Methods for treating parkinson's disease

Abstract

In some embodiments, a method is provided for the treatment of Parkinson's disease (Parkinson Disease), including a TRIM72 protein modulator. Also provided is a composition comprising a TRIM72 protein modulator and uses thereof.

Description

Background Art

[0001] Parkinson's disease is a voluntary movement disorder in which muscles become stiff and slow, movements become clumsy and difficult, and uncontrollable rhythmic twitching of muscle groups produces characteristic tremors or tremors. The condition is believed to be caused by the degeneration of presynaptic dopaminergic neurons in the brain. Therefore, the lack of sufficient release of the chemical transmitter dopamine during neuronal activity leads to Parkinson's syndrome. At present, the clinical therapy of PD is still in the "symptomatic treatment" stage, that is, PD patients with motor or non-motor symptoms are selected for treatment. There is a lack of drugs developed based on the pathogenesis or pathophysiology of the disease. Drugs used to treat motor system disorders mainly target the dopaminergic system pharmacologically, including levodopa, non-ergodopa agonists and monoamine oxidase inhibitors. Levodopa drugs have a significant effect on improving motor symptoms, with fast onset and fewer side effects. However, dyskinesias occur in subjects later, and the dosage and frequency of such drugs are also high. Moreover, treatment with dopamine receptor agonists may cause impulse control disorders and other behavioral disorders, or aggravate patients' psychiatric symptoms (such as anxiety, insomnia, etc.). Few drugs can target the non-motor symptoms of Parkinson's disease. Although a variety of psychiatric drugs can effectively improve some symptoms of PD patients, whether the use of these drugs will affect PD progression and safety remains to be verified. Therefore, disease-modifying therapies for PD are urgently needed. Oxidative stress is a key pathological mechanism of PD, but anti-oxidative stress drugs have not been successful so far. Endogenous antioxidant molecular targets are potential breakthroughs in drug development.

[0002] Therefore, there remains a need to explore safe and effective alternative approaches for treating Parkinson's disease. Summary of the invention

[0003] The present application provides a method for preventing and / or treating Parkinson's disease. The method can prevent and / or treat Parkinson's disease safely and effectively.

[0004] In one aspect, the present application provides a method for preventing and / or treating Parkinson's disease, comprising administering one or more TRIM72 protein modulators.

[0005] In some embodiments, a TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

[0006] In some embodiments, the TRIM72 modulator is selected from one or more of the group consisting of: a protein, a peptide, a peptide mimetic, a chemical compound, an antibody, a ribozyme, a small molecule chemical compound, a nucleic acid, a vector, and an antisense nucleic acid.

[0007] In some embodiments, the TRIM72 protein comprises a TRIM72 protein or a variant thereof or a functional fragment thereof.

[0008] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof.

[0009] In some embodiments, the TRIM72 protein comprises a full-length TRIM72 protein.

[0010] In some embodiments, the TRIM72 protein comprises a wild-type TRIM72 protein.

[0011] In some embodiments, the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:2.

[0012] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof.

[0013] In some embodiments, the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

[0014] In some embodiments, the PRYSPRY domain comprises amino acid positions 278aa-470aa of the TRIM72 protein.

[0015] In some embodiments, the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:6.

[0016] In some embodiments, the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0017] In some embodiments, the TRIM72 truncated protein does not comprise the coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0018] In some embodiments, the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

[0019] In some embodiments, the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:5.

[0020] In some embodiments, the TRIM72 truncated protein further comprises the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0021] In some embodiments, the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0022] In some embodiments, the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

[0023] In some embodiments, the B-box domain comprises the amino acid sequence shown in SEQ ID NO:4.

[0024] In some embodiments, the TRIM72 truncated protein further comprises the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0025] In some embodiments, the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0026] In some embodiments, the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

[0027] In some embodiments, the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:3.

[0028] In some embodiments, the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9, and 11.

[0029] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises an amino acid mutation at position C14.

[0030] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises the amino acid mutation C14A.

[0031] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof does not comprise an amino acid mutation at position C242.

[0032] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof is secreted via exosomes.

[0033] In some embodiments, the TRIM72 protein modulator comprises a nucleic acid molecule encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0034] In some embodiments, the TRIM72 protein modulator comprises a vector comprising a gene encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0035] In some embodiments, the vector is a plasmid or a virus.

[0036] In some embodiments, the vector is an adeno-associated virus (rAAV) expression vector.

[0037] In some embodiments, the universal promoter is selected from one or more of the group consisting of chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV), and simian virus 40 (SV40) promoters.

[0038] In some embodiments, the vector comprises a neuron-specific promoter.

[0039] In some embodiments, the promoter comprises a human promoter.

[0040] In some embodiments, the promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a neocortex and hippocampus excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

[0041] In some embodiments, the promoter is selected from the group consisting of human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

[0042] In some embodiments, the serotype of the AAV vector is selected from AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ, and AAVhull.

[0043] In some embodiments, the TRIM72 protein modulator comprises a cell, wherein the cell comprises the vector.

[0044] In some embodiments, the TRIM72 protein modulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0045] In some embodiments, TRIM72 protein modulators protect neurons by reducing oxidative stress, thereby preventing and / or treating Parkinson's disease.

[0046] On the other hand, the present application provides a composition for preventing and / or treating Parkinson's disease, comprising one or more TRIM72 protein regulators.

[0047] In some embodiments, a TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

[0048] In some embodiments, the TRIM72 modulator is selected from one or more of the group consisting of: a protein, a peptide, a peptide mimetic, a chemical compound, an antibody, a ribozyme, a small molecule chemical compound, a nucleic acid, a vector, and an antisense nucleic acid.

[0049] In some embodiments, the TRIM72 protein comprises a TRIM72 protein or a variant thereof or a functional fragment thereof.

[0050] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof.

[0051] In some embodiments, the TRIM72 protein comprises a full-length TRIM72 protein.

[0052] In some embodiments, the TRIM72 protein comprises a wild-type TRIM72 protein.

[0053] In some embodiments, the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:2.

[0054] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof.

[0055] In some embodiments, the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

[0056] In some embodiments, the PRYSPRY domain comprises amino acid positions 278aa-470aa of the TRIM72 protein.

[0057] In some embodiments, the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:6.

[0058] In some embodiments, the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0059] In some embodiments, the TRIM72 truncated protein does not comprise the coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0060] In some embodiments, the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

[0061] In some embodiments, the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:5.

[0062] In some embodiments, the TRIM72 truncated protein further comprises the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0063] In some embodiments, the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0064] In some embodiments, the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

[0065] In some embodiments, the B-box domain comprises the amino acid sequence shown in SEQ ID NO:4.

[0066] In some embodiments, the TRIM72 truncated protein further comprises the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0067] In some embodiments, the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0068] In some embodiments, the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

[0069] In some embodiments, the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:3.

[0070] In some embodiments, the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9, and 11.

[0071] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises an amino acid mutation at position C14.

[0072] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises the amino acid mutation C14A.

[0073] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof does not comprise an amino acid mutation at position C242.

[0074] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof is secreted via exosomes.

[0075] In some embodiments, the TRIM72 protein modulator comprises a nucleic acid molecule encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0076] In some embodiments, the TRIM72 protein modulator comprises a vector comprising a gene encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0077] In some embodiments, the vector is a plasmid or a virus.

[0078] In some embodiments, the vector is an adeno-associated virus (rAAV) expression vector.

[0079] In some embodiments, the universal promoter is selected from one or more of the group consisting of chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV), and simian virus 40 (SV40) promoters.

[0080] In some embodiments, the vector comprises a neuron-specific promoter.

[0081] In some embodiments, the promoter comprises a human promoter.

[0082] In some embodiments, the promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a neocortex and hippocampus excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

[0083] In some embodiments, the promoter is selected from the group consisting of human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

[0084] In some embodiments, the serotype of the AAV vector is selected from AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ, and AAVhull.

[0085] In some embodiments, the TRIM72 protein modulator comprises a cell, wherein the cell comprises the vector.

[0086] In some embodiments, the TRIM72 protein modulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0087] In some embodiments, TRIM72 protein modulators protect neurons by reducing oxidative stress, thereby preventing and / or treating Parkinson's disease.

[0088] On the other hand, the present application provides the use of a TRIM72 protein modulator in the preparation of a medicament for preventing and / or treating Parkinson's disease.

[0089] In some embodiments, a TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

[0090] In some embodiments, the TRIM72 modulator is selected from one or more of the group consisting of: a protein, a peptide, a peptide mimetic, a chemical compound, an antibody, a ribozyme, a small molecule chemical compound, a nucleic acid, a vector, and an antisense nucleic acid.

