCTGCTAA (Mouse Rab1a N124I ORF codon sequence, SEQ ID NO: 8), or ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGTCTTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGAACTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a D47N ORF codon sequence, SEQ ID NO: 17), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0047] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids comprise the following nucleic acid sequence: (SEQ ID NO: 19, MG-008 ORF DNA sequence with 5' luciferase tag), or (SEQ ID NO: 20, MG-008 ORF mRNA sequence with 5' luciferase tag), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0048] In another specific embodiment, the present disclosure provides the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences, and which preferentially binds GDP, The polypeptides provide polypeptides for use in increasing cell viability of dopaminergic neurons in an individual in need thereof, or for preventing or treating Parkinson's disease, or for increasing cell viability of dopaminergic neurons in vitro or in vivo.

[0049] In another specific embodiment, the present disclosure provides GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof, and Pharmaceutical compositions containing other anti-Parkinson's agents are provided.

[0050] In yet another specific embodiment, the present disclosure provides GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP one or more expressible nucleic acids encoding antiparkinsonian drugs, instructions for carrying out any one or more of the methods described herein; or Any combination of these,

[0023] A kit is provided comprising one or more of:

[0051] In yet another specific embodiment of any of the methods, uses or polypeptides used above, the symptom is resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behaviour, movement disorders, hallucinations, impulse control disorders and sleep disorders.

[0052] These and other features can be further understood by reference to the following figures: GDP Treatment was carried out using the method described above. [Brief explanation of the drawings]

[0053] [Figure 1] In a hemiparkinsonian rat model created by intracranial injection of 6-hydroxydopamine (also known as hydroxydopamine or 2,4,5-trihydroxyphenethylamine, 6-OHDA) as described in Example 1, MG-008 administration increases the latency to fall (in seconds) (A, C) and increases the velocity of the fall (B, D) at 3 weeks (AB) and 5 weeks (CD) after administration. [Figure 2] As described in Example 1, in a rat model of hemiparkinsonian disease created by intracranial injection of 6-OHDA, MG-008 administration reduced the time (in seconds) required for rats to cross the beam after 4 weeks of administration. [Figure 3] As described in Example 1, in a rat model of hemiparkinsonian disease created by intracranial injection of 6-OHDA, administration of MG-008 resulted in a decrease in the rats' turning rate, measured as an average of 360° / hour in a rotometer, 4 weeks after administration. [Figure 4] In a hemiparkinsonian rat model created by intracranial injection of 6-OHDA as described in Example 1, recovery of dopaminergic neurons in the substantia nigra pars compacta (SNpc) of hemiparkinsonian rats (B) compared to the control group (A), and recovery of dopaminergic neurons in the substantia nigra pars reticulata (SNr) of hemiparkinsonian rats compared to the control group (A) are shown. [Figure 5-1] Specific amino acid and nucleic acid sequences described herein are shown. [Figure 5-2] Same as above [Figure 5-3] Same as above [Figure 5-4] Same as above [Figure 5-5] Same as above [Figure 5-6] Same as above [Figure 5-7] Same as above [Figure 5-8] Same as above [Figure 5-9] Same as above [Figure 5-10] Same as above [Figure 5-11] Same as above [Figure 6] 1 shows an example of an experimental schedule for F11 administration in a hemiparkinsonian rat model created by intracranial injection of 6-OHDA. [Figure 7] 1 shows an example of an alternative experimental schedule for F11 administration in a hemiparkinsonian rat model produced by intracranial injection of 6-OHDA. [Figure 8] No significant differences in mean body weight (A) and mean body weight change (B) were observed between F11-treated rats and untreated (sham-treated) and 6-OHDA-treated rats. [Figure 9] Figure 1 shows that the latency to fall in the rotarod test was significantly improved in hemiparkinsonian rats at (A) 2 weeks, (B) 4 weeks, (C) 8 weeks, and (D) 12 weeks after the first intravenous injection of F11. [Figure 10] Decreased beam walking time in a hemiparkinsonian rat model at (A) 2 weeks, (B) 4 weeks, (C) 8 weeks, and (D) 12 weeks after the first intravenous injection of F11. [Figure 11] Figure 1 shows that rotational speed was reduced in a hemiparkinsonian rat model at (A) 2 weeks, (B) 4 weeks, (C) 8 weeks, and (D) 12 weeks after the first intravenous injection of F11. [Figure 12] Increased expression of tyrosine hydroxylase confirmed by immunohistochemical staining in the substantia nigra of a hemiparkinsonian rat model after administration of F11. [Figure 13] Representative images of tyrosine hydroxylase expression in the substantia nigra of a hemiparkinsonian rat model after administration of F11 are shown. DETAILED DESCRIPTION OF THE INVENTION

[0054] This specification relates to compounds, compositions, uses and methods used to increase cell viability of dopaminergic neurons in vitro or in vivo, and / or to prevent or treat symptoms of Parkinson's disease in dopaminergic neurons or in individuals in need thereof. It should be understood that specific examples and embodiments are provided for purposes of illustration to those of ordinary skill in the art and are not intended to be limiting in any way.

[0055] Parkinson's disease (PD), the second most common neurodegenerative disease affecting 2%–3% of people over the age of 65, is primarily caused by the death of dopaminergic neurons in the substantia nigra of the midbrain. Although the pathological mechanisms are not fully understood, both genetic and environmental factors are thought to contribute to the pathogenesis of PD (Gan-Or et al., 2015). A major obstacle in the development of neuroprotective therapies is our limited understanding of the key molecular events that trigger neurodegeneration, particularly the death of dopaminergic neurons.

[0056] A missense mutation (A53T) in α-synuclein, encoded by the SNCA / PARK1 gene, was the first pathogenic mutation identified in familial PD (Polymeropoulos et al., 1997). Subsequently, a series of missense point mutations in the SNCA gene were identified, demonstrating that α-synuclein plays a key role in the progression of PD. Indeed, mutations in α-synuclein cause toxicity to dopaminergic neurons by altering the intracellular signaling program. The A53T mutation has been shown to suppress autophagy at an early stage in the brains of transgenic mice, resulting in α-synucleinopathy (Pupyshev et al., 2017). This disease is characterized by the abnormal accumulation of α-synuclein aggregates in neurons, neurofilaments, or glial cells. These α-synuclein aggregates also trigger cell death pathways mediated by mitochondrial dysfunction and endoplasmic reticulum stress (Smith et al., 2005), and by binding to TrkB receptors, they suppress BDNF / TrkB signaling, leading to the death of dopaminergic neurons (Zharikov et al., 2015). Other mutations in α-synuclein have been shown to impair autophagy, such as the E46K mutation, which inactivates the JNK1-Bcl-2 pathway, or the A30P mutation, which is rescued by increased expression of HMGB1 or Beclin in PC12 cells (Wang et al., 2016).

[0057] Another PD pathogenic mutation has been found in the LRRK2 / PARK8 gene, which is the greatest risk factor for familial PD and causes autosomal dominant PD (Zimprich et al., 2004). LRRK2 mutations are associated with "classical" late-onset PD, accounting for 4% of hereditary PD cases (Ferreira and Massano, 2017). Leucine-rich repeat kinase 2 (LRRK2) is a multidomain protein with both Rab GTPase and kinase activities (Steger et al., 2017). The LRRK2G2019S mutation enhances kinase activity and selectively inhibits synaptic vesicle trafficking in neurons of the ventral midbrain, including dopaminergic neurons (Pan et al., 2017). This particular LRRK2 G2019S mutation has been shown to enhance α-synuclein accumulation (Volpicelli-Daley et al., 2016), induce ASK1-mediated apoptosis (Yoon et al., 2017), increase mitochondrial DLP1, leading to mitochondrial fragmentation and dysfunction (Wang et al., 2015), and disrupt autophagy (Su et al., 2015).

[0058] Mutations in the protein kinase PTEN-induced putative kinase 1 (PTEN-induced putative kinase 1) have been shown to be a key protein involved in the pathogenesis of mitochondrial dysfunction and PD. PINK1 mutations result in the accumulation of dysfunctional mitochondria due to activation of the mitochondrial unfolded protein response. Parkin, encoded by PARK2, is a ubiquitin E3 ligase that is activated by autophosphorylated PINK1, which accumulates on the membranes of dysfunctional mitochondria. This activates the E3 ubiquitin ligase activity of Parkin, recruiting it to damaged mitochondria and promoting their autophagic degradation (Narendra et al., 2009). Parkin mutations, such as R42P, R46P, K211N, C212Y, C253Y, C289G, and C441R, impair its recruitment to depolarized mitochondria and inhibit mitochondrial autophagy (Narendra et al., 2010).

[0059] Mutations in the vacuolar protein sorting 35 (VPS35) gene account for 1% of familial PD cases, and the VPS35 protein regulates the sorting process of transmembrane proteins between endosomes and the Golgi (Bonifacino and Hurley, 2008). Dopaminergic neurons lacking VPS35 exhibit impaired retrieval of Lamp2a from endosomes to the Golgi, which promotes reduced degradation of α-synuclein via chaperone-mediated autophagy (Tang et al., 2015).

