Pharmaceutical composition comprising 2-(4-(1-hydroxypropan-2-yl)phenyl)isoindolin-1-one compound for preventing or treating Parkinson's disease

Abstract

The present application relates to a pharmaceutical composition for preventing or treating Parkinson's disease, the pharmaceutical composition including a 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-ketone compound or a pharmaceutically acceptable salt thereof, the pharmaceutical composition for preventing or treating Parkinson's disease according to the present application can increase the protein level of PGC-1α in the brain of an individual by successfully passing through the blood-brain barrier (BBB).

Description

Technical Field

[0001] This disclosure relates to a pharmaceutical composition for the prevention or treatment of Parkinson's disease, the pharmaceutical composition comprising a 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound or a pharmaceutically acceptable salt thereof. Background Technology

[0002] Parkinson's disease (whose main symptoms are tremors, rigidity, ataxia (slowness of movement), and chronic postural instability) is a chronic disease caused by a deficiency of the neurotransmitter dopamine in the brain and is one of the degenerative diseases of the central nervous system. It begins with changes in the substantia nigra pars compacta in the midbrain and is accompanied by its pathophysiological symptoms, such as reduced brain volume and α-synuclein (αSyn) accumulation, as well as gait imperfections, hand tremors, and rigidity.

[0003] Most treatment strategies for Parkinson's disease are limited to managing motor function symptoms using medications (such as L-DOPA or dopamine receptor agonists) and deep brain stimulation. Furthermore, these treatments, at the current level of research, have not yet prevented the gradual death of dopaminergic neurons (DA).

[0004] Recently, on the other hand, when it comes to cell death and survival, studies have been reported on the function of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and various diseases that may be caused by PGC-1α dysregulation.

[0005] Neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and Lou Gehrig's disease) are caused by the gradual loss of neuronal function and death, and the overall symptoms of these diseases are due to the loss of certain parts of neurons. Unlike the hyperactivity due to neurodegeneration observed in PGC-1α knockout mice and the fewer damaged areas observed in the cerebral cortex, it is noteworthy that PGC-1α is directly associated with neurodegenerative diseases based on the clearly identified damaged areas in the striatum.

[0006] In addition to these findings, the identification of vacuolar damage in the central nervous system of PGC-1α knockout mice revealed that PGC-1α plays a key role in maintaining neuronal function.

[0007] Decreased PGC-1α expression increases BACE1 expression, leading to mitochondrial dysfunction and cell death. BACE1 produces β-amyloid protein and increases its quantity by degrading and cleaving the progenitor protein of amyloid protein, which causes Alzheimer's disease. Single nucleotide mutations in the PGC-1α gene are highly associated with increased risk factors for developing Parkinson's disease and Huntington's disease and are known to reduce PGC-1α gene expression in patients with Alzheimer's, Parkinson's, and Huntington's diseases.

[0008] Recently, there has been increasing interest in therapies for Parkinson's disease that pharmacologically activate PGC-1α by targeting its functional mechanisms (which are closely related to neurodegenerative diseases).

[0009] [Existing Technical Documents]

[0010] [Patent Literature]

[0011] (Patent Document 1) Korean Patent No. 10-1384642 Summary of the Invention

[0012] Technical Purpose

[0013] The purpose of this disclosure is to provide a composition that can prevent or treat Parkinson's disease in a subject because the expression of PGC-1α is enhanced in the brain by comprising a 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound.

[0014] Technical solution

[0015] This disclosure provides a pharmaceutical composition for the prevention or treatment of Parkinson's disease, the pharmaceutical composition comprising, as an active ingredient, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0016] [Chemical Formula 1]

[0017]

[0018] In addition, the pharmaceutical composition can increase the expression of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α).

[0019] In addition, the pharmaceutical composition may have a dosage form selected from the group consisting of solutions, suspensions, syrups, emulsions, liposomes, powders, granules, tablets, sustained-release formulations and capsules.

[0020] In addition, the pharmaceutical composition may be a composition for oral administration and has a dosage form including a drug carrier comprising liposomes or a sustained-release agent.

[0021] In addition, the pharmaceutical composition may be a composition for parenteral administration and has a dosage form comprising a drug carrier or a sustained-release agent including liposomes and ultrasound contrast agents.

