WO2018062837A1 - Chalcone derivative compound, optical isomer thereof, or pharmaceutically acceptable salt thereof, and pharmaceutical composition comprising same as effective ingredient for preventing or treating disease caused by reduction in mitochondrial oxygen consumption rate - Google Patents

Abstract

The present invention relates to a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same as an effective ingredient for preventing or treating diseases caused by reduction in a mitochondrial oxygen consumption rate. A pharmaceutical composition, when comprising the chalcone derivative compound, exerts excellent activity of increasing a mitochondrial oxygen consumption rate and can be usefully used for preventing or treating diseases caused by a reduction in the oxygen consumption rate of mitochondria, such as neurodegenerative diseases, attention deficit/hyperactivity disorder, schizophrenia, drug addiction, chronic renal failure, hepatic encephalopathy, and chronic liver disease.

Description

Pharmaceutical composition for the prevention or treatment of diseases caused by reduced oxygen consumption of the chalcone derivative compound, optical isomer thereof, or a pharmaceutically acceptable salt thereof and mitochondria comprising the same as an active ingredient

The present invention relates to a pharmaceutical composition for the prophylaxis or treatment of a disease caused by a reduction in oxygen consumption of mitochondria comprising a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof and an active ingredient thereof.

Mitochondria are one of the intracellular organelles that form the backbone of energy metabolism in cells and are surrounded by a nuclear membrane that is characteristic of eukaryotic cells. Mitochondria are also described as cell energy production plants because ATP, the most cellular energy required for cells, is produced. This ATP production process is also called cell breathing, mitochondria is activated as the respiration occurs actively.

Respiration refers to the action by which life breaks down nutrients and obtains the energy necessary for life. The respiration of cells is a complex process that is broken down through various stages of oxidation in the cell, through glycolysis, the TCA circuit, and the electron transport system. Respiration of mitochondria is an essential process for consuming oxygen and producing carbon dioxide through the electron transport chain (ETC), and about 85-90% of the total oxygen consumption is used for the oxidative phosphorylation required for the synthesis of ATP (Korzeniewski, B., Biochem Mol Biol Int., 39 (2), 415-419, 1996).

Because mitochondria not only supply energy for cells, but also are involved in various regulation such as signal transduction, cell differentiation, and cell death, mitochondrial dysfunction can cause various diseases, and mitochondrial dysfunction changes oxygen consumption rate. (Hu, Y. et al., Cell Res., 22 (2), 399-412, 2012; Kwon, J. et al., EMBO Rep., 13 (2), 150-156, 2012; Resseguie, EA et al., Redox Biol., 176-185, 2015).

As diseases related to mitochondrial dysfunction, Alzheimer's disease (Onyango, I. et al., J Alzheimers Dis., 9 (2), 183-193, 2006; Dumont, M. et al., J Alzheimers Dis., 20 (Suppl 2), S633-643, 2010), Parkinson's disease (Winklhofer, KF et al., Biochim Biophys Acta., 1802 (1), 29-44, 2010; Luo, Y. et al., Int J Mol Sci ., 16 (9), 20704-20730, 2015), chronic fatigue syndrome (Filler, K. et al., BBA Clin., 1, 12-23, 2014; Myhill, S. et al. , Int J Clin Exp Med., 6 (1), 1-15, 2013), fibromyalgia (Cordero, MD et al., Neuro Endocrinol Lett., 31 (2), 169-173, 2010; Cordero , MD et al., Arthritis Res Ther., 12 (1), R17, 2010), autoimmune diseases (Lopez-Armada, MJ et al., Mitochondrion, 13 (2), 106-118, 2013; Andrews, HE et al., Med Hypotheses., 64 (4), 669-677, 2005), diabetes mellitus (Green, K. et al., Diabetes, 53 (suppl 1), S110-S118, 2004; Kim, JA et al. , Circ Res., 102 (4), 401-414, 2008).