[0060] A common theme in the pathogenesis of PD is the accumulation of misfolded proteins and defects in the mechanisms responsible for their clearance, preventing cytotoxicity in dopaminergic neurons. In fact, autophagy is typically affected in the early stages of PD and plays a key role in the clearance of misfolded proteins and the degradation of damaged organelles, which is widely considered cytoprotective. The interaction of PINK1 with α-synuclein in the cytoplasm promotes the degradation of cytotoxic α-synuclein by activating autophagy (Liu et al., 2017). ER stressors activate the recruitment of MKK4 to lysosomes, triggering chaperone-mediated autophagy and providing a pro-survival mechanism (Li et al., 2017). However, autophagy is commonly defective or inhibited in PD, which has been proposed as a pathogenic mechanism in the progression of PD (Cai et al., 2015). Furthermore, inhibition of autophagy increases the release of α-synuclein from cells via extracellular vesicles and its translocation to other cells, accelerating the progression of synucleinopathy. Furthermore, α-synuclein mutations, such as A53T and A30P, allow α-synuclein to bind to receptors on lysosomal membranes and inhibit chaperone-mediated autophagy, leading to the accumulation of abnormal α-synuclein in Lewy bodies and subsequent parkinsonism. Furthermore, inhibition of chaperone-mediated autophagy leads to the progressive loss of dopaminergic neurons in the SNc, a significant decrease in dopamine content in the striatum, and motor impairment (Xilouri et al., 2016). Therefore, promoting autophagy is a rational strategy to target the early onset of PD.

[0061] Autophagy is the primary intracellular degradation pathway, deriving its degradative capacity from lysosomes. It is a constitutive and evolutionarily conserved self-digestion mechanism by which cells degrade and recycle long-lived proteins and excess or damaged organelles. Autophagy is a highly regulated process through which misfolded proteins and organelles reach the lysosome for degradation (Kim KH, Lee MS. Autophagy - a key player in cellular and body metabolism. Nat Rev Endocrinol. 2014 June, 10(6):322-37). There are three types of autophagy: macroautophagy (commonly referred to as autophagy), chaperone-mediated autophagy, and microautophagy. In chaperone-mediated autophagy and microautophagy, lysosomal degradation of substrates occurs directly inside the lysosome (Parzych KR, Klionsky DJ. An overview of autophagy: morphology, mechanism, and regulation. Antioxid Redox Signal. 2014 Jan 20;20(3):460-73).

[0062] Cell death can involve or be related to different factors, such as changes in cell morphology or function. There are three main types of cell death: apoptosis (type 1), autophagy (type 2), and necroptosis (type 3). (Fuchs Y, Steller H. Programmed cell death in animal development and disease. Cell. 2011, November 11, 147(4):742-58) Generally, autophagy acts as a protective mechanism for cells. However, autophagy can also be used as a cellular suicide mechanism, termed "autophagic cell death." Therefore, the autophagic cell death process is distinct from apoptosis or necroptosis. (Kroemer G, Levine B. Autophagic cell death: the story of a misnomer. Nat Rev Mol Cell Biol. 2008, December 9(12):1004-10)

[0063] In this regard, the present inventors have recognized that in addition to apoptosis (programmed cell death) and necroptosis, the process of autophagy-lysosome-mediated cell death, particularly microautophagy, could be utilized to develop effective strategies for the treatment of Parkinson's disease.

[0064] Stimulating microautophagy (which, in certain specific embodiments, may involve the stepwise engulfment of one or more target membranes while promoting lysosome movement toward the target membrane) may be a particularly desirable means for treating a variety of diseases and / or pathologies and / or conditions. Specific lysosomal localization within a cell may be associated with different types of lysosomal activity. Lysosomal localization is also associated with mTOR activity, regulating autophagic flux. During macroautophagy, mTORC1 is inactivated, potentially leading to lysosome accumulation in the perinuclear region of the cell, which stimulates fusion of the enveloped target membrane with the lysosome, thereby promoting macroautophagy. Meanwhile, during microautophagy, the stepwise engulfment of one or more target membranes by lysosomes occurs through lysosomal movement and direct interaction with one or more target membranes, without the need for the formation of one or more autophagosomes. Lysosomes move bidirectionally along microtubules, and this lysosomal movement is controlled by distinct motor proteins recruited by different mechanisms.

[0065] In certain specific embodiments, microautophagy involves the direct uptake of cytoplasmic cargo by autophagic tubes at the limiting membrane, which can mediate lysosomal invagination and vesicle scission (see Li, W.-W., Li, J., and Bao, .-K. Microautophagy: Lesser-known self-eating. Cell. Mol. Life Sci. 69, 1125-1136 (2011)). For example, DNA can undergo direct lysosomal degradation of target substrates (referred to as stepwise autophagy; see Fujiwara, Y. et al., Direct uptake and degradation of DNA by Lysosomes. - PubMed - NCBI. Autophagy 9, 1167-1171 (2014)).Furthermore, lysosomes move towards different organelles and / or membrane substrates (e.g., plasma membrane, mitochondria) and interact directly with them by recruiting motor and SNARE proteins (Andrews, NW Lysosomes and the plasma membrane. J. Cell Biol. 158, 389-394 (2002); Hofmann, I. and Munro, S. An N-terminally acetylated Arf-like GTPase is localized to lysosomes and affects their motility. J. Cell. Sci. 119, 1494-1503 (2006); Fraldi, A. et al., Lysosomal fusion and SNARE function are impaired by cholesterol accumulation in lysosomal storage disorders. The EMBO Journal 29, 3607-3620 (2010); and Pankiv, S. et al., FYCO1 is a Rab7 effector that binds to LC3 and (See PI3P to mediate microtubule plus end-directed vesicle transport. J. Cell Biol. 188, 253-269 (2010)). Lysosome movement to the cell periphery and their intracellular localization are associated with signal transduction (see R. and Bonifacino, J.S. Lysosome Positioning Influences mTORC2 and AKT Signaling. Molecular Cell 75, 26-38, e3 (2019)).In this regard, it is thought that activation of mTORC1, mTORC2 and AKT may be crucial for the peripheral distribution of lysosomes (see Pous, C. and Codogno, P. Lysosome positioning coordinates mTORC1 activity and autophagy. Nature Cell Biology 13, 342-344 (2011) and Cabukusta, B. and Neefjes, J. Mechanisms of lysosomal positioning and movement. Traffic 19, 761-769 (2018)).

[0066] In certain specific embodiments, microautophagy and / or stepwise degradation of target substrates and / or membranes involves lysosomes moving toward (e.g., directly interacting with) target membranes at the cell periphery and within the cytoplasm (see Pu, J., Guardia, C. M., Keren-Kaplan, T., and Bonifacino, J. S. MeChanisms and Functions of lysosome positioning. J. Cell. Sci. 129, 4329-4339 (2016); and Katherine R Parzych, D. J. K. An Overview of Autophagy: Morphology, MeChanism, and Regulation. Antioxid. Redox Signal. 20, 460-473 (2014)). In certain specific embodiments, lysosome migration (from the perinuclear region of the cell) toward the cell periphery and interaction with target membranes / substrates (e.g., glycogen, lipids, proteins) at the cell periphery may be associated with activation of mTORC1 / mTORC2 (see Rabanal-Ruiz, Y. and Korolchuk, V.I. mTORC1 and Nutrient Homeostasis: The Central Role of the Lysosome. Int J Mol Sci 19, 818 (2018); Jia, R. and Bonifacino, J.S. Lysosome Positioning Influences mTORC2 and AKT Signaling. Molecular Cell 75, 26-38. e3 (2019)).

[0067] Not wanting to be bound by theory, we consider that Rab1a DN (dominant negative form of Rab1a) mimics the peripheral distribution of lysosomes (from the perinuclear region) by activating mTORC1 / mTORC2 proteins within the cell, thus activating them (to support the peripheral distribution of lysosomes) without the need for activation by external (or extracellular) signals.

[0068] Rab1a in increasing cell survival of dopaminergic neurons and / or preventing cell death of dopaminergic neurons GDP or Rab1a GTP Use of : As detailed herein, GDP-bound Rab1a, e.g., Rab1a S25N , Rab1a N124I (mouse Rab1 sequence), Rab1a D41N , Rab1a D47N or another dominant-negative (DN) GDP-bound form of Rab1a, or these Rab1a GDP It has been determined that treatment with a microautophagy-enhancing agent comprising one or more expressible nucleic acids encoding is useful for increasing the survival rate and / or preventing the death of dopaminergic neurons.

[0069] As will be appreciated, in certain specific embodiments, the methods described herein are ex vivo methods, in vivo methods, or both.

[0070] In one specific embodiment, the present disclosure provides a method for alleviating symptoms of, or preventing or treating Parkinson's disease in an individual in need thereof, the method comprising: GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof; and This allows the Rab1a GDP and increasing the cellular content of the cells, thereby alleviating symptoms of Parkinson's disease in the individual.

[0071] In one specific embodiment of the method, the symptoms include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0072] In one specific embodiment, the present disclosure provides a method for increasing cell viability of dopaminergic neurons or preventing or treating dopaminergic neuron death in an individual in need thereof, the method comprising: GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP treating the individual's dopaminergic neurons with one or more expressible nucleic acids encoding: This results in Rab1a expression in dopaminergic neurons. GDP and increasing the cellular content of dopaminergic neurons, thereby reducing neuronal cell death of dopaminergic neurons.