[0022] Additionally, the compound represented by chemical formula 1 is 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one.

[0023] According to another aspect of this disclosure, a health functional food for alleviating Parkinson's disease is provided, the health functional food comprising a compound represented by chemical formula 1 as an active ingredient.

[0024] According to another aspect of this disclosure, a method for preventing or treating Parkinson's disease may be provided, the method comprising administering to an individual a composition comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0025] Beneficial effects

[0026] The pharmaceutical composition disclosed herein for the prevention or treatment of Parkinson's disease successfully crosses the blood-brain barrier (BBB) ​​and increases the expression of PGC-1α, which has neuroprotective capabilities in the individual's brain, thereby demonstrating an effect of preventing or treating Parkinson's disease in the subject. Attached Figure Description

[0027] Figure 1 A reaction scheme illustrating a method for synthesizing 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compounds according to this disclosure is shown.

[0028] Figure 2 NMR data used for structural identification following the synthesis of the 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound according to this disclosure are shown.

[0029] Figure 3 The illustrative compound screening experimental method in Example 2 is shown.

[0030] Figure 4 A graph showing the results of activity measurements against the PGC-1α promoter via luciferase detection in Example 2 is presented.

[0031] Figure 5 The figure shows the results of experiments conducted with 14 drugs that increased the activity of the PGC-1α promoter by 2.5 times or more.

[0032] Figure 6 Images of proteins from immunoblotting are shown after the SH-SY5Y cell line was treated with the compound (Example 4).

[0033] Figure 7An immunoblot of the PGC-1α protein in the substantia nigra (SN) of mice fed with the compound is shown (Example 4).

[0034] Figure 8 Immunohistochemistry and immunoblotting in brain tissue of a Parkinson's disease model mouse fed a diet containing compounds are shown (Example 5).

[0035] Figure 9 A pole-climbing experiment is shown in a Parkinson's disease model mouse fed a diet containing the compound (Example 6).

[0036] Figure 10 The expression of PGC-1α and its major target genes in Parkinson's disease model mice fed a diet containing the compounds is shown (Example 7). Detailed Implementation

[0037] Because this disclosure is subject to various modifications and has various exemplary embodiments, specific exemplary embodiments will be shown and described in detail in the accompanying drawings. However, this is not intended to limit this disclosure to the specific exemplary embodiments, and this disclosure should be understood to include all modifications, equivalents, and alternatives contained within the spirit and scope of this disclosure. In describing this disclosure, detailed descriptions of relevant known technologies will be omitted where it is determined that such detailed descriptions may obscure the gist of this disclosure.

[0038] This disclosure provides a pharmaceutical composition for the prevention or treatment of Parkinson's disease, the pharmaceutical composition comprising, as an active ingredient, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0039] [Chemical Formula 1]

[0040]

[0041] In the following, a pharmaceutical composition for the prevention or treatment of Parkinson's disease according to specific embodiments of the present disclosure will be described in more detail.

[0042] Conventional treatment strategies for Parkinson's disease have been limited to using medications (such as L-DOPA or dopamine receptor agonists) and deep brain stimulation to manage motor function symptoms, which have not yet prevented the gradual death of dopaminergic neurons (DA).

[0043] Therefore, the inventors have completed this disclosure by determining that when a specific derivative compound is administered, the compound successfully crosses the BBB to increase the activity of the PGC-1α promoter and the protein expression level of PGC-1α in the brain.

[0044] This disclosure provides a pharmaceutical composition for the prevention or treatment of Parkinson's disease, the pharmaceutical composition comprising, as an active ingredient, a 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound or a pharmaceutically acceptable salt thereof.

[0045] PGC-1α is a key regulator of mitochondrial function, co-regulating transcriptional programs that are essential for mitochondrial organisms and protect mitochondria from oxidative stress. This PGC-1α level is reduced in patients with Parkinson's disease, and the decrease in PGC-1α levels in Parkinson's disease is thought to be due to methylation on the PGC-1α promoter.

