Composition for prevention or treatment of parkinson's disease comprising alpha-synuclein epitope as active ingredient

Abstract

The present invention relates to a composition for the prevention or treatment of Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient. More specifically, the alpha-synuclein epitope, which is the active ingredient of the present invention, comprises an alpha-synuclein peptide VAEKTKEQVT, and has an acetylated N-terminal and a C-terminal that is conjugated with a carrier protein (keyhole limpet hemocyanin (KLH) or ovalbumin (OVA)) by using cysteine as a linker. The alpha-synuclein epitope was confirmed to have the effect of preventing and treating Parkinson's disease in an animal model with induced Parkinson's disease and an in vitro model, and therefore, the active ingredient of the present invention can be effectively used as a medicine and a vaccine for the prevention or treatment of Parkinson's disease.

Description

Composition for preventing or treating Parkinson's disease comprising alpha-synuclein epitope as an active ingredient

[0001] The present invention relates to a composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

[0002] This invention was carried out with the support of the "Biomedical Technology Development Project (Brain Disease Overcoming Research Project)" of the National Research Foundation of Korea under the Ministry of Science and ICT (or affiliated with it), under the project titled "Discovery of New Immune Control Target Substances and Verification of Efficacy; Application to Development of New Concept Parkinson's Disease Preventive Treatment" (Project Number: 2020M3E5D9080660).

[0003] Parkinson's disease is a representative degenerative brain disease that is the second most common after Alzheimer's disease. It is a disease in which dopamine, a neurotransmitter secreted by nerve cells in a specific part of the brain called the substantia nigra, located in the midbrain, is gradually lost for unknown reasons. Dopamine is one of the neurotransmitters secreted for signal transmission between brain nerve cells, and if there is a problem with the regulation of its secretion, symptoms such as tremors in the hands gradually occur, and walking difficulties, loss of balance, and depression are also experienced.

[0004] A typical pathological feature is the selective death of dopamine-secreting neurons, and the discovery of abnormal protein aggregates called Lewy bodies, the main component of which is alpha-synuclein. Various studies are being conducted to remove these abnormal protein aggregates, but there is still no known technology that can definitively remove them. Research is being conducted on a monoclonal antibody specific for alpha-synuclein produced in human B cells, rather than a general natural substance, but clinical trials have been discontinued due to side effects. Research on removing alpha-synuclein protein using humoral immunity using B cells in the body itself, rather than through purification, is receiving new attention, but is still in its initial stages.

[0005] To remove alpha-synuclein produced in the body, the principle of humoral immune-based protein removal is to administer alpha-synuclein to generate antibodies against it, which can then be rapidly processed when alpha-synuclein is produced in the future. However, administering the entire sequence to generate antibodies can stimulate multiple immune cells, potentially causing side effects. Therefore, it is necessary to identify a protein sequence that is specific to B cells. Because peptide sequences are relatively short, chemical modifications that enable stable binding to B cell receptors are also necessary.

[0006] Korean Patent No. 1517447 discloses the use of a mimotope of an alpha-synuclein epitope for treating Lewy body disease, and Korean Patent Publication No. 2020-0099160 discloses an anti-alpha synuclein antibody, but no mention has been made yet of a composition for preventing or treating Parkinson's disease comprising the alpha synuclein epitope of the present invention as an active ingredient.

[0007] The present invention was derived from the above-mentioned needs, and the present invention provides a composition for preventing or treating Parkinson's disease, which contains an alpha-synuclein epitope as an active ingredient, and the active ingredient of the present invention has been confirmed to have the effect of increasing the total moving distance in an open space test using an animal model induced with Parkinson's disease, increasing the time spent on a cylinder without falling off in a rotarod test, reducing the expression level of α-Syn, and increasing the expression levels of TH (Tyrosine hydroxylase), DAT (Dopamine transporter), and VMAT2 (Vesicular monoamine transporter 2), which is a dopamine transport protein, and increasing the production of IgG in the spleen and plasma, and reducing the production of IL-10 in plasma, thereby completing the present invention.

[0008] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating Parkinson's disease, which comprises an alpha-synuclein epitope as an active ingredient.