[0073] In another specific embodiment, the present disclosure provides a method for increasing the survival rate of dopaminergic neurons in vitro or in vivo, the method comprising: GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP treating dopaminergic neurons with one or more expressible nucleic acids encoding: This results in Rab1a expression in dopaminergic neurons. GDP and increasing the cellular content of the dopaminergic neurons, resulting in increased survival or decreased death of the dopaminergic neurons.

[0074] In another specific embodiment of any of the one or more methods described above, Rab1a GDP Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N , or another dominant-negative (DN) GDP-bound form of Rab1a.

[0075] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDPbut the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0076] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0077] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or comprises or consists of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0078] In yet another specific embodiment of any of the one or more methods described above, GDP is in the form of a fusion protein, and Rab1a GDP optionally via a linker, to a signaling or targeting peptide, a fluorescent peptide or other marker or tracer, or a compound that is useful for targeted delivery, enhanced cellular uptake, increased stability or in vivo half-life, or Rab1a GDPand / or fused or otherwise linked, directly or indirectly, to other peptide or non-peptide moieties for other therapeutic, diagnostic, or improved in vivo properties of the fusion protein. In one specific embodiment, not intended to be limiting in any manner, the fusion protein comprises any protein or tag, such as, for example, GFP, YFP, mCherry, a luciferase-specific antibody, an aptamer or the like for identification or delivery to a specific organ.

[0079] In yet another specific embodiment of any of the one or more methods preceding, the fusion protein has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0080] In yet another specific embodiment of any of the one or more methods described above, Rab1a GDP is in the form of a fusion protein, having the following amino acid sequence: MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDR DKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLD TGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKLMSSMNPEYDYLFKLLL IGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 21) or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0081] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are selected from the group consisting of Rab1a, Rab2a, Rab3a, Rab4a, Rab5a, Rab6a, Rab7a, Rab8a, Rab9a, Rab10a, Rab11a, Rab12a, Rab13a, Rab14a, Rab15a, Rab16a, Rab17a, Rab18a, Rab19a, Rab19b, Rab19c, Rab19d, Rab19e, Rab19f, Rab19g, Rab19i, Rab19i, Rab19i, Rab20a, Rab21a, Rab22a, Rab23a, Rab24a, Rab25b, Rab26a, Rab27a, Rab28a, Rab29b, Rab30a GDP The gene encoding the nucleotide sequence encoding one or more of the following:

[0082] In yet another specific embodiment of any of the preceding one or more methods, the one or more expressible nucleic acids are DNA-based or RNA-based.

[0083] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are transiently expressed in the cell as Rab1a GDP or one or more expressible nucleic acids are integrated into the genome of a cell and express Rab1a in the cell. GDP is expressed.

[0084] In yet another specific embodiment of any of the one or more methods described above, the one or more expressible nucleic acids are Rab1a GDP It encodes Rab1a GDP The present invention includes one or more expression vectors, plasmids or mRNAs capable of expressing the above.

[0085] In yet another specific embodiment of any of the one or more methods preceding, the one or more expressible nucleic acids comprise the following nucleic acid sequence: ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a S25N ORF codon sequence, SEQ ID NO: 5), ATGGGGGACTACAAGGACGACGATGACAAGGGGGGTAGCGGTGGATCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGATTCTGGGGTTGGAAAGTCCTGCCTTCTCCTTAGGTTTGCAGATGATACGTATACGGAAAGCTACATC AGCACAATTGGTGTGGATTTCAAGATACGAACTATAGAGTTAGATGGGAAAACAATCAAGCTACAGATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACTTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAG GAGTCCTTCAATAACGTTAAACAGTGGCTGCAGGAGATAGATCGCTACGCCAGTGAAAATGTCAACAAGTTGTTGGTAGGGATCAAATGTGACCTGACCACAAAGAAAGTAGTAGACTACACAACAGCAAAGGAATTTGCAGATTCCCTTGGAATTCCATTTTTG GAAACCAGTGCTAAGAACGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAAGCGAATGGGTCCTGGAGCTACAGCTGGTGGTGCCGAGAAGTCCAATGTTAAAATCCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGCTGCTGCTAA (Mouse Rab1a N124I ORF codon sequence, SEQ ID NO: 8), or ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGTCTTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGAACTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a D47N ORF codon sequence, SEQ ID NO: 17), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0086] In yet another specific embodiment of any of the one or more methods preceding, the one or more expressible nucleic acids comprise the following nucleic acid sequence: (SEQ ID NO: 19, MG-008 ORF DNA sequence with 5' luciferase tag), or (SEQ ID NO: 20, MG-008 ORF mRNA sequence with 5' luciferase tag), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0087] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or combinations thereof, for alleviating the symptoms of Parkinson's disease in an individual, or for preventing or treating Parkinson's disease in an individual in need thereof.

[0088] In one specific embodiment of the use, the symptoms include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0089] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or combinations thereof, for increasing cell survival of dopaminergic neurons or for preventing or treating dopaminergic neuron death in an individual in need thereof.

[0090] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDPThe present invention provides the use of one or more expressible nucleic acids encoding the following, or combinations thereof, for increasing cell survival of dopaminergic neurons in vitro or in vivo.

[0091] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or combinations thereof, for use in the manufacture of a medicament for alleviating the symptoms of Parkinson's disease in an individual, or for preventing or treating the symptoms of Parkinson's disease in an individual in need thereof.

[0092] In specific embodiments of the use, the symptoms include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0093] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof, for the manufacture of a medicament for increasing cell survival of, or preventing or treating death of, dopaminergic neurons in an individual in need thereof.

[0094] In another specific embodiment, the present disclosure provides a method for the production of GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or combinations thereof, for the manufacture of a medicament for increasing cell survival of dopaminergic neurons in vitro or in vivo.

[0095] In yet another specific embodiment of any of the one or more uses above, Rab1a GDPHowever, Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N , or another dominant-negative (DN) GDP-bound form of Rab1a.

[0096] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences and which preferentially binds GDP.

[0097] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with any of these sequences and which preferentially binds to GDP.

[0098] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP but the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or comprises or consists of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0099] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP is in the form of a fusion protein, and Rab1a GDP optionally, via a linker, to a signaling or targeting peptide, a fluorescent peptide or other marker or tracer, or a peptide or peptides for targeted delivery, enhanced cellular uptake, increased stability or in vivo half-life, or Rab1a GDP or are fused or otherwise linked, directly or indirectly, to other peptide or non-peptide moieties for the improvement of other therapeutic, diagnostic or in vivo properties of the compound.

[0100] In yet another specific embodiment of any of the one or more uses above, the fusion protein has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0101] In yet another specific embodiment of any of the one or more uses above, Rab1a GDP is in the form of a fusion protein and has the following amino acid sequence: MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDR DKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLD TGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKLMSSMNPEYDYLFKLLL IGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and which preferentially binds GDP.

[0102] In yet another specific embodiment of any of the one or more uses described above, the one or more expressible nucleic acids are one or more Rab1a as described herein. GDP Code the following.

[0103] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids are DNA-based or RNA-based.

[0104] In yet another specific embodiment of any of the one or more uses described above, the one or more expressible nucleic acids are GDP or one or more expressible nucleic acids are integrated into the cellular genome and express Rab1a in the cell. GDP is expressed.

[0105] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids are Rab1a GDP and Rab1a GDP The present invention includes one or more expression vectors, plasmids or mRNAs capable of expressing the above.

[0106] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids comprise the following nucleic acid sequence: ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a S25N ORF codon sequence, SEQ ID NO: 5), ATGGGGGACTACAAGGACGACGATGACAAGGGGGGTAGCGGTGGATCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGATTCTGGGGTTGGAAAGTCCTGCCTTCTCCTTAGGTTTGCAGATGATACGTATACGGAAAGCTACATC AGCACAATTGGTGTGGATTTCAAGATACGAACTATAGAGTTAGATGGGAAAACAATCAAGCTACAGATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACTTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAG GAGTCCTTCAATAACGTTAAACAGTGGCTGCAGGAGATAGATCGCTACGCCAGTGAAAATGTCAACAAGTTGTTGGTAGGGATCAAATGTGACCTGACCACAAAGAAAGTAGTAGACTACACAACAGCAAAGGAATTTGCAGATTCCCTTGGAATTCCATTTTTG GAAACCAGTGCTAAGAACGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAAGCGAATGGGTCCTGGAGCTACAGCTGGTGGTGCCGAGAAGTCCAATGTTAAAATCCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGCTGCTGCTAA (Mouse Rab1a N124I ORF codon sequence, SEQ ID NO: 8), or ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGTCTTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGAACTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a D47N ORF codon sequence, SEQ ID NO: 17), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0107] In yet another specific embodiment of any of the one or more uses above, the one or more expressible nucleic acids comprise the following nucleic acid sequence: (SEQ ID NO: 19, MG-008 ORF DNA sequence with 5' luciferase tag), or (SEQ ID NO: 20, MG-008 ORF mRNA sequence with 5' luciferase tag), or a Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity therewith and which preferentially binds to GDP. GDP a nucleic acid sequence encoding Alternatively, it includes nucleic acid sequences that are equivalent to any of the above sequences due to codon redundancy.

[0108] In another specific embodiment, the present disclosure provides the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 6, human Rab1a S25N ), MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 9, mouse Rab1 N124I ), Human Rab1a D41N or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (SEQ ID NO: 18, human Rab1a D47N ), or a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any of these sequences, and which preferentially binds GDP, The polypeptides are provided for use in increasing cell viability of dopaminergic neurons in an individual in need thereof, or for preventing or treating dopaminergic neuron death, or for increasing cell viability of dopaminergic neurons in vitro or in vivo.