[0046] On the other hand, PGC-1α knockout mice are more sensitive to the degenerative effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (a neurotoxin for Parkinson's disease). Overexpression of PGC-1α is known to have a protective effect against N-methyl-4-phenylpyridinium ion (MPP+) toxin (the active metabolite of MPTP). Overexpression of PGC-1α is also known to have a protective effect against α-synuclein, MPTP, oxidative stress and rotenone-induced degeneration.

[0047] The PGC-1α response gene is downregulated in dopaminergic neurons derived from patients with Parkinson's disease, suggesting that PGC-1α plays an important role in the pathogenesis of Parkinson's disease; and when PARIS (a substrate of Parkin), a pathogenic protein of Parkinson's disease, is overexpressed, the loss of dopaminergic neurons is inhibited by PGC-1α overexpression, which is believed to show that PGC-1α is a major target of Parkin in dopaminergic neurodegeneration.

[0048] Therefore, defects in PGC-1α signaling have become an important cause of dopaminergic degeneration in Parkinson's disease, and the reduction of PGC-1α due to Parkin's dysfunction may be a major target for the prevention or treatment of Parkinson's disease.

[0049] In Example 2 of this disclosure, the results of a promoter activity measurement of PGC-1α by luciferase detection are shown. Figure 4 The study also presented results from experiments with 14 drugs that increased the activity of the PGC-1α promoter by 2.5 times or more, among which the compound 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one, represented by Formula 1, exhibited the highest activity. Figure 5 ).

[0050] Furthermore, in Example 4 of this disclosure, based on image analysis of immunoblotting performed after treating the SH-SY5Y cell line with various concentrations of the compound (0 μM, 0.01 μM, 0.1 μM, 1.0 μM, 10 μM, 48 hours), it was found that even at low concentrations, 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one increased the amount of PGC-1α protein. Figure 6 ).

[0051] Additionally, as shown in the immunoblotting of the SN of mice fed with food, indobuprofen, or a YPD-01 (2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one) diet (0.5% w / w), it was found that 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one successfully crossed the BBB and statistically significantly increased the protein expression of PGC-1α in the SN of mice (Example 4). Figure 7 ).

[0052] Additionally, in Example 5 of this disclosure, after feeding Parkinson's disease model mice with food or a YPD-01 diet (0.5% w / w) and then slicing the brains using a slicer after brain collection, AAV-PARIS injection significantly killed dopamine neurons that were significantly inhibited by administration of the YPD-01 diet. Figure 8 ).

[0053] Furthermore, in Example 6 of this disclosure, as a result of a pole-climbing experiment performed on Parkinson's disease model mice after feeding them with food or a YPD-01 diet (0.5% w / w), the AAV-PARIS8-injected mice took twice as long to fall from the top of the pole as the AAV-GFP-injected mice, and it was found that the behavioral abnormalities were absent due to YPD-01 administration. Figure 9 ).

[0054] Furthermore, in Example 7 of this disclosure, as a result of measuring the expression of PGC-1α and its target genes after feeding Parkinson's disease model mice with food or a YPD-01 diet (0.5% w / w), overexpression of PARIS by AAV-PARIS led to the inhibition of PGC-1α expression and a decrease in the expression of its target genes. It was found that the inhibition of PGC-1α and the expression of its major target genes (NRF-1, Tfam) by PARIS in the substantia nigra of mice fed a YPD-01 diet was significantly restored. Figure 10 ).

[0055] On the other hand, the compound represented by the following chemical formula 1 is 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one.

[0056] [Chemical Formula 1]

[0057]

[0058] Figure 1 This is the synthetic reaction scheme for 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compounds. Figure 2 These are NMR data used to identify the structure of a compound after its synthesis.

[0059] In the pharmaceutical compositions disclosed herein, the active ingredient is a compound of formula I, a pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0060] As used herein, “pharmacologically acceptable salt” refers to a salt of a compound that induces the desired pharmacological effect (i.e., expression of PGC-1α). These salts can be formed using inorganic acids (such as hydrochlorides, hydrobroms, and hydroiodates) and organic acids (such as acetates, adipates, alginates, aspartates, benzoates, phenyl sulfates, p-toluenesulfonates, hydrogen sulfates, aminosulfonates, sulfates, naphthalates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconate, dodecyl sulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptarates, hexanoates, 2-hydroxyethanesulfonates, lactates, malates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, oxalates, toluenesulfonates, and undecanoates). As used herein, the term "pharmaceuticalally acceptable hydrate" refers to the hydrate of a compound having the desired pharmacological activity, and the term "pharmaceuticalally acceptable solvate" as used herein refers to the solvate of a compound having the desired pharmacological activity. Hydrates and solvates may also be prepared using the acids described above.