[0091] In some embodiments, the TRIM72 protein comprises a TRIM72 protein or a variant thereof or a functional fragment thereof.

[0092] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof.

[0093] In some embodiments, the TRIM72 protein comprises a full-length TRIM72 protein.

[0094] In some embodiments, the TRIM72 protein comprises a wild-type TRIM72 protein.

[0095] In some embodiments, the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:2.

[0096] In some embodiments, the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof.

[0097] In some embodiments, the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

[0098] In some embodiments, the PRYSPRY domain comprises amino acid positions 278aa-470aa of the TRIM72 protein.

[0099] In some embodiments, the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:6.

[0100] In some embodiments, the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0101] In some embodiments, the TRIM72 truncated protein does not comprise the coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

[0102] In some embodiments, the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

[0103] In some embodiments, the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:5.

[0104] In some embodiments, the TRIM72 truncated protein further comprises the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0105] In some embodiments, the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

[0106] In some embodiments, the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

[0107] In some embodiments, the B-box domain comprises the amino acid sequence shown in SEQ ID NO:4.

[0108] In some embodiments, the TRIM72 truncated protein further comprises the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0109] In some embodiments, the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

[0110] In some embodiments, the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

[0111] In some embodiments, the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:3.

[0112] In some embodiments, the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9, and 11.

[0113] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises an amino acid mutation at position C14.

[0114] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof comprises the amino acid mutation C14A.

[0115] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof does not comprise an amino acid mutation at position C242.

[0116] In some embodiments, the TRIM72 protein or its variant or functional fragment thereof is secreted via exosomes.

[0117] In some embodiments, the TRIM72 protein modulator comprises a nucleic acid molecule encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0118] In some embodiments, the TRIM72 protein modulator comprises a vector comprising a gene encoding the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0119] In some embodiments, the vector is a plasmid or a virus.

[0120] In some embodiments, the vector is an adeno-associated virus (rAAV) expression vector.

[0121] In some embodiments, the universal promoter is selected from one or more of the group consisting of chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV), and simian virus 40 (SV40) promoters.

[0122] In some embodiments, the vector comprises a neuron-specific promoter.

[0123] In some embodiments, the promoter comprises a human promoter.

[0124] In some embodiments, the promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a neocortex and hippocampus excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

[0125] In some embodiments, the promoter is selected from the group consisting of human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

[0126] In some embodiments, the serotype of the AAV vector is selected from AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ, and AAVhull.

[0127] In some embodiments, the TRIM72 protein modulator comprises a cell, wherein the cell comprises the vector.

[0128] In some embodiments, the TRIM72 protein modulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein or a variant thereof or a functional fragment thereof.

[0129] In some embodiments, TRIM72 protein modulators protect neurons by reducing oxidative stress, thereby preventing and / or treating Parkinson's disease.

[0130] Other aspects and advantages of the present application will be apparent to those skilled in the art from the following detailed description, in which only illustrative embodiments of the present application are shown and described. As will be appreciated, the present application is capable of other and different embodiments, and its several details are capable of modification in various obvious aspects, all without departing from the present application. Therefore, the drawings and description are to be regarded as illustrative in nature, and not restrictive.

[0131] Incorporated by Reference

[0132] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0133] The novel features of the present invention are specifically set forth in the appended claims. The features and advantages of the present invention may be better understood by reference to the following detailed description, which sets forth exemplary embodiments employing the principles of the present invention, and the accompanying drawings (also referred to herein as "figures"), in which:

[0134] Figure 1 Different TRIM72 vectors and TRIM40 vectors in the pAAV-ITR vector are illustrated.

[0135] Figure 2 The following schematic diagram is illustrated, which shows the timeline of mouse treatment and behavioral testing via MPTP modeling and scAAV-TRIM72 administration.

[0136] Figure 3 The performance of rotating rod behavior is illustrated. The residence time on the rotating rod was recorded in the experiment. (A) MPTP administration shortened the residence time. (B) Mice injected with scAAVs-TRIM72 had a significantly longer residence time. For all data points, the values ​​are expressed as mean ± SEM. Student t-test or one-way ANOVA was performed to compare data sets, ***p<0.001 (t-test), *p<0.05 (ANOVA), NS, not statistically significant.

[0137] Figure 4 The number of TH-positive dopaminergic (DA) neurons in the substantia nigra region is illustrated. (A) MPTP administration significantly reduced DA neurons, (B) while scAAV-TRIM72 injection effectively prevented the loss of these motor neurons. For all data points, the values ​​are expressed as mean ± SEM. Student's t-test was performed to compare data sets, ***p<0.001, *p<0.05, NS, not statistically significant.

[0138] Figure 5 The expression pattern of TRIM72 in exosomes was illustrated by Western blot analysis.

[0139] Figure 6 The secretion efficiency of full-length or different truncated TRIM72 in exosomes is illustrated. The values ​​of all data points are expressed as mean ± SEM. One-way ANOVA was performed to compare the data sets, ***p < 0.001, NS, not statistically significant.

[0140] Figure 7 TRIM72 domain annotation and key residues for TRIM72 function are illustrated.

[0141] Figure 8 The expression of domain-deleted TRIM72 fused to a Flag tag and EGFP is illustrated. GAPDH serves as a protein loading control. FL, full-length TRIM72.

[0142] Fig. 9 The effects of domain-deleted TRIM72 on cell viability after stress challenge are illustrated. Coiled-coil domain or PRYSPRY deletion, but not RING domain or B-box deletion, abolished the protective effect of TRIM72. Values ​​are expressed as mean ± SEM, and Student's t-test was performed to compare data sets, ***p < 0.001, **p < 0.01, NS, not statistically significant.

[0143] Fig.10 It shows that in H 2 O 2 Viability of cells expressing full-length TRIM72, various TRIM72 truncations, or full-length TRIM40 by scAAV9 infection under stress. For all data points, the values ​​are expressed as mean ± SEM. Student's t-test was performed to compare the data set with the control, **p<0.01, *p<0.05, NS, not statistically significant.

[0144] Fig.11 The effect of expression of TRIM72 mutants on cell viability after stress challenge is illustrated. The C242A mutation, but not the C14A mutation, abolished the anti-stress effect of TRIM72. These values ​​are generated from at least three independent experiments (n≥3). One-way ANOVA was performed to compare data sets, *p<0.05, **p<0.01, ***p<0.001. NS, not statistically significant. DETAILED DESCRIPTION

[0145] Although various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Without departing from the present invention, those skilled in the art may conceive of many variations, changes and substitutions. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0146] In this application, the term "TRIM72 protein modulator" generally refers to a substance or means that changes the expression, activity and / or biological function of the TRIM72 protein compared to the expression, activity and / or biological function of the TRIM72 protein in the absence of the modulator. Modulators may include (but are not limited to) chemical compounds, proteins, peptides, peptide mimetics, antibodies, ribozymes, small molecule chemical compounds, nucleic acids, vectors and antisense nucleic acids.

[0147] In this application, the term "Parkinson's disease" generally refers to a neurodegenerative disease. Parkinson's disease is a chronic progressive neurological disease characterized by neurodegeneration, especially degeneration of dopaminergic neurons. Symptoms include stooped posture, resting tremor, resting muscle weakness, unsteady gait, speech disorders, movement difficulties and eventually slowing of mental processes and / or dementia.

[0148] In the present application, the terms "TRIM72 protein" and "MG53" protein are used interchangeably, and generally include TRIM72 protein or its variants, functional fragments, analogs, homologs. TRIM72 protein may contain a ring finger, a B-box motif, a coiled coil region and / or a C-terminal PRYSPRY domain. For example, the ring finger domain may include amino acid sites of 14aa-56aa of TRIM72 protein or a functional fragment thereof. For example, the B-box domain may include amino acid sites of 86aa-117aa of TRIM72 protein or a functional fragment thereof. For example, the coiled coil domain may include amino acid sites of 135aa-232aa of TRIM72 protein or a functional fragment thereof. For example, the PRYSPRY domain may include amino acid sites of 278aa-470aa of TRIM72 protein. The term may also include TRIM72 proteins derived from any known species having TRIM72 protein.

[0149] In the present application, the term "truncated protein" generally refers to a protein having one or more amino acid deletions compared to a full-length protein. For example, a truncated protein may contain a major functional fragment of a protein. For example, a truncated protein also includes (but is not limited to) variants, functional fragments, analogs, and homologs thereof.

[0150] In the present application, "amino acid mutation Xn" refers to an amino acid mutation occurring in the amino acid residue X at position n of the amino acid sequence as shown in SEQ ID NO: 2, wherein n is a positive integer and X is an abbreviation for any amino acid residue. For example, "amino acid mutation C14" refers to an amino acid substitution occurring in the amino acid residue C corresponding to position 14 of the amino acid sequence as shown in SEQ ID NO: 2.