In particular, referring to the previous literature [Borghammer, P. et al., J Neurol Sci., 313 (1-2), 123-128, 2012], it was reported that the oxygen consumption rate in the cerebral of patients with Parkinson's disease was reduced. Kanai, H. et al., Am J Kidney Dis., 38 (4 Suppl 1), S129-133, 2001; Dam, G. et al., Hepatology, 57 (1), 258-265, 2013, reported reduced oxygen metabolism in the cerebral tract of patients with chronic renal failure and hepatic encephalopathy, prior art [Kamada, T. et al. , Dig Dis Sci., 31 (2), 119-124, 1986, reported reduced oxygen consumption of hepatocytes in patients with chronic liver disease. See also, Ben-Shachar, D., J Neurochem., 83 (6), 1241-1251, 2002; Flemmen, G. et al., Medicine (Baltimore), 94 (44), e1914, 2015] suggest that mental disorders such as attention deficit hyperactivity disorder and schizophrenia are associated with mitochondrial abnormalities. In the case of drug addiction caused by cannabis and amphetamines, it was reported that oxygen consumption was reduced. Therefore, as a method for treating diseases related to the dysfunction of mitochondria as described above, research on drugs that increase oxygen consumption rate is required.

Neurodegenerative disease, also known as neurodegenerative disease, is a disorder in which the brain or spinal cord causes abnormalities in the brain or spinal cord due to abnormal neuronal death, which decreases cognitive, gait, and motor skills. Diseases, Alzheimer's disease, stroke, etc. (Sagara, Y. et al., Free Radic Biol Med., 24 (9), 1375-1389, 1998; Jeong, GS et al., Biol Pharm Bull., 32 ( 5), 945-949, 2009; Parfenova, H. et al., Am J Physiol Cell Physiol., 290 (5), 1399-1410, 2006; Ha, JS et al., Neurosci Lett., 393, 165- 169, 2006).

Parkinson's disease, a representative neurodegenerative disease, is a chronic progressive degenerative disease of the nervous system caused by sudden degeneration or a significant decrease in the number of cells that produce the neurotransmitter dopamine in the substantia nigra area of the midbrain. Is a representative refractory disease. Parkinson's disease reduces the balance of the neurotransmitter by reducing dopamine in neurotransmitters, resulting in tremor, rigidity, bradykinesia, and postural instability. symptoms such as postural instability.

Chronic renal failure is a disease in which the kidneys are damaged or renal function continues for more than three months. Even if the kidney function decreases by 35-50% of normal due to various causes, it does not cause any systemic symptoms. However, if the kidney function is bad enough that even the most basic functions such as waste excretion and electrolyte concentration control do not work properly, it leads to kidney failure. If renal failure progresses and reaches end stage renal failure, the diet or drug therapy alone can not be treated, so hemodialysis, peritoneal dialysis, or kidney transplants can be substituted for kidney function.

Hepatic encephalopathy is an unconscious or behavioral change in patients with liver dysfunction. If your consciousness changes frequently or changes day and night, starting with a slight change in your personality or behavior, it can range from deep coma to severely unresponsive pain. Hepatic encephalopathy develops in many cases, but when it does not recover early and in a deep coma, it may recover very slowly or not respond to treatment. It is important to find and treat appropriately when you begin to sleep poorly or do something different than usual. When hepatic encephalopathy is suspected, it is a key treatment method to take diarrhea or enemas to get rid of ammonia, and to take a lot of stool.Amino acid preparations or other medications can be used, and chronic hepatic encephalopathy is not well recovered. You can think of a liver transplant for its treatment.

In chronic liver diseases, hepatitis, which usually causes inflammation of the liver, accounts for the majority of liver diseases. Depending on the pattern, acute hepatitis and chronic infection, depending on the cause, viral hepatitis, alcoholic hepatitis, drug hepatitis, etc. Can be divided. In addition, liver diseases caused by such abnormalities include fatty liver, hepatitis, cirrhosis, liver cancer, and the like. The mechanism of the progression of liver disease is not fully understood, but it is thought to develop into advanced liver disease such as fatty liver disease and cirrhosis due to secondary cell damage after primary fatty liver development. If controllable, the development of more serious liver disease could be prevented. In the treatment of liver disease, exercise, alcoholism, diet and treatment are treated in parallel, but there are no treatment methods established to date and fundamentally complete healing is difficult.

