KR101855087B1 - Chalcone derivatives, optical isomer thereof, or pharmaceutically acceptable salts thereof, and a pharmaceutical composition for preventing or treating mitochondrial disease induced by decrease of oxygen consumption rate comprising the same as an active ingredient - 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 chalcone derivative compound as an active ingredient, a neurodegenerative disease, attention deficit hyperactivity disorder, asthma, drug poisoning, chronic renal failure, The present invention relates to a pharmaceutical composition for preventing or treating a disease caused by a decrease in oxygen consumption rate of mitochondria selected from liver diseases, wherein the chalcone derivative compound has excellent activity of increasing the oxygen consumption rate in mitochondria, Or a pharmaceutically acceptable salt thereof.

Description

TECHNICAL FIELD The present invention relates to a chalcone derivative, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition for preventing or treating diseases caused by reduction of oxygen consumption rate of mitochondria containing the chalcone derivative, optical isomer thereof, or and pharmaceutically acceptable salts thereof, and a pharmaceutical composition for preventing or treating mitochondrial disease induced by decrease in oxygen consumption rate comprising the same as an active ingredient,

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 chalcone derivative compound or a pharmaceutically acceptable salt thereof as an active ingredient, a neurodegenerative disease, an Attention Deficit Hyperactivity Disorder, a Chronic Disease, The present invention relates to a pharmaceutical composition for preventing or treating diseases caused by reduction of oxygen consumption rate of mitochondria selected from liver diseases.

Mitochondria are one of the subcellular organelles that form the backbone of energy metabolism in cells and are surrounded by nuclear membranes that are characteristic of eukaryotic cells. Mitochondria are also described as the energy production plants of cells, because ATP, which is the most cellular energy required for cells, is produced. This process of ATP production is called cell respiration, and the active respiration activates the mitochondria.

Breathing refers to the action of living organisms to decompose nutrients to obtain the energy needed for living. The respiration of cells is a complex process in which cells are degraded by various stages of oxidation, and they undergo the process, the TCA circuit and the electron transfer system. Mitochondrial respiration is an essential process to consume oxygen through the electron transport chain (ETC) and produce carbon dioxide. Approximately 85-90% of the total oxygen consumption rate is used for oxidative phosphorylation, which is necessary for the synthesis of ATP (Korzeniewski, B., Biochem Mol Biol Int., 39 (2), 415-419, 1996).

Since mitochondria not only supply the energy required for cells but also are involved in various regulatory processes such as signal transduction, cell differentiation and cell death, mitochondrial dysfunction can be a cause of various diseases, and mitochondrial dysfunction changes the oxygen consumption rate Eesser 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).

(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 Myhill, S. et al., ≪ RTI ID = 0.0 > 1, < / RTI > 12-23, 2014), chronic fatigue syndrome (Filler, K. 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, Diabetes, 53 (suppl 1), S110-S118, 2004, Kim, JA et al., Circ Res., 102 (4), 401-414, 2008) have been reported in the prior art.

In particular, the oxygen consumption rate in the cerebrum of patients with Parkinson's disease was reported to be reduced by referring to the prior literature [Borghammer, P. et al., J Neurol Sci., 313 (1-2), 123-128, 2012] 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 decreased oxygen metabolism in the cerebral cortex of patients with chronic renal failure and hepatic encephalopathy [Kamada, T. et al , Dig Dis Sci., 31 (2), 119-124, 1986] reported that the oxygen consumption rate of hepatocytes in chronic liver disease patients was reduced. Also, the prior art [Ben-Shachar, D., J Neurochem., 83 (6), 1241-1251, 2002; (Eg, attention deficit hyperactivity disorder and schizophrenia) have been associated with mitochondrial dysfunctions (eg, Flemmen, G. et al., Medicine (Baltimore), 94 (44), e1914, 2015] And drug addiction decreased the oxygen consumption of drug addicts caused by cannabis and amphetamine in the case of drug addiction. Therefore, as a method for treating diseases related to dysfunction of mitochondria, studies on drugs that increase the oxygen consumption rate are required.