[0151] The amino acid mutation of the present application can be a non-conservative mutation. The non-conservative mutation can include changing the amino acid residues in the target protein or polypeptide in a non-conservative manner, for example, replacing an amino acid residue with a specific side chain size or a specific feature (e.g., hydrophilicity) with an amino acid residue with a different side chain size or a different feature (e.g., hydrophobicity).

[0152] The amino acid substitution can also be a conservative substitution. The conservative substitution can include changing the amino acid residues in the target protein or polypeptide in a conservative manner, such as replacing an amino acid residue with a specific side chain size or a specific feature (e.g., hydrophilicity) with an amino acid residue with the same or similar side chain size or the same or similar feature (e.g., still having hydrophilicity). Such conservative substitutions usually do not have a significant effect on the structure or function of the resulting protein. In the present application, an amino acid sequence variant is a mutant of a fusion protein, a fragment thereof, or a variant thereof that undergoes one or more amino acid substitutions, and may include conservative amino acid substitutions that do not significantly change the structure or function of the protein.

[0153] For example, in the present application, substitutions between the following amino acids in each group can be considered conservative substitutions:

[0154] The group of amino acids with nonpolar side chains: alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, and methionine.

[0155] The group of uncharged amino acids with polar side chains: glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine.

[0156] Negatively charged amino acids with polar side chains: aspartic acid and glutamic acid.

[0157] Positively charged basic amino acid group: lysine, arginine, and histidine.

[0158] Group of amino acids containing a phenyl group: phenylalanine, tryptophan and tyrosine.

[0159] In the present application, the term "fusion protein" generally refers to a composite polypeptide, that is, a single continuous amino acid sequence composed of two (or more) polypeptides. Fusion proteins can generally be artificially prepared by recombinant nucleic acid or chemical synthesis.

[0160] In the present application, the term "nucleic acid molecule" generally refers to a nucleotide, deoxyribonucleotide or ribonucleotide in an isolated form, or an analog thereof of any length isolated from its natural environment or artificially synthesized. The nucleic acid molecule of the present application can be isolated. For example, it can be produced or synthesized in the following manner: (i) in vitro amplification, such as polymerase chain reaction (PCR) amplification, (ii) cloning recombination, (iii) purification, such as by restriction enzyme digestion and gel electrophoresis fractionation, or (iv) synthesis, such as chemical synthesis. In some embodiments, the isolated nucleic acid is a nucleic acid molecule prepared by recombinant DNA technology. In the present application, the nucleic acid encoding the truncated protein or its functional fragment can be prepared by a variety of methods known in the art. These methods include but are not limited to overlapping extension PCR using restriction fragment manipulation or synthetic oligonucleotides. Specific operations can be found in Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989; and Ausube et al., Current Protocols in Molecular Biology, Greene Publishing and Wiley-Interscience, New York NY, 1993.

[0161] In this application, the term "vector" generally refers to a vector containing a recombinant polynucleotide, wherein the recombinant polynucleotide includes an expression control sequence operably linked to the nucleotide sequence to be expressed. The vector includes cis-acting elements sufficient for expression; other elements for expression can be provided by the host cell or can be provided in an in vitro expression system. The vector can include all expression vectors known in the art that can be incorporated into a recombinant polynucleotide, including cosmids, plasmids (e.g., naked plasmids or plasmids encapsulated in liposomes) and viruses (e.g., lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses).

[0162] In the present application, the term "coding" generally refers to the inherent properties of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA or mRNA) as a template for synthesizing other polymers and macromolecules in bioengineering, and the polymers and macromolecules have a specified nucleotide sequence (i.e., rRNA, tRNA and mRNA) or a specified amino acid sequence and the resulting biological properties. Therefore, if the transcription and translation of the mRNA corresponding to a gene produce a protein in a cell or other biological system, the gene encodes the protein. The coding strands that the nucleotide sequence is identical to the mRNA sequence and that are usually provided in the sequence table and the non-coding strands that are used as gene or cDNA transcription templates can all be referred to as proteins or other products of coding genes or cDNAs. In the present application, the term "coding element" generally refers to a nucleic acid (RNA or DNA molecule) comprising a nucleotide sequence encoding a protein.

[0163] In the present application, the terms "host cell", "cell" and "host" are used interchangeably, and generally refer to a plasmid or vector that may include or have included a nucleic acid molecule of the present application, or a single cell, cell line or cell culture that may express a protein, a fragment thereof or a variant thereof of the present application. The host cell may include progeny of a single host cell. Due to natural, unexpected or deliberate mutations, progeny cells and original parent cells do not have to be identical in morphology or genome, as long as they can express a protein or a fragment thereof of the present application. The host cell may be obtained by transfecting cells in vitro with a vector of the present application. The host cell may be a prokaryotic cell (e.g., Escherichia coli) or a eukaryotic cell (e.g., a yeast cell, such as a COS cell, a Chinese hamster ovary (CHO) cell, a HeLa cell, a HEK293 cell, a COS-1 cell, a NS0 cell or a myeloma cell). In the present application, the host cell may be a CHO cell.

[0164] As used herein, the term "treating" generally refers to slowing or ameliorating the progression, severity and / or duration of a proliferative condition, or ameliorating one or more symptoms (eg, one or more identifiable symptoms) of a proliferative condition by administering one or more therapies.

[0165] In this application, the term "subject" generally refers to any human or non-human animal. The term "non-human animal" can include all vertebrates, such as mammals and non-mammals, for example non-human primates, goats, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0166] In this application, the terms "peptide", "polypeptide" and "protein" are used interchangeably and generally refer to compounds consisting of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and there is no limit on the maximum number of amino acids that can be included in a protein or peptide sequence. A polypeptide may include any peptide or protein containing two or more amino acids linked to each other via peptide bonds. In this application, this term refers to two short chains, also commonly referred to as peptides, oligopeptides and oligomers in the art, such as long chains, which are commonly referred to as proteins in the art, and there are many types. "Polypeptide" includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, polypeptide variants, modified polypeptides, derivatives, analogs, fusion proteins, etc. Polypeptides include natural peptides, recombinant peptides, or combinations thereof.

[0167] In addition to the specific proteins and nucleotides mentioned herein, the present application may also include functional variants, derivatives, analogs, homologues and fragments thereof.

[0168] The term "functional variant" refers to a polypeptide having an amino acid sequence substantially identical to a naturally occurring sequence or being encoded by a nucleotide sequence substantially identical to a naturally occurring sequence and being capable of having one or more activities of a naturally occurring sequence. In the context of the present application, a variant of any given sequence refers to a sequence in which a specific residue (amino acid or nucleotide residue) sequence has been modified so that the polypeptide or polynucleotide substantially maintains a sequence of at least one endogenous function. The variant sequence can be obtained by adding, deleting, replacing, modifying, displacing and / or changing at least one amino acid residue and / or nucleotide residue present in a naturally occurring protein and / or polynucleotide, as long as the original functional activity is maintained. In the present application, the term "derivative" generally refers to a polypeptide or polynucleotide of the present application comprising any substitution, change, modification, displacement, deletion and / or addition of one (or more) amino acid residues of a sequence, provided that the resulting polypeptide or polynucleotide substantially maintains at least one function of its endogenous function.

[0169] In the present application, the term "analog" about a polypeptide or polynucleotide generally includes any mimetics of a polypeptide or polynucleotide, i.e., a compound having at least one endogenous function of a polypeptide or polynucleotide simulated by a mimetics. Generally speaking, an amino acid can be substituted, for example, at least one (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 20 or more) amino acid can be substituted, provided that the modified sequence substantially maintains the desired activity or ability. Amino acid substitutions can include the use of non-naturally occurring analogs. The protein or polypeptide used in the present application can also have a deletion, insertion or substitution of an amino acid residue, wherein the amino acid residue undergoes silent changes and produces a functionally equivalent protein. Intentional amino acid substitutions can be made based on the similarity of the polarity, charge, solubility, hydrophobicity, hydrophilicity and / or amphoteric properties of the residue, as long as the endogenous function is retained. For example, negatively charged amino acids include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; and amino acids containing uncharged polar head groups and having similar hydrophilicity values ​​include asparagine, glutamine, serine, threonine, and tyrosine.