[0109] In one specific embodiment of the polypeptide, the polypeptide is used to alleviate the symptoms of Parkinson's disease in an individual or to prevent or treat Parkinson's disease in an individual in need thereof.

[0110] In one specific embodiment of the polypeptide, the symptoms of Parkinson's disease include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, movement disorders, hallucinations, impulse control disorders, and sleep disorders.

[0111] In another specific embodiment, the present disclosure provides GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof, and Other antiparkinsonian drugs The present invention provides a pharmaceutical composition comprising:

[0112] In yet another specific embodiment, the present disclosure provides GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP one or more expressible nucleic acids encoding antiparkinsonian drugs, instructions for carrying out any one or more of the methods described herein; or Any combination of these, One or more of the following: A kit comprising:

[0113] As will be appreciated, in certain specific embodiments of any of the methods, uses, or polypeptides for use described herein, the dopaminergic activity may be any of a wide variety of dopaminergic types. As shown in Example 1 below, test results demonstrate that antiparkinsonian effects are observed across multiple different phenotypes, supporting broad antiparkinsonian applicability.

[0114] In yet another specific embodiment of any of the foregoing methods, uses or polypeptides for use, the dopaminergic neurons are localized in the midbrain.

[0115] In yet another specific embodiment of any of the methods, uses or polypeptides for use, the dopaminergic neurons are located in the substantia nigra pars compacta (SNpc) or the substantia nigra pars reticulata (SNr).

[0116] As understood, lysosome-mediated microautophagy refers to the cellular process by which cellular lipids or proteins or glycogen substrates, or portions thereof, are taken up and degraded by lysosomes. In this particular aspect, lysosome-mediated microautophagy is an essential component of cellular bioenergetic processes (i.e., ATP generation) and biosynthetic processes (i.e., synthesizing new biological substances by recycling existing "old" materials).

[0117] It will also be understood that reference to increased lysosomal-mediated microautophagy refers to an increase in the rate, extent, capacity, or effectiveness of the process of lysosomal-mediated microautophagy in a cell compared to baseline levels in the cell, or compared to treated or untreated control cells, or compared to levels in diseased cells or control cells having accumulation of protein, lipid, or glycogen substrates.

[0118] As will be further understood, increasing lysosome-mediated microautophagy includes restoring or increasing lysosomal motility or bidirectional lysosomal motility within cells, and / or restoring or rescuing lysosomal motility activity within cells through enhancement, increase, activation, or other methods. This restoration or rescue increases autophagy (e.g., microautophagy and / or macroautophagy) and / or lysosomal degradation capacity. As detailed herein, there are various diseases, pathologies, and cellular conditions in which cellular lysosomal motility is impaired, reduced, inhibited, or suppressed. Because lysosome-mediated microautophagy is involved in lysosomal motility and "kiss-and-run" events, microautophagy enhancers restore or increase lysosomal motility within cells. Lysosomal motility plays an important role in several cellular functions, including lysosome-mediated microautophagy, lysosome-mediated macroautophagy, lysosome regeneration, and lysosomal maturation processes. Restoring lysosomal motility and / or bidirectional lysosomal motility refers to adjusting the level of lysosomal motility / bidirectional motility of a cell to a level corresponding to that of normal or healthy control cells having a baseline level of lysosomal motility / bidirectional motility. In certain specific embodiments, this adjustment also refers to adjusting the level of degradative capacity (i.e., uptake) to that of normal or healthy cells.

[0119] As understood, a lysosomal binding-dissociation event between a lysosome and a lipid, protein, or glycogen substrate refers to an event in which the lysosome binds to a lipid, protein, or glycogen substrate (i.e., for example, a lipid droplet or protein aggregate), acquires at least a portion of the lipid, protein, or glycogen substrate, and then dissociates from the lipid, protein, or glycogen substrate. Lysosomal binding-dissociation (i.e., "on" and "off") events between a lysosome and a lipid, protein, or glycogen substrate are considered "kiss-and-run" events. As part of the binding (or "kiss") event, at least a portion of the substrate (i.e., lipid) may be "grabbed" or taken up by the lysosome from the substrate (i.e., for example, a cytosolic lipid droplet, or CLD). In the case of lipid droplet substrates, this is achieved by the formation of a fusion pore between the lysosome and the CLD. As part of the dissociation (or "run") event, the lysosome dissociates from the substrate (i.e., CLD). An increase in lysosome binding-dissociation events refers to an increase in the rate, extent, or effect of lysosome binding-dissociation events in a cell compared to baseline levels in treated or untreated control cells, e.g., identical cells under identical conditions but without a microautophagy-modulating agent, or treated with a compound or composition known not to affect the process.

[0120] As will be appreciated, in certain specific embodiments, microautophagy-enhancing agents are used to correct a microautophagy deficiency or reduced microautophagy state in cells.

[0121] Given the teachings herein, one skilled in the art will understand that a microautophagy-enhancing agent is any suitable agent that increases or promotes the rate, activity, extent, or effect of lysosome-mediated microautophagy in a cell, or that increases lysosomal movement or bidirectional movement. In a specific embodiment, a suitable microautophagy-enhancing agent is an agent that enhances GDP-bound Rab1a (Rab1a GDP ), Rab1a GDP or a combination thereof.

[0122] The Ras-related protein Rab-1A (i.e., Rab1a) is a protein encoded by the RAB1A gene in humans. The protein regulates the transport of vesicles from the endoplasmic reticulum (ER) through the Golgi compartment to the cell surface and plays an important role in the secretion of IL-8 and growth hormone. Furthermore, when the protein is in its GTP-bound form, it is involved in macroautophagy and the formation of autophagosomes in the cellular defense response against pathogenic bacteria. It also regulates the movement of one or more endocytic compartments.

[0123] As described in detail herein, lysosome-mediated microautophagy of target protein, lipid, or glycogen substrates in cells is mediated by GDP-bound Rab1a (e.g., Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N Alternatively, it has been confirmed that microautophagy is increased by using another microautophagy enhancer, including a dominant-negative (DN) GDP-bound form of Rab1a.

[0124] As will be understood, the specific amino acid or nucleic acid sequence of a particular gene varies between species. For example, the human Rab1a amino acid sequence exists in other species as a homolog with sequence variation from the human sequence. In some specific embodiments, even though the homolog sequence varies between species, the general effect (e.g., phenotypic effect) of the homolog sequence is substantially similar to the effect of the wild-type sequence in a particular cell or individual.

[0125] In certain specific embodiments, the microautophagy-enhancing agent is GDP-bound Rab1a (Rab1a GDP ), e.g., Rab1a S25N , Rab1a N124I , Rab1a D41N , Rab1a D47N or functional equivalents thereof, or another dominant-negative (DN) GDP-bound form of Rab1a. The sequences of human and / or mouse Rab1aWT and their specific GDP-bound forms are shown in Figure 41. Suitable GDP-bound forms of Rab1a include any suitable Rab1a mutant, which is "dominant negative," or binds GDP preferentially over GTP. These Rab1a GDPMutants are identified using techniques known to those skilled in the art (see, e.g., Chan, C.-C. et al., Systematic Discovery of Rab GTPases with Synaptic Functions in Drosophila. Current Biology 21, 1704-1715 (2011); Tabancay, AP et al., Identification of dominant negative mutants of Rheb GTPase and their use to implicate the involvement of human Rheb in the activation of p70S6K. J. Biol. Chem. 278, 39921-39930 (2003); and Dumas, JJ, Zhu, Z., Connolly, JL, and Lambright, DG, Structural basis of activation and GTP hydrolysis in Rab proteins. Structure 7, 413-s2 (1999), which are incorporated herein by reference in their entireties).

[0126] Figure 5 shows the specific sequences of the nucleic acids and amino acids / proteins described herein. In Figure 5, SEQ ID NOS: 1-3 correspond to human Rab1a, respectively. WT The DNA gene sequence, ORF codon sequence and amino acid sequence are provided. SEQ ID NOs: 4-6 are human Rab1a S25N The DNA gene sequence, ORF codon sequence and amino acid sequence are provided. SEQ ID NOs: 7-9 are mouse Rab1 N124I The DNA gene sequence, ORF codon sequence and amino acid sequence are provided. SEQ ID NOs: 10-12 are human Rab1a Q70L The DNA gene sequence, ORF codon sequence and amino acid sequence are provided. SEQ ID NOs: 13-15 are human Rab1a Q63L The DNA gene sequence, ORF codon sequence and amino acid sequence are provided. SEQ ID NOs: 16-18 are human Rab1a D47NThe DNA gene sequence, ORF codon sequence, and amino acid sequence are provided. SEQ ID NOs: 19-21 also provide the MG-008 ORF DNA sequence with a 5' luciferase tag, the MG-008 ORF mRNA sequence with a 5' luciferase tag, and the MG-008 protein sequence with an N-terminal luciferase tag, respectively (see Example 1 for further details of SEQ ID NOs: 19-21). Human Rab1a D41N The sequences of can be found at https: / / www.addgene.org / 49581 / (which are incorporated herein by reference in their entirety) and are commercially available. In certain specific embodiments, the present specification provides nucleic acids or amino acids comprising any of these sequences. In certain specific embodiments, the present specification provides nucleic acids or amino acids comprising any of these sequences (i.e., any of SEQ ID NOS: 1-21, or Rab1a). D41N or an active fragment thereof.