[0009] In addition, the present invention provides a veterinary composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

[0010] In addition, the present invention provides a method for preventing or treating Parkinson's disease, characterized in that it includes a step of administering the pharmaceutical composition for preventing or treating Parkinson's disease to an animal other than a human.

[0011] In addition, the present invention provides a peptide consisting of an amino acid sequence of sequence number 1.

[0012] In addition, the present invention provides a vaccine composition for preventing or treating Parkinson's disease, which comprises an alpha-synuclein epitope as an active ingredient.

[0013] The present invention relates to a composition for preventing or treating Parkinson's disease, which comprises an alpha-synuclein epitope as an active ingredient. More specifically, the alpha-synuclein epitope, which is an active ingredient of the present invention, comprises an alpha-synuclein peptide VAEKTKEQVT, and the N-terminus is acetylated, and the C-terminus is bound to a carrier protein [KLH (keyhole limpet hemocyanin) or OVA (ovalbumin)] using cysteine ​​as a linker.

[0014] The above alpha-synuclein epitope has a preventive and therapeutic effect on Parkinson's disease in animal models and in vitro models that induce Parkinson's disease, and therefore can be usefully utilized as a drug for preventing or treating Parkinson's disease; and as a vaccine.

[0015] Figure 1 shows the structure of the alpha-synuclein antigen PEP1 and the results of identifying alpha-synuclein protein residues interacting in B cells through Docking simulation.

[0016] Figure 2 is a schematic diagram showing the schedule for alpha-synuclein antigen animal testing.

[0017] Figure 3 shows the total moving distance of mice in an open field test according to the administration of the alpha-synuclein antigen of the present invention in an animal model of Parkinson's disease. Con is a normal group, α-Syn is a group in which Parkinson's disease was induced by administering alpha-synuclein protein, PDE-OVA is a group in which alpha-synuclein antigen (Ac-VAEKTKEQVTC-OVA) conjugated to ovalbumin was administered before inducing Parkinson's disease, and PDE-KLH is a group in which alpha-synuclein antigen (Ac-VAEKTKEQVTC-KLH) conjugated to KLH (keyhole limpet hemocyanin) was administered before inducing Parkinson's disease. The same was applied to the following drawings.

[0018] Figure 4 shows the time (sec) taken for a mouse to fall from a cylinder as a result of a rotarod test following administration of the alpha-synuclein antigen of the present invention in an animal model of Parkinson's disease.

[0019] Figure 5 shows the results of confirming the changes in the expression levels of α-Syn, TH (Tyrosine hydroxylase), DAT (Dopamine transporter), and VMAT2 (Vesicular monoamine transporter 2), a dopamine transport protein, by administration of selected PDE-OVA and PDE-KLH in a Parkinson's disease model induced by injection of alpha-synuclein. **, *** indicate that the difference between the comparison groups is statistically significant; ** is p<0.01, and *** is p<0.001.

[0020] Figure 6 shows the results of confirming changes in IgG production in the spleen (A) and IL-10 and IgG expression in the plasma (B) by administration of selected PDE-OVA and PDE-KLH in a Parkinson's disease model induced by injection of alpha-synuclein. **** indicates that the difference between the comparison groups is statistically significant, with p<0.0001.

[0021] The present invention relates to a pharmaceutical composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

[0022] The above alpha-synuclein epitope is preferably (a) a peptide consisting of the amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a), wherein one or more amino acid residues are substituted, deleted or inserted within the amino acid sequence of SEQ ID NO: 1, and wherein Parkinson's disease can be prevented or treated; more preferably, the (a) peptide consisting of the amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a), wherein one or more amino acid residues are substituted, deleted or inserted within the amino acid sequence of SEQ ID NO: 1, and wherein Parkinson's disease can be prevented or treated; is linked to a carrier protein by an amino acid linker having an acetylated N-terminus and a thiol group at the C-terminus, but is not limited thereto. The amino acid linker including the thiol group is preferably cysteine, but is not limited thereto.

[0023] The carrier protein is preferably, but not limited to, KLH (keyhole limpet hemocyanin) or OVA (ovalbumin).