Meanwhile, as a prior document related to a chalcone derivative compound, International Publication No. 2012-116362 discloses an Nrf2 activator including a chalcone derivative, and International Publication No. 2013-025498 discloses a polyphenol analog having a neuroprotective action. International Publication No. 2015-187934 discloses a functional heteroaryl enone showing Nrf2 activation and a method for preparing the same. However, there is no disclosure about the availability of a chalcone derivative having the same structure as the present invention as a therapeutic agent for diseases having a reduced oxygen consumption rate as a phenotype.

It is an object of the present invention to provide a pharmaceutical composition for the prophylaxis or treatment of a disease caused by a reduction in the oxygen consumption rate of mitochondria comprising a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof and an active ingredient thereof. .

The present invention provides a pharmaceutical for preventing or treating a disease caused by reducing oxygen consumption of mitochondria comprising a chalcone derivative compound represented by Formula 1 or Formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. It relates to a composition.

[Formula 1]

[Formula 2]

The term optical isomers include forms of R-form, S-form or racemic compounds, respectively.

Pharmaceutically acceptable salts of the chalcone derivative compounds of the present invention are preferably addition salts and citric acid formed by inorganic acids such as hydrochloride, sulfate, phosphate, hydrobromide, hydroiodide, nitrate, pyrosulfate, metaphosphate, etc. Addition salts formed by organic acids such as salts, oxalates, benzoates, acetates, trifluoroacetates, propionates, succinates, fumarates, lactates, maleates, tartarates, glutarates, sulfonates, or Metal salts, such as lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts, but are not limited thereto.

In addition, when the compound of Formula 1 or Formula 2 is more specifically exemplified, ( E ) -1- (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-ene -1-one (Compound 1), ( E ) -1- (3-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 2) , ( E ) -1- (3-hydroxyphenyl) -3- (4- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 3), ( E ) -1- ( 4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 4) and ( E ) -1- (4-hydroxyphenyl) -3 -(4- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 5), preferably ( E ) -1- (3-hydroxyphenyl) -3- (2- (Trifluoromethyl) phenyl) prop-2-en-1-one (Compound 1) and ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) It is 1 or more types chosen from prop-2-en-1-one (compound 4).

The disease caused by the reduced oxygen consumption rate of the mitochondria is selected from the group consisting of neurodegenerative diseases, attention deficit hyperactivity disorder, schizophrenia, drug addiction, chronic renal failure, hepatic encephalopathy and chronic liver disease.

In addition, the neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Lou Gehrig's disease, Pick disease, Huntington's disease, multiple sclerosis and dementia, the chronic liver disease is viral hepatitis, alcoholic hepatitis, drug hepatitis, Fatty liver, chronic hepatitis and cirrhosis.

In another aspect, the present invention provides ( E ) -1- (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-ene-1 represented by the following formula (3): -One (compound 1) and ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 4) The present invention relates to a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 3]

The pharmaceutical compositions according to the invention may be formulated in a suitable form with the pharmaceutically acceptable carriers generally used. Pharmaceutically acceptable refers to a composition that is physiologically acceptable and does not cause an allergic or similar reaction, such as gastrointestinal disorders, dizziness, or the like, when administered to a human. In addition, the compositions may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl paraoxybenzoate, propyl paraoxybenzoate, talc, magnesium stearate and mineral oil, but are not limited thereto. When formulated, diluents or excipients such as fillers, stabilizers, binders, disintegrants and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid form preparations include at least one excipient such as starch, microcrystalline cellulose, sucrose or lactose, It is prepared by mixing low-substituted hydroxypropyl cellulose, hypromellose and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used. The chalcone derivative compounds, optical isomers thereof, or pharmaceutically acceptable salts thereof are sterilized and / or preservatives, stabilizers, wetting or emulsifying accelerators, salts and / or buffers for the control of osmotic pressure for formulation into parenteral dosage forms. And adjuvant, and other therapeutically useful substances, may be mixed into water to prepare a solution or suspension, which may be prepared in ampoules or in vial unit dosage forms.