[0127] Rab1a S25N , Rab1a D41N , Rab1a D47N and Rab1a N124I Suitable functional equivalents of include, for example, suitable Rab1a variants such as: WT or Rab1a S25N or Rab1a N124I or Rab1a D41N or Rab1a D47N and binds GDP preferentially over GTP, and at the same time has at least 80% (or >85%, or >90%, or >95%, or >99%) sequence identity with Rab1a, as described in detail herein. S25N or Rab1a N124I or Rab1a D41N or Rab1a D47NAs will be appreciated in another specific embodiment, the microautophagy enhancer is one that maintains the associated cellular / biochemical functions of Rab1a. GDP The nucleic acid / expression vector may be or include one or more expressible nucleic acids, such as any suitable nucleic acid / expression vector (i.e., e.g., vector, cassette, mRNA, modified mRNA, plasmid) encoding GDP-bound Rab1a (Rab1a GDP ), e.g., Rab1a S25N or Rab1a N124I or Rab1a D41N or Rab1a D47N , or a functional equivalent thereof.

[0128] As will be understood, the present specification will be written primarily with reference to the sequences of the human and / or mouse homologs. Functional equivalents and / or variants may be found in multiple different species, for example, different mammalian species. For convenience, Rab1a is referred to herein as Rab1a. GDP (DN) or Rab1a GTP Specific sequence modifications and / or variants of (DA) forms are generally provided with reference to the location and modification / mutation information of the human and / or mouse homologues / sequences (e.g., S25N, D41N, D47N, N124I, Q70L, Q67L, Q63L). However, it will be understood that equivalent DN and / or DA forms may also be implemented for homologue sequences from other species and / or other sequences related to the human and / or mouse sequences provided herein. Depending on the particular sequence of interest, the location and / or nature of the modification / mutation may vary slightly. For example, Rab1 N124I is the drug / mutation of the mouse sequence. Further examples include Rab1 Q67L is a modification / mutation in the mouse sequence, but there is no Q at position 67 in human Rab1a. In fact, there is a Q at position 63 in the human sequence, so the modification / mutation in the human sequence is Rab1a. Q63L is.

[0129] As will be appreciated by those skilled in the art, a microautophagy-reducing agent is a GTP-bound form of Rab1a (Rab1a GTP ), Rab1a GTP one or more expressible nucleic acids encoding Rab1a wild type (Rab1a WT ), or Rab1a WT In certain specific embodiments, the nucleic acid is or comprises one or more expressible nucleic acids encoding Rab1a. GTP Rab1a Q70L , Rab1a (in mouse sequence) Q67L , Rab1a (in human Rab1a sequence) Q63L or a functional equivalent thereof, or another dominant active (DA) GTP-binding form of Rab1a. GTP-binding forms of Rab1a include any "dominant active" Rab1a mutants, or Rab1a mutants that bind GTP in preference to GDP. These Rab1a GTPMutants can be identified by those skilled in the art using known techniques (see, for example, Chan, C.-C. et al., Systematic Discovery of Rab GTPases with Synaptic Functions in Drosophila. Current Biology 21, 1704-1715 (2011); Tabancay, A.P. et al., Identification of dominant negative mutants of Rheb GTPase and their use to implicate the involvement of human Rheb in the activation of p70S6K. J. Biol. Chem. 278, 39921-39930 (2003); Dumas, J.J. Zhu, Z., Connolly, J.L. and Lambright, D.G. Structural basis of activation and GTP hydrolysis in Rab proteins. Structure 7, 413-s2 (1999), which are incorporated herein by reference in their entireties). For example, Rab1a Q70L and / or Rab1a Q67L and / or Rab1a Q63L These techniques are used to identify:

[0130] As will be appreciated, in certain specific embodiments, Rab1a GDP Treatment with microautophagy enhancers such as Rab1a GDP Introducing proteins into cells, Rab1a GDP or both.

[0131] As understood, expression of a specific protein in a cell refers to the production of a polypeptide from a nucleic acid sequence encoding the polypeptide. Gene expression includes both transcription and translation processes, and therefore, gene expression refers to the production of a nucleic acid sequence (i.e., transcription), for example, mRNA, the production of a protein (i.e., translation), or both. Furthermore, overexpression of a specific gene in a cell refers to increased expression of the specific gene in the cell compared to the wild-type, baseline, or untreated level. Overexpression of a mutant gene or introduction into a cell can be achieved using any of several methods known to those skilled in the art. For example, a vector (virus, plasmid, or other) can be introduced into a cell by transfection, electroporation, viral infection, or another suitable method known to those skilled in the art. The vector contains one or more copies of the specific gene, or mRNA or a chemically modified form thereof, each driven by an appropriate promoter sequence (e.g., a constitutive or inducible promoter). Suitable expression vector technology for overexpressing or introducing a specific gene into a cell is known to those skilled in the art (see, for example, Molecular Cloning: A Laboratory Manual (4th ed.), 2012, Cold Spring Harbor Laboratory Press). Given the teachings herein, one of skill in the art will be able to identify specific proteins, e.g., Rab1a GDP It will be appreciated that a wide variety of expressible nucleic acids encoding proteins can be prepared and introduced into cells (e.g., transiently or long-term by integration into the genome) to provide expression of a protein of interest.

[0132] As will be appreciated, compounds and / or compositions comprising or consisting of one or more of the nucleic acids and / or proteins described herein are used, which compositions further comprise one or more pharmaceutically acceptable diluents, carriers, excipients, or buffers, and which are used to administer one or more nucleic acids and / or proteins to cells in vitro or in vivo.

[0133] When inserting a nucleic acid sequence into a cell, gene introduction refers to "transfection," "transformation," or "transduction," which includes incorporating or introducing a nucleic acid sequence into a eukaryotic cell. The nucleic acid sequence is integrated into the cellular genome or transiently expressed (e.g., mRNA transfection), depending on the context. Proteins or enzymes are introduced into cells by delivering the protein or enzyme itself into the cell, or by expressing and translating mRNA encoding the protein or enzyme within the cell.

[0134] As known to those skilled in the art, expressible nucleic acids for expression of a particular gene encode or contain the features described in "Genes VII," Lewin, B., Oxford University Press (2000) or "Molecular Cloning: A Laboratory Manual," Sambrook et al., Cold Spring Harbor Laboratory, 3rd ed. (2001). The nucleotide sequence encoding the polypeptide or protein is incorporated into an appropriate vector, such as a commercially available vector. Vectors are individually constructed or modified using standard molecular biology techniques outlined in Sambrook et al. (Cold Spring Harbor Laboratory, 3rd ed. (2001)). Those skilled in the art will understand that a vector contains nucleotide sequences encoding desired elements, such that the elements are operably linked to the nucleotide sequence encoding the polypeptide or protein. The nucleotide sequence encoding the desired elements may include a transcriptional promoter, a transcriptional enhancer, a transcription termination sequence, a translation initiation sequence, a translation termination sequence, a ribosome binding site, a 5' untranslated region, a 3' untranslated region, a cap structure, a polyadenylic acid (polyA) tail, and / or an origin of replication. Selection of an appropriate vector will depend on several factors, including, but not limited to, the size of the nucleic acid to be incorporated into the vector, the type of transcriptional and translational control elements desired, the desired expression level, the desired copy number, whether chromosomal integration is required, the type of selection process desired, or the host cell or host range intended for transformation.

[0135] Those skilled in the art will appreciate that the biomolecules and / or compounds described herein may be provided in pharmaceutical compositions, together with a pharmaceutically acceptable diluent, carrier, or excipient, and / or together with one or more individual active agents or drugs as part of a pharmaceutical combination or composition. In certain specific embodiments, the biomolecules, compounds, and / or pharmaceutical compositions are administered simultaneously, sequentially, or in combination with other drugs or pharmaceutical compositions during a treatment regimen, separately, or as a combined formulation or combination.

[0136] The biomolecules, compounds, and / or compositions described herein may contain one or more pharmaceutically acceptable excipients, diluents, and / or carriers. Pharmaceutically acceptable carriers, diluents, or excipients include any suitable carriers, diluents, or excipients known to those skilled in the art. Examples of pharmaceutically acceptable excipients include, but are not limited to, cellulose derivatives, sucrose, and starch. Those skilled in the art will understand that pharmaceutically acceptable excipients include suitable fillers, binders, lubricants, buffers, glidants, and disintegrants known in the art (see, for example, Remington: The Science and Practice of Pharmacy (2006)). Examples of pharmaceutically acceptable carriers, diluents, and excipients are described, for example, in Remington's Pharmaceutical Sciences (2000, 20th edition) and the United States Pharmacopeia: The National Formulary (USP24 NF19), 1999.

[0137] It is also understood that one or more conservative amino acid substitutions are possible. Conservative amino acid substitutions include the following: Amino acids are substituted with other amino acids having similar properties without significantly affecting protein folding, activity, or other functions. Examples of aromatic amino acids that can be substituted include phenylalanine, tryptophan, and tyrosine. Examples of interchangeable hydrophobic amino acids that can be substituted include leucine, isoleucine, methionine, and valine. Examples of interchangeable polar amino acids that can be substituted include glutamine and asparagine. Examples of interchangeable basic amino acids that can be substituted include arginine, lysine, and histidine. Examples of interchangeable acidic amino acids that can be substituted include aspartic acid and glutamic acid. Finally, examples of interchangeable small amino acids that can be substituted include alanine, serine, threonine, cysteine, and glycine.