[0024] The most preferred example of the alpha-synuclein epitope, which is an effective ingredient of the present invention, is (a) a peptide consisting of the amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a), wherein one or more amino acid residues are substituted, deleted or inserted within the amino acid sequence of SEQ ID NO: 1, and which is capable of preventing or treating Parkinson's disease; wherein the N-terminus of the peptide is acetylated and the C-terminus is bound to KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) protein via a cysteine ​​linker containing a thiol group, but is not limited thereto.

[0025] The above active ingredient is characterized by being an antigen that induces anti-alpha synuclein antibodies.

[0026] The pharmaceutical composition of the present invention may further include a carrier, excipient, or diluent in addition to the above-mentioned effective ingredient. Specifically, in addition to the above-mentioned effective ingredient, it may further contain one or more carriers selected from pharmaceutically acceptable saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, and in addition to the above-mentioned effective ingredient, it may further contain one or more auxiliary agents selected from pharmaceutically acceptable antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, and preservatives.

[0027] The above pharmaceutical composition may be administered orally or parenterally in a conventional manner, and when formulated, may be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the composition with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cacao butter, laurin, and glycerogelatin.

[0028] In addition, the present invention relates to a veterinary composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

[0029] The veterinary composition of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil. The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using a method well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, etc. The effective amount of the veterinary composition according to the present invention may be appropriately selected depending on the individual animal. It may be determined according to factors including the severity of the disease or condition, the sensitivity to the active ingredient of the present invention depending on the age, weight, health condition or sex of the individual, the route of administration, the period of administration, other compositions combined with or used simultaneously with the composition, and other factors well known in the physiological or veterinary fields.

[0030] In addition, the present invention relates to a method for preventing or treating Parkinson's disease, characterized in that it includes a step of administering the pharmaceutical composition for preventing or treating Parkinson's disease to an animal other than a human.

[0031] In the above method for preventing or treating Parkinson's disease, the active ingredient of the pharmaceutical composition for preventing or treating Parkinson's disease is (a) a peptide consisting of an amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a), wherein one or more amino acid residues are substituted, deleted or inserted within the amino acid sequence of SEQ ID NO: 1 and capable of preventing or treating Parkinson's disease; the N-terminus of the peptide is acetylated, and the C-terminus is characterized in that KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) protein is bound to it by a cysteine ​​linker containing a thiol group.

[0032] In addition, the present invention relates to a peptide consisting of an amino acid sequence of SEQ ID NO: 1. The peptide is preferably linked to KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) protein by a cysteine ​​linker having an acetylated N-terminus and a thiol group at the C-terminus, but is not limited thereto.

[0033] In addition, the present invention relates to a vaccine composition for preventing or treating Parkinson's disease, which comprises an alpha-synuclein epitope as an active ingredient.

[0034] In the vaccine composition of the present invention, the alpha-synuclein epitope may be, but is not limited to, (a) a peptide consisting of the amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a) in which one or more amino acid residues are substituted, deleted, or inserted within the amino acid sequence of SEQ ID NO: 1 and which is capable of preventing or treating Parkinson's disease.

[0035] In addition, in one embodiment of the present invention, the vaccine composition may further comprise at least one selected from the group consisting of pharmacologically acceptable carriers, diluents, preservatives, and immunoadjuvants according to techniques known to those skilled in the art.

[0036] The above pharmacologically acceptable carrier means a carrier medium that does not interfere with the effectiveness of the biological activity of the active ingredient, is chemically inert, and is not toxic to the subject to which it is administered. Various carriers are known in the art, and may include distilled water, saline solution, or mineral water, and preservatives may include, for example, EDTA salts, methiolate, and the like, and immunoadjuvants may include, but are not limited to, mineral gels such as aluminum hydroxide, surfactants such as lysolecithin, glycosides such as saponins or saponin derivatives, lantidine, avridine, carbopol, polyphosphazenes, dimethyldioctadecylammonium bromide (DDA), quil A, amphigen, mineral oil, and the like.

[0037]

[0038] Hereinafter, the present invention will be described in more detail using examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0039]

[0040] Example 1. Selection of a novel alpha-synuclein antigen using the IEDB (The Immune Epitope Database) system.

[0041] In this Example 1, a novel alpha-synuclein antigen capable of producing B cell-specific alpha-synuclein was selected using three theoretical analyses.