Neurodegenerative disease is also called neurodegenerative disease. Unlike the normal aging process, abnormal neuron death causes brain or spinal cord abnormalities, resulting in decreased cognitive, walking, and athletic ability. (Sagara, Y. et al., Free Radic Biol Med., 24 (9), 1375-1389, 1998; Jeong, GS et al., Biol Pharm Bull., 32 JS et al., Neurosci Lett., 393,165-945 (2005), 945-949, 2009; Parfenova, H. et al., Am J Physiol Cell Physiol., 290 169, 2006).

Parkinson's disease, a typical neurodegenerative disease, is a chronic progressive degenerative disease of the nervous system resulting from sudden degeneration or a decrease in the number of cells that produce dopamine, a neurotransmitter in the substantia nigra area of the midbrain , Is a representative refractory disease. Parkinson's disease is characterized by a decrease in dopamine in the neurotransmitter, resulting in a complete balance of the neurotransmitter system, resulting in tremor, rigidity, bradykinesia, instability of posture postural instability).

Chronic renal failure is a condition in which kidneys are damaged for more than 3 months or in which kidney function is continuously decreased. Even if kidney function is reduced by 35 ~ 50% due to various causes, it does not cause any systemic symptoms. However, kidney function is exacerbated by wasting waste and electrolyte concentration control, even the most basic functions that do not do well enough to get worse when the kidney failure. If kidney function deteriorates and end-stage renal failure is reached, treatment can not be done by diet or medication alone. Therefore, hemodialysis, peritoneal dialysis or kidney transplantation should be performed to replace kidney function.

Hepatic encephalopathy refers to the loss of consciousness or changes in behavior in patients with liver dysfunction. It can range from a little change in personality or behavior to a deep coma in which the state of consciousness changes from day to day, day and night, and seriously unresponsive to pain. When hepatic encephalopathy develops, it is recovered in many cases, but it is not recovered early, and deep coma becomes very slow or does not respond to treatment. It is important to find out and treat them appropriately when they begin to sleep early or start doing something different from usual. If suspected hepatic encephalopathy is suspected, it is a key treatment method to eat ammonia or take a laxative so that the ammonia escapes and enema is seen more often. It is possible to administer amino acid formulation or other medication, or if chronic hepatic encephalopathy does not recover well Liver transplantation can be considered for its treatment.

In chronic liver diseases, hepatitis, which usually causes inflammation of the liver, is the most common cause of liver disease. According to the symptoms, acute hepatitis and chronic infection, viral hepatitis, alcoholic hepatitis, Can be divided. In addition, liver abnormalities caused by abnormalities include fatty liver, hepatitis, liver cirrhosis and liver cancer. Although the mechanism of progression of liver disease is not fully understood, it is thought that primary liver injury is accompanied by secondary cell damage after the occurrence of fatty liver, which leads to progressive liver disease such as hepatitis and liver cirrhosis. Therefore, The possibility of more serious liver disease can be prevented. Although the treatment of liver disease is combined with exercise, diet, and medication in combination, there is no established treatment until now, and it is difficult to completely heal.

On the other hand, as a prior art related to chalcone derivative compounds, International Publication WO-A-2012-116362 discloses an Nrf2 activator including a chalcone derivative, and International Publication No. 2013-025498 discloses a neuroprotective polyphenol analogue And International Publication No. 2015-187934 discloses a functional heteroaryl enone that exhibits Nrf2 activation and a method for its production. However, the possibility of using chalcone derivatives having the same structure as the present invention as a therapeutic agent for a disease having a decrease in oxygen consumption rate as a phenotype has not been disclosed.