[0170] In the present application, the term "homologue" generally refers to an amino acid sequence or nucleotide sequence with a specific homology to a wild-type amino acid sequence and a wild-type nucleotide sequence. The term "homology" may be equivalent to the "identity" of a sequence. A homologous sequence may include an amino acid sequence that is at least 80%, 85%, 90%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical to the target sequence. Typically, a homologue contains an active site identical to the target amino acid sequence, etc. Homology may be considered based on similarity (i.e., amino acid residues with similar chemical properties / functions), or homology may be represented by sequence identity. In the present application, a sequence having a certain identity percentage with any SEQ ID NO of the amino acid sequence or nucleotide sequence mentioned refers to a sequence having the identity percentage over the entire length of the SEQ ID NO mentioned. To determine sequence identity, sequence alignment can be performed by a variety of methods known to those skilled in the art, for example, by using BLAST, BLAST-2, ALIGN, NEEDLE or Megalign (DNASTAR) software, etc. Those skilled in the art can determine appropriate parameters for alignment, including any algorithms required to achieve optimal alignment over the full length of the sequences being compared.

[0171] In this application, the term "about" generally refers to a variation within a range of 0.5%-10% above or below a specified value, for example, a variation within a range of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% above or below a specified value.

[0172] In this application, the term "comprising" generally means including, containing, having or covering. In some cases, it also means "being" or "consisting of...".

[0173] In this application, the term "does not include" generally refers to excluding the possibility of a certain behavior, structure or structure. For example, "A does not include B" generally means excluding the possibility of B appearing in A.

[0174] Methods, uses and preparation

[0175] In one aspect, the present application provides a method for preventing and / or treating Parkinson's disease, comprising administering one or more TRIM72 protein modulators.

[0176] On the other hand, the present application provides a composition for preventing and / or treating Parkinson's disease, comprising one or more TRIM72 protein regulators.

[0177] On the other hand, the present application provides the use of a TRIM72 protein modulator in the preparation of a medicament for preventing and / or treating Parkinson's disease.

[0178] In the present application, the TRIM72 protein regulator can increase the expression and / or activity of the TRIM72 protein.

[0179] In the present application, the TRIM72 modulator may be selected from one or more of the following groups: proteins, peptides, peptide mimetics, chemical compounds, antibodies, ribozymes, small molecule chemical compounds, nucleic acids, vectors and antisense nucleic acids.

[0180] In the present application, the TRIM72 protein may include the TRIM72 protein or a variant or a functional fragment thereof.

[0181] In the present application, the TRIM72 protein regulator may include a vector. For example, the vector may include a recombinant adeno-associated virus (rAAV) expression vector, which includes a gene encoding a TRIM72 protein or a variant thereof or a functional fragment thereof.

[0182] In the present application, the TRIM72 protein or a functional fragment thereof may include a human TRIM72 protein or a functional fragment thereof.

[0183] In the present application, the TRIM72 protein may include a full-length TRIM72 protein. In the present application, the TRIM72 protein may include a wild-type TRIM72 protein. In the present application, the human TRIM72 protein includes the amino acid sequence shown in SEQ ID NO: 2.

[0184] In the present application, the TRIM72 protein may include a TRIM72 truncated protein.

[0185] In the present application, the TRIM72 truncated protein comprises a PRYSPRY domain or a functional fragment thereof. For example, the TRIM72 truncated protein may comprise the amino acid sites 278aa-470aa of the TRIM72 protein. For example, the TRIM72 truncated protein may comprise the amino acid sequence shown in SEQ ID NO:6.

[0186] In the present application, the TRIM72 truncated protein may also contain other domains of the TRIM72 protein.

[0187] For example, the TRIM72 truncated protein may include a PRYSPRY domain and a coiled-coil domain. For example, the TRIM72 truncated protein may include a deletion of the B-box domain and the RING finger domain of the TRIM72 protein. For example, the TRIM72 protein may include an amino acid sequence as shown in SEQ ID NO:11.

[0188] For example, the TRIM72 truncated protein may comprise a PRYSPRY domain and a B-box domain. For example, the TRIM72 truncated protein may comprise a deletion of the coiled-coil domain and the RING finger domain of the TRIM72 protein.

[0189] For example, the TRIM72 truncated protein may comprise a PRYSPRY domain and a RING finger domain. For example, the TRIM72 truncated protein may comprise a deletion of the B-box domain and the coiled-coil domain of the TRIM72 protein.

[0190] For example, the TRIM72 truncated protein may comprise a PRYSPRY domain, a coiled-coil domain, and a RING finger domain. For example, the TRIM72 truncated protein may comprise a deletion of the B-box domain of the TRIM72 protein.

[0191] For example, the TRIM72 truncated protein may comprise a PRYSPRY domain, a coiled-coil domain, and a B-box domain. For example, the TRIM72 truncated protein may comprise a deletion of the RING finger domain of the TRIM72 protein.

[0192] For example, the TRIM72 truncated protein may comprise a PRYSPRY domain, a RING finger domain, and a B-box domain. For example, the TRIM72 truncated protein may comprise a deletion of the coiled-coil domain of the TRIM72 protein.

[0193] In the present application, the TRIM72 protein or its fragment may include a variant thereof. For example, compared with the corresponding wild-type sequence, the TRIM72 protein may include one or more amino acid mutations.

[0194] In the present application, the TRIM72 protein may include an amino acid mutation at position C14. For example, the amino acid mutation may be C14A.

[0195] According to the applicant's research, the amino acid cysteine ​​at position 242 is important for the oligomer formation of TRIM72 protein. The substitution of amino acid C242 will block the protective function of neurons. Therefore, the amino acid substitution at position C242 (e.g., C242A) may not be included in the TRIM72 protein of the present application.

[0196] Exosomes are small extracellular biological vesicles released into the surrounding body fluids via multivesicular bodies and plasma membrane fusion, which contain proteins, nucleic acids, lipids and other biologically active substances. In the present application, TRIM72 protein or its variants or fragments thereof can be secreted via exosomes. For example, a vector comprising a gene encoding TRIM72 protein or its variants or fragments thereof can be constructed and expressed in a host cell. TRIM72 protein or its variants or fragments thereof can affect the biological processes of surrounding cells via the exosome secretion pathway.

[0197] In the present application, rAAV can include AAV genome or its derivative, and / or AAV capsid protein or its derivative. In the present application, rAAV can be chimeric AAV, reorganized AAV or capsid modified AAV. In the present application, AAV genome or AAV capsid protein can be from any one of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh, AAVDJ and AAVhull. In the present application, rAAV can be a heterozygous AAV (e.g., AAV-DJ, AAV-DJ / 8 or AAV-DJ / 9). In the present application, rAAV can be produced via directed evolution and / or rational design (e.g., AAV7m8 or AAV-PHP.eB).

[0198] In the present application, the rAAV expression vector may comprise a universal promoter. In the present application, the universal promoter may be selected from: chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1α (EF1A), mouse or human phosphoglycerate kinase (PGK), mouse stem cell virus (MSCV), spleen focus forming virus (SFFV) or simian virus 40 (SV40) promoter.

[0199] In the present application, the rAAV expression vector may comprise a neuron-specific promoter. For example, the promoter may comprise a human promoter.

[0200] In the present application, the promoter can be one or more of the following groups: an excitatory neuron-specific promoter, a neocortex and hippocampus excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

[0201] In some embodiments, the promoter can be selected from the group consisting of: human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

[0202] In the present application, the TRIM72 protein may include a recombinant protein containing the TRIM72 protein or a variant or a functional fragment thereof.

[0203] In the present application, the TRIM72 protein regulator may include one or more nucleic acid molecules capable of encoding the TRIM72 protein or a variant or functional fragment thereof.

[0204] In the present application, the TRIM72 protein modulator may include one or more vectors, which may include one or more nucleic acid molecules of the present application. In the present application, the TRIM72 protein modulator may include a cell (eg, a host cell), which may include a nucleic acid molecule of the present application or a vector of the present application.

[0205] In the present application, the vector may be a polynucleotide, which, when introduced into a suitable host cell, may be transcribed and translated into a polypeptide. Typically, the vector may produce a desired expression product by culturing a suitable host cell containing the vector. In the present application, the vector may include one or more of the nucleic acid molecules. For example, the vector may include all nucleic acid molecules required to encode the TRIM72 protein or its variant or its functional fragment.

[0206] In addition, the vector may also include other genes, such as marker genes that allow the vector to be selected in a suitable host cell and under suitable conditions. In addition, the vector may also include expression control elements that allow the coding region to be correctly expressed in a suitable host. Such control elements are well known to those skilled in the art. For example, it may include promoters, ribosome binding sites, enhancers and other control elements that regulate gene transcription or mRNA translation. In some embodiments, the expression control sequence is a regulatory element. The specific structure of the expression control sequence may vary according to the function of the species or cell type, but generally includes 5' non-transcribed sequences and 5' and 3' non-translated sequences that participate in transcription and translation initiation, such as TATA boxes, capping sequences, CAAT sequences, etc. For example, a 5' non-transcribed expression control sequence may include a promoter region, and the promoter region may include a promoter sequence for transcription control of a functionally connected nucleic acid.