The pharmaceutical composition comprising the chalcone derivative compound disclosed in the present invention as an active ingredient may be administered to mammals such as rats, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections. Dosage may include the age, sex, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the time of administration, the route of administration, the absorption, distribution and excretion of the drug, the type of drug used and the prescriber's It will vary depending on judgment. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosage is 1 mg / kg / day to 500 mg / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The present invention relates to a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition for preventing or treating a disease caused by a reduced oxygen consumption rate of mitochondria including the same as an active ingredient. When the compound is treated, the activity of increasing oxygen consumption in the mitochondria is excellent, and the oxygen consumption rate of mitochondria in neurodegenerative diseases, attention deficit hyperactivity disorder, schizophrenia, drug addiction, chronic renal failure, hepatic encephalopathy and chronic liver disease is reduced. It can be usefully used as a pharmaceutical composition for preventing or treating a disease caused.

1 is a graph showing the measurement of oxygen consumption rate change (FIG. 1A and 1B) when Compound 4 of the present invention is treated with dopaminergic neurons in a short time.

FIG. 2 is a graph showing the change in oxygen consumption rate (FIGS. 2A and 2B) when Compound 4 of the present invention is treated with dopaminergic neurons for a long time.

FIG. 3 shows the relative dopaminergic neuron density when the compound 4 of the present invention is treated in a Parkinson's disease-induced mouse model. FIG. 3A is a result of immunostaining dopaminergic neurons of the mouse model. FIG. 3B Is a graph that quantifies this.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the information provided herein is to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

Example 1 Synthesis of Chalcone Derivative Compound

Referring to Scheme 1 below, a chalcone derivative compound of the present invention was prepared.

Scheme 1

To refer to reaction conditions shown in Table 1, the compound 1-a or a-2 (1.0 equiv) was dissolved in ethanol, addition of LiOH and H ₂ O (0.2 ~ 1.0 equivalent) and stirred at room temperature for 15 minutes. After 15 minutes, Compound 1-b, 2-b or 3-b (1.0-1.2 equivalents) was added thereto, stirred for 2 hours, and distilled water (100 mL) was added to terminate the reaction. Thereafter, the mixture was diluted with ethyl acetate (200 mL), washed with distilled water (200 mL × 2 times) and brine (200 mL), and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified using silica gel chromatography or recrystallization to obtain compounds 1 to 5 of the present invention.

Compound number	Reaction conditions	Reaction yield
Compound 1	Compound 1-a (0.5 g, 3.67 mmol), LiOH.H 2 O (0.18 mg, 4.41 mmol), Compound 1-b (0.58 mL, 4.41 mmol), ethanol (15 mL)	0.61 g, 57%
Compound 2	Compound 1-a (0.5 g, 3.67 mmol), LiOH.H 2 O (0.18 mg, 4.41 mmol), Compound 2-b (0.59 mL, 4.41 mmol), ethanol (15 mL)	0.75 g, 50%
Compound 3	Compound 1-a (0.5 g, 3.67 mmol), LiOH.H 2 O (0.18 mg, 4.41 mmol), compound 3-b (0.60 mL, 4.41 mmol), ethanol (15 mL)	0.78 g, 73%
Compound 4	Compound 1-b (5.0 g, 36.7 mmol), LiOH.H 2 O (1.85 g, 41.9 mmol), Compound 2-b (5.9 mL, 44.0 mmol), ethanol (150 mL)	1.6 g, 15%
Compound 5	Compound 1-b (1.0 g, 7.34 mmol), LiOH.H 2 O (0.37 g, 8.81 mmol), Compound 3-b (1.2 mL, 8.81 mmol), Ethanol (30 mL)	0.6g, 27%

< Example  2. Calcon  Determination of Physicochemical Properties of Derivative Compounds>

Example 2-1. ( E ) -1- (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 1)

Light yellow solid;

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.10-7.12 (m, ArH), 7.41 (t, J = 7.88 Hz, ArH), 7.50 (s, ArH), 7.57-7.69 (m, ArH, trans H), 7.74-7.86 (m, ArH, trans H), 7.97 (s, 2ArH), 8.33 (d, J = 7.8 Hz, ArH), 9.90 (s, ArH).

Example 2-2. ( E ) -1- (3-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 2)

White solid;

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.08 (d, J = 7.88 Hz, ArH), 7.39 (t, J = 7.88 Hz, ArH), 7.51 (s, ArH), 7.69-7.82 (m , 3ArH, trans H), 8.05 (d, J = 15.7 Hz, trans H), 8.18 (d, J = 7.6 Hz, ArH), 8.34 (s, ArH), 9.84 (s, OH).