[0061] On the other hand, it may also include suitable carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. Additionally, it can be formulated according to conventional methods into oral dosage forms (such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols) as well as external preparations, suppositories, and sterile injectable solutions.

[0062] The carriers, excipients, and diluents that may be included in the composition are lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. When used in formulation, the composition may be prepared using commonly used diluents or excipients (such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc.).

[0063] The pharmaceutical compositions according to this disclosure can be administered in pharmaceutically effective amounts. As used herein, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dose level can be determined by factors such as the patient's disease type, severity, drug activity, drug sensitivity, time of administration, route of administration, excretion rate, duration of treatment, factors including concomitant drugs, and other factors known in the medical field.

[0064] The pharmaceutical compositions according to this disclosure can preferably be administered simultaneously, separately, or sequentially with concomitant drugs to enhance therapeutic effects, and can be administered by single or multiple doses. Taking all the foregoing factors into account, it is important to administer the drug in the minimum amount that yields the maximum effect without side effects, which can be readily determined by those skilled in the art. Specifically, the effective amount of the pharmaceutical composition according to this disclosure can vary depending on the patient's age, sex, condition, and weight, the absorption, inactivation rate, and excretion rate of the active ingredient in the body, the type of disease, and concomitant drugs.

[0065] The pharmaceutical compositions disclosed herein can be administered to an individual via various routes. All routes of administration (including, for example, oral administration, intranasal administration, bronchial administration, arterial injection, intravenous injection, subcutaneous injection, intramuscular injection, or intraperitoneal injection) are predictable.

[0066] The pharmaceutical compositions disclosed herein are determined based on the type of drug as the active ingredient and various relevant factors such as the disease to be treated, route of administration, patient's age, sex, weight, and severity of the disease.

[0067] In another aspect of this disclosure, a method for inhibiting neuroinflammation is provided, comprising administering a pharmaceutical composition to an individual. As used herein, the term “individual” refers to a subject requiring treatment for a disease, and more specifically, a human or a non-human primate and a mammal (such as a mouse, dog, cat, horse, and cattle).

[0068] In addition, the pharmaceutical compositions according to the exemplary embodiments of this disclosure may be dosage forms selected from the group consisting of solutions, suspensions, syrups, emulsions, liposomes, powders, granules, tablets, sustained-release agents and capsules.

[0069] Furthermore, the composition can be a composition for oral administration and has a dosage form comprising a drug carrier including liposomes or a sustained-release agent. Alternatively, the composition can be a composition for parenteral administration and has a dosage form comprising a drug carrier including liposomes and an ultrasound contrast agent or a sustained-release agent.

[0070] The pharmaceutical compositions disclosed herein can be encapsulated in liposomes to provide stability in dosage forms used for drug delivery. The liposomes used herein can be prepared from a mixture of polyols, surfactants, phospholipids, fatty acids, and water.

[0071] There are no particular limitations on the polyols used in liposomes, and they preferably include propylene glycol, dipropylene glycol, 1,3-butanediol, glycerol, methylpropylene glycol, isopentyl glycol, pentylene glycol, erythritol, xylitol and sorbitol, but propylene glycol is the most preferred.

[0072] Any surfactant known in the art can be used to prepare liposomes, such as anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants, with anionic and nonionic surfactants being preferred. Specific examples of anionic surfactants include alkyl acylglutamate esters, alkyl phosphate esters, acetylated alkyl esters, dialkyl phosphate esters, and trialkyl phosphate esters. Specific examples of nonionic surfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters, alkyl polyglycosides, polyglycerol esters, and glycolipids.