[0138] As described in detail herein, dominant-negative (DN) Rab1a (e.g., GDP-bound Rab1a, Rab1a GDP ) can be used to increase microautophagy, which provides for the degradation of cellular targets of interest, such as disease-related proteins, glycogen, or lipids. As will be appreciated, any suitable Rab1a DN or Rab1a GDP In light of the teachings herein, one of skill in the art will already be aware of several Rab1a DN proteins. In certain specific embodiments, generally, any suitable dominant-negative form of Rab1a (i.e., Rab1a constitutively / anchored in its GDP form) is used to promote the translocation of lysosomes (from the perinuclear region of the cell) to the cytoplasm and cell periphery, and to stimulate, for example, direct interaction of lysosomes with target substrates.

[0139] In certain specific embodiments, Rab1a DN( GDP Expression of mTORC1 / mTORC2 and AKT is thought to stimulate the activation of mTORC1 / mTORC2 and AKT, based on its effects in promoting lysosome localization and trafficking of lysosomes to the periphery and target substrates (see also Jia, R. and Bonifacino, JS, Lysosome Positioning Influences mTORC2 and AKT Signaling. Molecular Cell 75, 26-38. e3 (2019)).

[0140] It is contemplated herein that various genetic modifications of Rab1a to lock the protein in its constitutive GDP-bound state can be used to promote direct interaction with lysosomes and the stepwise uptake of one or more different target substrates (e.g., one or more protein, lipid, and / or glycogen targets). The constitutive GDP-bound form (DN) of Rab1a is generally not obtained under normal physiological conditions; the native protein, Rab1a, is typically constantly converted between the GTP and GDP forms. In certain specific embodiments, genetic mutations and / or amino acid substitutions / modifications are used to lock the GTPase in its GDP-bound form (or constitutive GTP form) and prevent it from entering the GTP state (or GDP state). In certain specific embodiments, any one or more appropriate modifications that substantially maintain the integrity of the GDP form / state of the GTPase (which affect signaling and lysosome trafficking to the cell periphery / cytoplasm) promote microautophagy and degradation of one or more target substrates via stepwise lysosomal uptake.

[0141] In certain specific embodiments, Rab1a GDP can generally be administered to a particular cell type or individual in need thereof by any suitable method, with the method being selected to suit the particular cell type, individual, and / or indication. In certain specific embodiments, Rab1a is administered by any suitable transfection or nucleic acid delivery method known to those of skill in the art in view of the teachings herein. GDPA nucleic acid sequence encoding and capable of expressing the protein is administered to an individual or introduced into a cell type. In certain specific embodiments, delivery is based on DNA or RNA transfection (e.g., using one or more common transfection reagents, such as Lipofectamine (Invitrogen), Fugene (Roche)), or using DNA adenovirus (gene therapy), or using modified RNA (i.e., stabilized RNA) to deliver it into the body (e.g., using viruses or microvesicles, exosomes, or ectosomes). In certain specific embodiments, the protein is administered or delivered to cells in need, assisted by any suitable technique or delivery vehicle to facilitate protein delivery to one or more cells, as the case may be.

[0142] In certain specific embodiments, lipid nanoparticles (LNPs) are used to detect Rab1a GDP or Rab1a DN The LNP is administered to a specific cell type or to a patient. GDP or Rab1a DN , Rab1a GDP or Rab1a DN It is envisioned that the vector may encapsulate a nucleic acid capable of expressing a nucleic acid sequence of the invention, a nucleic acid sequence capable of expressing a nucleic acid sequence of the invention, or a combination thereof.

[0143] LNPs are an advanced non-viral gene delivery system that enables practitioners to safely and effectively deliver nucleic acids to cells ex vivo or in vivo, with applications including, but not limited to, gene editing, rapid vaccine development, immuno-oncology, and treatment of rare genetic and drug-resistant diseases.

[0144] LNPs are being used more frequently in the art due to their advantageous properties, including controllable and sustained release properties, low toxicity, biocompatibility with tissues and cells, low immune response, increased gene size that can be delivered, cell-free manufacturing, high nucleic acid encapsulation efficiency, potent transfection, and improved tissue penetration for therapeutic agent delivery.

[0145] LNP formulations are combinations of multiple lipids / molecules, including, but not limited to, ionizable cationic lipids, neutral lipids, helper lipids, phospholipids, poly(lactic-co-glycolic acid) (PLGA), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, and polyethylene glycol (PEG)-lipid conjugates. The LNPs used can be customized or purchased from commercial sources, such as, but not limited to, TriLink BioTechnologies or Precision Nano System.

[0146] It is also envisioned that LNPs will be replaced by liposomes or polymeric nanoparticles. [Example]

[0147] Rab1a DN (GDP form) reduces the symptoms of Parkinson's disease, reduces cell death of dopaminergic neurons, and stimulates microautophagy. Rab1a DNA series of experiments were conducted to investigate the antiparkinsonian effects of MG-008 (also referred to herein as MG-008) on a wide range of Parkinson's disease symptoms and dopaminergic neuronal cell death. 6-OHDA was intracranially injected into the medial forebrain bundle (MFB) of male Sprague-Dawley rats (8 weeks old, N = 16) to create a hemiparkinsonian rat model. Four weeks after 6-OHDA injection, motor deficits appeared, and half of the rats (N = 8) were administered MG-008 (encoded by mRNA and encapsulated in an ANM formulation) via unilateral stereotaxic injection.

[0148] First, motor performance was assessed using a rotarod apparatus. As shown in Figure 1, rotarod tests were conducted at different time intervals. At 3 weeks (A and B) and 5 weeks (C and D) after MG008 treatment, untreated (6-OHDA) and treated (+MG008) rats were subjected to rotarod testing according to a previously described standard operating procedure (https: / / med.stanford.edu / sbfnl / services / bm / sm / rotor-rod.html). Briefly, rats were placed on a horizontal rotor (rod) suspended above the cage bottom at a height low enough to avoid injury but high enough to induce an attempt to avoid falling. Rats naturally attempted to stay on the rotarod and avoid falling to the ground. The length of time the rats remained on the rotarod was measured as a measure of their balance, coordination, physical condition, and motor planning ability. The rotarod apparatus was set to accelerate from 4 rpm to 40 rpm over a 300-second period. The test began when acceleration began and ended when the rat fell from the rotarod. This procedure was repeated for a total of three tests, with a 15-minute inter-test interval. The latency to fall (in seconds) (A and C) and the speed (rpm) during the fall (B and D) were recorded. Statistical analysis was performed using one-way ANOVA, **p<0.01, N=8. Figure 1 shows that administration of MG-008 increased the latency to fall and the rotarod speed during the fall compared to untreated (6-OHDA) rats. This indicates that administration of MG-008 improves motor performance in a hemiparkinsonian rat model.

[0149] Next, motor ability was assessed using a beam-walking test, which evaluates motor coordination, particularly in the hindlimbs. Rats were placed on one end of a narrow beam and asked to walk from one end to the other at least three times. The narrow beam was 1–3 cm wide and spanned the space between the rod and the cage (to attract the rats to the end point). This training step was used to obtain a stable baseline measurement. The number of foot slips and the time required to cross the beam were recorded for each trial. Figure 2 shows the beam-walking test performed 4 weeks after MG008 treatment. The prevalence of motor impairment was assessed by measuring the time (in seconds) required for the rats to cross the beam. This procedure was repeated for a total of three trials, with a 30-second intertrial interval. A decrease in the time required to cross the beam indicates an improvement in motor ability. Statistical analysis was performed using one-way ANOVA; *p<0.05, N=8. Figure 2 shows that MG-008 treatment improved motor coordination in hemiparkinsonian rats. This is evidenced by a reduction in the time required to traverse the beam compared to untreated rats (6-OHDA).

[0150] Next, motor performance was assessed using the amphetamine rotation test. The rotation test is commonly used to measure the degree of motor impairment induced by lesions such as 6-OHDA injection. This test is a standard tool for demonstrating the effectiveness of neuroprotective interventions aimed at functional recovery or preserving or restoring dopaminergic neuron function. Figure 3 shows the apomorphine (APO)-induced contralateral rotation test performed 4 weeks after MG008 treatment to evaluate the effect of MG008 on the motor performance of hemiparkinsonian rats that had unilateral damage to dopaminergic neurons in the substantia nigra induced by injection of 6-OHDA into the midbrain ventricular fibula (MFB). The rats' rotational speed in the rotator was automatically recorded throughout the entire test period (typically 60 min) and displayed as the average 360° / hour. A decrease in rotation speed indicates the recovery of the rats' dopaminergic neurons. Statistical analysis was performed using one-way ANOVA; **p<0.01, N=8. FIG. 3 shows that MG-008 treatment in hemiparkinsonian rats reduced rotational speed and improved motor performance compared to untreated rats (6-OHDA).