International Publication No. 2012-116362 (Chalcone derivatives as Nrf2 activators, published on Aug. 30, 2012) International Patent Publication No. 2013-025498 (Neuroprotective polyphenol analogs, published Feb. 21, 2013) International Publication No. 2015-187934 (Functionalized hetroarylenones exhibiting Nrf2 Activation and their method of use, published on Dec. 10, 2015)

Andrews, H. E. et al., Mitochondrial dysfunction plays a key role in the progressive axonal loss in Multiple Sclerosis, Med Hypotheses., 64 (4), 669-677, 2005. Ben-Shachar, D., Mitochondrial dysfunction in schizophrenia: a possible linkage to dopamine, J Neurochem., 83 (6), 1241-1251, 2002. Borghammer, P. et al., Cerebral oxygen metabolism in patients with early Parkinson's disease, J Neurol Sci., 313 (1-2), 123-128, 2012. Cordero, M. D. et al., Mitochondrial dysfunction and mitophagy activation in blood mononuclear cells of fibromyalgia patients: implications in the pathogenesis of the disease, Arthritis Res Ther., 12 (1), R17, 2010. Cordero, M. D. et al., Oxidative stress and mitochondrial dysfunction in fibromyalgia, Neuro Endocrinol Lett., 31 (2), 169-173, 2010. Dam, G. et al., Hepatic encephalopathy associated with decreased cerebral oxygen metabolism and blood flow, not increased ammonia uptake, Hepatology, 57 (1), 258-265, 2013. Dumont, M. et al., Mitochondria and antioxidant targeted therapeutic strategies for Alzheimer's disease, J Alzheimers Dis., 20 (Suppl 2), S633-643, 2010. Filler, K. et al., Association of Mitochondrial Dysfunction and Fatigue: A Review of the Literature, BBA Clin., 1, 12-23, 2014. Flemmen, G. et al., Impaired Aerobic Endurance and Muscular Strength in Substance Use Disorder Patients: Implications for Health and Premature Death, Medicine (Baltimore), 94 (44), e1914, 2015. Green, K. et al., Prevention of mitochondrial oxidative damage as a therapeutic strategy in diabetes, Diabetes, 53 (suppl 1), S110-S118, 2004. Ha, J. S. et al., Glutamate-induced oxidative stress, but not cell death, is largely dependent upon extracellular calcium in mouse neuronal HT22 cells, Neurosci Lett., 393, 165-169, 2006. Hu, Y. et al., K-ras (G12V) transformation leads to mitochondrial dysfunction and a metabolic switch from oxidative phosphorylation to glycolysis, Cell Res., 22 (2), 399-412, 2012. Jeong, G. S. et al., Cytoprotective constituents of the heartwood of Caesalpinia sappan on glutamate-induced oxidative damage in HT22 cells, Biol Pharm Bull., 32 (5), 945-949, 2009. Kamada, T. et al., Estimated hepatic oxygen consumption in patients with chronic liver diseases as assessed by organ reflectance spectrophotometry, Dig Dis Sci., 31 (2), 119-124, 1986. Kanai, H. et al., Depressed cerebral oxygen metabolism in patients with chronic renal failure: a positron emission tomography study, Am J Kidney Dis., 38 (4 Suppl 1), S129-133, 2001. Kim, J. A. et al., Role of mitochondrial dysfunction in insulin resistance, Circ Res., 102 (4), 401-414, 2008. Korzeniewski, B., What regulates respiration in mitochondria ?, Biochem Mol Biol Int., 39 (2), 415-419, 1996. Kwon, J. et al., Assurance of mitochondrial integrity and mammalian longevity by p62-Keap1-Nrf2-Nqo1 cascade, EMBO Rep., 13 (2), 150-156, 2012. Lopez-Armada, M.J. et al., Mitochondrial dysfunction and the inflammatory response, Mitochondrion, 13 (2), 106-118, 2013. Luo, Y. et al., Mitochondria: A Therapeutic Target for Parkinson's Disease ?, Int J Mol Sci., 16 (9), 20704-20730, 2015. Myhill, S. et al., Targeting mitochondrial dysfunction in the treatment of Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME / CFS) -a clinical audit, Int J Clin Exp Med., 6 (1), 1-15, 2013. Onyango, I. et al., Mitochondrial genomic contribution to mitochondrial dysfunction in Alzheimer's disease, J Alzheimers Dis., 9 (2), 183-193, 2006. Parfenova, H. et al., Glutamate induces oxidative stress and apoptosis in cerebral vascular endothelial cells: Contributions of HO-1 and HO-2 to cytoprotection, Am J Physiol Cell Physiol., 290 (5), 1399-1410, 2006. Resseguie, E. A. et al., Hyperoxia activates ATM independent from mitochondrial ROS and dysfunction, Redox Biol., 176-185, 2015. Sagara, Y. et al., Cellular mechanisms of resistance to chronic oxidative stress, Free Radic Biol Med., 24 (9), 1375-1389, 1998. Winklhofer, K. F. et al., Mitochondrial dysfunction in Parkinson's disease, Biochim Biophys Acta., 1802 (1), 29-44, 2010.