[0073] Phospholipids, including natural and synthetic phospholipids (preferably lecithin) (another component used in the preparation of liposomes), are used as amphiphilic lipids. The fatty acids used in the preparation of liposomes are higher fatty acids, preferably saturated or unsaturated fatty acids with C12-C22 alkyl chains, including, for example, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid. The water used for the preparation of liposomes is typically deionized distilled water.

[0074] Liposomes can be prepared by various methods known in the art, but are most preferably prepared by applying a mixture comprising the components to a high-pressure homogenizer. The liposome system prepared in this way has the advantage of dissolving various types of poorly soluble substances and stabilizing unstable substances, thereby maximizing drug delivery.

[0075] The pharmaceutical compositions disclosed herein can be prepared as sustained-release formulations to increase drug compliance by continuously maintaining an effective blood concentration of the active ingredient and reducing the number of drug doses.

[0076] In addition to the active ingredient disclosed herein, the sustained-release agent is prepared by including a sustained-release carrier and other adjuvants. Various sustained-release carriers known in the art can be used as sustained-release carriers herein, but polyethylene oxide is preferred.

[0077] In addition to other adjuvants, diluents commonly used in the pharmaceutical field may also be included. Examples of diluents for this purpose include lactose, dextrin, starch, microcrystalline cellulose, dicalcium phosphate, calcium carbonate, sugar, and silicon dioxide, and may include other flow aids (such as zinc stearate or magnesium stearate) to increase flowability, or may include other adjuvants available in the pharmaceutical field.

[0078] The compositions disclosed herein can be used to administer to subjects in whom the expression of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) is reduced.

[0079] According to another aspect of this disclosure, a health functional food for improving Parkinson's disease may be provided, the health functional food comprising a compound represented by chemical formula 1 as an active ingredient.

[0080] When the compositions of this disclosure are prepared as food compositions or functional food compositions, they may include not only compounds of Formula I as active ingredients, but also components typically added during food preparation, including, for example, proteins, carbohydrates, fats, nutrients, flavorings, and flavor enhancers. Examples of carbohydrates include: monosaccharides such as glucose and fructose; disaccharides such as maltose, sucrose, and oligosaccharides; and polysaccharides such as conventional sugars (e.g., dextrin and cyclodextrin); and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavor enhancers, natural and synthetic flavor enhancers (saccharin, aspartame, etc.) may be used. Furthermore, when the food compositions of this disclosure are prepared as beverages, in addition to the compounds of Formula I of this disclosure, they may also include citric acid, liquid fructose, sugars, glucose, acetic acid, malic acid, fruit juice, Eucommia ulmoides bark extract, jujube extract, licorice extract, etc.

[0081] According to another aspect of this disclosure, a method for preventing or treating Parkinson's disease is provided, the method comprising administering to a subject an preparation comprising a compound represented by chemical formula 1.

[0082] Methods for implementing the invention

[0083] Preferred exemplary embodiments of the present disclosure will be described in detail below with reference to the accompanying drawings. However, these exemplary embodiments are intended to illustrate the present disclosure only, and the scope of the present disclosure should not be construed as being limited by these exemplary embodiments.

[0084] Example 1: Preparation of reporter cell lines and human cell lines

[0085] To prepare a lentivirus for the production of stable reporter SH-SY5Y cells (SH-PGC-1α-Luc), a 1-kbp GL3-PGC-1α promoter-luciferase was cloned into pGreenFire (System Biosciences). The lentivirus construct was first prepared (provided by Akyosi Fukamizu from the University of Tsukuba, Japan).

[0086] HEK293T cells cultured in 15cm culture dishes were transfected with 32μg of pGreenFire vector, 9μg of VSVg envelope, 6.25μg of Prev and 12.5μg of pMDL.

[0087] After 48 hours, the viral supernatant was collected and concentrated using ultracentrifugation, and the viral globules were dissolved in PBS. SH-SY5Y cells were treated with the concentrated virus and screened with puromycin (1 μg / mL) one day later.

[0088] To culture human neuronal SH-SY5Y cells (ATCC, Manassas, VA), 10% FBS (vol / vol, Welgene Gold Serum, cat#S 001-07) and DMEM containing antibiotics (Welgene fresh mediaDMEM, cat#LM 001-05) were used, and cultured in an incubator at 37°C in the presence of 5% carbon dioxide (Forma DirectHeat CO2 incubator, Thermo Scientific).