[0207] In the present application, the composition may include one or more TRIM72 modulators.

[0208] In the present application, the composition may include one or more TRIM72 modulators and a pharmaceutically acceptable adjuvant.

[0209] In some embodiments, pharmaceutically acceptable adjuvants may include buffers, antioxidants, preservatives, low molecular weight polypeptides, proteins, hydrophilic polymers, amino acids, sugars, chelating agents, counterions, metal complexes and / or nonionic surfactants, and the like.

[0210] In some embodiments, the pharmaceutically acceptable adjuvant may include a drug, a toxin, a cytokine, a radioactive element, a carrier protein, an enzyme, a lectin, a fluorescent quantum dot, and / or a chromophore with a high absorption coefficient.

[0211] In the present application, the composition can be formulated with a pharmaceutically acceptable carrier or diluent and any other known adjuvants and excipients according to conventional techniques in the art, such as following the procedures in: Remington: The Science and Practice of Pharmacy, 19th edition, Gennaro, ed., Mack Publishing Co., Easton, PA, 1995.

[0212] In the present application, the composition can be formulated for oral administration, intravenous administration, intramuscular administration, in situ administration at the tumor site, inhalation, rectal administration, vaginal administration, transdermal administration, or administration of the drug via a subcutaneous reservoir.

[0213] In the present application, the composition can be used to protect neurons. For example, the composition of the present application can inhibit or delay the development or progression of Parkinson's disease, and / or can alleviate and / or stabilize the disease state.

[0214] The composition of the present application may contain a therapeutically effective amount of the TRIM72 protein regulator. The therapeutically effective amount is the dose required to prevent and / or treat (at least partially treat) Parkinson's disease and / or any complications thereof in a subject suffering from Parkinson's disease or at risk of suffering from the disease.

[0215] Example

[0216] The following examples are set forth in order to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor to represent that the following experiments are all or the only experiments performed. Efforts have been made to ensure the accuracy of the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be considered. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. Standard abbreviations may be used, such as bp, base pairs; kb, kilobase; pl, picoliter; s or sec, seconds; min, minutes; h or hr, hours; aa, amino acids; nt, nucleotides; im, intramuscular (ly); ip, intraperitoneal; sc, subcutaneous; etc.

[0217] method

[0218] MPTP modeling

[0219] 5-week-old C57Bl6 / J male mice were used for MPTP-induced PD modeling. ScAAV was injected into mice by retro-orbital intravenous injection 14 days before MPTP administration. Mice were injected intraperitoneally (ip) with 0.9% NaCl or saline (0.9% NaCl) containing MPTP-HCl using a long-term dosing regimen of 20 mg / kg per day for 14 days.

[0220] Behavioral testing

[0221] Rotarod test was performed 8 days after MPTP induction.

[0222] Rotarod performance was measured by an automated system (Med Associates Inc.). Briefly, animals were placed on an accelerating spindle (5-40 rpm) for 5 min per trial, with three consecutive trials per day. A 20-min rest period was set between each trial. When mice fell off the spindle during a 5-min interval, the system automatically calculated the time they fell off the spindle. The residence time was calculated by subtracting the time they fell from 5 min, and the mean of the residence time of three consecutive trials per day was used for statistical analysis.

[0223] Immunofluorescence staining of mouse brain

[0224] On the 8th day after MPTP modeling was completed, mice were killed to obtain brain samples for immunostaining research. For tissue preparation, anesthetized mice were perfused with PBS and 4% PFA in sequence. Before embedding, the brain and spinal cord were post-fixed overnight in 4% PFA. All tissues were sliced ​​with 40 μm using a vibrator (Leica VT1000S, Germany). In order to observe the TH (tyrosine hydroxylase) positive dopaminergic neurons in the substantia nigra, floating slices were incubated in PBS containing 5% bovine serum albumin (BSA) and primary and secondary antibodies. DAPI (1:1000, Beyotime C1002) was included in the secondary antibody incubation medium for nuclear staining, and then washed. After staining, the slices were sealed with Fluoromount-g (southrenbiotech 0100-01). Fluorescent images were collected by confocal microscopy (Nikon A1, Japan).

[0225] Plasmids and lentiviral vectors

[0226] A DNA fragment corresponding to the full length of TRIM72 was amplified by PCR from a mouse cDNA library and inserted between the SalI and XhoI sites of the pCMV-N-3×Flag expression vector using a seamless cloning kit (Beyotime) to generate Flag-tagged TRIM72. A series of Flag-tagged TRIM72 domain deletions were generated from the full-length TRIM72 expression vector: Δ loop domain (domain 14-69 deleted); Δ B box domain (domain 81-122 deleted); Δ coiled-coil domain (domain 135-232 deleted); Δ PRYSPRY domain (domain 278-470 deleted) constructs.

[0227] To construct a TRIM72 stable expression cell line, the DNA fragment of the Flag-tagged TRIM72 mutant or domain-deleted or single-domain series was amplified from the corresponding TRIM72 expression vector by PCR and inserted between the BsrGI and EcoRI sites of the pLJM1-EGFP lentiviral vector to generate a fusion of the Flag-tagged TRIM72 mutant or domain-deleted or single-domain series with EGFP in the lentiviral vector.

[0228] Lentivirus expressing Flag-tagged TRIM72 constructs was generated from the corresponding EGFP fused to the Flag-tagged TRIM72 vector.

[0229] Cell culture, lentiviral packaging, and lentiviral infection

[0230] HEK293FT cells were maintained in a cell culture incubator (37°C, 5% CO 2 ) in DMEM (Invitrogen) with 10% fetal bovine serum (Gemini). For lentiviral packaging, HEK293FT cells were seeded in growth medium in three 10-cm culture dishes. When approximately 90% confluence was reached, cells were co-transfected with VSVG (10 μg), pxPAX2 (15 μg) and pLJM1-EGFP lentiviral vectors or pLentiCRISPRv2 (Addgene) or pLenticas9-Blast (Addgene) (20 μg) using PEI (Sigma) according to the manufacturer's instructions, and the medium was replaced with fresh growth medium 5-6 h after transfection. The culture medium was harvested 72 hours after transfection and centrifuged at 20,000 rpm, 4°C for 2 hours. After centrifugation, the lentivirus was enriched in the precipitate. The lentivirus was resuspended using 100 μl DPBS and stored at -80°C.

[0231] HEK293FT cells or Hela cells were infected with the indicated lentiviruses. Three days after infection, infected cells were selected with 2 μg / ml puromycin or 10 μg / ml blasticidin, depending on the resistance plasmid, for at least one week. Puromycin- or blasticidin-selected cells were used for further analysis.

[0232] Cell viability assay

[0233] Cell viability was assessed using CCK-8. 3 Cells were seeded into 96-well plates at a density of 7 × 10 cells / well for arsenite treatment or 7 × 10 3 Cells were seeded at a density of 10 cells / well for H 2 O 2 Arsenite (Sigma) was added to each well at a concentration of 0.125 mM, 0.25 mM, or 0.5 mM, and the cells were washed after incubation at 37°C for 2 hours. 2 O 2 For treatment experiments, scAAV9 infection (estimated multiplicity of infection (MOI): 10,000 vg / cell) was performed 17 hours after cell inoculation for 24 hours. 2 O 2 Add to each well at a concentration of 300 μM and incubate at 37 °C for 1 h before washing the cells. 2 O 2 After treating the cells, a total of 10 μL of CCK-8 solution (Yeasen) was added to each well. After incubation for another 2 hours at 37°C, the optical density (OD) value of each well was measured using a microplate reader with an excitation wavelength of 450 nm. The cell viability of 293FT was calculated. The experiment was repeated at least three times to obtain an average value.

[0234] Western blot analysis

[0235] The total protein content in the cells was extracted using RIPA lysis buffer (50mM Tris-HCl, pH 8.0, 150mM NaCl, 1mM EDTA, 0.1% SDS, 1% TritonX-100, 0.5% sodium deoxycholate) supplemented with phenylmethylsulfonyl fluoride (PMSF) and a protease inhibitor cocktail (Bimake), and the lysate was incubated on ice for 30 minutes. After centrifugation at 12000rpm for 10 minutes, the supernatant was extracted and incubated at 95°C for 10 minutes after mixing with SDS loading buffer. Then, proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) membrane. Afterwards, the membrane was blocked for 1 hour at room temperature using 5% skim milk. Then incubate with diluted primary antibodies overnight at 4°C, including GAPDH (Ameribio), tubulin (Ameribio), TRIM72 antibody (kind gift from Dr. Jianjie Ma), TSG101 (Abcam), ITGAV (Abcam), H3 (Abcam). The membrane was then incubated with HRP-conjugated secondary antibodies for 1 hour at room temperature. Images were analyzed using Fiji ImageJ to obtain grayscale values.