It is an object of the present invention to provide a chalcone derivative compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof and a neurodegenerative disease comprising the same as an active ingredient, attention deficit hyperactivity disorder, asthma, drug addiction, chronic renal failure, And a pharmaceutical composition for preventing or treating a disease caused by a decrease in oxygen consumption rate of mitochondria selected from chronic liver diseases.

The present invention relates to a neurodegenerative disease comprising attention-deficit hyperactivity disorder (CHD) comprising the chalcone derivative compound represented by the following general formula (1) or (2), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient, hyperactivity disorder, schizophrenia, drug addiction, chronic renal failure, hepatic encephalopathy, and chronic liver diseases, by reducing the oxygen consumption of mitochondria And to a pharmaceutical composition for the prevention or treatment of diseases caused thereby.

[Chemical Formula 1]

(2)

The term " optical isomer " includes forms of R-form, S-form or racemic compounds, respectively.

The pharmaceutically acceptable salt of the chalcone derivative compound of the present invention is preferably an addition salt formed by an inorganic acid such as a hydrochloride, a sulfate, a phosphate, a hydrobromide, a hydroiodide, a nitrate, a pyrosulfate or a metaphosphate, An addition salt formed by an organic acid such as a salt, an oxalate, a benzoate, an acetate, a trifluoroacetate, a propionate, a succinate, a fumarate, a lactate, a maleate, a tartrate, a glutarate, Lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts and the like, but are not limited thereto.

( E ) -1- (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-ene (Compound 2), ( E ) -1- (3-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop- , (E) -1- (3- hydroxyphenyl) -3- (4- (trifluoromethyl) phenyl) prop-2-en-1-one (compound 3), (E) -1- ( En-1-one (compound 4) and ( E ) -1- (4-hydroxyphenyl) -3- En-1-one (Compound 5), preferably ( E ) -1- (3-hydroxyphenyl) -3- (3- (Compound 2) and ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop- 2-en-1-one (Compound 4).

The neurodegenerative diseases of the present invention are selected from the group consisting of Parkinson's disease, Alzheimer's disease, stroke, Lou Gehrig's disease, Pick's disease, Huntington's disease, multiple sclerosis and dementia. Chronic liver disease includes viral hepatitis, Hepatitis, fatty liver, chronic hepatitis, and liver cirrhosis.

( 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- , Or an optically isomer thereof, or a pharmaceutically acceptable salt thereof.

(3)

The pharmaceutical composition according to the present invention can be formulated into a suitable form together with a commonly used pharmaceutically acceptable carrier. &Quot; Pharmaceutically acceptable " refers to a composition that is physiologically acceptable and, when administered to humans, does not normally cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. In addition, the compositions can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method.

Examples of carriers, excipients and diluents that can be contained in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl parahydroxybenzoate, propyl parahydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, stabilizers, binders, disintegrants, surfactants and the like which are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, microcrystalline cellulose, sucrose or lactose, Low-substituted hydroxypropylcellulose, hypromellose, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used. In order to formulate the formulation for parenteral administration, the chalcone derivative compound of Formula 1 or Formula 2, its optical isomer, or a pharmaceutically acceptable salt thereof may be sterilized and / or sterilized and preserved, stabilized, hydrated or emulsified, And other therapeutically useful substances in water to prepare solutions or suspensions, which may be prepared in ampoules or vial unit dosage forms.