[0089] Example 2: Compound screening and data analysis to identify compounds that increase PGC-1α expression.

[0090] Considering the properties of drugs, a library of 8,320 compounds was classified from 230,000 drugs (Korean Chemical Library, Daegeon). This library is a combined library of 6,000 drugs and 2,320 spectroscopic acquisitions (Microsource, http: / / www.msdiscovery.com / spectrum.html), and its quality is controllable by liquid chromatography-mass spectrometry (LC-MS) (60% clinically approved drugs, 25% natural products, and 15% bioactive drugs).

[0091] To determine the applicability of this experiment to high-throughput screening (HTS), standard parameters (including signal-to-background (S / B) ratio, daily rate of change, and rate of change per plate) as well as Z' factor and coefficient of variation (CV) were measured.

[0092] All HTS experiments followed the NIH guidelines (High-Throughput Screening Assay Guidance Criteria).

[0093] (http: / / www.ncats.nih.gov / research / reengineering / ncgc / assay / criteria / criteria.html)

[0094] SH-PGC-1α-Luc were distributed into white flat-bottomed 96-well plates, where cells were cultured to contain 10,000 cells per well in 100 μL of DMEM (10% FBS + penicillin / streptomycin (P / S)) and stabilized for 12 hours in an incubator at 37°C in the presence of 5% CO2.

[0095] On the second day, each drug was added to 50 μL of warm DMEM to a final concentration of 20 μM. Then, 50 μL of DMEM was removed from the cell-containing plate, and 50 μL of drug-containing DMEM was added (final concentration: 10 μM). The cells were then exposed to the drugs for 48 hours, and luciferase activity was measured using SteadyGlo reagent (Promega).

[0096] Each plate contained three internal controls: daidzein (positive control, 10 μM) and two negative controls (treated with DMSO and untreated with DMSO). Luciferase measurements for each well are shown as a proportion based on the mean of the untreated controls.

[0097] The Z' factor for this experiment showed values ​​between 0.5 and 1. Differences between each well, plate, and experimental day were measured by comparing changes in the control. DMSO resistance, reagent stability, and experimental conditions were also examined to demonstrate validity.

[0098] Activity measurements of the PGC-1α promoter, detected by luciferase assay, showed that... Figure 4 In the middle. Luciferase activity was measured by reading the first data, where comparisons were made based on DMSO, and Figure 4 The blue square boxes in the diagram represent drugs that have been activated more than 1.5 times.

[0099] Experimental results using 14 drugs that increased the activity of the PGC-1α promoter by 2.5 times or more showed... Figure 5 middle( Figure 5 YPD-01 is 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one, YPD-02 is 2-[4-(3-oxo-1H-isoindoline-2-yl)phenyl]propionic acid, and YPD-03 is isopropyl 2-(4-(1-oxoisoindoline-2-yl)phenyl)propionic acid).

[0100] Example 3: Sampling of SH-SY5Y cells and tissues for immunoblotting

[0101] RIPA buffer was added to SH-SY5Y cells and SNs of C57L / 6N mice, followed by homogenization. The freeze-thaw process was then repeated three times, and total protein levels were determined using a BSA-based BCA kit.

[0102] Immunoblotting was performed by adding 2×SDS sample buffer and heating at 95°C for 10 minutes, followed by antibody treatment of the desired protein to determine the soluble residue.

[0103] In the immunoblotting experiments, the band density was measured using ImageJ (NIH, Bethesda, MO, USA, http: / / rsb.info.nih.gov / ij / ), and statistical analysis was performed using protein band densities proportional to those of the control. Statistical analysis was performed using GraphPad Prism version 7 (GraphPad Software). Unpaired two-tailed Student's t-tests were applied to the data, where p < 0.05 was considered statistically significant.

[0104] In Example 4 below, samples that show statistical significance compared to controls by Student's t-test are marked with an asterisk (*p<0.05, **p<0.01, ***p<0.001).

[0105] Example 4: The role of 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one in the SH-SY5Y cell line and mouse brain

[0106] As shown in Example 3 Figure 6 The image shown is an image of an immunoblot performed after the SH-SY5Y cell line was treated with the compound (10 μM, 48 h).