[0151] Next, we detected dopaminergic neuronal death in hemiparkinsonian rats, which were unilaterally damaged in the substantia nigra by injecting 6-OHDA into the midbrain. Figure 5 shows hematoxylin-eosin staining in the substantia nigra. Eight weeks after MG008 treatment, rats were euthanized, and brain tissue was extracted and subjected to stereological analysis using an optical splitter. Figures show the substantia nigra pars compacta (SNpc) and substantia nigra pars reticulata (SNr) regions of MG008-treated (A) and control (B) rats. Compared to the significant neuronal degeneration observed in control rats (6-OHDA, B), neurons in MG008-treated rats (A) displayed normal cytoplasmic staining and healthy cell morphology, suggesting the recovery of dopaminergic neurons in hemiparkinsonian rats.

[0152] Although one or more illustrative embodiments have been described by way of example, those skilled in the art will recognize that various changes and modifications can be made without departing from the scope of the invention as set forth in the claims. [Example]

[0153] Rab1a encapsulated in lipid nanoparticles DN (GDP format) reduces symptoms of Parkinson's disease A series of experiments were conducted to study the effects of F11 treatment on Parkinson's disease symptoms and dopaminergic neuron survival. F11 was administered to Rab1a encapsulated in lipid nanoparticles (LNPs), as described in the following paragraphs and Figures 6 and 7. DN is.

[0154] Two experimental schedules outlined in Figures 6 and 7 illustrate various dosing schedules for F11 in a hemiparkinsonian rat model. Both schedules include pre-dose measurements, including ambulation, blood sampling and testing (complete blood count (CBC) and BCS), beam walking test, rotarod test, open field test, and metabolic cage test. The principles of the beam walking test and rotarod test are described in Example 1. The open field test is an experimental test for assessing anxiety and general locomotor activity and is well known to those skilled in the art. The metabolic cage test consists of a specialized housing unit for small animals, such as rodents, designed to allow researchers to closely monitor and measure metabolic rate, food and water intake, and other physiological parameters of the animals. Cages are typically used to study the effects of various factors, such as diet, drugs, and environmental conditions, on animal metabolism and behavior and are well known to those skilled in the art.

[0155] After collecting the pre-dose baseline measurements shown above, Sprague-Dawley rats were intracranially injected with 8 μg of 6-OHDA per rat into the substantia nigra pars compacta (SNpc) to establish a hemiparkinsonian rat model, as described in Example 1, to evaluate the effects of F11.

[0156] One week after administration of 6-OHDA, rats were intravenously injected according to one of the following administration schedules.

[0157] [Table 1]

[0158] Next, at 2, 4, 8, and 12 weeks after treatment with F11, the animals underwent the beam walking test, rotarod test, rotation test, and open field test (Figures 6 and 7). The beam walking test, rotarod test, and rotation test were described in Example 1.

[0159] In the rotarod test, the latency to fall was significantly increased at 2, 4, 8, and 12 weeks after the first intravenous administration of F11 compared with rats administered 6-OHDA alone, across almost all dosing schedules shown in Table 1 (Figure 9). These results indicate that F11 improves balance and motor coordination in hemiparkinsonian rats. Similarly, compared with rats administered 6-OHDA, hemiparkinsonian rats administered F11 generally showed improved beam walking time (i.e., reduced time required to cross the beam) at 2, 4, 8, and 12 weeks after the first administration of F11 (Figure 10). Therefore, the beam walking test also indicates improved motor coordination in hemiparkinsonian rats administered F11. Finally, compared with rats administered 6-OHDA, hemiparkinsonian rats administered F11 showed reduced rotational speed and improved motor performance in the rotation test (Figure 11).

[0160] Twelve weeks after the start of the experiment, rats were euthanized, and brain, liver, and kidney tissues were collected for immunohistochemistry and H&E staining. Brain sections were stained to confirm the expression of tyrosine hydroxylase (TH) in the substantia nigra of hemiparkinsonian rats treated with or without F11. While the mechanism of Parkinson's disease (PD) pathogenesis remains unclear, symptoms appear to be related to the degeneration of dopaminergic neurons. As the disease progresses, neuronal degeneration gradually reduces dopamine synthesis, leading to abnormal discharges in the cerebral cortex. TH is the rate-limiting enzyme in dopamine synthesis, plays a key role in dopamine synthesis, and is associated with the progression of PD. Therefore, interfering with TH expression and synthesis may improve neurological symptoms in PD rats. Figures 12 and 13 show that intravenous administration of F11 increased TH expression in the substantia nigra of hemiparkinsonian rats. In particular, the most significant effects were observed in the PD+IV*2 (once a week), PD+IV*4 (once a week), PD+IV*2 (every 3 weeks), and PD+IV*2 (every 4 weeks) groups.

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[0162] All references cited herein and throughout the specification are hereby incorporated by reference in their entirety.

Claims

1. A drug for reducing the symptoms of Parkinson's disease in individuals where it is needed, or for preventing or treating Parkinson's disease in individuals where it is needed, for increasing the cell viability of dopaminergic neurons in individuals where it is needed, or for preventing or treating the death of dopaminergic neurons in individuals where it is needed, or for increasing the cell viability of dopaminergic neurons in or outside the body, GDP-bound Rab1a (Rab1a ＧＤＰ ), Rab1a ＧＤＰ A drug comprising one or more expressible nucleic acids that encode a substance, or a combination thereof.

2. The agent according to claim 1, wherein the symptoms include at least one of resting tremor, postural instability, bradykinesia, muscle rigidity, stereotypic behavior, motor disorder, hallucinations, impulse control disorder, and sleep disorder.

3. The aforementioned Rab1a ＧＤＰ However, Rab1a Ｓ２５Ｎ Rab1a Ｎ１２４Ｉ Rab1a Ｄ４１Ｎ Rab1a Ｄ４７Ｎ The agent according to claim 1 or 2, which is or another dominant-negative (DN) GDP-bound Rab1a, or comprises the same.

4. The aforementioned Rab1aGDP has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (Sequence ID 6, Human Rab1a) Ｓ２５Ｎ ); MGDYKDDDDKGGSGGSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTD QESFNNVKQWLQEIDRYASENVNKLLVGIKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (Accession No. 9, Mouse Rab1 Ｎ１２４Ｉ ); Human Rab1a Ｄ４１Ｎ The amino acid sequence of; or MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVNFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (Sequence ID 18, Human Rab1a) Ｄ４７Ｎ ); The agent according to claim 1 or 2, comprising a polypeptide that includes, or is composed of, or has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with any of these sequences and preferentially binds to GDP.

5. The aforementioned Rab1a ＧＤＰ This is the form of a fusion protein, and the Rab1a ＧＤＰ However, if necessary, via the linker, signaling peptides or targeted peptides, fluorescent peptides or other markers or tracers, or targeted delivery, enhanced uptake into cells, increased stability or in vivo half-life, or the aforementioned Rab1a ＧＤＰ The agent according to claim 1 or 2, which is fused with, or otherwise directly or indirectly conjugated with, another peptide or non-peptide moiety for other therapeutic, diagnostic or improvement of bio-characteristics.

6. The aforementioned fusion protein has the following amino acid sequence: MSSMNPEYDYLFKLLLIGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNV KQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (Sequence ID 6, Human Rab1a) Ｓ２５Ｎ ); Alternatively, the agent according to claim 5, comprising a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the polypeptide and preferentially binding to GDP.

7. The aforementioned fusion protein has the following amino acid sequence: MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDR DKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLD TGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKLMSSMNPEYDYLFKLLL IGDSGVGKNCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC (Sequence No. 21); Alternatively, the agent according to claim 6, comprising a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the polypeptide and preferentially binding to GDP.

8. The agent according to claim 1, wherein the one or more expressible nucleic acids are DNA-based or RNA-based.

9. The one or more expressible nucleic acids are temporarily expressed within the dopaminergic neuron, and the Rab1a ＧＤＰ Either expresses Rab1a, or one or more of the expressible nucleic acids are incorporated into the cell genome, and Rab1a is present in the dopaminergic neurons. ＧＤＰ The drug according to claim 1, which expresses [the specified expression].

10. The one or more expressible nucleic acids described above are expressed within the cell, and the Rab1a ＧＤＰ The agent according to claim 1, comprising one or more expression vectors, plasmids, or mRNAs that encode and are expressible.

11. The one or more expressible nucleic acids mentioned above have the following nucleic acid sequences: ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a Ｓ２５Ｎ ORF codon sequence, SEQ ID NO: 5); ATGGGGGACTACAAGGACGACGATGACAAGGGGGGTAGCGGTGGATCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGATTCTGGGGTTGGAAAGTCCTGCCTTCTCCTTAGGTTTGCAGATGATACGTATACGGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAGATACGAACTATAGAGTTAGATGGGAAAACAATCAAGCTACAGATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACTTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAACGTTAAACAGTGGCTGCAGGAGATAGATCGCTACGCCAGTGAAAATGTCAACAAGTTGTTGGTAGGGATCAAATGTGACCTGACCACAAAGAAAGTAGTAGACTACACAACAGCAAAGGAATTTGCAGATTCCCTTGGAATTCCATTTTTGGAAACCAGTGCTAAGAACGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCTGGAGCTACAGCTGGTGGTGCCGAGAAGTCCAATGTTAAAATCCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGCTGCTGCTAA (Mouse Rab1a) Ｎ１２４Ｉ ORF codon sequence, SEQ ID NO: 8); or ATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGTCTTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGAACTTCAAAATAAGAA CTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAA CAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTG CTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Human Rab1a Ｄ４７Ｎ ORF codon sequence, SEQ ID NO: 17); Alternatively, Rab1a having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with these, and preferentially binding to GDP. ＧＤＰ nucleic acid sequence encoding; Alternatively, the agent according to claim 10, comprising a nucleic acid sequence equivalent to any of the above sequences due to codon redundancy.