The pharmaceutical composition comprising the compound of formula (I) or (II) as an active ingredient of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. The dosage will depend on the age, sex, body weight, the particular disease or condition being treated, the severity of the disease or condition, the time of administration, the route of administration, the absorption, distribution and excretion of the drug, It depends on judgment. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several doses. The dose is not intended to limit the scope of the invention in any way.

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 chalcone derivative compound as an active ingredient, a neurodegenerative disease, attention deficit hyperactivity disorder, asthma, drug poisoning, chronic renal failure, The present invention relates to a pharmaceutical composition for preventing or treating a disease caused by a decrease in oxygen consumption rate of mitochondria selected from liver diseases, wherein the chalcone derivative compound has excellent activity of increasing the oxygen consumption rate in mitochondria, Or a pharmaceutically acceptable salt thereof.

1 is a graph showing changes in oxygen consumption rate (FIGS. 1A and 1B) when Compound 4 of the present invention is treated with neurons for 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 was treated with neuronal cells for a long time. Fig.
FIG. 3 is a graph (FIG. 3B) showing the result of immuno-staining (FIG. 3A) of dopamine cell changes when Compound 4 of the present invention is treated in a mouse model in which Parkinson's disease is induced.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

≪ Example 1: Synthesis of chalcone derivative >

Referring to the following Reaction Scheme 1, the chalcone derivative compounds of the present invention were prepared.

[Reaction Scheme 1]

Compound 1-a or 2-a (1.0 eq.) Was dissolved in ethanol, LiOH.H ₂ O (0.2-1.0 eq.) Was added with reference to the reaction conditions shown in Table 1 below, and the mixture was stirred at room temperature for 15 minutes. After 15 minutes, the compound 1-b, 2-b or 3-b (1.0-1.2 equivalents) was added and stirred for 2 hours. Distilled water (100 ml) was added to terminate the reaction. Then, the mixture was diluted with ethyl acetate (200 ml), washed with distilled water (200 ml x 2 times) and brine (200 ml), and then the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography or recrystallization to obtain Compounds 1 to 5 of the present invention.

Compound No. 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) 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) 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 (5.0g, 36.7mmol) , LiOH · H 2 O (1.85g, 41.9mmol), compound 2-b (5.9㎖, 44.0mmol) , ethanol (150㎖) 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) 0.6 g, 27%

≪ Example 2: Identification of physicochemical properties of chalcone derivative compounds >

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

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 Propan-2-en-1-one (Compound 2) was prepared in the same manner as in Example 1,

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).

Examples 2-3. ( E Prop-2-en-1-one (Compound 3) [0157] < EMI ID =

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 , 3 ArH, trans H), 8.03 (d, J = 15.7 Hz, trans H), 8.12 (d, J = 7.8 Hz, 2ArH), 9.86 (s, OH).

Examples 2-4. ( E Propan-2-en-1-one (Compound 4) was prepared in the same manner as in Example 1,

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 C -F = 31.6 Hz, CCF ₃ ), 131.8, 133.1, 136.5, 141.3, 162.9 (ArC, CH), 187.4 (s, CO).

Examples 2-5. ( E Propan-2-en-1-one (Compound 5) was obtained in the same manner as in Example 1,

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 CF = 270.5 Hz, CF 3), 126.1, 129.3, 129.7, 129.9, 130.0, 130.7 (q, J C -F = 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 rate in nerve cells >

Example 3-1. Nerve cell manufacturing

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

Example 3-2. Measurement of oxygen consumption rate during short-time treatment

As a method for evaluating the ability of intracellular mitochondria, measurement of oxygen consumption rate through XF-analyzer of Seahorse is known. (A port: target compound, B port: Oligomycin A, an ATPase inhibitor, C port: uncoupler, CCCP (carbonyl cyanide m-chlorophenyl hydrazone)) to measure the change of oxygen consumption rate using XF- D port: mitochondrial complex 1 inhibitor rotenone), and the mitochondrial function was evaluated from the results of changes in oxygen consumption rate for each drug.