[0107] like Figure 6As shown, it was found that YPD-02 (2-[4-(3-oxo-1H-isoindol-2-yl)phenyl]propionic acid) and YPD-03 (isopropyl 2-(4-(1-oxoisoindolin-2-yl)phenyl)propionic acid) increased the amount of PGC-1α protein with increasing drug concentration (0.01 μM, 0.1 μM, 1 μM, 10 μM), while YPD-01 (2-(4-(1-hydroxypropane-2-yl)phenyl)isoindolin-1-one) increased the amount of PGC-1α protein regardless of concentration.

[0108] exist Figure 7 Immunoblots of proteins of SN in mice fed a diet derived from self-consumption food, YPD-02 (2-[4-(3-oxo-1H-isoindol-2-yl)phenyl]propionic acid) or YPD-01 (2-(4-(1-hydroxypropane-2-yl)phenyl)isoindolin-1-one) diet (0.5% w / w) for 1 week are shown (quantitative plots quantified with β-actin / data are presented as mean ± SEM / statistical significance was measured by applying an unpaired two-tailed Student's t-test, *p<0.05, **p<0.01.).

[0109] pass Figure 7 The results showed that YPD-01 (2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one) successfully crossed the BBB and increased the protein level of PGC-1α in the SN of mice.

[0110] Example 5: Inhibitory effect of YPD-01 on dopamine neuron death in a mouse model of Parkinson's disease.

[0111] To establish a mouse model of Parkinson's disease, AAV-PARIS was stereotactically injected into the substantia nigra region. Mice were fed with food or a YPD-01 (2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one) diet (0.5% w / w) for 4 weeks, after which brains were collected and sectioned using a microtome. Dopaminergic neurons were visualized by reacting the mouse brain tissue, which was cut into 35 μm sections, with TH antibody and neuronal dopamine markers, followed by exposure to vestestatin ABC (Vectorbiolabs) and DAB (Sigma) solutions. Six rats were used in each experiment.

[0112] - Introducing adeno-associated virus (AAV) via stereotactic injection.

[0113] Eight-week-old mice were anesthetized by injecting 100 μL of pentobarbital (10 mg / mL) into their abdomen. The cranial periosteum was dissected, and the left (X: 1.2, Y: -3.2, Z: -4.5) and right (X: 1.2, Y: 3.2, Z: -4.5) brain regions were marked based on the anterior fontanelle. Holes were drilled into the marked areas, and the virus was slowly injected via syringe (0.2 μL every 30 seconds). After injecting the virus into the left brain and waiting 2 minutes, the same procedure was performed on the other side. The mice were carefully monitored after surgical suturing and subsequently multiplied in cages until recovery.

[0114] - Immunoblot (Western Blot, protein blotting)

[0115] Midbrains were collected from mice that had been stereotactically injected with AAV-PARIS and fed either a diet or a YPD-01 diet (0.5% w / w) for 4 weeks, and proteins were extracted using RIPA dissolution buffer. The protein concentration was adjusted to 4 mg / mL using a BCA-validated method. Electrophoresis was performed on a 7% polyacrylamide gel after mixing with 2x Laemmli sample buffer. Following transfer, a secondary antibody bound to primary and HRP was attached, followed by color development using ECL solution. AAV-GFP was used as a control for AAV-PARIS.

[0116] Immunohistochemistry

[0117] Mouse brain tissue, cut into 35µm ice slices, was reacted overnight at 4°C with tyrosine hydroxylase (TH) antibody (a neuronal dopamine marker). The next day, it was reacted with a biotin-bound secondary antibody, and then the dopamine neurons were shaped by exposure to vesstatin ABC (Vector Biolabs) and DAB (Sigma) solutions. The developed brain tissue was placed on a glass slide and examined under a microscope.

[0118] like Figure 8 As shown, AAV-PAIS injection significantly killed dopamine neurons, and in mice fed a YPD-01 diet, dopamine neuron death was intentionally inhibited. Consistent with the results shown in immunohistochemistry, as a result of immunoblotting in the substantia nigra of mice using TH antibody, the dopamine neuron markers (quantitative plots quantified with β-actin / data are presented as mean ± SEM / statistical significance measured by applying one-way ANOVA, *p<0.05, ***p<0.001) reaffirmed that AAV-PARIS inhibited dopamine neuron death by YPD-01, as confirmed by the results found in immunohistochemistry.