[0042] (1) Selection of antigen sequences that recognize B cells;

[0043] (2) Analysis of binding affinity, stability and toxicity of antigen sequences and

[0044] (3) A novel alpha-synuclein antigen was adopted through biochemical and behavioral verification in an animal model of Parkinson's disease.

[0045] To find the antigenic region that binds to B cells and stimulates humoral immunity among the alpha-synuclein sequences, the sequence was predicted using IEDB (The Immune Epitope Database).

[0046] The full alpha-synuclein sequence [MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTK EGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA (SEQ ID NO: 4)] was input into the software, and three representative B-cell epitope prediction algorithms in IEDB (Bepipred Linear Epitope Prediction 2.0, Bepipred Linear Epitope Prediction, and Emini Surface Accessibility Prediction) were used.

[0047] As a result, among the 140 amino acid sequences of alpha-synuclein, the middle sequence was predicted to be an antigen site that can bind to B cells, and three alpha-synuclein sequences (PEP1, PEP2, PEP3) were primarily selected as candidates. Through Gold (Genetic optimization for ligand docking), clustering formation information of binding modes was obtained as Gold scores, and binding affinity and stability were calculated through molecular docking simulation, and the top three peptides (PEP1, PEP2, PEP3) were selected (Table 1), and the binding affinity of the epitope was predicted on the Galaxypepdock web server for cross-validation.

[0048] GOLD (Genetic optimization for ligand docking) scores of the three selected candidate alpha-synuclein antigens PEP1: VAEKTKEQVT (SEQ ID NO: 1) PEP2: AEKTKEQVTN (SEQ ID NO: 2) PEP3: EKTKEQVTNV (SEQ ID NO: 3)1122.65106.8996.842119.29105.2196.463115.51100.0093.504106.8099.6388.535104.5995.9788.376101.9795.9787.257101.4395.4083.598101.1491.6383.059100.3891.5382.761090.2690.4681.13

[0049] After 10 GOLD attempts, PEP1 with the highest average GOLD score was confirmed to be non-toxic using the Toxinpred server, and PEP1 was selected as the alpha-synuclein antigen.

[0050]

[0051] Example 2. Stability and toxicity tests

[0052] Stability and toxicity tests were conducted based on the PEP1 sequence selected in Example 1 above.

[0053] Toxicity of selected alpha-synuclein antigen PEP1 Peptide sequence (PEP1) VAEKTKEQVT Mutation site No mutation QM score - 10.9 Toxicity prediction Non-toxic Hydrophobicity - 0.32 Hydrophobicity - 0.95 Hydrophilicity 0.79 Carbohydrate 0 Molecular weight 1132.42

[0054]

[0055] Example 3. Modification of selected candidate alpha-synuclein peptides Although in vivo alpha-synuclein is specific for B cell receptors, the process until it is delivered in vivo is very complex, and a peptide modification process is necessary to stably induce B cell-based humoral immunity. Since the immune system reacts to the entire peptide-protein conjugate, some antibodies are always generated and induced against the peptide, linker, and carrier protein, which serve as a trigger point for further promoting immunity. Therefore, each candidate alpha-synuclein peptide of the present invention was conjugated to a carrier protein.

[0056] In this Example 3, the PEP1 sequence (VAEKTKEQVT) was acetylated at the N-terminus, and a cysteine ​​(C) containing a thiol group was added to the C-terminus to bind a carrier protein, and then the carrier protein KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) was bound, thereby increasing the stability of the peptide against various decomposing enzymes and the biological immunological activity in vivo. Thereafter, the modified PEP1 was named PDE-OVA (Ac-VAEKTKEQVTC-OVA) and PDE-KLH (Ac-VAEKTKEQVTC-KLH), and used to analyze the efficacy against Parkinson's disease.

[0057] Example 4. Molecular docking simulation between the PEP1 sequence (VAEKTKEQVT) and B cells.

[0058] Docking simulations between the PEP1 sequence (VAEKTKEQVT) and B cells were performed to determine which residues on B cells PEP1, selected as an alpha-synuclein antigen, binds to.

[0059] As a result, the binding relationship and residues that interact with the alpha synuclein antigen of the present invention and B cells were confirmed (Fig. 1).

[0060] Example 5. Confirmation of behavioral changes in an animal model of Parkinson's disease caused by alpha-synuclein antigen consisting of the PEP1 sequence.