First, the dopamine cell line SN4741 cultured in Example 3-1 was divided into 20,000 cells per well in an XF-plate. Thereafter, the compound of the present invention was not treated, and 3 ratios (one rate was carried out for 7 minutes) were measured. Then, the compounds 1 to 5 of the present invention, which are the target compounds, were dissolved in DMSO in a port A, , Respectively, and 5 rates were measured. As a control group, DMSO was used instead of the compound of the present invention. As a comparative group, Comparative Examples 1 to 3 were each dissolved in DMSO and injected at a concentration of 5 μM, respectively. After that, oligo-A was injected to B port, CCCP was injected to C port, and rottenone was injected to D port. The results of measuring the change in the oxygen consumption rate are shown in Table 2 with reference to the following formula, and in particular, in the case of treating the compound 4, the results are shown in FIGS. 1A and 1B.

[Equation]

Condition potency (fold) The control (DMSO) - Compound 1

1.2 Compound 2

1.2 Compound 3

1.2 Compound 4

1.3 Compound 5

1.2 Comparative Example 1 (No. 10)

no effect Comparative Example 2 (No. 14)

no effect Comparative Example 3 (No. 17)

no effect

Referring to Table 2 and FIG. 1, when the compounds 1 to 5 of the present invention were treated with dopamine cells, it was confirmed that the oxygen consumption rate was increased as compared with the control or Comparative Examples 1 to 3.

Particularly, the compound 1 of the present invention had a higher effect of increasing the oxygen consumption rate as compared with the comparative example 1, and it was confirmed that the same tendency was also obtained with the results of the compound 4 of the present invention and the comparative example 2. That is, from the above results, it was found that, in the structure of the chalcone derivative compound, the compound having a hydroxyl group substituent, rather than having a methoxy group substituent, was more effective in increasing the oxygen consumption rate in neurons.

In addition, in the structure of the chalcone derivative compound, compounds having a hydroxyl group substituent and having a hydroxy group substituent at the para or meta position rather than the ortho position are more excellent in the effect of increasing the oxygen consumption rate. In the compounds 2 and 4 of the present invention and Comparative Example 3 The results showed.

Example 3-3. Measurement of oxygen consumption rate during long-term treatment

Compound 4 of the present invention was pre-treated at a concentration of 2.5 μM for 48 hours in the dopamine cell line SN4741 cultured in Example 3-1 to observe a change in oxygen consumption rate at the time of long-term treatment. To the XF-plate, 20,000 cells. Thereafter, the 3 rate was measured, and the A port was injected sequentially with CCCP at Oligomycin A and B ports, and the ROTONON was injected into the C port sequentially, and the rate was measured at 3 rates (each rate was 7 minutes) . 2A and 2B show the measurement results of the oxygen consumption rate for the case of treating the compound of the present invention for a long time by measuring a change in oxygen consumption rate at a total of 12 rates.

Referring to FIG. 2, it was confirmed that when the compound 4 of the present invention was treated for a long time in neurons, the oxygen consumption rate was increased. In particular, comparing FIGS. 1 and 2, it was confirmed that treatment of the compound of the present invention for a long time rather than a short-time treatment of the compound of the present invention showed a higher maximum oxygen consumption rate change.

Example 4. Confirmation of dopaminergic neuronal cell death in a mouse model of Parkinson's disease.

10-week-old B27 mice were treated with a dose of 20 mg / kg of MPTP (1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine), a neurotoxic substance used in the model of Parkinson's disease mice, After intraperitoneal injection, 4 doses of Compound 4 were orally administered at a dose of 10 mg / kg per day for 3 days after injection of MPTP. As a control group, B27 mice were treated with MPTP without treatment. As a control group, B27 mice were treated with only MPTP and treated with MPTP alone and B27 mice without MPTP Compound 4 alone treated with Compound 4 alone was used.