12. The one or more expressible nucleic acids mentioned above have the following nucleic acid sequences: ATGGAAGACGCCAAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTAGAGGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGTGAACATCACGTACGCGGAATACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGAACATTTCGCAGCCTACCGTAGTGTTTGTTTCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAATTACCAATAATCCAGAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTACCAGAGTCCTTTGATCGTGACAAAACAATTGCACTGATAATGAACTCCTCTGGATCTACTGGGTTACCTAAGGGTGTGGCCCTTCCGCATAGAACTGCCTGCGTCAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTTTACGATCCCTTCAGGATTACAAAATTCAAAGTGCGTTGCTAGTACCAACCCTATTTTCATTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGGGGCGCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCAAAACGCTTCCATCTTCCAGGGATACGACAAGGATATGGGCTCACTGAGACTACATCAGCTATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGCGAATTATGTGTCAGAGGACCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAACACTTCTTCATAGTTGACCGCTTGAAGTCTTTAATTAAATACAAAGGATACCAGGTGGCCCCCGCTGAATTGGAGTCGATATTGTTACAACACCCCAACATCTTCGACGCGGGCGTGGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCCAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGTCCAAATTGATGTCCAGCATGAATCCCGAATATGATTATTTATTCAAGTTACTTCTGATTGGCGACTCAGGGGTTGGAAAGAATTGCCTTCTTCTTAGGTTTGCAGATGATACATATACAGAAAGCTACATCAGCACAATTGGTGTGGATTTCAAAATAAGAACTATAGAGTTAGACGGGAAAACAATCAAGCTTCAAATATGGGACACAGCAGGCCAGGAAAGATTTCGAACAATCACCTCCAGTTATTACAGAGGAGCCCATGGCATCATAGTTGTGTATGATGTGACAGATCAGGAGTCCTTCAATAATGTTAAACAGTGGCTGCAGGAAATAGATCGTTATGCCAGTGAAAATGTCAACAAATTGTTGGTAGGGAACAAATGTGATCTGACCACAAAGAAAGTAGTAGACTACACAACAGCGAAGGAATTTGCTGATTCCCTTGGAATTCCGTTTTTGGAAACCAGTGCTAAGAATGCAACGAATGTAGAACAGTCTTTCATGACGATGGCAGCTGAGATTAAAAAGCGAATGGGTCCCGGAGCAACAGCTGGTGGTGCTGAGAAGTCCAATGTTAAAATTCAGAGCACTCCAGTCAAGCAGTCAGGTGGAGGTTGCTGCTAA (Sequence ID 19, MG-008 ORF DNA sequence with 5' luciferase tag); or AUGGAAGACGCCAAAAACAUAAAGAAAGGCCCGGCGCCAUUCUAUCCGCUAGAGGAUGGAACCGCUGGAGAGCAACUGCAUAAGGCUAUGAAGAGAUACGCCCUGGUUCCUGGAACAAUUGCUUUUACAGAUGCACAUAUCGAGGUGAACAUCACGUACGCGGAAUACUUCGAAAUGUCCGUUCGGUUGGCAGAAGCUAUGAAACGAUAUGGGCUGAAUACAAAUCACAGAAUCGUCGUAUGCAGUGAAAACUCUCUUCAAUUCUUUAUGCCGGUGUUGGGCGCGUUAUUUAUCGGAGUUGCAGUUGCGCCCGCGAACGACAUUUAUAAUGAACGUGAAUUGCUCAACAGUAUGAACAUUUCGCAGCCUACCGUAGUGUUUGUUUCCAAAAAGGGGUUGCAAAAAAUUUUGAACGUGCAAAAAAAAUUACCAAUAAUCCAGAAAAUUAUUAUCAUGGAUUCUAAAACGGAUUACCAGGGAUUUCAGUCGAUGUACACGUUCGUCACAUCUCAUCUACCUCCCGGUUUUAAUGAAUACGAUUUUGUACCAGAGUCCUUUGAUCGUGACAAAACAAUUGCACUGAUAAUGAACUCCUCUGGAUCUACUGGGUUACCUAAGGGUGUGGCCCUUCCGCAUAGAACUGCCUGCGUCAGAUUCUCGCAUGCCAGAGAUCCUAUUUUUGGCAAUCAAAUCAUUCCGGAUACUGCGAUUUUAAGUGUUGUUCCAUUCCAUCACGGUUUUGGAAUGUUUACUACACUCGGAUAUUUGAUAUGUGGAUUUCGAGUCGUCUUAAUGUAUAGAUUUGAAGAAGAGCUGUUUUUACGAUCCCUUCAGGAUUACAAAAUUCAAAGUGCGUUGCUAGUACCAACCCUAUUUUCAUUCUUCGCCAAAAGCACUCUGAUUGACAAAUACGAUUUAUCUAAUUUACACGAAAUUGCUUCUGGGGGCGCACCUCUUUCGAAAGAAGUCGGGGAAGCGGUUGCAAAACGCUUCCAUCUUCCAGGGAUACGACAAGGAUAUGGGCUCACUGAGACUACAUCAGCUAUUCUGAUUACACCCGAGGGGGAUGAUAAACCGGGCGCGGUCGGUAAAGUUGUUCCAUUUUUUGAAGCGAAGGUUGUGGAUCUGGAUACCGGGAAAACGCUGGGCGUUAAUCAGAGAGGCGAAUUAUGUGUCAGAGGACCUAUGAUUAUGUCCGGUUAUGUAAACAAUCCGGAAGCGACCAACGCCUUGAUUGACAAGGAUGGAUGGCUACAUUCUGGAGACAUAGCUUACUGGGACGAAGACGAACACUUCUUCAUAGUUGACCGCUUGAAGUCUUUAAUUAAAUACAAAGGAUACCAGGUGGCCCCCGCUGAAUUGGAGUCGAUAUUGUUACAACACCCCAACAUCUUCGACGCGGGCGUGGCAGGUCUUCCCGACGAUGACGCCGGUGAACUUCCCGCCGCCGUUGUUGUUUUGGAGCACGGAAAGACGAUGACGGAAAAAGAGAUCGUGGAUUACGUCGCCAGUCAAGUAACAACCGCCAAAAAGUUGCGCGGAGGAGUUGUGUUUGUGGACGAAGUACCGAAAGGUCUUACCGGAAAACUCGACGCAAGAAAAAUCAGAGAGAUCCUCAUAAAGGCCAAGAAGGGCGGAAAGUCCAAAUUGAUGUCCAGCAUGAAUCCCGAAUAUGAUUAUUUAUUCAAGUUACUUCUGAUUGGCGACUCAGGGGUUGGAAAGAAUUGCCUUCUUCUUAGGUUUGCAGAUGAUACAUAUACAGAAAGCUACAUCAGCACAAUUGGUGUGGAUUUCAAAAUAAGAACUAUAGAGUUAGACGGGAAAACAAUCAAGCUUCAAAUAUGGGACACAGCAGGCCAGGAAAGAUUUCGAACAAUCACCUCCAGUUAUUACAGAGGAGCCCAUGGCAUCAUAGUUGUGUAUGAUGUGACAGAUCAGGAGUCCUUCAAUAAUGUUAAACAGUGGCUGCAGGAAAUAGAUCGUUAUGCCAGUGAAAAUGUCAACAAAUUGUUGGUAGGGAACAAAUGUGAUCUGACCACAAAGAAAGUAGUAGACUACACAACAGCGAAGGAAUUUGCUGAUUCCCUUGGAAUUCCGUUUUUGGAAACCAGUGCUAAGAAUGCAACGAAUGUAGAACAGUCUUUCAUGACGAUGGCAGCUGAGAUUAAAAAGCGAAUGGGUCCCGGAGCAACAGCUGGUGGUGCUGAGAAGUCCAAUGUUAAAAUUCAGAGCACUCCAGUCAAGCAGUCAGGUGGAGGUUGCUGCUAA (Sequence ID 20, MG-008 ORF mRNA sequence with 5' luciferase tag); Alternatively, nucleic acid sequences encoding Rab1aGDP that have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with these, and preferentially bind to GDP; Alternatively, the agent according to claim 10, comprising a nucleic acid sequence equivalent to any of the above sequences due to codon redundancy.

13. GDP-bound Rab1a (Rab1a ＧＤＰ ), Rab1a ＧＤＰ A pharmaceutical composition comprising one or more expressible nucleic acids encoding a certain substance, or a combination thereof, and other antiparkinsonian agents.

14. The agent according to claim 1, wherein the dopaminergic neurons are located in the midbrain.

15. The agent according to claim 14, wherein the dopaminergic neurons are located in the substantia nigra pars compacta (SNpc) or substantia nigra pars reticularis (SNr).

16. A drug for treating or preventing Parkinson's disease, or for increasing the cell viability of dopaminergic neurons in or outside the body, Rab1a ＧＤＰ A drug containing encapsulated lipid nanoparticles.

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