[0236] Exosome purification

[0237] The protocol for purifying exosomes from 100 ml supernatant of 293FT cells includes two steps: ultrafiltration and polyethylene glycol (PEG) precipitation. First, pour the collected supernatant into a centrifuge tube and centrifuge at 3000 × g for 20 minutes to remove cell debris. Then, filter the supernatant once with a 0.45 μm filter. After that, take a new Amicon ULTRA-15 ultrafiltration tube and wash it with PBS or autoclaved water. Then immediately add the supernatant to the ultrafiltration tube and centrifuge at 3000 × g for 5-10 minutes. Discard the filtrate, continue to add supernatant, and centrifuge at 3000 × g for 5-10 minutes until all the supernatant is introduced into the ultrafiltration tube. Then transfer the concentrate to a 50 mL centrifuge tube. Add separation reagent to the concentrate. Mix the sample thoroughly by vortexing or aspirating. Finally, place the sample at 2-8 ° C overnight. The next day, centrifuge the sample at 10,000 × g for 1 hour at 4 ° C. The supernatant was discarded and the exosomes were in the pellet.

[0238] Example 1 AAV-TRIM72 packaging

[0239] The AAV packaging system is a commonly used three-plasmid system. By transfecting these three plasmids into mammalian cells (e.g., HEK293) at the same time, all components required for AAV packaging can be expressed in this cell and assembled into viral particles. In this article, we use a modified three-plasmid system from PackGene (Guangzhou PackGene Biotech Co., Ltd). This system consists of three plasmids: pAAV-ITR containing target genes, serotype vector pRepCapX and auxiliary vector pADHelper. The target vector pAAV-ITR contains eukaryotic promoters and other components required for high-level gene expression in mammalian cells when foreign sequences are cloned into the multiple cloning site (MCS). The vector also contains AAV reverse terminal repeats (ITR) that guide viral replication and packaging. The vector pRepCapX contains AAV rep and CAP genes encoding replication proteins and viral capsid proteins. The stability of rep and CAP gene expression levels is a key step in obtaining the desired high-titer viral product. The vector pADHelper contains a set of adenoviral genes VA, E2A and E4, which are essential for cells to produce high titers of virus. Figure 1 The construction of the pAAV-ITR vector of scAAV is shown.

[0240] Example 2 scAAV-TRIM72 alleviates motor dysfunction and dopaminergic neuron loss in MPTP-treated mice

[0241] In this paper, the MPTP-induced PD model was used according to Figure 2 The efficacy of scAAV-hSyn1-TRIM72 was evaluated by the administration procedure shown in the figure. The MPTP model group showed obvious behavioral defects and dopamine neuron loss, which were determined by rotating rod performance and TH (tyrosine hydroxylase) immunostaining ( Figure 3 A and 4A). Using the established MPTP-induced PD model, 14 days before MPTP administration, a dose of 10 11 vg / mouse AAV (PHP.eB) vector or a dose of 8×10 12vg / mouse AAV9 vector was injected into mice. The mice were equally divided into 3 groups. (1) MPTP group; (2) MPTP-scAAV(PHP.eB)-TRIM72 group; (3) MPTP-scAAV9-TRIM72 group. First, we examined their motor ability on the rotating rod. The MPTP group mice showed impaired rotating rod performance, while MPTP-scAAV(PHP.eB)-TRIM72 mice or MPTP-scAAV9-TRIM72 mice showed significantly reduced abnormal motor function of rotating rod behavior, indicating that scAAV-TRIM72 rescued the motor function of MPTP-treated mice ( Figure 3 B).

[0242] In addition, injection of scAAV(PHP.eB)-TRIM72 or scAAV9-TRIM72 effectively prevented these motor neuron losses (MPTP vs. MPTP-scAAV(PHP.eB)-TRIM72 mice or MPTP vs. MPTP-scAAV9-TRIM72 mice), suggesting that TRIM72 has a protective effect against motor neuron damage. Figure 4 B).

[0243] Example 3. TRIM72 can be secreted via exosomes

[0244] Exosomes are small extracellular biological vesicles that are released into the surrounding body fluids through the fusion of multivesicular bodies and plasma membranes, and contain proteins, nucleic acids, lipids and other bioactive substances. Exosomes play an important role in the exchange of information between cells by releasing bioactive substances that fuse with receptor cell membranes or bind to cell surface receptors. Full-length TRIM72 or TRIM72 mutants with different domains destroyed were constructed and stably overexpressed in the 293FT cell line by lentiviral infection. We found that TRIM72 was enriched in TSG101-labeled exosomes, which means that TRIM72 can affect the biological processes of surrounding cells through the exosome secretion pathway ( Figure 5 ). Further analysis revealed that both the coiled-coil domain and the PRYSPRY domain are essential for its exosome secretion, and the absence of either domain prevents TRIM72 from being secreted via exosomes ( Figure 6 In contrast, removal of the RING domain or the B-box domain had only a limited effect on the secretion efficiency of TRIM72 ( Figure 6 ).

[0245] Example 4 Protective effect of truncated TRIM72 in cultured cells

[0246] TRIM72 protein contains a RING finger motif, a B-box domain, a coiled-coil domain, and a PRYSPRY domain ( Figure 7To investigate which domain of TRIM72 performs the protective function against oxidative stress, full-length TRIM72 or truncated TRIM72 mutants were constructed and stably overexpressed in the 293FT cell line by lentiviral infection followed by puromycin selection ( Figure 8 ). CCK-8 was used to measure cell viability after arsenite treatment. The results showed that TRIM72 mutants lacking the coiled-coil domain and the PRYSPRY domain abolished the protective effect of TRIM72, while TRIM72 mutants lacking the RING domain or the B-box domain retained a protective effect similar to that of the wild type ( Fig. 9 ).

[0247] To further investigate which key domains of TRIM72 protein are required for protecting neurons, TRIM72 lacking different domains was constructed and overexpressed in N2a cell line by scAAV9 infection ( Figure 1 ). CCK-8 was used to measure H 2 O 2 In fact, H 2 O 2 After treatment, cell viability decreased ( Fig.10 ). Notably, overexpression of either the coiled-PRYSPRY or PRYSPRY single domains was sufficient to protect cells from oxidative stress, but overexpression of the coiled-coil single domain was not, suggesting that the coiled-PRYSPRY construct or PRYSPRY alone was able to elicit protection ( Fig.10 ).

[0248] Example 5. TRIM72, but not other TRIM proteins, protects cells from oxidative stress

[0249] To investigate whether other TRIM proteins could also elicit protection against oxidative stress, TRIM40 was overexpressed in the N2a cell line by scAAV9 infection (estimated multiplicity of infection (MOI): 10,000 vg / cell) ( Figure 1 ). CCK-8 was used to measure H 2 O 2 Interestingly, treatment with TRIM72 but not TRIM40 triggered protection against oxidative stress, demonstrating that only TRIM72 but not other TRIMs can protect cells from oxidative stress ( Fig.10 ).

[0250] Example 6. Key sites of TRIM72 protein that protect cells from oxidative stress

[0251] A series of Flag-tagged TRIM72 mutants: C14A (cysteine ​​at position 14 was substituted by alanine) and C242A (cysteine ​​at position 242 was substituted by alanine) were generated from the wild-type TRIM72 construct by point mutation.

[0252] Previous studies have determined that TRIM72 senses changes in the oxidative environment and forms oligomeric complexes to complete membrane repair. The cysteine ​​residue (C242) plays a key role in the formation of TRIM72 oligomers. In addition, the cysteine ​​residue (C14) is essential for the activity of the TRIM72 E3 ligase. Mutation of C242 to alanine (C242A) blocked the protective effect of TRIM72, while its E3 ligase-inactive mutant (C14A) retained the same protective effect as the wild type ( Fig.11 ). It can be concluded that TRIM72 protects cells from oxidative stress and mainly depends on its oligomerization rather than E3 ligase activity.

[0253] Although preferred embodiments of the present invention have been shown and described herein, it is apparent to those skilled in the art that these embodiments are provided by way of example only. This does not mean that the present invention is limited by the specific implementation scheme provided in the specification. Although the present invention has been described with reference to the foregoing description, the description and illustration of the embodiments herein are not meant to be interpreted in a limiting sense. Without departing from the present invention, those skilled in the art will now expect many variations, changes and substitutions. In addition, it should be understood that all aspects of the present invention are not limited to the specific description, configuration or relative proportions set forth herein, which depend on various conditions and variables. It should be understood that various alternatives to the embodiments of the present invention described herein can be adopted to practice the present invention. Therefore, it is expected that the present invention will also cover any such substitution, modification, variation or equivalent. The following claims are intended to define the scope of the present invention, and thus cover methods and structures and their equivalents within the scope of these claims.