Subsequently, the brain was extracted through a perfusion process one week after MPTP injection, and the extracted brain tissue was sectioned. Substantia nigra and striatum regions were treated with the dopaminergic neuron marker, tyrosine hydroxylase (TH, tyrosine hydroxylase) antibody, and the result is shown in FIG. 3B.

Referring to FIG. 3, in the mouse model of Parkinson's disease caused by the MPTP drug, it was confirmed that the compound 4 of the present invention was inhibited more than twice as much as that of the compound 4 when dopamine was killed I could.

That is, from the results shown in Table 2 and FIG. 3, when the chalcone derivative of the present invention was treated in vitro, the potency value was increased by 1.3 times or more. In a practical Parkinson's disease model (in vivo) To 3 times or more. Thus, the chalcone derivative compound can be effectively used as a pharmaceutical composition for preventing or treating Parkinson's disease.

<Example 5: Toxicity test>

Example 5-1. Acute toxicity

( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) of the present invention in a short period of time (Within 24 hours) to determine toxicity to the animal body, and to determine the mortality rate. Twenty ICR mice were prepared, and 10 mice were assigned to each group. In the control group, 30% PEG-400 alone was administered, and in the experimental group, Compound 4 was orally administered at a concentration of 1.0 g / kg each. After 24 hours of administration, the respective mortality rates were examined. As a result, both the control group and the test group to which the compound 4 was administered survived.

Example 5-2. Organ organs toxicity test in experimental group and control group

( E ) -1- (4-hydroxyphenyl) -3- (3-tert-butoxycarbonylphenyl) -3- (3- (Trifluoromethyl) phenyl) prop-2-en-1-one) was administered at a concentration of 1.0 g / kg and the control group, -pyruvate transferase) and BUN (blood urea nitrogen) were measured by Select E (vital scientific NV, Netherland). As a result, GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to renal toxicity, showed no significant difference compared to the control group. In addition, liver and kidney were cut from each animal and histological observation was performed with an optical microscope through a conventional tissue section production process, but no abnormal abnormalities were observed.

&Lt; Formulation Example 1 >

Formulation Example 1-1. Manufacture of Powder

2 g of ( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en-1-one) Mixed and airtightly packed to prepare powder.

Formulation Example 1-2. Manufacture of tablets

100 mg of the compound 4 of the present invention (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop- Mg of lactose, 60 mg of lactose hydrate, 20 mg of low-substituted hydroxypropylcellulose, and 2 mg of magnesium stearate were mixed, and tablets were prepared by tableting according to a conventional preparation method.

Formulation Example 1-3. Preparation of capsules

100 mg of the compound 4 of the present invention (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop- Mg of lactose, 60 mg of lactose hydrate, 20 mg of low-substituted hydroxypropylcellulose, and 2 mg of magnesium stearate were mixed, and the above components were mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  1-4. Injection preparation

10 mg of the compound 4 of the present invention (( E ) -1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop- Distilled water, and pH adjuster were mixed, and the contents were adjusted to the above contents per ampoule (2 ml) according to the usual injection preparation method.

Claims (4)

(E) - (3-hydroxyphenyl) -3- (2- (trifluoromethyl) phenyl) prop-2-en- Neurodegenerative diseases and neurodegenerative diseases comprising 1- (4-hydroxyphenyl) -3- (3- (trifluoromethyl) phenyl) prop-2-en- A pharmaceutical composition for the prevention or treatment of diseases caused by a reduction in the oxygen consumption rate of selected mitochondria.
[Chemical Formula 1]

delete The method according to claim 1,
Wherein the neurodegenerative disease is a disease selected from the group consisting of Parkinson's disease, Alzheimer's disease, stroke, Lou Gehrig's disease, Huntington's disease, multiple sclerosis and dementia, by reduction of the oxygen consumption rate of mitochondria selected from neurodegenerative diseases and asthma A pharmaceutical composition for the prevention or treatment of a disease caused thereby. delete

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