Example 2-3. ( E ) -1- (3-hydroxyphenyl) -3- (4- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 3)

Light yellow solid;

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.10 (d, J = 7.4 Hz, ArH), 7.40 (t, J = 7.76 Hz, ArH), 7.51 (s, ArH), 7.67-7.83 (m , 3ArH, trans H), 8.03 (d, J = 15.7 Hz, trans H), 8.12 (d, J = 7.8 Hz, 2ArH), 9.86 (s, OH).

Example 2-4. ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 4)

White solid;

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 6.91 (d, J = 8.6 Hz, 2ArH), 7.69-7.65 (m, 2ArH, trans H), 8.06-8.17 (m, 3ArH, trans H), 8.32 (s, ArH), 10.46 (s, OH);

¹³ C NMR (75 MHz, DMSO- d ₆ ): δ 115.8, 124.5 (q, J _{C -F} = 270.8 Hz, CF ₃ ), 124.6, 125.3, 126.7, 129.4, 130.2, 130.3 (q, J _CF = 31.6 Hz, CCF ₃ ), 131.8, 133.1, 136.5, 141.3, 162.9 (ArC, CH), 187.4 (s, CO).

Example 2-5. ( E ) -1- (4-hydroxyphenyl) -3- (4- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 5)

Yellow oil;

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 6.91 (d, J = 8.5 Hz, 2ArH), 7.71-7.81 (m, 2ArH, trans H), 8.03-8.11 (m, 4ArH, trans H), 10.5 (s, OH);

¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 115.9, 124.5 (q, J _{C -F} = 270.5 Hz, CF ₃ ), 126.1, 129.3, 129.7, 129.9, 130.0, 130.7 (q, J _CF = 31.6 Hz , CCF ₃ ), 131.8, 139.4, 141.1, 162.9 (ArC, CH), 187.4 (s, CO).

Example 3 Measurement of Oxygen Consumption in Dopaminergic Neurons

Example 3-1. Dopaminergic Neuron Production

The dopaminergic neuron line SN4741 was incubated at 33 ° C. and 5% CO ₂ in d-MEM medium containing 10% FBS, 1% glucose, penicillin / streptomycin, and l-glutamine.

Example 3-2. Oxygen consumption rate measurement in short time treatment

As a method for evaluating the ability of mitochondria in cells, oxygen consumption rate measurement by Seahorse's XF-analyzer is known. In order to measure the change in oxygen consumption rate using XF-analyzer, a total of four ports (port A: target compound, port B: oligomycin A, an ATPase inhibitor, port C: carbonyl cyanide m-chlorophenyl hydrazone, uncoupler) Port D: Four different drugs were administered to rotenone, a mitochondrial complex 1 inhibitor, at different time intervals, and mitochondrial function was evaluated from the oxygen consumption rate change for each drug.

First, SN4741, a dopaminergic neuron cultured in Example 3-1, was dispensed at 20000 cells per well on an XF-plate. Then, after measuring the 3 rate (but one rate proceeds for 7 minutes) without drug treatment, the compound 1 to 5 of the present invention, the target compound is dissolved in DMSO and injected into the A port at a concentration of 5 μM, respectively, and 5 rate. Was measured. In this case, DMSO was used instead of the compound of the present invention as a control, and Comparative Compounds 1 to 5 were used as the comparative group.

Next, oligomycin A in port B, CCCP in port C, and rotenone in the port D were measured sequentially, and then measured by 3 rates, respectively, to measure a change in oxygen consumption rate of a total of 17 rates. The result of measuring the change in oxygen consumption rate was calculated by referring to the following formula and is shown in Table 2, in particular, the results for the compound 4 is shown in Figures 1A and 1B.