Claims

1. A method for preventing and / or treating Parkinson's disease, comprising administering one or more TRIM72 protein modulators. 2 . The method according to claim 1 , wherein the TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

3. The method according to any one of claims 1 to 2, wherein the TRIM72 modulator is selected from one or more of the group consisting of: proteins, peptides, peptide mimetics, chemical compounds, antibodies, ribozymes, small molecule chemical compounds, nucleic acids, vectors and antisense nucleic acids. 4 . The method according to any one of claims 1 to 3 , wherein the TRIM72 protein modulator comprises a TRIM72 protein or a variant thereof or a functional fragment thereof. The method according to claim 4 , wherein the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof. 6 . The method according to any one of claims 4 to 5 , wherein the TRIM72 protein comprises a full-length TRIM72 protein. 7 . The method according to any one of claims 4 to 6 , wherein the TRIM72 protein comprises a wild-type TRIM72 protein. 8 . The method according to any one of claims 5 to 7 , wherein the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:

2. 9 . The method according to any one of claims 4 to 8 , wherein the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof. 10 . The method according to claim 9 , wherein the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

11. The method of claim 10, wherein the PRYSPRY domain comprises amino acid sites 278aa-470aa of the TRIM72 protein.

12. The method according to any one of claims 10 to 11, wherein the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:

6. 13 . The method according to any one of claims 9 to 12 , wherein the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof. 14 . The method according to any one of claims 9 to 12 , wherein the TRIM72 truncated protein does not comprise a coiled-coil domain of the TRIM72 protein or a functional fragment thereof. 15 . The method according to claim 13 , wherein the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

16. The method according to any one of claims 13 to 14, wherein the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:

5. 17 . The method according to any one of claims 9 to 16 , wherein the TRIM72 truncated protein further comprises a B-box domain of the TRIM72 protein or a functional fragment thereof. 18 . The method according to any one of claims 9 to 16 , wherein the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

19. The method according to any one of claims 17 to 18, wherein the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

20. The method according to any one of claims 17 to 19, wherein the B-box domain comprises the amino acid sequence shown in SEQ ID NO:

4. 21 . The method according to any one of claims 9 to 20, wherein the TRIM72 truncated protein further comprises a RING finger domain of a TRIM72 protein or a functional fragment thereof. 22 . The method according to any one of claims 9 to 20 , wherein the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

23. The method according to any one of claims 21 to 22, wherein the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

24. The method according to any one of claims 21 to 23, wherein the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:

3. 25 . The method according to any one of claims 9 to 24 , wherein the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9 and 11.

26. The method according to any one of claims 4 to 25, wherein the TRIM72 protein or a variant thereof or a functional fragment thereof comprises an amino acid mutation at position C14.

27. The method according to any one of claims 4 to 26, wherein the TRIM72 protein or a variant thereof or a functional fragment thereof comprises the amino acid mutation C14A.

28. The method according to any one of claims 4 to 27, wherein the TRIM72 protein or its variant or functional fragment thereof does not comprise an amino acid mutation at position C242.

29. The method according to any one of claims 4 to 27, wherein the TRIM72 protein or a variant thereof or a functional fragment thereof is secreted via exosomes. 30 . The method according to any one of claims 1 to 29 , wherein the TRIM72 protein modulator comprises a nucleic acid molecule encoding the TRIM72 protein according to any one of claims 4 to 29 or a variant thereof or a functional fragment thereof. 31 . The method according to any one of claims 1 to 30 , wherein the TRIM72 protein regulator comprises a vector comprising a gene encoding the TRIM72 protein or a variant thereof or a functional fragment thereof according to any one of claims 4 to 30 .

32. The method of claim 31, wherein the vector is a plasmid or a virus.

33. The method of any one of claims 31 to 32, wherein the vector is an adeno-associated virus (rAAV) expression vector.

34. The method according to any one of claims 31 to 33, wherein the vector comprises a universal promoter.

35. The method of claim 34, wherein the universal promoter is selected from one or more of the group consisting of chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV) and simian virus 40 (SV40) promoter.

36. The method according to any one of claims 31 to 35, wherein the vector comprises a neuron-specific promoter.

37. The method of claim 36, wherein the neuron-specific promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a cerebral neocortex and hippocampal excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

38. according to the method described in any one of claims 36 to 37, wherein the neuron-specific promoter is selected from the group consisting of: human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

39. The method of any one of claims 34 to 38, wherein the promoter comprises a human promoter.

40. The method of any one of claims 33 to 39, wherein the serotype of the AAV vector is selected from the group consisting of AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ and AAVhull.

41. The method according to any one of claims 1 to 40, wherein the TRIM72 protein modulator comprises a cell, wherein the cell comprises the vector according to any one of claims 31 to 40.

42. The method according to any one of claims 1 to 41, wherein the TRIM72 protein modulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein according to any one of claims 4 to 41, or a variant thereof, or a functional fragment thereof.

43. The method according to any one of claims 1 to 42, wherein the TRIM72 protein modulator protects neurons by reducing oxidative stress, thereby preventing and / or treating the Parkinson's disease.

44. A composition for preventing and / or treating Parkinson's disease, comprising one or more TRIM72 protein regulators.

45. The composition of claim 4, wherein the TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

46. ​​The composition of any one of claims 44 to 45, wherein the TRIM72 modulator is selected from one or more of the group consisting of proteins, peptides, peptide mimetics, chemical compounds, antibodies, ribozymes, small molecule chemical compounds, nucleic acids, vectors, and antisense nucleic acids.

47. The composition according to any one of claims 44 to 46, wherein the TRIM72 protein modulator comprises a TRIM72 protein or a variant thereof or a functional fragment thereof.

48. The composition according to claim 47, wherein the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof.

49. The composition of any one of claims 47 to 48, wherein the TRIM72 protein comprises a full-length TRIM72 protein.

50. The composition of any one of claims 47 to 49, wherein the TRIM72 protein comprises a wild-type TRIM72 protein.

51. The composition according to any one of claims 48 to 50, wherein the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:

2.

52. The composition according to any one of claims 47 to 51, wherein the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof.

53. The composition of claim 5, wherein the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

54. The composition of claim 53, wherein the PRYSPRY domain comprises amino acid positions 278aa-470aa of the TRIM72 protein.

55. The composition of any one of claims 53 to 54, wherein the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:

6.

56. The composition according to any one of claims 52 to 55, wherein the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

57. The composition according to any one of claims 52 to 55, wherein the TRIM72 truncated protein does not comprise the coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

58. The composition of any one of claims 56 to 57, wherein the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

59. The composition of any one of claims 56 to 58, wherein the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:

5.

60. The composition according to any one of claims 52 to 59, wherein the TRIM72 truncated protein further comprises a B-box domain of the TRIM72 protein or a functional fragment thereof.

61. The composition of any one of claims 52 to 60, wherein the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

62. The composition of any one of claims 60 to 61, wherein the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

63. The composition of any one of claims 60 to 62, wherein the B-box domain comprises the amino acid sequence shown in SEQ ID NO:

4.

64. The composition according to any one of claims 52 to 63, wherein the TRIM72 truncated protein further comprises a RING finger domain of the TRIM72 protein or a functional fragment thereof.

65. The composition of any one of claims 52 to 63, wherein the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

66. The composition of any one of claims 64 to 65, wherein the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

67. The composition of any one of claims 64 to 66, wherein the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:

3.

68. The composition of any one of claims 52 to 67, wherein the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9 and 11.

69. The composition according to any one of claims 47 to 68, wherein the TRIM72 protein or a variant thereof or a functional fragment thereof comprises an amino acid mutation at position C14.

70. The composition according to any one of claims 47 to 69, wherein the TRIM72 protein or its variant or its functional fragment comprises the amino acid mutation C14A.

71. The composition according to any one of claims 47 to 70, wherein the TRIM72 protein or its variant or its functional fragment does not comprise an amino acid mutation at position C242.

72. The composition according to any one of claims 47 to 71, wherein the TRIM72 protein or its variant or functional fragment thereof is secreted via exosomes.

73. The composition according to any one of claims 44 to 72, wherein the TRIM72 protein regulator comprises a nucleic acid molecule encoding the TRIM72 protein according to any one of claims 47 to 72, or a variant thereof, or a functional fragment thereof.

74. The composition according to any one of claims 44 to 72, wherein the TRIM72 protein regulator comprises a vector comprising a gene encoding the TRIM72 protein according to any one of claims 47 to 72 or a variant thereof or a functional fragment thereof.

75. The composition of claim 74, wherein the vector is a plasmid or a virus.

76. A composition according to any one of claims 74 to 75, wherein the vector is an adeno-associated virus (rAAV) expression vector.