[0061] [Animal model]

[0062] Eight-week-old male C57BL / 6N mice weighing 25–30 g were purchased from Samtaco Bio and acclimatized for 1 week under a 12-h light / dark cycle and a temperature of 23°C. Animal experiments were conducted with the approval of the Animal Ethics Committee of Gyeongsang National University (Approval No. 125).

[0063] Using the synthesized alpha-synuclein antigen, in vivo experiments were conducted as follows. 15 μg of alpha-synuclein antigen per g of mouse body weight was administered intraperitoneally (IP) once a week for 4 weeks (6–10 weeks of age). After 4 weeks, 2 μl of a 2 μg / μl alpha-synuclein (α-Syn) solution dissolved in 1× PBS was injected into the ventricle, followed by a 1-week recovery period, to establish a Parkinson's disease animal model (Fig. 2).

[0064] (1) Open field test

[0065] Locomotor activity was measured in a white Plexiglas box consisting of 16 identically sized cubes (40 × 40 × 40 cm), 4 boxes in the center, and 12 surrounding boxes. Mouse movements were recorded using Tracking Software 3.0, and each mouse was placed in the center of a box for several hours before the experiment to acclimate to the new environment. To prevent abnormal behavior, the experiment was performed in a quiet, lighted room, and the chamber was deodorized using 10% ethanol before the experiment.

[0066] As a result, in the open field test, the animal group induced with Parkinson's disease showed lower levels of motor activity compared to the normal group. However, in contrast, the group administered alpha-synuclein antigen showed statistically significant improvements in motor activity levels. In particular, the PDE-KLH treatment group showed the most dramatic recovery (Fig. 3).

[0067] (2) Rotarod test

[0068] The rotarod test was performed to determine motor control ability by measuring limb coordination and balance ability. Since the rotarod test requires walking and balance on a rotating cylinder, the Parkinson's disease-induced group (α-Syn) with motor nerve abnormalities, compared to the normal group (Con), lost balance more quickly and fell off the rotating cylinder more quickly. However, in contrast, it was confirmed that the group administered the alpha-synuclein antigen (PDE-OVA or PDE-KLH) of the present invention showed a statistically significant increase in the time taken to fall off the cylinder (Fig. 4).

[0069] Example 6. Confirmation of changes in the expression levels of alpha-synuclein proteins and dopamine-related proteins according to treatment with the alpha-synuclein antigen of the present invention.

[0070] (1) Protein isolation from mouse brain

[0071] After completing the behavioral analysis experiment, mice were anesthetized with ketamine / xylazine and sacrificed. Brain tissue was removed, and the substantia nigra (SNpc) and striatum were isolated and disrupted in PRO-PREP™ extraction solution. The samples were then centrifuged at 13,000 rpm for 25 minutes at 4°C to obtain the supernatant. The obtained supernatant was stored at -80°C.

[0072] (2) Western blot

[0073] Protein concentration was measured using the Bradford assay, and equal amounts of protein samples were electrophoresed on SDS-PAGE and then transferred to PVDF membranes. To prevent nonspecific binding, the PVDF membranes were blocked with 5% skim milk, and incubated overnight at 4°C after primary antibody treatment. The membranes were then washed with 1× TBST (3 times × 10 min), blocked with horseradish-peroxidase-conjugated secondary antibodies, diluted in 1× TBST for 1 h, and then washed with 1× TBST (4 times × 2 min). Proteins were detected using ECL detection reagents according to the manufacturer's instructions. Immunoblot bands were acquired using X-ray film. The X-ray films were scanned, and the optical densities of the bands were analyzed by densitometry using the computer-based Sigma Gel program version 1.0 (SPSS, Chicago, IL, USA).

[0074] As a result, the expression level of α-Syn in the mouse group induced with Parkinson's disease increased compared to the normal group, and in contrast, the expression level of alpha-synuclein protein (α-Syn) in the mouse group administered with the alpha-synuclein antigen (PDE-OVA or PDE-KLH) according to the present invention decreased.

[0075] When the above alpha-synuclein antigen (PDE-OVA or PDE-KLH) was administered, the decrease in the expression level of α-Syn was judged to be due to the removal of alpha-synuclein by a humoral immune response in mice induced with Parkinson's disease.