[Equation]

Condition	potency (fold)
Control (DMSO)	One
Compound 1	1.2
Compound 2	1.2
Compound 3	1.2
Compound 4	1.3
Compound 5	1.2
Comparative Compound 1	no effect
Comparative Compound 2	no effect
Comparative Compound 3	no effect
Comparative Compound 4	no effect
Comparative Compound 5	no effect

Referring to Table 2 and FIG. 1, when the dopaminergic neurons were treated with the compounds 1 to 5 of the present invention, it was confirmed that the oxygen consumption rate was increased more than when the control or the comparative compounds 1 to 5 were treated. there was.

In particular, the compound 1 or 4 of the present invention having a hydroxy group substituent in the structure of the chalcone derivative compound has a higher effect of increasing the oxygen consumption rate than the comparative compound 1 or 2 having a methoxy group substituent, and also has a hydroxyl group substituent in the para or meta position. Compounds 1 to 5 of the present invention were found to have an excellent effect of increasing the oxygen consumption rate compared to the comparative compounds 3 to 5 having a hydroxy group substituent in the ortho position, and it was found that there was a large difference in the oxygen consumption activity according to the substituents and positions of the chalcone derivatives. Could.

Example 3-3. Oxygen consumption rate measurement at long time treatment

In order to confirm the change in oxygen consumption rate after long time treatment, the compound 4 of the present invention was pretreated to SN4741, the dopaminergic neurons cultured in Example 3-1, at a concentration of 2.5 μM for 48 hours, and then placed on an XF-plate. Dispense at 20000 cells per well. Thereafter, after measuring 3 rates, oligomycin A in port A, CCCP in port B, and rotenone in C port were sequentially injected and measured by 3 rates (but one rate proceeded for 7 minutes). By measuring the change in oxygen consumption rate of a total of 12 rates, the oxygen consumption rate measurement results for the case of long-term treatment of the compound of the present invention is shown in Figures 2A and 2B.

Referring to FIG. 2, when the compound 4 of the present invention was treated for a long time with dopaminergic neurons, the oxygen consumption rate was increased. Particularly, the compound of the present invention was treated for a long time rather than a short time. It was confirmed that a high maximum oxygen consumption rate change.

Example 4 Confirmation of Dopaminergic Neuronal Death in Parkinson's Disease Mouse Model

10-week-old B27 mice were dosed at 20 mg / kg with MPTP (1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine), a neurotoxic substance used to make Parkinson's mouse A total of four injections were given intraperitoneally, and compound 1, 4 or 5 was orally administered once daily at a dose of 10 mg / kg for 3 days after the injection of MPTP. At this time, as a control group (normal group) was used a group that did not process both MPTP and compound in B27 mice, the comparison group was treated with only MPTP only in B27 mice and compound 1 without treatment with MPTP in B27 mice A group treated with only 4, 5 or 5 compounds alone was used.

A week after MPTP injection, the brain was extracted through perfusion, and the extracted brain tissue was sectioned, and then the substantia nigra and striatum regions were labeled with tyrosine hydroxylase, a dopaminergic neuron marker. Immunostaining with TH, tyrosine hydroxylase) antibody, the dopaminergic neuron density is shown in Figure 3 and Table 3. In this case, the relative density of TH-positive cell fibers of Fig. 3B and Table 3 means that dopaminergic neuronal cell death increases as the value is lower.

Condition	Relative density of TH-positive cell fibers (fold)
Control	1.0
MPTP	0.25
MPTP + Compound 1	0.65
MPTP + Compound 4	0.66
MPTP + Compound 5	0.30
MPTP + Comparative Compound 3	0.28
MPTP + Comparative Compound 4	0.28
MPTP + Comparative Compound 5	0.29

Referring to Figure 3 and Table 3, the relative dopaminergic neuron density of the group induced Parkinson's disease by MPTP on the basis of the control group (normal group) showed a value of 0.25, MPTP and the compound 1 or 4 of the present invention Relative dopaminergic neuron density of the group treated with showed a value of 0.65, 0.66, confirming that the death of dopaminergic neurons more than two times.

On the other hand, the relative dopaminergic neuron density of the group treated with MPTP and Compound 5 of the present invention showed a value of 0.3 so that the potency values of dopaminergic neurons (in vitro) (see Table 2) were similar to those of Compound 1 or 4. Parkinson's disease-induced animal model (in vivo) shows similar dopaminergic neuron killing effect as MPTP-induced Parkinson's disease-induced animal model. It was found that the use shows a better therapeutic effect.