77. A composition according to any one of claims 74 to 76, wherein the vector comprises a universal promoter.

78. The composition of claim 77, wherein the universal promoter is selected from one or more of the group consisting of chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV), and simian virus 40 (SV40) promoters.

79. A composition according to any one of claims 74 to 78, wherein the vector comprises a neuron-specific promoter.

80. The composition of claim 79, wherein the neuron-specific promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a cerebral neocortex and hippocampal excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a γ-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

81. according to the composition described in any one of claims 79 to 80, wherein the neuron-specific promoter is selected from the group consisting of: human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

82. A composition according to any one of claims 77 to 81, wherein the promoter comprises a human promoter.

83. The composition of any one of claims 76 to 82, wherein the serotype of the AAV vector is selected from AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ and AAVhull.

84. according to the composition described in any one of claims 44 to 83, wherein the TRIM72 protein regulator comprises a cell, wherein the cell comprises the vector described in any one of claims 72 to 81.

85. The composition according to any one of claims 44 to 84, wherein the TRIM72 protein modulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein according to claims 47 to 84 or a variant thereof or a functional fragment thereof.

86. The composition of any one of claims 44 to 85, wherein the TRIM72 protein modulator protects neurons by reducing oxidative stress, thereby preventing and / or treating the Parkinson's disease.

87. Use of a TRIM72 protein regulator in the preparation of a medicament for preventing and / or treating Parkinson's disease.

88. The use according to claim 87, wherein the TRIM72 protein modulator increases the expression and / or activity of the TRIM72 protein.

89. The use according to any one of claims 87 to 88, wherein the TRIM72 modulator is selected from one or more of the group consisting of: proteins, peptides, peptide mimetics, chemical compounds, antibodies, ribozymes, small molecule chemical compounds, nucleic acids, vectors and antisense nucleic acids.

90. The use according to any one of claims 7 to 89, wherein the TRIM72 protein modulator comprises a TRIM72 protein or a variant thereof or a functional fragment thereof.

91. The use according to claim 90, wherein the TRIM72 protein or a functional fragment thereof comprises a human TRIM72 protein or a functional fragment thereof.

92. The use according to any one of claims 90 to 91, wherein the TRIM72 protein comprises a full-length TRIM72 protein.

93. The use according to any one of claims 90 to 92, wherein the TRIM72 protein comprises a wild-type TRIM72 protein.

94. The use according to any one of claims 90 to 93, wherein the human TRIM72 protein comprises the amino acid sequence shown in SEQ ID NO:

2.

95. The use according to any one of claims 90 to 94, wherein the TRIM72 protein or a functional fragment thereof comprises a TRIM72 truncated protein or a functional fragment thereof.

96. The use according to claim 95, wherein the TRIM72 truncated protein comprises the PRYSPRY domain of the TRIM72 protein or a functional fragment thereof.

97. The use according to claim 96, wherein the PRYSPRY domain comprises amino acid sites 278aa-470aa of the TRIM72 protein.

98. The use according to any one of claims 96 to 97, wherein the PRYSPRY domain comprises the amino acid sequence shown in SEQ ID NO:

6.

99. The use according to any one of claims 95 to 98, wherein the TRIM72 truncated protein further comprises a coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

100. The use according to any one of claims 95 to 98, wherein the TRIM72 truncated protein does not comprise the coiled-coil domain of the TRIM72 protein or a functional fragment thereof.

101. The use according to any one of claims 99 to 100, wherein the coiled-coil domain comprises amino acid positions 135aa-232aa of the TRIM72 protein.

102. The use according to any one of claims 99 to 101, wherein the coiled-coil domain comprises the amino acid sequence shown in SEQ ID NO:

5.

103. The use according to any one of claims 95 to 102, wherein the TRIM72 truncated protein further comprises the B-box domain of the TRIM72 protein or a functional fragment thereof.

104. The use according to any one of claims 95 to 102, wherein the TRIM72 truncated protein does not comprise the B-box domain of the TRIM72 protein or a functional fragment thereof.

105. The use according to any one of claims 103 to 104, wherein the B-box domain comprises amino acid positions 86aa-117aa of the TRIM72 protein.

106. The use according to any one of claims 103 to 105, wherein the B-box domain comprises the amino acid sequence shown in SEQ ID NO:

4.

107. The use according to any one of claims 95 to 106, wherein the TRIM72 truncated protein further comprises the RING finger domain of the TRIM72 protein or a functional fragment thereof.

108. The use according to any one of claims 95 to 107, wherein the TRIM72 truncated protein does not comprise the RING finger domain of the TRIM72 protein or a functional fragment thereof.

109. The use according to any one of claims 107 to 108, wherein the RING finger domain comprises amino acid positions 14aa-56aa of the TRIM72 protein.

110. The use according to any one of claims 107 to 109, wherein the RING finger domain comprises the amino acid sequence shown in SEQ ID NO:

3. 111 . The use according to any one of claims 95 to 110, wherein the TRIM72 truncated protein comprises an amino acid sequence as shown in any one of SEQ ID NOs: 6, 7, 8, 9 and 11.

112. The use according to any one of claims 95 to 111, wherein the TRIM72 protein or its variant or its functional fragment comprises an amino acid mutation at position C14.

113. The use according to any one of claims 95 to 112, wherein the TRIM72 protein or its variant or its functional fragment comprises the amino acid mutation C14A.

114. The use according to any one of claims 95 to 113, wherein the TRIM72 protein or its variant or its functional fragment does not comprise an amino acid mutation at position C242.

115. The use according to any one of claims 95 to 114, wherein the TRIM72 protein or its variant or its functional fragment is secreted via exosomes. 116 . The use according to any one of claims 95 to 115 , wherein the TRIM72 protein regulator comprises a nucleic acid molecule encoding the TRIM72 protein according to any one of claims 95 to 115 , or a variant thereof, or a functional fragment thereof.

117. The use according to any one of claims 87 to 116, wherein the TRIM72 protein regulator comprises a vector comprising a gene encoding the TRIM72 protein or its variant or functional fragment thereof according to any one of claims 95 to 116.

118. The use according to claim 117, wherein the vector is a plasmid or a virus.

119. The use according to any one of claims 117 to 118, wherein the vector is an adeno-associated virus (rAAV) expression vector.

120. The use according to any one of claims 117 to 119, wherein the vector comprises a universal promoter.

121. The use according to claim 120, wherein the universal promoter is selected from one or more of the group consisting of: chicken β-actin (CBA), cytomegalovirus (CMV), CMV immediate enhancer / β-actin (CAG), truncated CBA hybrid (CBh), ubiquitin C (UBC), elongation factor 1 alpha (EF1A), mouse or human phosphoglycerate kinase (PGK), murine stem cell virus (MSCV), spleen focus forming virus (SFFV) and simian virus 40 (SV40) promoter.

122. The use according to any one of claims 117 to 121, wherein the vector comprises a neuron-specific promoter.

123. The method of claim 122, wherein the neuron-specific promoter is selected from one or more of the group consisting of an excitatory neuron-specific promoter, a cerebral neocortex and hippocampal excitatory neuron-specific promoter, a short neuron-specific promoter, a dopaminergic neuron-specific promoter, a glutamatergic neuron-specific promoter, a gamma-aminobutyric acid neuron-specific promoter, a cholinergic neuron-specific promoter, and a serotonergic neuron-specific promoter.

124. The use according to any one of claims 122 to 123, wherein the neuron-specific promoter is selected from the group consisting of: human synaptophysin (hSyn), calcium / calmodulin-dependent kinase IIa (CamKIIa), c-fos, methylated CpG binding protein 2 (Mecp2), neuron-specific enolase (NSE), somatostatin (SST), human vesicular GABA (γ-aminobutyric acid) transporter (hVGAT), choline acetyltransferase (ChAT), serotonin transporter (SERT) and tyrosine hydroxylase (TH).

125. The use according to any one of claims 120 to 124, wherein the neuron-specific promoter comprises a human promoter.

126. The use according to any one of claims 119 to 125, wherein the serotype of the AAV vector is selected from AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, AAVrh, AAVDJ and AAVhull.

127. The use according to any one of claims 87 to 126, wherein the TRIM72 protein modulator comprises a cell, wherein the cell comprises the vector according to any one of claims 117 to 126.

128. The use according to any one of claims 87 to 127, wherein the TRIM72 protein regulator comprises a fusion protein, wherein the fusion protein comprises the TRIM72 protein or its variant or functional fragment thereof according to any one of claims 90 to 127.

129. The use according to any one of claims 87 to 128, wherein the TRIM72 protein modulator protects neurons by reducing oxidative stress, thereby preventing and / or treating the Parkinson's disease.

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