[0076] In addition, the expression levels of dopamine synthetase TH (Tyrosine hydroxylase), DAT (Dopamine transporter), and VMAT2 (Vesicular monoamine transporter 2), a dopamine transport protein, were decreased in the Parkinson's disease-induced group compared to the normal group, and in contrast, when the alpha-synuclein antigen of the present invention was administered, the expression levels of TH (Tyrosine hydroxylase), VMAT2 (Vesicular monoamine transporter 2), and DAT (Dopamine transporter) were statistically significantly increased (Fig. 5).

[0077] Example 7. Confirmation of IgG levels in the spleen and plasma of a Parkinson's disease model.

[0078] When administering the alpha synuclein antigen (PDE-OVA, PDE-KLH) of the present invention, the degree of antibody production was evaluated by confirming the amount of IgG produced in the spleen and plasma of a Parkinson's disease model, and the degree of inflammation according to antibody production was evaluated by confirming the level of IL-10 in the plasma.

[0079] As a result, as disclosed in Fig. 6, it was confirmed that when the antigens selected in the present invention (PDE-OVA, PDE-KLH) were administered, the IgG levels in the spleen and plasma increased. Through this, it was determined that the selected antigens bound to B cells and produced alpha-synuclein antibodies, and it was determined that the inflammation level was reduced by antibody production through a decrease in the IL-10 level in the plasma.

Claims

1. A pharmaceutical composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

2. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that in claim 1, the alpha-synuclein epitope is (a) a peptide consisting of the amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a) in which one or more amino acid residues are substituted, deleted, or inserted within the amino acid sequence of SEQ ID NO: 1 and capable of preventing or treating Parkinson's disease.

3. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that the peptide comprising the amino acid sequence of sequence number 1 in the second paragraph; or (b) a peptide derived from (a) in which one or more amino acid residues are substituted, deleted, or inserted within the amino acid sequence of sequence number 1 and capable of preventing or treating Parkinson's disease; is bound to a carrier protein by an amino acid linker having an acetylated N-terminus and a thiol group at the C-terminus.

4. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that the amino acid linker containing the thiol group in the third paragraph is cysteine.

5. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that the carrier protein in the third paragraph is KLH (keyhole limpet hemocyanin) or OVA (ovalbumin).

6. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that the N-terminus of the peptide comprising the amino acid sequence of sequence number 1 in paragraph 2; or (b) a peptide derived from (a) in which one or more amino acid residues are substituted, deleted, or inserted within the amino acid sequence of sequence number 1 and capable of preventing or treating Parkinson's disease; is acetylated, and the C-terminus is bound to KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) protein by a cysteine ​​linker containing a thiol group.

7. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that the effective ingredient in claim 1 is an antigen that induces anti-alpha synuclein antibodies.

8. A pharmaceutical composition for preventing or treating Parkinson's disease, characterized in that, in addition to the effective ingredient in paragraph 1, it further comprises a carrier, excipient or diluent.

9. A veterinary composition for preventing or treating Parkinson's disease, comprising an alpha-synuclein epitope as an active ingredient.

10. A method for preventing or treating Parkinson's disease, characterized by comprising a step of administering a pharmaceutical composition for preventing or treating Parkinson's disease according to any one of claims 1 to 8 to an animal other than a human.

11. A peptide consisting of the amino acid sequence of sequence number 1.

12. In claim 11, the peptide is characterized in that the N-terminus of the peptide is acetylated and the C-terminus is bound to a KLH (keyhole limpet hemocyanin) or OVA (ovalbumin) protein by a cysteine ​​linker containing a thiol group.

13. A vaccine composition for the prevention or treatment of Parkinson's disease comprising an alpha-synuclein epitope as an active ingredient.

14. A vaccine composition for preventing or treating Parkinson's disease, characterized in that in claim 13, the alpha-synuclein epitope is (a) a peptide consisting of an amino acid sequence of SEQ ID NO: 1; or (b) a peptide derived from (a), wherein one or more amino acid residues are substituted, deleted, or inserted within the amino acid sequence of SEQ ID NO: 1, and is capable of preventing or treating Parkinson's disease.

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