Example 5. Toxicity Test

Example 5-1. Acute Toxicity

Ingestion of the compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) of the present invention in a short time This experiment was carried out to investigate the toxicity in animals acutely (within 24 hours) and to determine mortality. Twenty ICR mice, which are common mice, were prepared, and 10 were assigned to each group. Only 30% PEG-400 was administered to the control group, and the experimental group orally administered Compound 4 at a concentration of 1.0 g / kg. Each mortality rate was examined 24 hours after administration, and both the control group and the experimental group administered with Compound 4 survived.

Example 5-2. Long-term and tissue toxicity experiments in experimental and control groups

Long-term toxicity experiments were carried out in C57BL / 6J mice to investigate the effect on the organs (tissues) of animals in order to investigate the effects of compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (Trifluoromethyl) phenyl) prop-2-en-1-one) was collected after 8 weeks from the animals of the experimental group and the control group administered only the solvent at a concentration of 1.0 g / kg GPT (glutamate) Blood concentrations of pyruvate transferase (BUN) and blood urea nitrogen (BUN) were measured using a Select E (vital scientific NV, Netherland) instrument. As a result, in the case of GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to nephrotoxicity, there was no difference in the experimental group compared to the control group. In addition, liver and kidneys were excised from each animal, and histological observations were performed by optical microscopy through a conventional tissue fabrication process. No abnormalities were observed.

Preparation Example 1. Pharmaceutical Formulations

Formulation example  1-1. Powder  Produce

2 g of Compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) of the present invention and 1 g of lactose The powder was prepared by mixing and filling in an airtight cloth.

Formulation Example 1-2. Manufacture of tablets

Compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) 100 mg of the present invention, 100 microcrystalline cellulose Tablets were prepared by mixing mg, lactose monohydrate, 20 mg of low-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearate, followed by compression according to a conventional method for preparing tablets.

Formulation Example 1-3. Preparation of Capsule

Compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) 100 mg of the present invention, 100 microcrystalline cellulose MG, lactose monohydrate 60mg, low-substituted hydroxypropyl cellulose 20mg and magnesium stearate 2mg were mixed, and the above ingredients were mixed and filled in gelatin capsules according to a conventional capsule preparation method to prepare a capsule.

Formulation Example 1-4. Preparation of Injectables

10 mg of compound 4 (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) of the invention, sterilized for injection After distilled water and pH adjuster were mixed, the amount of the above-mentioned ingredient was prepared per ampoule (2 ml) according to a conventional injection method.

Claims (5)

1. A pharmaceutical composition for preventing or treating a disease caused by a reduction in oxygen consumption of mitochondria comprising a chalcone derivative compound represented by the following Chemical Formula 1 or Chemical Formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

   [Formula 1]

   

   [Formula 2]

   
2. The method of claim 1,

   The compound of formula (I) or formula (II) (E) -1- (3- hydroxyphenyl) -3- (2- (methyl) phenyl trifluoromethyl) prop-2-en-1-one (Compound 1) And ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 4). A pharmaceutical composition for the prophylaxis or treatment of diseases caused by a reduction in the oxygen consumption rate of mitochondria comprising a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
3. The method according to claim 1 or 2,

   The disease caused by the reduced oxygen consumption rate of the mitochondria is selected from the group consisting of neurodegenerative diseases, attention deficit hyperactivity disorder, schizophrenia, drug addiction, chronic renal failure, hepatic encephalopathy and chronic liver disease Pharmaceutical compositions for the prevention or treatment of diseases caused by reduced consumption rate.
4. The method of claim 3,

   The neurodegenerative disease is a disease selected from the group consisting of Parkinson's disease, Alzheimer's disease, Lou Gehrig's disease, Pick disease, Huntington's disease, multiple sclerosis and dementia for the prevention or treatment of diseases caused by reduced oxygen consumption of mitochondria. Pharmaceutical composition.
5. ( E ) -1- (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-en-1-one (Compound 1) and ( E ) ) 1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 4) Derivative compounds, optical isomers thereof, or pharmaceutically acceptable salts thereof.

   [Formula 3]

   

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