[0119] Example 6: The inhibitory effect of YPD-01 on behavioral abnormalities in Parkinson's disease

[0120] To determine whether YPD-01 effectively inhibits dopamine neuron death and Parkinson's-like behavioral abnormalities in a Parkinson's disease model, a pole climbing experiment was performed, as shown in Example 5 above, which is the most reliable behavioral experiment.

[0121] - Pole Climbing Test

[0122] To investigate behavioral abnormalities, Parkinson's disease model mice were transferred to a pole-climbing cage and allowed to acclimatize for 3 minutes. The mice were then raised by their tails and placed on the tip of a vertically upright pole. The time from when the mouse removed its hind paw from the tip of the pole until it reached the floor was measured.

[0123] like Figure 9 As shown, due to the potential AAV-PARIS-induced dopamine neuron death, mice injected with AAVPARIS took approximately twice as long to descend from the top of the rod as mice injected with AAV-GFP (used as a control). Symptoms of behavioral abnormalities disappeared upon YPD-01 ingestion, consistent with the inhibition of dopamine neuron death described in Example 5 above (data are presented as mean ± SEM / statistical significance measured by applying one-way ANOVA, *p<0.05).

[0124] Example 7: YPD-01 confirmed that it increased the expression of PGC-1α in a mouse model of Parkinson's disease.

[0125] In the animal model experiment set up as in Example 5, the expression of PGC-1α and its major target gene (NRF-1, Tfam) was measured by RT-qPCR to determine whether the expression was increased due to YPD-01 uptake.

[0126] -Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR)

[0127] Midbrains were collected from mice that had been stereotactically injected with AAV-PARIS and fed a diet of food or YPD-01 for 4 weeks, and RNA was extracted using a total RNA extraction kit (Intron Biotechnology). cDNA was synthesized using a cDNA synthesis kit (Enzynomics) and oligo-dT from the extracted RNA. Gene expression in the rotor gene Q (Qiagen) was quantified by qPCR using primers for the gene to be analyzed and SYBRgreen (Qiagen) reagent.

[0128] like Figure 10As determined in the study, AAV-PARIS overexpression of PARIS led to the inhibition of PGC-1α expression and the reduction of expression of its target genes. The PARIS-induced inhibition of PGC-1α and the expression of its major target genes (NRF-1, Tfam) were significantly restored in the substantia nigra of mice fed with YPD-01 (quantitative plots quantified with β-actin / data are presented as mean ± SEM / statistical significance was measured by applying one-way ANOVA, *p<0.05, **p<0.01; ns, not significant).

[0129] As specific portions of this disclosure have been described in detail above, it will be apparent to those skilled in the art that these specific techniques are merely preferred embodiments, and that the scope of this disclosure is not limited thereto. Therefore, the essential scope of this disclosure will be defined by the appended claims and their equivalents.

Claims

1. Use of a pharmaceutical composition in the preparation of a medicament for the prevention or treatment of Parkinson's disease, said pharmaceutical composition comprising, as an active ingredient, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] 2. The application according to claim 1, wherein, The pharmaceutical composition increases the expression of peroxisome proliferator-activated receptor-γ coactivator-1α.

3. The application according to claim 1, wherein, The pharmaceutical composition has a dosage form selected from the group consisting of solutions, suspensions, syrups, emulsions, liposomes, powders, granules, tablets, sustained-release formulations, and capsules.

4. The application according to claim 3, wherein, The pharmaceutical composition is a composition for oral administration and has a dosage form comprising a drug carrier or sustained-release agent including liposomes.

5. The application according to claim 3, wherein, The pharmaceutical composition is a composition for parenteral administration and has a dosage form comprising a drug carrier or sustained-release agent including liposomes and an ultrasound contrast agent.

6. The application according to claim 1, wherein, The pharmaceutical composition is administered to subjects in whom the expression of peroxisome proliferator-activated receptor-γ coactivator-1α is reduced.

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